Acquity Operators Manual

8/20/2019 Acquity Operators Manual

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ACQUITY UPLC SystemOperator’s Guide

71500082502 / Revision C

Copyright © Waters Corporation 2006.

All rights reserved.



Copyright notice

© 2004−06 WATE RS CO RP ORATION . P RI NTE D I N TH E U NI TE D S TATE SOF AMERI CA AND I RELAND. ALL RI G HTS RE SE RVED . THI SDOCU MENT OR P ARTS THE REOF MAY NOT B E REP RODU CE D I N ANYFORM WITHOU T THE WRITTEN P ER MISS ION OF THE P U B LIS HE R.

The informa tion in th is document is subject to cha nge w ithout notice an dshould not be constru ed as a commit ment by Wa ters C orpora tion. Wa tersCorpora tion a ssumes no responsibil ity for a ny errors tha t ma y a ppear in this

document . This document is believed t o be complete a nd a ccura te a t th e timeof publica tion. In no event sha ll Wa ters Corpora tion be liable for incident a l orconsequ entia l dam a ges in connection w ith, or ar ising from, its use.

Wa ters C orporat ion34 Ma ple Str eetMilford, MA 01757U S A

Trademarks

Millennium a nd Wa ters a re registered tra demarks, a nd ACQU ITY UP LC,eCord, Empow er, MassLy nx, and na noACQU ITY UP LC a re tra demarks ofWa ters C orporat ion.

P EE K is a tra demark of Victrex Corpora tion.

P ha rMed and Tygon a re registered tra demarks of Sa int-G obain Cera mics &P last ics, Inc.

P hill ips is a registered tra demark of P hil lips S crew C ompan y.

TORX is a registered t ra dema rk of Textron Corpora tion.

Windows is a registered tra dema rk of Microsoft C orpora tion.

Other t ra demarks or registered tra demarks a re the sole property of theirrespective owners.



iii

Customer comments

Wa ters’ Technical C ommunicat ions depa rt ment invites you to tell us of a ny

errors you encount er in th is document or t o suggest ideas for oth erwiseimproving it. P lease help us better und ersta nd w ha t y ou expect from ourdocument a tion so th a t w e ca n continuously improve its a ccuracy a ndusabil i ty.

We seriously consider every cust omer comm ent w e receive. You ca n r ea ch usat [email protected].

Wa ters C orporat ion34 Ma ple Str eetMilford, MA 01757U S A



iv

Safety considerations

Some rea gents a nd sa mples used wit h Wa ters

®

instru ment s ca n posechemica l, biologica l, and ra diologica l haz a rds. B e sure you are a w a re of thepotent ially ha za rdous effects of all substa nces you work with . Alwa ys observeG ood La bora tory P ra ctice (G LP ) guidelines, published by th e U .S. Food an dDrug Administra tion, and consult your organizat ion’s sa fety representa tivefor guida nce.

When y ou develop meth ods, follow th e “P rotocol for t he Adopt ion of Ana lyt ical

Methods in th e Clinica l Chemistry La bora tory,” Amer ican Journ al of M edi calTechnology , 44, 1, pa ges 30–37 (1978). This prot ocol a ddr esses g ood opera t ingprocedures a nd t he techniques necessary to va lidat e system a nd m ethodperformance.

Safety advisories

Consult Appendix A for a comprehensive list of w a rning a nd cau tionadvisories.

Operating the ACQUITY UPLC system

When opera ting t he ACQU ITY U P LC system, follow sta nda rd q ua lity contr olprocedures a nd t he guidelines presented in th is section.



v

Symbols

Intended use

The Wa t ers®

ACQU ITY UP LC™ system is intended for a na lyzing compoundsa nd intr oducing sepa ra ted sa mples into a ma ss spectrometer.

When y ou develop met hods, follow t he “P rotocol for t he Adoption of Ana lyt icalMethods in th e Clinica l Chemistry La bora tory,” Amer ican Journ al of M edi cal

Technology , 44, 1, pa ges 30–37 (1978). This protocol covers good opera t ingprocedures a nd t echniques necessary to va lida te syst em a nd m ethodperformance.

Calibration

To calibrat e UP LC sy st ems, follow a ccepta ble calibrat ion meth ods using a tlea st f ive sta nda rds t o generat e a sta nda rd curve. The concentra tion ra nge for

Symbol Definition

Aut horized Representa tive of theEuropea n Community.

The CE sym bol serves as confirma -tion of the conformity of a productwit h a l l Europea n Community direc-tives applica ble to tha t product.

For in vitr o diagnostic use.

The ACQU ITY UP LC syst em is CE -ma rked a ccording to theE uropean U nion in vitr o Dia gnost ic Device Directive 98/79/E C.

Warning: The ACQU ITY U P LC syst em is to be used for in vitrodiagn ostic use only by t ra ined, qua lified la bora tory personnel.

EC REP

IVD



vi

sta nda rds should cover t he entire ra nge of qua lity-control sam ples, ty pica lspecimens, a nd a ty pica l specimens.

To ca librat e ma ss spectr ometers, consult t he calibra tion section of t heoperat or’s guide of the instr ument you ar e ca librat ing.

Quality control

Routinely run three quality-control samples that represent subnormal,norma l, an d a bove-norma l levels of a compound. Ensu re th a t q ua lity-cont rolsa mple results fa ll w ithin a n a ccepta ble ra nge, and eva lua te precision fromday to day a nd run t o run. Dat a collected wh en qua lity control samples a re outof range might n ot be valid. Do not report t hese da ta unti l you a re certa in tha tthe inst rument performs sa tisfactorily.

When a na lyzing sa mples from a complex mat rix such a s soil, tissue,serum/plasm a , w hole blood, etc., note t ha t th e ma tr ix components ca na dversely a ffect L C/MS r esults , enh a ncing or suppressing ioniza t ion. Tominimize th ese mat rix effects, Wa ters r ecommends you a dopt th e follow ingmeasures:

• P r i or t o th e in st r umen ta l a n a l ys is , u se a ppr opr ia te sa mplepretrea tm ent such a s protein precipita tion, liquid/liquid extra ction(LLE ), or solid pha se extra ction (SP E ) to remove ma tr ix interferences.

• Whenever possible , ver i fy method accuracy and precision usingma trix-ma tched calibra tors an d QC sa mples.

• Use one or more in terna l s t andard compounds , preferab lyisotopica lly-labeled a na lytes.



vii

IVD authorized representative information

IVD authorized representative

Wa ters Corpora tion (Micromas s U K Limited) isregistered in t he Un ited Kingdom w ith t heMedicines a nd H ealthcare P roducts Regula toryAgency (MH RA) a t M a rket Towers , 1 Nine E lmsLa ne, London, S W8 5NQ. The reference num beris IVD000167.

Wa ters C orporat ion (Microma ss U K Lt d.)

Floa ts Road

Wythenshawe

Ma nchester M23 9LZU nited Kingdom

Telephone: + 44-161-946-2400

Fax: + 44-161-946-2480

Contact: Qual ity ma nager



viii



Table of Contents ix

Safety considerations .......................................................................................... iv S a fety a dvisories ................................................................................................. iv

Operating the ACQUITY UPLC system ........................................................... iv Sym bols ................................................................................................................ v

In t ended use ......................................................................................................... v

Ca libra t ion ........................................................................................................... v Qua lity contr ol .................................................................................................... vi

IVD authorized representative information ................................................ vii IVD a ut horized represent a tive .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. vii

1 System Overview .................................................................................... 1-1

Instruments, components, and data systems ............................................. 1-2

UPLC system guidelines .................................................................................. 1-4 ACQ U I TY U P LC Column s C a lcula tor............................................................ 1-4

Binary solvent manager .................................................................................. 1-5 H ow t he bina ry solvent m a na ger w orks .. . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . 1-6

Sample manager ................................................................................................ 1-7 H ow sam ple flow s ............................................................................................ 1-7

Column heater ................................................................................................... 1-8

Optional sample organizer ............................................................................. 1-9

Optical detectors ............................................................................................. 1-10 TU V detector .................................................................................................. 1-10

P D A det ect or .................................................................................................. 1-10

EL S detector ................................................................................................... 1-10

Media n ba seline filter .................................................................................... 1-11

Data systems .................................................................................................... 1-11

E mpower softw a re ......................................................................................... 1-11 Ma ssLy nx softw a re ........................................................................................ 1-11

Table of Contents



x Table of Contents

Columns ............................................................................................................. 1-12 eCord column chip ......................................................................................... 1-12

FlexCart ............................................................................................................. 1-13

For more information .................................................................................... 1-13

2 System Setup ........................................................................................... 2-1

Before you begin ............................................................................................... 2-2 Tools. ........... ........... ............. ........... ............. ............. ........... ............ ............. ..... 2-3

Assembling the FlexCart ................................................................................. 2-3 Adjust ing t he FlexC a rt ’s height ...................................................................... 2-6

Locking th e FlexC a rt in place ......................................................................... 2-8

Moving t he a ssembled FlexC a rt ..................................................................... 2-9

Unpacking and opening the sample organizer ........................................ 2-10

Installing the binary solvent manager ...................................................... 2-10

Installing the sample manager .................................................................... 2-11

Installing the detector ................................................................................... 2-12

Plumbing the system ...................................................................................... 2-13 In st a lla t ion recomm enda t ions for fitt ings .................................................... 2-13

P lumbin g a TU V det ector .............................................................................. 2-15 P lumbin g a P DA det ect or .............................................................................. 2-17

P lumbin g a n E LS detector ............................................................................ 2-17

P lumbing the bina ry solvent ma na ger a nd sam ple ma na ger . . .. . .. . .. . .. . .. . .. . . 2-18

Rout ing system dra ina ge th rough the FlexCa rt .. . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. 2-25

Inst a lling th e column ..................................................................................... 2-27

Conn ect ing to t he solvent supply .................................................................. 2-34

Making Ethernet and signal connections ................................................. 2-37 E t hern et connections ..................................................................................... 2-37

B ina ry solvent ma na ger I/O signa l conn ect ors ............................................ 2-38

S a mple ma na ger I/O sign a l conn ect ors ........................................................ 2-40

TU V detector I /O sign a l conn ect ors .............................................................. 2-41

P D A det ect or signa l conn ect ors .................................................................... 2-42 E LS det ect or signa l conn ectors ..................................................................... 2-42



Table of Contents xi

S igna l conn ections ......................................................................................... 2-42

Connecting to the electricity source .......................................................... 2-45

Calibrating the XYZ mechanism using the teach block ......................... 2-47

3 Preparing System Hardware ............................................................... 3-1

Powering-on the system .................................................................................. 3-2

Monitoring startup tests ................................................................................. 3-2

Monitoring system instrument LEDs ........................................................... 3-3 P ower LE D ....................................................................................................... 3-3

S t a t us LE Ds ..................................................................................................... 3-3

Preparing the binary solvent manager ....................................................... 3-5 P erforming a seal w a sh prime ... .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . . 3-5

P riming t he bina ry solvent ma na ger .. . .. . . .. . . .. . . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . . .. . 3-8

P riming a dry bina ry solvent ma na ger.. . . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . 3-8 P riming a w etted bina ry solvent m a na ger .. . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . . . 3-10

Preparing the sample manager ................................................................... 3-14 Selecting wea k wa sh a nd st rong wa sh solvents .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . . 3-14

P rimin g th e sa mple ma na ger ........................................................................ 3-16

Wa shin g th e sa mple ma na ger needle ........................................................... 3-18

Ch a ra cterizing th e needle sea l...................................................................... 3-20

Ch a ra cterizing th e needle a nd sa mple loop volumes................................... 3-21

Loading sa mple plat es in th e sam ple ma na ger.. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. 3-22

Select ing th e opt imum sa mple inject ion mode............................................. 3-23

Inst a lling t he sam ple ma na ger sha de ... .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . 3-27

Preparing the sample organizer ................................................................. 3-29 In itia tin g comm unica t ions ............................................................................ 3-29

Loa ding sa mple pla tes ................................................................................... 3-30

D ispla ying sa mple plat e informa tion ... . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . 3-34

Preparing the detector .................................................................................. 3-35 P repa rin g t he TU V det ector .......................................................................... 3-35



xii Table of Contents

Conditioning the column .............................................................................. 3-38

Shutting down the system ............................................................................ 3-39

B etw een a na lyses ........................................................................................... 3-39 S hut t ing dow n for fewer th a n 72 hours ........................................................ 3-40

S hut t ing down for more th a n 72 hours ......................................................... 3-40

4 Configuring System Software ............................................................. 4-1

Configuring Empower ..................................................................................... 4-2 S t a rt ing E mpow er a nd logging in ................................................................... 4-2

S electing sys t em inst rum ent s ......................................................................... 4-2

About th e binar y solvent ma na ger cont rol pa nel.. . . . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . 4-4

About th e sa mple ma na ger cont rol pa nel ....................................................... 4-7

About t he TU V det ect or contr ol pa nel ............................................................ 4-9

Starting the ACQUITY UPLC Console from Empower .......................... 4-11

Configuring MassLynx software ................................................................. 4-12

Starting the ACQUITY UPLC Console from MassLynx software ........ 4-13

5 Verifying System Operation ................................................................ 5-1

Preparing the system ....................................................................................... 5-2

Creating the test methods ............................................................................... 5-5 C rea ting th e inst rument meth od ... .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. 5-5

Cr ea tin g th e sa mple set method ..................................................................... 5-8

Performing the gradient performance test ................................................ 5-9

6 Maintaining the System ........................................................................ 6-1

Maintenance schedule ..................................................................................... 6-2

Contacting Waters technical service ............................................................ 6-5

Maintenance considerations .......................................................................... 6-6 S a fety an d ha ndlin g ......................................................................................... 6-6

P roper opera tin g procedures ........................................................................... 6-6



Table of Contents xiii

Configuring maintenance warnings ............................................................. 6-7

Maintaining the binary solvent manager ................................................... 6-8

Replacing th e mixer ......................................................................................... 6-9 Repla cing t he check va lve ............................................................................. 6-10

Repla cing solvent filters ................................................................................ 6-14

Cleaning t he air filters in the bina ry solvent ma na ger door .. . .. . . .. . . . .. . . .. . . .. . 6-14

Replacing th e a ir filters in the bina ry solvent ma na ger door .. . .. . . .. . . . .. . . .. . . . 6-15

Removing an d repla cing th e solvent ma na ger head sea ls.. . . . . .. . . . .. . . .. . . .. . . .. . . 6-16

Replacing t he solvent ma na ger plungers .. . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . 6-20

Repla cing t he vent valve car tr idge ............................................................... 6-24

Replacing th e fuses ........................................................................................ 6-27

Clea ning th e inst rum ent ’s exterior ............................................................... 6-28

Maintaining the sample manager ............................................................... 6-29 Defrost ing th e sa mple compa rt ment ............................................................ 6-29

Repla cing t he sa mple needle a ssembly ........................................................ 6-30

Ca libra t ing the needle Z-a xis ........................................................................ 6-40

Ch a ra cterizing th e needle sea l...................................................................... 6-41

Ch a ra cterizing th e needle a nd sa mple loop volumes................................... 6-42

Repla cing t he punct ur e needle holder .......................................................... 6-43

Replacing t he sa mple loop............................................................................. 6-45

Repla cing t he sa mple syr inge ....................................................................... 6-49

Repla cing t he w a sh syrin ges ......................................................................... 6-53 Modifying sa mple syringe configura t ion pa ra meters .. . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. 6-56

Repla cing t he injection valve car tr idge ........................................................ 6-56

Testing t he sam ple ma na ger backpressure regulat or .. . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . 6-59



Maintaining the column heater ................................................................... 6-61 Repla cing t he filter in the colum n in-line filter unit ................................... 6-61

Maintaining the TUV detector ..................................................................... 6-64 Flus hing th e flow cell..................................................................................... 6-65

P urg ing th e flow cell ...................................................................................... 6-67

Reverse-flush ing t he flow cell ....................................................................... 6-68

Replacing t he flow cell ................................................................................... 6-68



xiv Table of Contents

Repla cing t he la mp ........................................................................................ 6-73



Maintaining the sample organizer .............................................................. 6-77 Defrost ing the sa mple orga nizer ................................................................... 6-77

Clea ning th e a ir filter .................................................................................... 6-77



7 Diagnostics and Troubleshooting ...................................................... 7-1

Troubleshooting with Connections Insight ................................................ 7-2

Proper operating procedures ......................................................................... 7-4 Cont a cting Wa t ers technical service............................................................... 7-4

Locat ing syst em seria l nu mbers ..................................................................... 7-5

G enera t ing a service profile report ................................................................. 7-5 Resett ing system inst rument s to their initia l sta te .. . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. 7-6

Reviewin g logs t o ident ify problems ............................................................... 7-6

Monitoring performance from the ACQUITY UPLC Console ................. 7-8 Det ermin ing syst em st a t us ............................................................................. 7-9

Monitoring da ta w ith rea l-tim e plots .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. 7-10

Exa mining sm a ller or la rger segments of da ta plots .. . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. 7-17 Modifying plot d ispla ys .................................................................................. 7-18

Cr ea tin g log ent ries ....................................................................................... 7-18

Displa ying the sta tus of bina ry solvent ma na ger components . . .. . .. . .. . .. . .. . .. 7-20

Modifying binar y solvent ma na ger component sett ings .. . .. . . .. . . .. . . . .. . . .. . . .. . . .. 7-21

Displa ying the sta tu s of sa mple ma na ger component s .. . .. . . .. . . .. . . .. . . .. . . . .. . . .. . 7-22

Modifying sa mple ma na ger component sett ings .......................................... 7-23

Displaying t he sta tu s of sam ple orga nizer component s .. . .. . . .. . . .. . . . .. . . .. . . .. . . .. 7-24

Modifying sa mple orga nizer component sett ings ......................................... 7-25

Dis play ing th e st a t us of TU V det ect or component s..................................... 7-27

Modifying TU V det ector component set tin gs ............................................... 7-28

Displa ying t he sta tu s of P DA detector component s .. . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . 7-29

Modifying P DA det ect or component sett ings ............................................... 7-30

Displaying t he sta tu s of ELS detector component s .. . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . 7-31



Table of Contents xv

Modifying ELS det ector component set tin gs ................................................ 7-32

Monitoring column history ........................................................................... 7-33

Reviewin g column hist ory ............................................................................. 7-34 F inding column ma nufa ctu ring informa tion... . . . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . . .. . . .. . 7-35

Det ermin ing column sta tu s ........................................................................... 7-36

Solving baseline noise and drift problems ................................................ 7-38

Binary solvent manager troubleshooting ................................................. 7-43 P ower LED ..................................................................................................... 7-43

Flow LE D ....................................................................................................... 7-43

Ala rm messa ges ............................................................................................. 7-47

P erforming t he st a t ic decay test ................................................................... 7-54

P erforming th e dyn a mic leak test ................................................................. 7-55

Repa iring leaks .............................................................................................. 7-56

Display ing the binar y solvent ma na ger’s rear pa nel int erface connections 7-57

Ch a ngin g th e rea r pa nel int erfa ce conn ect ions............................................ 7-59

Sample manager troubleshooting ............................................................... 7-60 P ower LED ..................................................................................................... 7-60

Run LE D ......................................................................................................... 7-60

Ala rm messa ges ............................................................................................. 7-62

P erforming th e sa mple syr inge leak test ...................................................... 7-84

P erforming t he w a sh s yringe lea k t est . . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . .. . . 7-85 P erforming th e needle sea l leak test ............................................................ 7-85

Repa iring leaks .............................................................................................. 7-86

Displaying t he sam ple ma na ger’s rear pan el interfa ce connections .. . .. . . .. . . 7-86

Ch a ngin g rea r pa nel int erfa ce conn ect ions .................................................. 7-88

Sample organizer troubleshooting ............................................................. 7-89 P ower LED ..................................................................................................... 7-89

Run LE D ......................................................................................................... 7-89

Ala rm messa ges ............................................................................................. 7-92

TUV detector troubleshooting ................................................................... 7-102 P ower LE D ................................................................................................... 7-102

La mp LE D .................................................................................................... 7-102

Clea ring bubbles from th e flow cell ............................................................ 7-102



xvi Table of Contents

Ala rm messa ges ........................................................................................... 7-105

D ispla ying TU V detector rear pa nel int erfa ce conn ect ions....................... 7-111

Ch a ngin g th e rea r pa nel int erfa ce conn ect ions.......................................... 7-113

PDA detector troubleshooting ................................................................... 7-114 P ower LE D ................................................................................................... 7-114

La mp LE D .................................................................................................... 7-114

Ala rm messa ges ........................................................................................... 7-118

Displa ying P DA detector rear pan el int erface connect ions .. . .. . . . .. . . .. . . .. . . .. . 7-125

Ch a ngin g th e rea r pa nel int erfa ce conn ect ions.......................................... 7-127

ELS detector troubleshooting .................................................................... 7-128 P ower LE D ................................................................................................... 7-128

La mp LE D .................................................................................................... 7-128

Ala rm messa ges ........................................................................................... 7-129

Displaying E LS det ector rear pa nel int erface connections .. . .. . . .. . . .. . . .. . . .. . . 7-138

Ch a ngin g th e rea r pa nel int erfa ce conn ect ions.......................................... 7-140Chromatography troubleshooting ............................................................ 7-141

A Safety Advisories .................................................................................. A-1

Warning symbols ............................................................................................... A-2 Ta sk-specific ha za rd w a rnin gs ........................................................................ A-2

Wa rnings tha t a pply to part icular instruments, instrum ent components, a ndsa mple t ypes............................................................................................... A-3

Caution symbol .................................................................................................. A-6

Warnings that apply to all Waters instruments ......................................... A-7

Electrical and handling symbols ................................................................. A-14

E lectr ical s ym bols .......................................................................................... A-14 Ha ndlin g symbols .......................................................................................... A-15



Table of Contents xvii

B Specifications ........................................................................................ B-1

Binary solvent manager specifications ...................................................... B-2

Sample manager specifications .................................................................... B-7

Column heater specifications ..................................................................... B-11

TUV detector specifications ........................................................................ B-13

Optional sample organizer specifications ............................................... B-17

eCord column chip specifications .............................................................. B-19

C Spare Parts ............................................................................................ C-1

ACQUITY UPLC system part numbers ....................................................... C-2

Binary solvent manager spare parts ........................................................... C-2

Sample manager spare parts ......................................................................... C-4TUV detector spare parts ............................................................................... C-5

Optional sample organizer spare parts ...................................................... C-6

D Solvent Considerations ....................................................................... D-1

Introduction ...................................................................................................... D-2

P revent ing cont a mina tion ............................................................................... D -2 Clea n solvents .................................................................................................. D -2

S olvent q ua lity ................................................................................................. D -2

S olvent prepa ra t ion ......................................................................................... D -2

Wa t er ................................................................................................................ D -3

Solvent recommendations ............................................................................. D-4 G enera l solvent gu idelines .............................................................................. D -4

Solvent s to use ................................................................................................. D -4

Solvent s to a void .............................................................................................. D -6

ACQ U ITY U P LC sys tem recommend a tions ................................................... D -7

B ina ry solvent ma na ger recomm enda tions .. . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. . . .. . . .. . . .. . . . .. D-8

S a mple ma na ger recomm enda tions ................................................................ D -8

Detector recommenda tions .............................................................................. D -8



xviii Table of Contents

Solvent miscibility ........................................................................................... D-9 U sing m iscibility n um bers (M-nu mbers )...................................................... D -10

Solvent stabilizers ......................................................................................... D-10Solvent viscosity ............................................................................................. D-11

Wavelength selection .................................................................................... D-11 U V cut offs for common solvents .................................................................... D -11

Mixed mobile pha ses...................................................................................... D -12

Mobile pha se a bsorba nce............................................................................... D -13

Index ..................................................................................................... Index-1



1-1

1 System Overview

This section describes the components and features of the ACQUITYU P L C ™ s ys t em .

Contents:

Topic PageInst ruments, component s, a nd da t a systems 1-2

U P LC syst em guidelines 1-4

B ina ry solvent ma na ger 1-5

Sa mple ma na ger 1-7

C olumn hea ter 1-8

Opt iona l sa mple orga n izer 1-9

Opt ica l det ect ors 1-10

D a t a systems 1-11

C olumns 1-12

F lexCa rt 1-13

For more informa t ion 1-13



1-2 System Overview

Instruments, components, and data systems

ACQU ITY UP LC syst ems include a binary solvent ma na ger, sam ple mana ger,column hea ter, detectors (t una ble ultr a violet, photodiode a rra y, orevaporat ive light scat tering), an d a s pecia lized ACQU ITY UP LC column .

Sma ll-part icle chemistr ies a s ut i lized in U P LC system chroma tographygenera te na rrow peaks. To mainta in these narr ow pea ks, extraba ndsprea ding m ust be controlled by low er detector cell volume, minimizedtubing volumes, a nd specialized fitt ings. Narr ow pea k widt hs ca n sometimesrequire higher da ta ra tes. The TU V, P DA, a nd E LS det ect ors ca n sa mple up to80 da ta points per second.

The bina ry solvent m a na ger a nd injector can produce pressures up t o103421 kP a (1034 ba r, 15000 psi) a nd can generat e high-pressure gra dientsw ith m inima l gra dient delay. The upper limit of the flow ra te ra nge is 2mL/min .

The sa mple ma na ger ca n a ccommodat e tw o plat es in a microti ter plat e forma t

or 2-mL vials in full-height plat e forma t. An optional sa mple orga nizerincrea ses the ca pacity of the syst em to a s ma ny a s 22 microtit er pla tes (21 inth e sample orga nizer an d one in th e sa mple ma na ger), or eight via l racks(seven in t he sam ple organ izer a nd one in t he sa mple ma na ger).

Waters® Empower™ chroma tography sof twa re or MassLynx™ ma ss

spectr ometry softw a re controls the systems.



Instruments, components, and data systems 1-3

Waters ACQUITY UPLC System

Solvent tray

Detector (optional)

Column heater

Sample manager

Binary solvent manager

Sample organizer(optional)



1-4 System Overview

UPLC system guidelines

Tip: ACU ITY UP LC system guidelines dif fer from st an da rd H P LC practices.

• When per forming fas t a na lyses , note tha t a peak of in terest can be asna rrow a s 0.5 second. Wa ters r ecommends a sa mpling ra te of 25 to 50points a cross th e peak, wh ich provides good qua nt ita tion an d pea krepresenta tion. S am pling ra tes fa ster t ha n 20 points per pea k yieldhigher ba seline noise and filter t ime consta nt s should be a djustedaccordingly.

• The opt ima l ACQUITY UP LC f low ra t e d if fer s from tha t o f a typ ica lHPLC column. The table below offer operating guidelines for ACQUITYU P LC columns under both isocra tic and gr a dient conditions. Note tha tth e guidelines a ssume sma ll-molecule an a lyt es. Also note tha t t hey a rea pproxima tions an d th a t optimum performa nce for your molecule orsepara tion ca n occur a t a different flow ra te a nd/or pressure.

ACQUITY UPLC Columns Calculator

The ACQU ITY U P LC column s calcula tor estima tes t he plat e count (N) of a nisocra tic separ a tion or th e peak capa city (Pc) of a gra dient separa tion based onyour current H P LC conditions. It th en offers y ou a choice of one or moreACQU ITY UP LC column s th a t can provide increa sed resolving power in the

sa me a mount of time or simila r resolving power in less tim e. Thechroma togra phic conditions provided a re a st a rt ing point a nd ca n be furth eroptim ized ba sed on your par ticular req uirements. After you insta ll theACQU ITY UP LC softw a re, you can f ind the insta l ler for th e ACQU ITY UP LCColumns C a lcula tor in th e Ut ilities folder of the destina tion comput er: forexa mple, c :\empow er\instruments\ util it ies or c:\progra m fi les\w at ersinstruments\util i t ies.

Optimal flow rates for molecular weight range

Column size Molecular weight Flow rate

2.1 × 50 mL <500 600 µL/min

2.1 × 50 mL 1000 300 µL/min

2.1 × 50 mL 1500 150 µL/min

2.1 × 50 mL 2000 100 µL/min



Binary solvent manager 1-5

Binary solvent manager

The binar y solvent ma na ger is a high -pressure pump t ha t m oves solvent

th rough th e system. It provides steady (pulse-free) solvent flow a t a na lyticalflow ra tes. The binar y solvent ma na ger delivers solvent a t flow ra tes of 1mL /min a t 103421 kP a (1034 ba r, 15000 psi) a nd up t o 2 mL /min a t redu cedpressur es t o 62053 kP a (621 bar , 9000 psi). The solvent ma na ger ca n pum ptwo solvents simultaneously.

Pressure flow envelope



1-6 System Overview

How the binary solvent manager works

Ea ch of the solvent ma na ger’s tw o independent pump syst ems, A (left) and B(right ), conta ins tw o linear-drive actua tors (left a nd right ). Ea ch left a nd righta ctu a tor pa ir comprises a single reciprocat ing “serial” pump th a t deliversprecise flow of a single solvent . The tw o pump sys tem s combine th eir tw osolvents a t a filter/tee mixer. From t here, the solvent mixtur e flows t o thesample mana ger .

The chroma togra phy softw a re cont rols th e tw o solvents’ mixing ra tio byva rying t he flow of pump A relat ive to th a t of pump B. A pressure tra nsducer

in each pump head relays pressure data to the solvent ma na ger, whosefirmw a re measu res pump head pressures during t he pumping cycle. Thus t hesolvent m a na ger independent ly pre-compresses the solvent s in both th e A a ndB port ions t o ensure consist ent solvent d elivery a nd m inimize pump-induceddetector ba seline distur ba nces.



Sample manager 1-7

Sample manager

The ACQU ITY U P LC sam ple man a ger injects the sa mples i t dra w s from

microtit er pla tes or vials into th e chroma togra phic flow str eam . A locat ingmecha nism uses a probe to access sam ple locat ions a nd dra w sa mple fromth em. In t he needle overfill loa d-a hea d mode, the sa mple man a ger ca nperform a n inject ion in a pproxima tely 15 seconds . The first inject ion req uiresa dditiona l overhead time.

The sa mple man a ger a ccepts st a nda rd footprint plat es, 5.03 ±0.02 inch ×3.365 ±0.02 inch, th a t conform to ANSI sta nda rds (ma ximum height = 2.2inches, including covers). You m a y progra m a ny combina tion of these plat esa nd via l holders for a ut omat ed sam ple processing. Sa mples are loa ded int oth e sam ple ma na ger via th e front d oor or the optiona l sam ple orga nizer, whichtra nsfers sa mples ba ck a nd forth betw een t he tw o instrum ents. The sam plema na ger ca n m a inta in sa mples a t a ny t empera tu re betw een 4º to 40º C (39.2ºto 104º F ) in 25º C (77º F) or less a mb ient cond it ions.

How sample flows

When t he default mode, pa rt ia l loop with needle overfill , is requested, t hesa mple man a ger needle ca rria ge moves t o the specified w ell locat ion a nddra w s in a n a ir ga p. A st a inless steel puncture needle pierces the w ell covera nd lowers int o the w ell. The sa mple needle emerges from w ithin th e punctur eneedle a nd protrudes into the sa mple, a nd dra ws in a sam ple volume equa l to

the specified in ject ion volum e plus 3.0 µL (2.0 µL pr e-sa mple volum e a nd 1.0µL post-sa mple volume). The sa mple syr inge cont inues t o pull the sa mplea liquot th rough the sa mple needle and t hrough th e injection va lve unt il th epre-sa mple an d sa mple injection volume passes t hrough th e injection va lve.The va lve actua tes, sw itching t he sa mple loop to the load position. The sa mpleis pushed ba ck tow a rd t he needle and t he sam ple volume is then pushed intoth e sa mple loop. The sa mple loop moves to t he injection position a nd t he

sa mple is carr ied by the pump t o the column.



1-8 System Overview

Column heater

The column h eat er is of a m odular design a nd its footprint is ident ica l to tha t

of the sa mple man a ger. Thus i t a tt a ches to the top of the sam ple man a ger andserves as t ha t inst rum ent’s top cover. The column h eat er’s front compa rt mentca n a ccommodat e any Wa ters column up to 4.6 mm ID a nd 150 mm long. Thecolumn rests in a U -sha ped t ra y t ha t swivels outwa rd t o receive the columnfrom either side.

To reduce dispersion a ssocia ted w ith dead volume a nd m inimize the lengt h oftubing betw een system instruments, the column tra y swings outw a rd to anyposition betw een 0° a nd 180° . In t he 0° , “home”, position, the column tr a y isdirectly a bove the sa mple ma na ger a nd connected to the optica l detector (ontop of the column h eat er). In t he 180°, “a w a y”, position, the column hea ter canbe plumbed into a ma ss spectr ometer (loca ted on t he syst em’s right ).

The column hea ter hea ts t he column compa rt ment t o a ny t empera tu re from 5ºC (9º F) ab ove am bient t o 65º C (149º F). A film element ins ula ted to minim ize

power consumption a nd facil ita te therma l sta bil ity is a tt a ched to the tra y a ndproduces hea t. A pa ssive column st a bilizer, inside the tr a y, reduces sensitivityto am bient t emperat ure swings an d minimizes bandspreading.

A recepta cle on t he column hea ter’s r ight side receives t he column's eCord™chip. The eCord colum n chip stores column informa t ion wh ich ca n be a ccessedfrom the ACQU ITY UP LC Console.

The column hea ter drip tra y captures a ny leaka ge, routing i t to the sa mple

ma na ger via a drip tube.



Optional sample organizer 1-9

Optional sample organizer

The optional sa mple orga nizer stores microtiter or vial plat es and t ra nsfers

them t o a nd from t he sample mana ger, a utoma ting th eir processing andincreasing thr oughput.

The sa mple orga nizer’s st ora ge shelf compart ment can hold a selection ofANSI pla tes. Sa mple plates a re loa ded into the orga nizer t hrough a large,sw ing-open front door. The shelf compa rt ment is t herma lly conditioned bysa mple organ izer hea ter/coolers tha t, t ogeth er with th e sam ple ma na gerhea t er/cooler, cont rol t he t empera t ur e betw een 4º a nd 40º C (39.2º a nd 104º F)in 25º C (77º F ) or less a mbien t cond it ions.

Three suba ssemblies move pla tes w ithin th e sa mple orga nizer: th e Z-Dr ive,the sa mple orga nizer t ra nsfer shut tle (Y-a xis), a nd t he sa mple ma na gertr a nsfer sh ut t le (X-a xis). The Z-Dr ive moves th e Y-a xis to th e ta rget s helf,w here t he Y-a xis picks t he plat e. Then t he Z-D rive moves th e Y-a xis to t hesa me eleva tion a s t he X-a xis. The Y-a xis shut tles t he plat e into th e X-a xis,

w hich tra nsfers th e plat e int o th e sam ple ma na ger for processing. When th esam ple ma na ger f inishes with the plat e, the Y-axis pulls i t back into thesa mple orga nizer. The process is reversed to retur n t he plat e to th e shelf itca me from.



1-10 System Overview

Optical detectors

The syst em can be configured w ith a n TU V, PD A, or ELS optical det ector or

a ny combina tion of the th ree.

TUV detector

The TU V (tu na ble ultra violet ) opt ical det ect or is a t w o-cha nn el,ult ra violet/visible (U V/Vis) a bsorb a nce detector design ed for use in t heACQU ITY U P LC syst em. The detector, controlled by E mpower softw a re for

LC a pplica tions or Ma ssLy nx softw a re for LC /MS a pplica tions, operat es as a nintegral part of the system.

The detector offers t w o flow cell options. The a na lyt ica l flow cell, wit h avolume of 500 nanoliters a nd a path length of 10 mm, a nd t he high sensitivityflow cell, wit h a volume of 2.4 microliters a nd a 25 mm pa th length, bothutilize th e Wa ters pat ented light -guiding flow cell technology.

The TU V detector opera tes a t w a velengt hs ra nging from 190 to 700 nm.

PDA detector

The P DA (photodiode ar ra y) optical det ect or is a n ult ra violet/visible light(U V/Vis) spect rophotometer t ha t opera tes betw een 190 an d 500 nm .

The detector offers t w o flow cell options. The a na lyt ica l flow cell, w ith a

volume of 500 nanoliters a nd a path length of 10 mm, a nd t he high sensitivityflow cell, wit h a volume 2.4 microliters a nd a 25 mm pa th lengt h, both u tilizeth e Wa ters pat ented light -guiding flow cell t echnology.

ELS detector

The ACQU ITY UP LC E LS d etector is an eva pora tive light s ca tt ering detector

designed for use in the ACQU ITY U P LC syst em. This detector can becontrolled by Empower or Ma ssLy nx softw a re.

The detector incorporates a flow-type nebulizer that is optimized forACQU ITY U P LC system performa nce.



Data systems 1-11

Median baseline filter

The media n ba seline filter is int ended to decrea se th e effects of gradientsepara tions on the chromat ographic baseline. While the filter is a va ilable forth e TU V, P DA, a nd EL S detectors, it ha s most a pplica bility in the a bsorba ncedetectors. The media n bas eline filter enha nces t he a bsorba nce detector’ssta bility by decrea sing its curva tur e, ma king the development of integra tionmeth ods easier.

See also: ACQU ITY U P LC Console online help.

Data systems

The syst em ca n run un der Em pow er or Ma ssLyn x softw a re control.

Empower software

Em pow er provides a gra phica l, icon-ba sed user interfa ce tha t a cquires,

processes, ma na ges, report s, an d stores chroma togra phic da ta . It su pport sWindows® 2000 and Windows XP a nd t heir multit a sking opera tions, allowingyou to simulta neously open ma ny w indows. Windows multit a sking lets y ouview real-time da ta a cquisit ion a s E mpow er produces summa ry r esults ofpreviously a cquired dat a or refines integra tion para meters for a previousinjection.

The base version of Em pow er support s da ta from TU V, P DA, and E LS

detectors, and single quadrupole mass spectrometers. Popular softwareoptions for ACQU ITY UP LC sy stem users include Syst em Suita bility ,Chemica l St ructures, and Method Va lida tion Ma na ger.

See also: Em power Softwar e Gett i ng Star ted Gui de .

MassLynx software

Ma ssLy nx is a high -performa nce ma ss spectrometry a pplica tion tha t a cquires,a na lyzes, mana ges, a nd distr ibutes UV and ma ss spectrometry dat a . I t offersintell igent instrum ent contr ol a nd can a cquire nominal m a ss, exact m a ss,MS/MS, a nd exa ct ma ss MS /MS da ta .

See also: M assL ynx Gett i ng Start ed Gu id e .




Columns

ACQU ITY UP LC columns a re pa cked with 1.7-µm, bridged, eth ylsiloxa ne,

hybrid pa rt icles t ha t can m echa nically endure high -pressure conditions. Thecolumn ha rdwa re an d t he ma tched outlet tu bing ca n w ithsta nd up t o 103421kP a (1034 ba r, 15000 psi). The column dimens ions a llow optima lMS-compa tible flow ra tes, a nd m a tched out let t ubing minimizes t he effect ofextra -column volume.

Although the system w orks with a ny a na lytica l HP LC column, speciallydesigned ACQU ITY U P LC columns m a ximize its high-pressure ca pabilities.

Compared wit h t ra ditiona l HP LC columns, ACQU ITY U P LC columns deliversuperior resolution an d sensitivity in th e same run t ime or equiva lentresolution, great er sensitivity, a nd fa ster run times.

eCord column chip

ACQU ITY UP LC columns include an eCord column chip th a t t ra cks the usa gehistory of the column. The eCord column chip intera cts w ith t he syst emsoftw a re, recording informa tion for up to 50 sam ple qu eues run on t he column.In regula ted environments, t he eCord column chip provides document a tion ofth e column u sed in t he valida tion method.

In a ddition to the var iable column us a ge dat a , the eCord column chip alsostores fixed column manufacturing data, including:

• U n iq ue colum n id en t ifica t ion

• C ert ifica t e of a na lysis

• QC test da t a

Once the eCord column chip is a tt a ched t o the recepta cle on t he columnheat er, informa tion is a ut oma tically recorded by the syst em. No user a ction isrequired. This informa tion is stored out side of the E mpower da ta ba se.



FlexCart 1-13

FlexCart

The optional FlexCa rt provides the ACQU ITY UP LC syst em a mobile

pla tform. It ca n hold the system instruments a s well a s the P C a nd monitora nd provides electr ical outlets for system inst ruments a nd integra ted w a stema na gement. U sed with a ma ss spectrometer, the ca rt ’s adjusta ble height letsyou position t he column outlet close t o the inlet probe, minimizing sy stemdead volume.

For more information

See also:

ACQUITY UPLC Syst em Qu i ck St ar t Gui de (par t nu mb er 71500082503)

• ACQUITY UP LC S ys tem B ookshel f (par t number 71500082521)

• ACQU I TY UPLC Photodiode Ar ray D etector Gett i ng Star ted Gu id e

(pa rt nu mber 71500108703)• ACQU I TY UPLC Evaporat i ve L ight Scatter in g Detector Gett i ng

Star ted Guide (pa rt nu mber 71500109303)

• AC QU I TY U P L C C on sole on lin e H elp

• AC QU I TY U P L C S ys tem relea s e n ot es

• Contr ol l in g Contami nat ion in L C/ M S Systems (par t

num ber 715001307) on the ACQU ITY U P LC Sy stem B ookshelf CD



2-1

2 System Setup

Contents:

Topic Page

B efore you begin 2-2Assembling t he F lexCa rt 2-3

U npa cking a nd opening t he sa mple orga nizer 2-10

Inst a lling t he bina ry solvent ma na ger 2-10

Inst a lling t he sa mple ma na ger 2-11

Inst a lling t he det ect or 2-12

P lumbing t he syst em 2-13

Ma king E t hernet a nd signa l connect ions 2-37

C onnect ing t o the elect r icit y source 2-45

C a libra t ing t he XYZ mecha nism using t he t ea ch block 2-47



2-2 System Setup

Before you begin

Requirement: To inst a ll the syst em, you should generally kn ow how to set up

a nd opera te labora tory inst rum ents a nd comput er-cont rolled devices an d howto ha ndle solvents.

B efore insta l ling the syst em, ensure tha t

• it i s n ot s itua ted un d er a h ea t i n g or coolin g ven t

• t h e r eq u ir ed com pon en t s a r e pr es en t

• none of the sh ipping conta iners or unpacked items a re damaged

Recommendation: I f y our syst em includes th e optiona l FlexCa rt , t he systemma y be pla ced on t he ca rt , eith er before or aft er it is as sembled. B eca use thesys tem w eighs a t lea st 140.6 kg (310 pounds), Wa t ers recomm ends you pla ceth e system on th e ca rt before you as semble it.

If you discover an y da ma ge or discrepan cy wh en you inspect th e contents ofth e ca rt ons, immedia tely conta ct the shipping agent a nd your local Wa tersrepresentative.

If you a re loca ted in t he US A or C an a da , report ma lfunctions or otherproblems to Wa t ers Technica l S ervice (800 252-4752). I f you a re locat eda nyw here else, phone Wa ters’ corpora te hea dqu a rt ers in Milford,Ma ssa chuset ts (U SA), or conta ct your loca l Wa ters s ubsidiar y. Our Web siteincludes phone num bers a nd e-ma il a ddresses for Wa ters loca tions w orldwide.G o to ww w .wa ters.com, a nd click About Wa ters > Worldw ide Offices.

For complete informa tion on report ing shipping da ma ges an d submitt ingclaims, see Wat er s L i censes, War r an ti es, and Su ppor t Ser vi ces .

Warning:

• To a void ba ck injuries, two or more people must unpack thesa mple orga nizer a nd t ra nsfer it t o its fina l position. Also, if onlyone person is to insta ll th e sample man a ger, bina ry solventma na ger, or other syst em instr ument , he or she should do sousing a mecha nica l l if t .

• To a void overhea ting, and t o provide clea ra nce for ca bleconn ections, m a ke sur e th ere is a t leas t 15.24 cm (6 inches) of

clea ra nce at the rear of the system.• To ma inta in proper dra inage and lea k control, the system must be

w ithin 1° of ground level.

T l



Assembling the FlexCart 2-3

Tools

You w ill need the follow ing tools a nd ma teria ls to inst a ll the Wa tersACQUI TY U P LC™ system.

• 5/16-inch open-end wrench (2)

• 5/8-in ch open -en d w r en ch


• P h illips®

screwdr iver

• Sma ll mirror

• T25 TORX® driver

Assembling the FlexCart

If y our syst em includes th e optional F lexCa rt , follow th e procedure below tounpack and a ssemble it .

Recommendation: You ma y set t he assem bled syst em on th e ca rt , or place itsinstruments a nd components on the ca rt a nd t hen assemble them. B eca useth e ass embled syst em weigh s a t lea st 140.6 kg (310 poun ds), you shouldchoose th e lat ter option.

To assemble the FlexCart:

1. Remove the contents f rom the box pa ckaged in the FlexCar t .Tip: The box cont a ins th e monit or arm , the bolts u sed to at ta ch themonitor a rm to the ba se, and a n I B M instruction book for converting themonitor from st a nd-a lone to a rm-mounted.

FlexCart components



2-4 System Setup

FlexCart components

TP02496

Top tray

Powerconnectors (2)

Monitor mountingbracket

Shelf mountingholes

Lock knobs (2)

Height adjusting

screw

T25 TORX screws

2 Remove the four P hi ll ips screws from the support pla te a t the rear of the




2. Remove the four P hi ll ips screws f rom the support pla te a t the rear of themonitor, an d mount t he monitor ont o th e mova ble a rm a ssembly. Referto th e IB M instr uction book, if necessa ry.

Movable monitor arm assembly components

3. Loosen the tw o T25 TORX screws, at the bott om left-ha nd side of theca rt , tha t secure the bra cket for the CP U .

4. Atta ch the power and v ideo cables to the CP U , and place i t in posi t ion onth e bott om shelf of th e ca rt .

5. Route the monitor ’s power a nd v ideo cables through the plast ic channelprovided, a nd plug th em into the monitor. Refer t o th e IB M instr uctionbook, if necessa ry .

Movable arm

bracket

Screws (4)

Line cord Video cable

Plastic channel

6 Inser t the keyboard shelf ’s tw o ca ptive thumbscrews into slots on the



2-6 System Setup

6. Inser t the keyboard shelf s tw o ca pt ive thumbscrews into s lots on theca rt ’s front panel a t a level tha t a f fords comforta ble a nd sa fe opera tion.Turn th e t hum bscrews 1/4-tu rn clockwise t o lock th e posit ion of th eshelf.

Location of keyboard shelf support screws

Adjusting the FlexCart’s height

To adjust the cart’s height:

1. Loosen the side lock knobs before raising or lowering th e top portion ofthe car t .

2. Remove the crank ha ndle f rom i ts s torage brackets on the lowerright -ha nd side of the ca rt .

TP02499

Keyboard shelf support screws

3. Inser t the crank handle into the bot tom front o f the car t , a nd turn it t o




3. Inser t the crank hand le into the bot tom fron t o f the ca r t , a nd turn it t ora ise or low er the cart .

Crank handle

Tip: If your syst em includes a m a ss spectr ometer, a nd it is positioned tothe r ight of the system sta ck, set t he ca rt t o a height t ha t m inimizes thelength of tubing needed betw een t he instruments sta cked on t he ca rta nd th e ma ss spectrometer.

4. Tighten the side lock knobs a f ter reaching the desired height .

5. Loosen t he keyboard shel f’s tw o capt ive thumbscrews on t he car t ’s f ront

panel.6. Move the keyboard t o a level tha t provides comfor ta ble and sa fe

opera t ion. Turn th e t hum bscrews 1/4-tu rn clockwise t o lock th e posit ionof the shelf.

Crank handle

Locking the FlexCart in place



2-8 System Setup

Locking the FlexCart in place

To lock the FlexCart in place:

Lock th e ca rt by depressing t he lock peda l loca ted a t t he front of th e ca rt .

To release the FlexCart floor lock brake:

Relea se the ca rt by depressing the bra ke relea se bar loca ted a t t he front of thecar t .

Floor lock brake

Lock pedal

Release bar

Moving the assembled FlexCart




g

Once it is a ssembled, you ca n move the ca rt t o other a reas of a la b to minimizetubing runs between the ACQU ITY U P LC instrum ents and a m a ss

spectr ometer. U se the lip on t op of th e ca rt to pull it.

Pulling the cart using the top lip

Caution:

• To a void spills, remove all solvent reservoirs from t he solvent tra ybefore moving t he cart .

• To avoid str iking low doorwa ys—part icular ly those wi th thresholdsbenea th th em—low er th e ca rt fully before moving it.

• To avoid toppl ing the instruments s t acked on t he car t , do not m ovethe cart by pushing on t hem.

Unpacking and opening the sample organizer



2-10 System Setup

p g p g p g

The optional sa mple orga nizer is ava ilable for situa tions requiring increased

th roughput capa city . If your syst em includes the sam ple organ izer, a Wa tersservice technicia n unpa cks a nd inst a lls it.

Installing the binary solvent manager

To install the binary solvent manager:

1. Li ft the b inary solvent ma nager on to the bench top .

Alternative: If your syst em includes the optional FlexCa rt , lift t hebinary solvent ma na ger onto the ca rt .

2. I f your system includes the opt iona l sample organizer , unscrew theexisting 7/8-inch high feet from t he bott om of the bina ry solventma na ger, an d replace them w ith t he 1/4-inch high feet from t he sta rt up

kit .3. Atta ch the two sel f-adhesive rubber pads from the sta r tup ki t to the two

front feet on the binary solvent ma na ger.

4. P lace the binary solvent mana ger onto the sample organizer 's ba seplate.

Warning: To avoid ba ck injuries, t w o or m ore people must unpa ckthe sa mple orga nizer a nd t ra nsfer i t to i ts f inal position.

Warning: To avoid injury, Wa ters recommends t ha t tw o people liftthe bina ry solvent ma na ger.

Installing the sample manager



Installing the sample manager 2-11

g p g

To install the sample manager:

1. U npack the sample ma nager , and place i t on top of the binary solventmanager .

Alternative: If your sy stem includes the optiona l sam ple organ izer, placethe sa mple ma na ger on t he sam ple orga nizer shelf.

2. P lace the 8-por t netw ork swi tch on the bench, beside the system.

3. Open the f ront a ccess door of the sample mana ger , and remove the foa mblock from th e sam ple compart ment .

4. Open both t he sample mana ger f lu id ics t r ay and the b inary solvent

ma na ger door t o ensure tha t the drip man a gement system is properlyaligned.

Tip: P EE K™ f it t ings, sample syr inges, and wa sh syr inges inside thesa mple man a ger ma y ha ve loosened during sh ipping. To prevent leaks,ensure a l l P EE K fi t t ings, sample syringes, and w a sh syringes a re t ight .

Warning: To avoid injury, Wa ters recommends t ha t tw o people lift t he

sample mana ger .

Installing the detector



2-12 System Setup

To install the detector:

1. P lace the detector a t op the column heater , ensur ing that the feet areproperly positioned in th e indenta tions of th e column hea ter. This a lignsth e detector's drip t ra y over the dra in routing h ole, on t he top left side ofth e column h eat er.

Proper placement for drip management system

2. P lace the solvent t r ay a top the detector .

TP02465

Indentation

Guides for feetplacement

Drain routing hole for drip

management system

Plumbing the system



Plumbing the system 2-13

When a ll the component s a re st a cked, ma ke the plumbing connections.

Compression f it t ings a nd ferrules a re a lrea dy f i t t ed to tubing a ssemblies, butth ey must be properly set.

Installation recommendations for fittingsThe sy st em uses gold-plat ed compression screw s a nd t w o-piece ferru les. Seethe dia gra m below for a ssembly orienta tion.

Compression screw ferrule assembly

Recommendations:

• To prevent bandspread ing, ensure the tube i s fu lly bot tomed in thefitt ing h ole before t ight ening t he compression screw .

• For easier accessibi li ty , use long compression screws to a t t ach tubes toth e injector a nd vent va lve.

Caution: To prevent contamination, wear particle-free, powder-free,non-lat ex gloves wh en plumbing t he syst em.

TubingCompression screw

Ferrule with

locking ring

When tight ening syst em fittings, consult th e follow ing ta ble.



2-14 System Setup

Installation recommendations for ACQUITY UPLC fittings

Fitting Recommended tightening1/4-28 fla ngeless w it h ferrule S nug plus 1/4-t urn

10-32 L T135 P E E K wit h fer rule S nug plus 1/4-t ur n; if lea kin g,

t ight en a noth er 1/8-t urn

10-32 one-piece P E E K Finger-t ight

St a inless steel wit h 2-piece sta inlessst eel ferrule (first use)

Fin ger-t ight plus 3/4-tu rn w ithwrench

St a inless steel with 2-piece sta inlessst eel ferr ule (re-inst a lled)

Fin ger-t ight plus 1/4-tu rn w ithwrench

Reusa ble finger-t igh t (first use) S nug plus 1/4-t urn

Installation recommendations for ACQUITY UPLC fittings (Continued)




Plumbing a TUV detector

P lumbing a TU V detector involves connecting t he flow cell an d inst a lling a

ba ckpressure regulat or.Alth ough t he in-line degas ser removes most of the ga s (a ir) from solvents,some ga s is una voida bly introduced into the syst em during pa rt ial loopinjections. U nder pressure, this ga s rema ins in solution. H owever, because th epost-column pressure is norma lly mu ch low er t ha n th e pre-column pressure,th e gas m a y come out of solut ion a nd produce an un sta ble ba selinecha ra cterized by lar ge, unexpected spikes. The ba ckpressure regulat or

ma int a ins a min imum post -column pressur e of 1724 kP a (17 ba r, 250 psi),elimina ting post-column outga ssing a nd ensuring a smooth ba seline.

Tip: When the ba ckpressure regulat or is insta l led, the system ma inta ins a tlea st 1724 kP a (17 ba r, 250 psi) ba ckpressure, regard less of th e out let tu bingconfigura tion a nd flow ra te, provided tha t t here is some positive flow .

To plumb a TUV detector:

1. Open the front pa nel door of the TU V detector , and insta l l the f low cellso tha t t he three thumbscrews a lign w ith t heir holes in th e bulkhead.

Requirement: En sure th e dust cover ha s been removed from th ebulkhead before you inst a ll the flow cell.

R eusa b le f in ger -t i gh t (r e-in s t a lled ) Sn ug pl us 1/4-tur n; i f lea k in g,t ight en a noth er 1/8-t urn

Caution: To prevent contamination, wear particle-free, powder-free,non-lat ex gloves w hen plumbing th e detector.

Fitting Recommended tightening

Collet

removal tool

2. F in ger -t i gh ten th e th umbscr ew s .



2-16 System Setup

TUV detector flow cell

3. Remove the protect ive cover f rom t he PE EK cel l in let tubing, andconnect th e tu bing t o the flow cell inlet.

Flow cell

assembly

Outlet tubing

Inlet tubing

Thumb screw

4. Atta ch the shor t length of out let t ubing f rom the backpressure regula torto t he outlet of th e flow cell.




Rule: Do not insta ll the backpressure regula tor if you a re connecting t o a

second detector or m a ss spectr ometer.

Backpressure regulator

5. Route the long end of the out let tubing f rom t he backpressure regula tor ,th rough th e cha nnel clips a long th e front right side of th e system, a ndinto the closed wa ste ma na gement t ra y on the binary solvent m a na ger.

6. Open the f ront panel door of the TU V detector .

Plumbing a PDA detector

If your syst em includes a P DA detector, see the ACQUI TY UPLC Photod iode

Ar r ay Detector Gett i ng Star ted Gui de for informa tion on plumbing it.

Plumbing an ELS detector

If your syst em includes an E LS d etector, see th e ACQU ITY U PLC

Evaporat ive L igh t Scatter i ng Detector Gett i ng Star ted Gui de for informa tionon plumbing it .

Warning: To a void spills, empty th e wa st e conta iner at regular

intervals.

From detector

outlet

To waste

Plumbing the binary solvent manager and sample manager



2-18 System Setup

To plumb the binary solvent manager and sample manager:

1. Route the solvent in let tubing a t ta ched to the in-l ine degasser and sealw a sh pump through the channel cl ips of the sa mple ma na ger.

2. Continue rout ing the lines betw een the column heat er door a nd hinge,th rough th e detector's clip, a nd t hen place th e lines into the solvent t ra y.

Routing solvent inlet lines

Exception: If your syst em uses a m a ss spectr ometer positioned to th eright of the syst em sta ck, ensure tha t th e column heat er door is in t he“a w a y” (sw ung t o the right ) position. Then route t he lines in front of thecolumn heat er door, not betw een t he door a nd h inge.

Caution: To prevent contamination, wear particle-free, powder-free,

non-lat ex gloves w hen plumbing th e bina ry solvent m a na ger andsample mana ger .

Clip

Solvent lines

3. Remove the tw o protective ca ps from the ora nge and w hite color codedw a sh l ines.




4. Remove the tw o protective screws from the corresponding wa sh ports on

th e in-line degass er of the binar y solvent ma na ger.5. From the sample manager, route the weak a nd s t rong need le wash l ines ,

indica ted by t he ora nge an d w hite la bels, to their corresponding ports onth e in-line degass er in th e binar y solvent ma na ger. Finger tight en theknurled nut .

6. Loca t e the pre-insta l led Tygon®

t ubing running from t he process wa ste

por t , a nd the P harMed

®

tu bing runn ing from th e needle-clean syst emw a ste port (found on th e low er drip dray of the sa mple ma na ger), a ndroute them t hrough th e pa ss-th rough of the upper bina ry solventma nager dr ip tray.

Pre-installed Tygon and PharMed tubing

TP02590

Process waste fromsample manager

(Tygon)

Needle clean systemdrainage waste

(PharMed)

Drip tray

7. Connect the P harMed tubing to the f ront barbed fi t t ing and the Tygon toth e front boss fitt ing, both of wh ich a re on t he low er binary solvent

d i t



2-20 System Setup

ma nager dr ip tray.

Location of various tubings and binary solvent manager mixer

8. Route the sea l wa sh and solvent in let l ine tubing a t ta ched to the in-l inedegas ser through th e chan nel clips of both t he bina ry solvent ma na geran d sample mana ger .

9. Continue rout ing the l ines betw een the column heat er door a nd hingea nd t hrough t he detector’s clips. Then place the lines in th e solvent t ra y.

10. Remove th e protective O-ring from the sta inless steel pump out let

tubing.

TP02589

Tygon tubing from

process waste

From backpressureregulator outlet

Mixer

PharMed tubing from

needle clean system

waste

11. Sea t t he end of the tubing w ith t he shorter compression screw into thebina ry solvent ma na ger mixer outlet, a nd tight en the compressionfit t ing us ing t he 1/4 inch a nd 5/8 inch open end w ren ches




fit t ing us ing t he 1/4-inch a nd 5/8-inch open-end w ren ches.

Tip: When us ing new fitt ings, t ight en 3/4-t urn beyond finger-t ight . Forolder or previously u sed fitt ings, t ight en 1/4-t urn beyond fing er-tigh t .

Needle wash and pump outlet connections

12. Route the other end of the tubing th rough t he cha nnel cl ips to thesa mple ma na ger injection valve.

TP02480

Strong needle wash

Weak needle wash

Binary solvent

manager mixer outlet

13. Remove the O-ring, and seat the end of the tubing with the longercompression screw int o port 5 on th e injection va lve. Tight en t hecompr ession fit t ing u sin g t he 1/4-inch open-end w ren ch



2-22 System Setup

compr ession fit t ing u sin g t he 1/4 inch open end w ren ch.

Injection valve connections

14. Remove th e protective cover from t he inject or out let tube, and t hen seatth e tube a nd ferrule int o port 6 of th e injection va lve. Tighten th ecompr ession fit t ing u sin g a 1/4-inch open-end w ren ch.

Pump outlet tubing

into port 5

Injector

outlet/columnstabilizer tubing

into port 6

15. Slide a w ast e line over the ba rbed dra in f it t ing loca ted at the bott om ofthe binary solvent m a na ger, a nd route i t to a suita ble wa ste conta iner.




Attaching the waste line to the binary solvent manager

16. Route the degasser vent l ine to a suita ble wa ste conta iner .

Caution: To a void distorting t he drip tra y or ca using th e dra in cup

to lea k, restra in the dra in cup when a tt a ching or removing thewa ste l ine.

Warning: To avoid relea sing solvent va pors int o the room, r oute th edegass er vent line

• to a fume hood or other su ita b le exhaust sys tem.

• to a suita ble wa ste conta iner , ensur ing the tubing's d ischarge end

is a t a ll times a bove the fluid level.

Warning: To a void spills, empty th e wa st e conta iner at regularintervals.

TP02479

Drip tray

Barbed drain fitting

Waste line

Drain cup

Degasser vent line

Caution: To avoid fluid backup, you must ensure proper dra ina ge ofwas te :



2-24 System Setup

Correct positioning of waste and degasser vent lines

• P l ace the wa s te conta iner below the sys tem s tack .

• Ensure tha t the was te and degasser ven t l ines do not c rimp orbend. A crimp or bend m a y impede flow to th e wa st e conta iner.

• Ensure the ex it o f the was te and degasser ven t l ines i s notimmersed in wa ste solvent . If necessa ry, shorten each line so th a tno port ion of it dr ops below th e top of th e w a ste cont a iner (see nextfigure).

TP02709

Waste line

Degasservent line

Waste line

Degasser

vent line

Correct Incorrect

17. Atta ch a wa ste l ine to the barbed f it t ing on t he f il ter dra inage assembly(on the lower front side of th e sam ple organ izer), a nd rout e it to asuita ble w a ste conta iner.




Sample organizer drain

18. Route a wa ste l ine from th e barbed fi t t ing on the rear of the solvent t ra y

to a suita ble wa ste conta iner.

19. Close the sa mple organ izer door, sample ma na ger door, sam ple mana gerfluidics tra y, a nd th e bina ry solvent ma na ger door.

Routing system drainage through the FlexCart

To route system drainage through the FlexCart:

1. P lace a sui ta ble waste conta iner in the bot t om r ight s ide of the car t .

2. Open the sam ple organizer , detector , sample mana ger , and binary

solvent manager doors.

3. Route the long end of the outlet tubing from the detector ’s backpressureregulat or, th rough th e cha nnel clips a long t he front right side of thesystem, a nd down through t he ca rt ’s top access hole to the wa stecontainer.


4. Route the wa ste and degasser vent l ines, loca t ed a t the front , bot tomright of the binary solvent ma na ger, dow n t hrough the cart ’s top a ccesshole to the w a ste conta iner.



2-26 System Setup

Waste and degasser vent lines through top access hole

5. Route the wa ste line, loca ted a t the f ront , bot t om r ight of the sampleorgan izer, down t hrough t he ca rt ’s top access hole to the w a stecontainer.

6. Close the sam ple organizer , detector , sample mana ger , and binarysolvent manager doors.


Access hole

Waste line

Degasser vent line

Installing the column

Ins ta lling t he column involves insta lling t he column st a bilizer assembly,column in line filter unit and the column itself




column in-line filter unit, and the column itself.

To install the column (optical detection):

1. Open the column heat er door , li ft t he meta l la t ch , and t i l t the columnhea ter t r a y downw ard .

Opening the column heater tray

2. Fold the thermal gasket f irmly around the column sta bil izer tubingblock.

Positioning the thermal gasket

Metal latchColumn heater tray

Column stabilizer

tubing blockThermal gasket

3. P osit ion the assembly block into the column heat er t ra y a s far t o the le fta s possible to a llow space for t he in-line filter a nd column.

Installing the column stabilizer tubing block



2-28 System Setup

Installing the column stabilizer tubing block

4. Ca refully route the column stabi lizer tubing through the groove on theright side of the column h eat er.

Column heater opening

Column stabilizer tubing block

5. Remove the pla st ic reta iner f rom th e inlet end of the column sta bil izertu bing, a nd t hen connect t he tu be to port 6 of th e injector.

Connecting the column stabilizer tubing assembly




Connecting the column stabilizer tubing assembly

6. Remove the column in-line fi l ter unit from its pla stic bag, a nd pla ce it ona clea n, f lat surface.

TP02575

o in-line filter unit

Injector port 6

7. P ick up the column in-l ine fi lter unit , a nd inser t t he male end of thecolumn sta bilizer tube a ssembly into th e filter unit ’s inlet side.Finger-tight en th e compression fitt ing unt il it is snug.



2-30 System Setup

Connecting the column in-line filter unit

8. Remove the O-ring from th e male end of the column in-line f il ter unit .

9. Remove the plug from the column in let .

10. P ush t he ma le end of the column in-line f il ter unit int o the column inletunt il it can go no fart her. Then t ighten t he hex nut using t he 1/4-incha nd 5/16-inch open-end w ren ches.

11. Sna p the plastic column clips onto the body of the column.

12. P lace the a ssembled component s (column, in-line filter unit, a nd columnsta bil izer tubing assembly) in the column tr a y. P ush down t he metal

lat ch a nd t i lt the t ra y t o the closed position.Tip: When using 100-mm ACQU ITY U P LC columns, y ou might need tomove th e assembled component s slightly t o th e right, in th e column t ra y,to a ccommoda te t he column in-line filter unit .

13. Remove the protective cover from the P EE K tubing from the f low cellinlet.

14. Att a ch the 0.0625-inch OD in let tubing included w ith t he flow cell to th ecolumn outlet. Ensu re the la bel on the flow cell inlet t ubing ma tches the

TP02544

Flow

Column inlet In-line filter unit Column stabilizer tubing assembly

Place wrenches here to connect the

in-line filter unit to the column inlet.

Reusable

finger-tight fitting

ty pe of detector (TU V or P D A) a nd flow cell (a na lyt ical or highsensitivity) in your syst em.

Connecting a column




Connecting a column

15. Close th e column heater door.

16. Check th e sea l ar ound t he front panel to ensure correct al ignment. Thisis cri t ica l to ensure that the t hermal environment is sta ble.

17. Att a ch the eCord column chip holder to the recepta cle on t he side of th ecolumn hea t er.

Connecting the eCord column chip

TP02717

Detector inlet

tubing

Sample manager

outlet tubing

Column stabilizer

tube

In-line filter unitColumn

eCord column chip

receptacle

eCord column

chip holder

To install the column (MS detection):

If your syst em includes a m a ss spectr ometer positioned to th e right of thesystem st a ck, swing t he column hea ter door t o the “aw a y” position (th a t is,



2-32 System Setup

sw ing it rightw a rd). In th is wa y, you connect to the ma ss spectr ometer’s inletwith out increa sing the tubing length . You must a lso insta l l the MS drip tra yto ensure a ny solvent leaked from th e column or column connections is r outedinto the drip mana gement syst em.

1. Open th e column hea te r door .

2. Remove any solvent lines tha t a re routed betw een the column heaterdoor a nd hinge.

3. P ush down on the meta l c lip a t the bot tom, le ft-hand side of the columnhea ter t o release th e door, an d th en pull the door t ow a rd you. The doorswings from left t o r ight. S wing i t fully t o the r ight.

Metal clip on left side of column heater

TP02596

Metal clip

Column heater door swung to right




4. P ull of f the rear cover , and then t i lt the column t r ay downw ard .

Column heater door with rear cover removed and column tray open

5. Sna p the plast ic column cl ips onto the body of the column.

6. Atta ch the column inlet to the sta inless s teel tubing f rom por t 6 of the

inject ion va lve.7. Atta ch the mass spectrometer inlet tubing included wi th th e system to

th e column outlet.

8. Set the column into the column tra y. Ensure tha t the sta inless s teeltubing from the sa mple mana ger is routed th rough t he front notch onth e left side of the column t ra y.

Columnheater door

Rear cover

Columnheater door

Column tray open

9. Tilt t he column tray upward, to the closed posi t ion, and refi t the rearcover. The cover sn a ps int o place.

10. Att a ch the eCord column chip holder to the recepta cle on t he side of th ecolumn h eat er See th e figure tit led “C onnectin g t he eCord column chip”



2-34 System Setup

column h eat er. See th e figure tit led C onnectin g t he eCord column chipon page 2-31.

11. Att a ch t he column outlet t ubing to the ma ss spectrometer’s inlet .Consult t he inst rum ent’s document a tion for more informa tion aboutconnecting to t he inlet.

Connecting to the solvent supply

The solvent t ra y loca ted on t op of the syst em is capa ble of holding up t o tw oliters of spilled solvent . You w ill need a suita ble wa ste cont a iner to collect a nyspil l from the wa ste l ine at the rear of the tra y.

To connect the solvent supply:

1. Choose solvent reservoirs tha t snugly f it t he reservoir ca ps supplied inth e sta rt up kit. Wa ters r ecommends 1-L r eservoirs.

Caution: To prevent contamination, wear particle-free, powder-free,non-lat ex gloves w hen connecting to t he solvent supply.

Warning: To avoid spills, do not pla ce solvent r eservoirs a top th esample organizer.

Caution: To maint a in a dequat e solvent head pressure and ensureproper solvent delivery, position the solvent reservoirs in the solventtra y a t th e top of the system sta ck.

2. Remove the solvent f il ter f rom t he shor t piece of P TFE tubing.

Caution: Wea r pa rt icle-free, powder-free, non-la tex gloves w henha ndling th e solvent filter. Skin oils ca n cont a mina te th e solvent




Removing the solvent filter

3. Inser t the solvent tubing through the solvent reservoir cap.

4. Re-insta l l the solvent f i lter on the short piece of PTFE t ubing.

filter.

TP02484

Solvent filter

PTFE tubing

Solvent tubing

5. Inser t the f il tered end of the solvent tubing into the solvent bot t le , in thetr a y, on top of the sa mple ma na ger or optiona l detector.

Solvent tubing in bottles



2-36 System Setup

6. Repeat s t eps 2 through 5 for the remaining solvent supply l ines.

Solvent tray

Solvent tubing

Solvent bottles

Making Ethernet and signal connections

Recommendation: The ACQU ITY UP LC syst em workst a tion should bepow ered-on before pow ering-on th e instr um ent s.



Making Ethernet and signal connections 2-37

Ethernet connections

To make Ethernet connections:

1. Unpack and ins ta l l the preconf igured ACQUITY UP LC sys temworkstation.

2. Connect one end of one shielded Et hernet cable to the netw ork swi tch,a nd th en connect t he other end to the E thernet card on th e workstat ion.

Tip: On preconfigured syst ems, the Et hernet car d is ident ified as t heInst rument LAN ca rd.

3. Connect one end of one shielded Et hernet cable to the solvent ma na ger ,a nd t hen connect the other end to the netw ork sw itch.

4. Connect one end of one shielded Et hernet cable to the sam ple mana ger ,a nd t hen connect the other end to the netw ork sw itch.

5. Connect one end of one shielded Et hernet cable to the detector , an d thenconnect th e oth er end t o th e netw ork sw itch.

6. If you are using a sa mple orga nizer , connect one end of one shielded

E th ernet cable to th e sam ple organ izer, a nd t hen connect t he oth er endto the netw ork switch.

Binary solvent manager I/O signal connectors

The rear pa nel of the bina ry solvent ma na ger includes tw o removableconn ectors t ha t h old t he screw t ermin a ls for I/O signa l ca bles. These



2-38 System Setup

connectors a re keyed so th a t t hey can be insert ed only one w a y.

Binary solvent manager I/O signal connectors

See also: For elect rical specifica t ions, see the ta ble t i t led “Bina ry solventma na ger electrical specifica tions” on pag e B -3.

Binary solvent manager analog-out/event-in connections

Signal connection Description

Auxilia ry 1 In Reserved for future use.

Auxilia ry 2 In Reserved for future use.Run S topped Out I ndica t es (w it h a cont a ct closure) t he bina ry

solvent ma na ger ha s ceased opera tion beca use ofa n error condition or operat or request.

S w it ch 1 Out U sed t o send t ime-ba sed cont a ct closure signa lsto externa l devices.

0−2V An a log 1 Out An a log ch a rt out put fun ct ion a lit y.

1

2

3

4

5

6

7

89

10

11

12

+

−

+

−

+

−

Auxiliary 1 In

Auxiliary 1 In

Auxiliary 2 In

Auxiliary 2 In

Ground

Run Stopped Out

Run Stopped Out

Switch 1 OutSwitch 1 Out

Ground

0-2V Analog 1 Out

0-2V Analog 1 Out

Connector I

1

2

3

4

5

6

7

89

10

11

12

+

−

+

−

+

−

Start Gradient In

Start Gradient In

Stop Flow

Stop Flow

Ground

Switch 2 Out

Switch 2 Out

Switch 3 OutSwitch 3 Out

Ground

0-2V Analog 2 Out

0-2V Analog 2 Out

Connector II

G ra dient In Init ia tes the pumps to begin gra dient opera t ion

b h l l

Binary solvent manager analog-out/event-in connections (Continued)





by eith er cont a ct closure input or 0-volt input .S top F low In Allow s you t o st op the flow from t he bina ry

solvent ma na ger when a n error condition orha rdwa re fai lure occurs on an oth er instrument.



0−2V An a log 2 Ou t Ou tput s t he a n a log sig na l t o a device su ch a s a nint egra tor or st rip-char t recorder. You ca n selectone of th e follow ing signa ls a s th e cha rt outsignal :

• F low ra t e

• S yst em pressure• Composi t ion (%A, %B )

Chart-out signal conditions

Signal Parameter Setting at 0Volts (Minimum) Parameter Setting at2.000 Volts (Maximum)

Flow Ra t e 0.000 mL/min 2 mL/min

S yst em P ressure –345 kP a (–3.45 ba r,–50 psi)

103421 kP a (1034 ba r,15000 psi)

Composit ion 0.0% 100.0%

Sample manager I/O signal connectors

The rear pa nel of the sa mple ma na ger includes tw o removable connectors th a thold th e screw t ermin a ls for I/O signa l cables. These connectors a re keyed so

th t th i i l bl i t d l



2-40 System Setup

tha t they can receive a signal cable inserted only one wa y.Requirement: A cont a ct closure input connection from t he sa mple ma na ger isrequired t o trigger a ma ss spectr ometer, an ACQU ITY 2996 P DA detector, ora n ACQUI TY EL S detector running under Ma ssLynx softwa re contr ol tos t a r t .

Sample manager I/O signal connectors

See also: For elect rical specifica t ions, see the ta ble t i t led “Sa mple ma na gerelectrica l specifica tions” on pa ge B -7.

Sample manager analog-out/event-in connections

Signal connections Description

I nject S ta r t Ou t I ndica t es (w it h a con t a ct closu re) t ha t a n in ject ionhas s ta r ted .

Auxilia ry In Reserved for fut ure use.

Inject Hold In Input signa ls from ot her syst em inst rument s t odelay th e injection.

0−2V An a log 2 Ou t An a log ch a r t ou t pu t fu nct ion a lit y .

Connector I

1

2

3

4

5

6

7

8

9

10

+

−

+

−

+

−

+

−

Inject Start

Inject Start

Auxiliary In

Auxiliary In

Ground

Inject Hold In

Inject Hold In

Ground

0-2V Analog Out

0-2V Analog Out

1

2

3

4

5

6

7

8

9

10

Switch 1 Out

Switch 1 Out

Switch 2 Out

Switch 2 Out

Switch 3 Out

Switch 3 Out

Switch 4 Out

Switch 4 Out

Run Stopped Out

Run Stopped Out

Connector II

S wit ch 1 Out U sed t o send t ime-ba sed cont act closure signa ls to

external devices

Sample manager analog-out/event-in connections (Continued)





TUV detector I/O signal connectors

The rea r pa nel of t he TU V detector includes tw o remova ble connectors t ha thold th e screw t ermin a ls for I/O signa l cables. These connectors a re keyed sotha t t hey ca n be inserted only one wa y.

TUV detector I/O signal connectors

external devices.S wit ch 2 Out U sed t o send t ime-ba sed cont act closure signa ls to

external devices.

S wit ch 3 Out U sed t o send t ime-ba sed cont act closure signa ls toexternal devices.

S wit ch 4 Out U sed t o send t ime-ba sed cont act closure signa ls to

external devices.R un S t opped O ut I n dica t es (w i th a con t a ct clos ur e) t h a t t he s a m ple

ma na ger ceased opera tion due to an error condit ionor a n operat or request.

1

2

3

4

56

7

8

9

10

+

−

+

−

Analog 1 Out

Analog 1 Out

Ground

Analog 2 Out

Analog 2 OutSwitch 1 Out

Switch 1 Out

Ground

Switch 2 Out

Switch 2 Out

Connector I

1

2

3

4

56

7

8

9

10

+

−

+

−+

−

Inject Start In

Inject Start In

Ground

Lamp Off In

Lamp Off InChart Mark In

Chart Mark In

Ground

Auto Zero In

Auto Zero In

Connector II

See also: For elect rical specifica t ions, see the ta ble tit led “TU V detectorelectrica l specifica tions” on pa ge B-14.

TUV detector analog-out/event-in connections




2-42 System Setup

PDA detector signal connectors

If your syst em includes a P DA detector, see the ACQUI TY UPLC Photod iode

Ar r ay Detector Gett i ng Star ted Gui de for informa tion on signa l connectors.

ELS detector signal connectors

If your syst em includes an E LS d etector, see th e ACQU ITY U PLC

Evaporat ive L igh t Scatter i ng Detector Gett i ng Star ted Gui de for informa tionon signal connectors.

Signal connectionsRefer to the sign a l connection locat ion shown on t he silk-screened label on t herear pa nel of each instrument.

Requirement: To meet t he regulat ory requirements of immunit y fromexterna l electr ica l disturba nces, you must insta ll connection covers over th esigna l connectors.


Ana log 1 (Out ) Not used.


S w it ch 1 (Out ) Cont rolled by t hreshold a nd t imed event s.


In ject S t a r t (In) Not used.La mp Off (In) When t r iggered, it ext inguishes t he la mp.

C ha rt Ma rk (I n) When t riggered, ca uses bot h a na log out put cha n-nels and t he digita l da ta sent t o the dat a systemto increase t heir va lue for a period of time.

Aut o Zero (In) Ca lcula tes a n offset va lue t ha t , w hen a dded t othe sa mple signal , ma kes the resulting ba selinesigna l zero.

To make signal connections:

1. Atta ch the posit ive and negat ive leads of the s ignal cable to theconnector.

Attaching signal cable to the connector




Attaching signal cable to the connector

2. Slide the clam p (wit h the bend facing dow n) into the protective shield.

Signal cable

Connector

3. Inser t the clamp and shield (wi th th e bend facing down) into theconnection cover and loosely tighten with one self-tapping screw.

Clamp and shield in the connection cover



2-44 System Setup

4. Inser t the connector wi th t he s ignal cable into the connect ion cover a ndposition th e clam p over t he cable lea ds. Tight en t he cla mp int o pla cew ith th e second self-ta pping screw.

Clamp positioned over cable leads

Connection cover

Shield

Clamp

Cable leads

Clamp

5. P lace th e second connection cover over the first cover, and sna p it intoplace.

Signal connector with connection cover



Connecting to the electricity source 2-45

Connecting to the electricity source

Ea ch syst em instru ment requires a sepa ra te, grounded pow er source. Theground connection in a ll pow er outlets m ust be common a nd phy sica lly close tothe system.

To connect to the electricity source:

Recommendation: U se a line condit ioner a nd un interru ptible pow er supply(U P S) for optimum long-term input volta ge sta bility .

1. Connect t he female end of the power cord to the recepta cle on the rearpanel of each instrument .

2. Connect t he male end of the power cord to a suita ble wal l out let .

Alternative: If your syst em includes the optional FlexCa rt , connect t he

female end of t he F lexca rt 's electr ica l cables (included in t he st a rt up kit)to th e recepta cle on t he rear pa nel of the each inst rum ent. C onnect t hehooded, ma le end of the F lexcart 's electrica l cables to t he power st rips on

Signal connector

Connection cover

th e back of the cart . Fina lly, connect ea ch pow er strip's cable to a w a lloutlet operat ing on its own circuit.

FlexCart power connections

To circuit A



2-46 System Setup

To circuit A

To circuit B

1 m e t e r

2 me ters

1 met er

1 m e t e r

1 m e t e r

1 m e t e r

2 me t e r s

Univ er sal IE C c oupler

AC line

AC line

Network switch

Detector

Sample manager

Solvent manager

Sample

organizer

LCD/monitor

CPU

FlexCartpower strips

Calibrating the XYZ mechanism using the teach block

B efore performing a ny chromat ogra phy, you must verify tha t the XYZmecha nism positions t he needle properly in th e sa mple cha mber.

Tip: If your system ha s a sam ple orga nizer , you must insta l l the organizer



Calibrating the XYZ mechanism using the teach block 2-47

Tip: I f your system ha s a sam ple orga nizer , you must insta l l the organizerbefore ca librat ing t he XYZ mecha nism.

To calibrate the XYZ mechanism:

1. Open th e s a mple ma n a g er door .

2. Remove the sample p la te f rom the sample t r ay .

3. Loosen the screw holding the sample tray in place by turning it

counterclockwise one qu a rt er-tur n. Then slide the sa mple tra y out. I fyour syst em does not include a sa mple orga nizer, you must remove tw osa mple tra ys from the sa mple cha mber.

4. With your f inger , remove the teach block from t he sample mana gerchassis floor.

Warning: To avoid punctu re w ounds, keep ha nds or loose clot hingclear of the needle assembly mecha nism w hile it is moving. Note th a tth e sam ple ma na ger beeps three times w henever t he door is opena nd th e needle a ssembly mecha nism is about t o move.

5. Inser t the teach block into the locat ing holes .

Teach block in position



2-48 System Setup

6. In the ACQU ITY UP LC Console, select Sa mple Mana ger from thesystem t ree.

Teach block

in position

7. Cl ick Troubleshoot > Cal ibra t e XY and Zp axes.

Calibrate XY and Zp Axes dialog box




8. In t he Cal ibra t e XY and Zp Axes dia log box, cl ick Sta r t t o begin .

Result: A w a rning a ppea rs, prompting you to keep clear of the sa mplecha mber. Confirm t ha t a l l sa mple pla tes an d tra ys a re removed, andensure th e tea ch block is insta lled.

9. Cl ick OK. The sample ma nager beeps 3 t imes to indica t e the needlea ssembly mecha nism is about t o move, an d the XYZ mecha nismpositions the puncture needle above the teach block.

Positioning of puncture needlePuncture needle



2-50 System Setup

10. E xam ine the loca tion of the punctur e needle relat ive to the needle holein t he tea ch block.

11. Select t he 1.0 millimeter displacement increment, a nd th en click the +Z

butt on to position th e XYZ mecha nism foot close to the t op of t he t eachblock. E a ch click of th e mouse moves th e needle 1 millimet er.

12. Select t he 0.1 millimeter displa cement increment , an d t hen f ine tune t hea djust ment s o th a t t he XYZ mecha nism foot is just a bove the tea chblock, but not t ouching it .

13. To la tera lly position t he puncture n eedle, use the +X and −X but tons t omove it left or right , respectively.

Caution: To avoid t rigger ing t he t op-of-of pla t e sensor prema tu rely, do

not a llow th e mecha nism foot t o touch t he tea ch block.

Caution: To avoid dam a ging t he puncture needle, do not m ove it left orright w hile it is in th e teach block.

Teach block

Puncture needle

Needle hole

XYZ mechanism

foot

Alternative: You can press t he keyboa rd a rrow keys ins tea d of clickingth e on-screen butt ons to ad just t he needle posit ion. Doing so lets y oua djust th e position wit hout t a king your eyes off the needle.

Caution: Avoid holding down a n a rrow key, a s doing so storeskeystr okes a nd cau ses the needle to cont inue moving, even aft er youl it




14. P osition sma ll mirror so tha t you ca n see the loca tion of the punctureneedle, an d th en use th e +Y and −Y butt ons to a djust t he front-to-ba cklocat ion of th e punctu re needle so th a t it is cent ered over th e needlehole.

Determining the front-to-back position

15. U sing the 0.1 millimeter displa cement increment , low er the needle into

th e teach block hole to confirm a n a ccura te X an d Y alignment .

16. Click Ca librat e. A w a rning a ppears, indicat ing you must keep clea r ofthe sa mple cha mber.

relea se it.

17. Click OK. The puncture needle fully lowers into th e tea ch block tocalibra t e th e top-of-pla te sensor.

18. Click S a ve. When th e confirmat ion w indow a ppears, click Yes.

19. Remove th e teach block, pla ce it in t he stora ge locat ion in t he floor of th esa mple compart ment, a nd re-insert the sa mple tra ys.



2-52 System Setup

20. Click Mainta in > Ca librat e needle Z a xis.

Calibrate Needle Z Axis dialog box

21. In the C a librat e Needle Z Axis dialog box, cl ick St a rt , a nd t hen click OKin t he confirmat ion w indow .

22. U s e t h e +Z but ton to drive the sa mple needle down t o w ithin 1millimeter of the tra y surfa ce.

23. Sw itch t he displa cement increment to 0.1 millimeter , a nd low er thesa mple needle unt il it a lmost touches the surfa ce of th e sam ple tr a yholder.

Tip: To ea sily a nd efficient ly a chieve t he correct needle sett ing, slide a

business card under t he needle. Then lower t he needle until it touchesth e ca rd lightly but does not restr ict t he ca rd’s free movement .




Adjusting the sample needle

24. Click Sa ve. The confirmat ion w indow a ppears.

25. Cl ick Yes .

Sample needle almost

touching surface of sampletray holder or just touching

the business card



2-54 System Setup

3 Preparing System Hardware



3-1

Contents:

Topic Page

P ow ering-on t he system 3-2

Monit or ing st a r tup t est s 3-2

Monitor ing syst em inst rument LE D s 3-3

P repa ring the bina ry solvent ma na ger 3-5

P repa ring the sa mple ma na ger 3-14

P repa ring the sa mple orga nizer 3-29

P repa ring the det ect or 3-35C ondit ioning the column 3-38

Shut t ing dow n t he syst em 3-39

Powering-on the system

P owering-on t he system enta ils st a rting ea ch syst em instrument individually,a s well a s the ACQU ITY UP LC sy stem worksta tion a nd the Em power or

MassLy nx opera ting softw a re.

1. P ress the power swi tch on the top, lef t-hand side of each instrument ’s



3-2 Preparing System Hardware

p p,door. Ea ch syst em instrument beeps th ree times a nd runs a series ofsta r tup tests .

The pow er and st a tus L ED s change a s follows:

• E a ch system i ns t rumen t ’s pow e r LE D sh ow s s t ea d y g reen . Th e

binary solvent m a na ger’s f low LE D, th e sample ma na ger’s run LE D,a nd th e detector’s lam p LED a ll show r ed for a few seconds.

• Dur ing in it i a l iza t ion , each system ins t rument ’s power LED showsstea dy green. The bina ry solvent ma na ger’s flow L E D, th e sa mplema na ger’s run LE D, a nd th e detector ’s lamp LE D show fla shinggreen. Full initia lizat ion of th e system usua lly requires about sevenminutes.

• Af ter the instruments are successful ly powered-on, each one’s powerLED shows stea dy green. The bina ry solvent ma na ger’s f low LE Da nd th e sample man a ger’s run LE D a re unlit. The detector’s lam pLED shows st eady green, indica ting tha t t he la mp is ignited.

2. P ower-on the ACQUITY UP LC system worksta t ion .

3. S ta r t the Empower or MassLynx opera t ing sof tware . You can monitorthe ACQUITY UP LC Console for messages a nd LE D indica tions.

Monitoring startup tests

These sta rt up tests run w hen you pow er-on th e ACQU ITY UP LC sy stemworkstation:

• C P U boa rd

• Memory (RAM a nd ROM)

• E xt er na l com m un ica t ion s ys t em (E t h er n et )

• C lock

If th e sta rtup t ests indica te a ma lfunction, see C h a pt er 7.

Monitoring system instrument LEDs

Light emitting diodes on ea ch syst em instrument indica te t he instrument’ssta te of functioning. The LE Ds a re specific to th eir instrum ents, so th e

significa nce of th eir var ious colors a nd m odes can differ from one instru mentto another.



Monitoring system instrument LEDs 3-3

Power LED

The pow er LE D, on th e left-ha nd side of a n inst rument ’s front pa nel, indicat esth e pow er-on or power-off sta tus of the instr ument . This LE D is green w hen

pow er is on a nd un lit w hen pow er is off.Tip: To provide adequa te venti lat ion, th e sa mple ma na ger a nd sa mpleorgan izer fa ns a re alw a ys runn ing, even w ith t he power off. These fa ns sw itchoff only w hen t he pow er ca ble is removed from th e back of th e instrum ent.

Status LEDs

Flow LED (solvent manager)

The flow LE D, t o th e right of the pow er LE D on th e solvent ma na ger’s frontpanel, indica tes the f low sta tus. A steady green f low LED indica tes th at thereis a f low through the solvent ma na ger.

Run LED (sample manager and sample organizer)

The run LE D, t o the r ight of the pow er LED on t he sam ple ma na ger’s a ndsam ple orga nizer ’s front panel, indica tes t he run sta tus. A steady green runLED indica tes th a t injections are being run.

Lamp LED (detector)

The lam p LED , to the right of th ey pow er LE D on t he detector’s front pa nel,indica tes the lamp sta tus. A stead green la mp LED indica tes tha t t he la mp ison .

Status LED indications




LED mode and color Description

U nlit S olvent Ma na ger, S a mple Ma na ger, a nd S a mpleOrga nizer - Ind ica tes the instru ment is current lyidle.

Det ector - Indicat es th e detector lam p isextinguished.

S tea dy green S olvent Ma na ger - Indica tes the bina ry solventma na ger is flow ing.

Sa mple Ma na ger - Indica tes the sam ple mana geris opera ting norma lly, at tempt ing to complete

a ny outsta nding sa mples or diagnostic requests.When sa mple and diagnostic requests a refinished, th e LED reverts to the unlit m ode.

Det ector - Indicat es the detector lamp is ignited.

Sa mple Organ izer- Indica tes t he sa mple orga -nizer is operat ing norma lly, a tt empting t ocomplete a ny outst a nding sa mples or diag nostic

request s. When sa mple a nd dia gnostic request sa re f inished, the LE D reverts to the unlit m ode.

F la sh ing green S a mple Ma na ger - Indica tes t he syst em isw a iting for a t lea st one instrument to becomeopera ble. Detector lamp w a rm-up an d columntempera ture equil ibra tion times t ypica lly ca usesuch a dela y.

Det ector - In dica tes t he detector is initia lizing orcalibrating.

F la shing red Indica t es a n error ha s st opped t he inst rument .Informat ion regarding t he error tha t caused thefa ilure ca n be found in t he ACQU ITY UP LCConsole.

S tea dy red Indica t es a n inst rument fa ilure tha t prevent s

furt her opera tion. Pow er-off th e instr ument , andth en power-on. If the LED is still steady r ed,cont a ct your Wa ters s ervice representa tive.

Status LED indications (Continued)




Preparing the binary solvent manager 3-5

Preparing the binary solvent manager

For optimal performa nce of the ACQU ITY UP LC™ sys tem , you must prepare

th e solvent ma na ger for opera tion.

To prepa re the solvent m a na ger for opera tion, you mus t perform a seal w a shprime an d th en prime the solvent ma na ger.

Requirement: To maint a in the efficiency of th e bina ry solvent m a na ger, andto obta in a ccura te, reproducible chroma togra ms, use only MS-gra de solvent s,wa ter , a nd a dditives. For deta ils, see Appendix D .

Performing a seal wash prime

P rime the sea l wa sh in the binary solvent ma na ger to lubrica te the plungers,f il l the tubing pat hs w ith solvent, a nd f lush a wa y solvent a nd/or a nyprecipita ted sa lts th a t h a ve been dra gged past the plunger seals from t hehigh-pressure side of the piston chambers.

P rime the plunger seal w a sh under t hese conditions:

• Aft er us in g bu ffer ed m obile ph a se

• When the b inary solvent mana ger has been inac t ive for a few hours orlonger

Warning: Observe sa fe labora tory pra ctices when you han dlesolvents. S ee the Ma terial Sa fety Da ta Sheets for th e solvents y ouuse.

Caution: Ch loroform a nd m ethy lene chloride should not be used.

• Wh en t he bin a r y s olv en t ma n a g er is d ry

Tip: The seal w a sh w ill self-prime, but y ou ca n use a syringe t o hast en th eprocess.

Caution: To a void da ma ge to the solenoid va lve seat s a nd sea ls in thesolvent pa th , do not use a nonvola tile buffer as t he seal wa sh solvent.




Rule: To prevent cont a mina tion, do not recycle seal w a sh.

Recommendations:

• Sea l wa sh should conta in 10% organic solvent . This prevents microbia l

grow th a nd ensures tha t t he sea l wa sh can solubolize the mobile pha se.• B efore pr iming the plunger sea l s , ensure the volume of sea l wa sh is

a dequat e for priming.

See also: Contr ol l ing Contami nat ion in L C/ M S Systems (par tnumber 715001307) on the ACQU ITY U P LC Sy st em B ookshelf CD.

Required materials

• Tu bing a da pt er (st a rt up kit )

• 30-m L sy rin ge (st a r t up kit )

• Sea l w ash solut ion

To perform a seal wash prime:

1. Ensure the sea l wash inlet tube is in the solvent reservoir .

2. Remove the sea l wa sh out let tube from the r ight s ide of the dr ip tra y.

Binary solvent manager seal wash outlet tube




3. P ush the syr inge plunger fully into the syr inge barrel.

4. Connect the tubing ada pter to the syr inge, and then connect the syr ingea ssembly to the outlet tubing from t he sea l wa sh system.

5. In the ACQU ITY UP LC Console, select B inary Solvent Ma nager f rom

the system t ree.6. Cl ick Control > P r ime seal w ash, and t hen cl ick Yes to begin the sea l

w a sh priming process.

7. Slowly draw back on the syr inge plunger to pull sea l wash solventthrough the system.

8. When the sea l w ash solut ion begins to flow into the syr inge wi thout

ma jor air bubbles, disconnect t he tubing a nd reinsta ll it on th e fitt ing onthe dr ip tra y.

9. Click (St op f low ) to stop the priming process.

Drip tray

Seal wash

outlet tube

Priming the binary solvent manager

P riming is used to prepare a new syst em or binary solvent ma na ger for use,cha nge reservoirs or solvents, a nd run th e system a fter it ha s been idle formore tha n four hours. Dur ing priming, t he vent va lve moves to Vent position

to both ensure minima l backpressure a nd direct t he flow to w a ste. The flowra t e during pr iming is 4 mL /min for ea ch pump being primed.

Tip: If you are priming a dry binary solvent m a na ger using a syringe will




Tip: I f you are priming a dry binary solvent m a na ger, using a syringe willshorten t he t ime required to complete priming.

Recommendation: E nsure t he solvent r eservoirs h a ve enough solvent fora dequat e priming an d th e wa ste conta iner has suff icient ca pacity for usedsolvent . The prim ing flow ra te is 4 mL/min for ea ch pump, or 8 mL /min t ota l.For exam ple, priming both solvents for 5 minutes r equires a pproxima tely 20

mL of each solvent .

Priming a dry binary solvent manager

To prime a dry binary solvent manager:

1. Open the ins t rument ’s fron t door .

2. Loca t e the appropria te solvent ven t l ine.

3. In the ACQU ITY UP LC Console, select B inary Solvent Ma nager f romthe system t ree.

4. In the binary solvent ma nager informat ion window, cl ick Control >P rime A/B S olvent s. The P rime A/B Solvent s dia log box a ppear s.

5. In t he P rime A/B Solvents dialog box, select solvent A and/or B .

6. In t he Time box, specify the number of minutes from 0.1 thr ough 60.0.

Default: 1.0 minut e

Caution: To prevent sa lts from precipita ting in t he syst em, intr oducea n intermediate solvent, such as w a ter , wh en cha nging from buffers to

high-organ ic-content solvents. B e sur e to consult th e solventmiscibility ta bles on page D-9.

Warning: To a void spills, empty t he wa ste cont a iner at regularintervals.

Recommendations: P rime the bina ry solvent ma na ger unti l a steadyflow exits the vent tube (typically 7 to 10 minutes).

7. Cl ick St ar t . When solvent f lows out of the vent l ine wi thout bubbles, thepat h is primed.

8. Repeat s tep 3 through 7 to pr ime the other solvents .

Requirement: There mu st be solvent in t he A1, A2, B1, a nd B 2i f t h d t f ti tl




reservoirs for t he dega sser t o function correctly.

To prime a dry binary solvent manager using a syringe:

1. Open the ins t rument ’s fron t door .

2. Loca t e the appropria te solvent ven t l ine.

• I f you a re pr iming solvent A, follow t he s ta in less s teel ven t l ine tha tis la beled “A-VENT” from port 4 on th e vent va lve, an d lift it out ofthe drip tra y.

• I f you a re pr iming solvent B , follow t he s ta in less steel ven t l ine tha tis labeled “B -VE NT” from port 1 on t he vent va lve, a nd lift it out of

the drip tra y.

Binary solvent manager vent lines

3. P ush the syr inge plunger fully into the syr inge barrel.

4. Connect the tub ing adapter to the syr inge.

Drip tray

Vent valve

Solventvent lines

5. Connect the syr inge a ssembly to the shor t length of Pha rMed tubing,a nd t hen connect the short length of P ha rMed tubing to the solvent ventline you loca t ed in st ep 2.

6. In the ACQU ITY UP LC Console, select B inary Solvent Ma nager f rom

the system t ree.

7. In the binary solvent ma nager informat ion window, cl ick Control >P rime A/B S olvent s. The P rime A/B Solvent s dia log box a ppear s.




g pp

8. In t he Pr ime A/B Solvents dialog box, select solvent A1.

9. In t he Time box, specify the number of minutes from 0.1 thr ough 60.0.

Default: 1.0 minut eRecommendation: P rime the bina ry solvent ma na ger unti l a steadyflow exits th e vent t ube (ty pica lly 3 minut es).

10. C lick S t a r t .

11. Slowly dra w back on t he syringe plunger to pull solvent through thesolvent pa th . When solvent flow s out of th e vent line wit hout bubbles,

the pat h is primed.12. Remove the syringe from t he vent l ine, and reconnect the vent l ine to the

dr ip tra y.

13. Repeat steps 6 through 12 for solvent A2, B 1, a nd B 2.

Requirement: The reservoirs for solvent s A1, A2, B1, a nd B 2 must notbe empty . Otherw ise the degasser w ill not function correctly.

Priming a wetted binary solvent manager

Tw o functions h elp prepar e th e syst em for opera tion. The length of time t hesyst em ha s been idle determines w hich is the bett er.

• R efr esh sy st em (S ys P r ep)

• S t a r t up

Refreshing the system

U se the Refresh (Sy s P rep) function a fter th e system ha s been idle a shortperiod of time (a few hours t o overnight ) a nd w hen you plan to use the sa mesolvents t ha t y ou used previously.

You can invoke th e Sys P rep function from th e cont rol pa nel or by a dding it a sa l ine in a sam ple set .

Recommendations:

• P r ime the b inary solvent mana ger for 1 minute i f the sys tem has beenidle for 4 or more hours a nd you will use the solvents th a t a re alrea dy inthe system.

• P r ime the binary solvent mana ger for 4 minutes if you wi ll use new




P r ime the b inary solvent mana ger for 4 minutes if you wi ll use newsolvents tha t a re of the sa me composition of wha t is a lrea dy in t hesystem.

To refresh the system:

1. In t he ACQU ITY UP LC C onsole, cl ick Control > Refresh system (SysP rep). The Refresh S yst em (Sy s P rep) dialog box appears.

2. Review the sett ings and select a different option i f needed. The systemprimes y our curr ent solvent select ions (A1 or A2, B 1 or B 2).

• S olven t lin e A on ly (d efa u lt )

• Solvent line B only• B oth A a nd B

3. C lick OK.

Result: The syst em primes t he selected solvents, primes t he sa mplema na ger with one wea k wa sh prime (wa sh a nd sam ple syringes), andignites the la mp in the detector.

Starting up the system

U se the Sta rt up function to prime the bina ry solvent m a na ger af ter cha ngingth e mobile pha se, aft er cha ngin g t he sa mple needle a nd/or sam ple loop, ora fter t he syst em ha s been idle a long period of time (overnight or over aw eekend). B efore you begin this procedure, ensure t ha t th e syst em is correctlyconfigu red for us e.

Recommendation: P rime the binary solvent ma na ger for 5 minutes i f you a recha ngin g t o solvent s w hose compositions differ from t he compositions ofsolvents a lrea dy in t he system.

To start up the system:

1. In the ACQU ITY UP LC C onsole, cl ick Control > Sta r t up. The SystemSt a rtup dialog box appea rs.

2. In the P r ime Solvents ta b of the System Sta r tup dia log box, review thesett ings for t he A /B Solvent s. In t he A /B Solvents a rea , you ca n selector clear a ny or all of the solvent s: A1, A2, B1, or B 2. You ca n cha nge t helength of time to prime solvents A a nd B by entering a different n umber




in Du ra tion of P rime. All selected solvent s a re primed for t he sa meduration.

Allowed values: 0.1 to 60.0 minutes

Tip: I f you wa nt t o return settings t o their original values, on a ny t a b,click Set D efault s.

Defaults: Solvent s A1, A2, B 1, a nd B 2 prime for 1.0 minu t e ea ch.

3. You can select or clear pr iming of the sea l wa sh, s trong wa sh, weakw a sh, a nd/or sa mple syringe.

Default: The seal w a sh is primed for 1.0 minut e, the w eak w a sh once,

a nd th e sam ple syringe once.

4. You can change the number of cycles to pr ime the strong wa sh byentering a different w hole num ber in th e Cycles field.

Default: 10

5. Select the Equi l ibra te to Method ta b to review t he set t ings for the fina lflow r a te, mobile pha ses, composition, temperat ures, an d lam p sta te.

Cha nge the values as needed to ma tch your requirements a tequilibration.

Equilibrate to Method tab values

System startupparameters

Default Allowed values

Met hod init ia l flow ra t e 1 m L/m in 0.1 t o 2.0 m L/m inComposition of A a nd B(su m m us t be 100%)

A1, 100

B 1, 0%

A1, A2; 0 to 100%

B 1, B 2; 0 t o 100%

C olumn tempera t ure Off Off or 5.0 ° C (9 ° F) a bovea mbient to 65.0 ° C(149 ° F)

S a mple t empera t ure Off Off, or 4.0 t o 40.0 ° C (39.2t o 104 ° F) in 25 ° C (77 ° F)a mbient conditions

La mp On On or off

Equilibrate to Method tab values (Continued)

System startupparameters

Default Allowed values




6. I f you cha nged the sample needle, cl ick the Optional : Cha racter izeVolume ta b, and t hen select “C ha ra cterize seal” and “Cha ra cterize

needle a nd loop volumes.”7. I f you changed the sample loop, in the Optiona l : Cha racter ize Volume

ta b, select C ha ra cterize needle an d loop volumes.

8. If either the needle or loop is a different size, in the Configurat ion a rea,click Ch a nge. In th e Volume C onfigura tion dia log box, select th e newsize of loop a nd/or needle, a nd th en click OK .

9. C lick St a rt .Result: The lam p in th e optica l detector ignites, t he ACQU ITY U P LCsystem sets t he column sa mple tempera tures, a nd a ll priming sta rts.After priming finishes, th e sam ple ma na ger cha ra cterizes the needleseal if tha t fun ction is selected a nd t hen logs th e results of th echa ra cterizat ions into the dat a base. Finally, the system esta blishes themeth od flow ra te, solvent selections, a nd composition. The defau lt

sett ings for t he met hod init ia l flow a re 100% S olvent A1 at 1.000mL/min a nd 0% B 1, the column a nd sa mple temperat ures a re Off, an dth e detector lamp is ignited.

La mp On On or off

Preparing the sample manager

P repa re the sa mple ma na ger for opera tion a fter you prepa re the binarysolvent ma na ger. Preparing t he sam ple ma na ger involves t hese steps:

• P r iming

• C ha ra ct er izing the sea l

• Ch a r a cter iz in g th e n eed le a n d sa mple loop volumes




Ch a r a cter iz in g th e n eed le a n d sa mple loop volumes

• Loa ding sa mple pla tes

Selecting weak wash and strong wash solvents

For best performa nce, follow th ese guidelines w hen selecting w a sh solvent s.Oth erwise, performa nce ma y be reduced, specifica lly Area /H eight RS D a ndLinea rity . The guidelines do n ot prohibit a ll other solvent combinat ions,however. Other combina tions can be run w ith low er performa nce expecta tionsor by ma nipula ting default injection para meters.

U se a w eak w a sh solvent based on the sa mple a nd mobile pha se chemistr ies ofyour a pplicat ion, ma king sur e all solutions/buffers a re miscible a nd soluble.

Recommendation: F or buffered a qu eous, reversed-pha se chroma togra phic

conditions a nd MS a pplica tions, use a w eak w a sh solvent of 100% w a ter or 0to 25% met ha nol or a cet onitrile a nd a st rong w a sh s olvent of 50 to 100%meth a nol or a cetonitrile. High sa mple concent ra tions ma y require oth er wea kwa sh solvents. I f y our separa tion permits, Wa ters recommends a dding a sma lla mount of a n orga nic solvent (~ 10%) t o prevent m icrobia l growt h.

See also: “Water” on page D-3.

Warning: To avoid solvent spills a nd t o ma inta in proper leak

dra inage, a lwa ys close the sa mple ma na ger f luidics tr a y beforeopera ting t he system.

Caution: To a void da ma ge to the solenoid va lve seat s a nd sea ls in thesolvent pat h, do not use a nonvolat i le buffer as the w eak w a sh orstrong wa sh solvent.

Tip: For best performa nce, the w eak w a sh solvent sh ould be similar oridentica l to your isocra tic or initia l gra dient solvent conditions, excludingbuffers. Do not use salt buffers in wa sh solvents.

Wash solvent effects

Property Effect

Orga nic species As a genera l principle, st rong a nd w ea k solvent sshould include the sa me orga nic species Note t ha t



Preparing the sample manager 3-15

should include the sa me orga nic species. Note t ha tthis ma y not a lwa ys be pra ctica ble. You ma y,however, use a 100% organic st rong w a sh solvent.

Solv en t compos it ion Th e w e a k w a sh solv en t sh ou ld r eflect a s closely a spossible the sa me composition a s t he initia lgra dient mobile pha se.

pH Adjust t he pH of st rong a nd w ea k solvent s for bestpea k sha pe a nd carr yover performa nce.

Concent ra tion of strongsolvent

St rong solvent sh ould be no stronger tha n th econcent ra tion needed to reduce ca rry over to a n

a ccepta ble level.Solubili ty of a na lyteand sa mple

The an a lyt e and sa mple must be soluble in boththe wea k and strong wa sh solvents.

Caution: P roteins (in pla sma , for example) donot dissolve in solvents whose organic componentis grea t er th a n 40%.

S ample diluent The w ea k w a sh solvent ca n cont act t he sa mple, so

ma t ch th ese as closely a s possible. To offseta dverse effects on pea k sha pe ca used by t hema trix’s composition, a djust the w eak w a shcomposition w hen using t he instrument in part ialloop mode.

Wa sh volume ra tio(w eak t o str ong)

Within a meth od, th is should be a bout 3:1, w eakw a sh to strong–sufficient t o ensure the w eak w a sh

flushes th e strong from t he needle a nd sa mple loopbefore sa mpling.

Cycle t imes H igher viscosit y w a sh solvent s lengthen w a shcycles.

Priming the sample manager

The priming process fills the s a mple needle w ith solvent, flushes n ew solventth rough t he inject or lines, an d/or purges a ir from t he lines. You prime t hesa mple needle a nd/or sa mple syringe t o accomplish t hese ta sks:

• P r epa r in g a n ew s a m ple m a n a g er for oper a t ion

• P r e pa r i ng a s a mple ma n a g er for oper a t i on a f t er h a s b een i dle for a nextend ed period




• C h a ngin g t h e solven t in t he sy rin ges

• R em ovin g bu bb les fr om t he lin es

Guidelines: En sure tha t t he priming solvent is correctly composed an d th a t itis high in qua lity a nd miscible with a ny other solvents used in your syst em.U se filters in a ll solvent reservoirs, an d ensure th e volumes of solvents a resufficient for priming.

Requirement: The sa mple man a ger must be primed before you at tempt tocha ra cterize the seal .

To prime the sample manager:


Sample manager information window




2. C l ick Con t r ol > P r i me sy rin ges .

Alternative: Right -click in the Em pow er or Ma ssLy nx sa mple ma na gercontrol panel, an d th en click P rime syringes.

3. In the P r ime Syr inges d ia log box , select S ample syr inge and w ashsyringes.

Rule: If you wa nt only t o remove a ir bubbles from the sa mple syringe,but do not w an t to prime the w a sh syringes, select Sa mple syringe only.H owever, do not select th is option routinely. P riming a ll components a tth e sam e time is good pra ctice.

4. Type the number of primes in the Number of cycles text box.

Default: 1

Recommendation: Wa ters recommends 5 to 7 primes w hen you a rechanging solvents.

5. Cl ick OK to s ta r t pr iming. When the system sta tus is “ Id le ,” pr iming isfinished.

Tip: Ea ch prime ta kes approxima tely 2 to 4 minut es.

Caution: Do not a bort the sa mple mana ger priming sequence.Doing so ma y leave str ong solvent in th e sam ple needle which ca n




Washing the sample manager needleWa shing t he needle is a n optiona l procedure tha t flushes str ong a nd/or wea kw a sh solvent th rough t he needle an d injection port . Wa shing t he needleremoves cont a mina nt s from t he inside a nd outside of the needle, the extern a lpiercing n eedle, and th e injection port . You can a lso perform a needle w a sh t oa scerta in proper f low through t he wa ste t ubing a nd t o confirm tha t the needlewa sh syst em is primed a nd properly operat ing.

Rule: Do not use buffered solvents a s w a sh solvent s.

Tip: P riming t he system w a shes the sa mple needle, so w henever you primeth e system, you can omit t his procedure.

Observing wash solvent recommendations

Wa ters recomm ends th a t you observe these guidelines for w a shing t he needle:

• To ensure tha t the s t rong wash solvent i s complete ly removed , thesystem w a shes the needle with 500 µL of wea k wa sh solvent a f ter youuse strong w a sh solvent. You can increase, but not decrea se, th e defa ultva lue of 500 µL.

• Th e a n a l y tes a n d sa mple ma t r ix mus t be soluble in b oth w e a k a n dst rong solvents. P roteins do not dissolve in solvent s w hose orga nic

component is grea ter t ha n 40%. B uffers should not be used in a ny w a shsolvent.

Example: I f th e weak w a sh solvent is 30% a cetonitrile and 70% w a t er ,the st rong w a sh solvent should conta in a greater concentra tion ofa cetonitr i le in w a ter .

a dversely a ffect chromat ography.

• The weak wa sh solvent should be the same as the in it i a l elut ing solvent ,a nd t he strong wa sh solvent should a t lea st equa l the composition of thefina l eluting solvent.

• Use a w ea k w a sh solv en t b a sed on th e s a mple a n d mob ile ph a se

chemistr ies of your a pplica tion. Ma ke sure all solutions a re miscible andsoluble. For best r esults, w eak w a sh solvent should ma tch t he initialgradient conditions and mobile phase composition (isocratic). Highsa mple concentra tions can require additiona l wea k w a sh solvents.

F b f f d d h h h k h




• For buf fered aqueous , reversed-phase chromatography , use weak wa shsolvent consist ing of 100% w a t er or up t o 25% met ha nol or a cetonit rile.For st rong w a sh solvent , use 50 t o 100% met ha nol or a cet onitrile.

B efore you begin, ensure tha t t he solvents a re compat ible w ith y oura pplica tion, tha t their volumes are suff icient, a nd t ha t the w a ste reservoir islar ge enough to conta in the w a ste solvent.

To wash the sample manager needle:

1. In the ACQU ITY UP LC Console, select Sa mple Mana ger from thesystem t ree. The sam ple ma na ger informa tion window a ppears.

2. Cl ick Control > Wash Needle.

Alternative: Right -click in the Em pow er or Ma ssLy nx sa mple ma na gercont rol pan el, an d t hen click Wa sh N eedle. The Wa sh N eedle dialog boxappears.

3. In t he Strong Wash box, speci fy the volume for the strong wa sh solvent .

Range: 0.0 th roug h 99999 µLException: To omit st rong w a sh solvent , enter 0 in the S tr ong Wa sh box,or lea ve it blank.

Default: 0.0 µL

Recommendation: 100 th rough 500 µL

Tip: U sing both a wea k and st rong wa sh solvent increa ses the w a sh time

a nd solvent consumption beca use the sy stem m ust be fully clea nsed ofth e strong solvent before sta rt ing t he next injection.

4. In t he Weak Wash box, speci fy t he volume for the w ea k wa sh solvent .

Range: 1.0 th roug h 99999 µL

Default: 200.0 µL w ithout strong wa sh or 500 µL with s trong wa sh

Recommendation: 200 th rough 500 µL or three times the strong wa shvolume

5. Cl ick OK. The need le wash beg ins .

6. When needle wa shing is complete , the sta tus returns to Id le.

Caution: If you do not use a suff icient qua ntit y of weak w a sh solvent,the st rong w a sh solvent ma y conta ct t he sam ple, corrupting i t .




To stop a needle wash routine before it finishes:

From th e sample ma na ger inform a tion wind ow , click Control > Reset SM.

Alternative: Right -click in th e Empower or Mass Lyn x sa mple ma na ger cont rolpan el, a nd t hen click Reset SM.

Characterizing the needle seal

The needle seal cha ra cteriza tion procedure finds th e position a t w hich th eneedle obta ins a seal w ithin th e wa sh st a tion block. The sam ple ma na germust be primed before sta rt ing t his procedure.

Requirements:

• P er form this procedure before ca l ibra t ing the needle and sample loopvolumes.

• P erform th is procedure a f te r priming the sample mana ger or a f ter yourepla ce a nd/or a djust t hese items :

• The needle

• An y pa r t of t h e n eed le a s sem bly

• The needle (Z) or piercing needle (Zp) f lags (home and top-of-plate)

• A h ome or t op-of-pl a te sen sor

• The inject port sea l

• The w a sh st a tion

• Th e N VR a m ba t t er y on th e C P U 2000

Caution: Do not abort t he sam ple needle wa sh sequence. Doing so mayleave strong solvent in th e sam ple needle w hich can a dversely a ffectchromatography.

To characterize the needle seal:

1. In the ACQU ITY UP LC Console, select Sa mple Mana ger from thesystem t ree. The sam ple ma na ger informa tion window a ppears.

2. Cl ick Mainta in > Chara cter ize > Needle sea l . The Cha racter ize Needle

Sea l dialog box appears.

3. Cl ick Sta r t . The ca l ibra t e sea l opera t ion begins, and the samplema nager s ta tus d isplays “Ca l ibra t ing sea l .”




4. When ca l ibra t ion ends, the sample ma nager s ta tus d isplays “ Id le. ”

5. Cl ick Results to v iew the resul ts of the needle sea l chara cter iza t ion

operation.

Characterizing the needle and sample loop volumes

Whenever you r eplace the sa mple loop a nd/or th e sa mple needle, you mu st setth e system t o cha ra cterize the volume of the repla cement pa rt s. Do thisregardless of wheth er the sizes of the repla cement part s a re nominally th esa me a s th ose of th e origina l part s or differ from t hem. Also perform th isprocedure wh en the composition of th e wea k w a sh solvent cha nges, beca usesolvent char a cteristics such as viscosity , surface tension, a nd polar ity cancha nge. Durin g sa mple injection, the w eak w a sh solvent precedes a nd follow sth e sa mple in t he fluid-ca rry ing lines, so th e sam ple is directly a ffected by t hewea k wa sh solvent.

Ch a ra cterizing t he loop volume compa res th e loop’s nomina l volume (in µL) to

its mea sured volume.Ch a ra cterizing t he needle volume compa res t he needle’s n ominal volume (inµL) to its mea sured volume.

Tip: Cha ra cterizing t he system volume is critical to accepta ble sa mplema na ger performa nce.

Requirements:

• Speci fy the s izes of the sam ple needle, loop, and syr inge in the Volumesdia log box before cha ra cterizing t he volumes.

• P r i me th e s a mple ma n a g er , a n d ch a r a cter ize th e sea l bef or echa ra cterizing th e volumes.

• Cr ea te a meth od (usin g E mpow e r or M a ssL yn x sof tw a r e) th a t h a s th esame a ir gap and sa mple draw ra t e tha t you w i ll be using.

To characterize the needle and sample loop volumes:

1. In the ACQUI TY UP LC Console, select Sa mple Mana ger from thesystem t ree.

2. Click Mainta in > Cha ra cterize> Needle a nd loop volumes. The

Ch a ra cterize Needle a nd L oop Volumes dia log box appears.

3. Click Sta rt . The needle a nd loop volumes cha ra cterizat ion opera tionbegins.

Tip: This procedure ta kes at least 5 minutes




Tip: This procedure ta kes at least 5 minutes.

4. Click Results to view t he results of the needle a nd loop volumescha ra cterizat ion opera tion.

Result: I f t he needle fa ils th e test , suspect it is bent, broken, or blocked.If t he sam ple loop fa ils th e test, suspect t ha t it is blocked or leaking, th a ti t ha s a loose fi t t ing, or th a t t he draw ra te is too high.

Loading sample plates in the sample manager

The ACQU ITY UP LC sam ple man a ger holds up to two ANSI pla tes tha t y ouloa d t hrough t he front door. The left plat e is referred t o as position 1, the rightplat e a s position 2.

Exception: If t he optional sa mple orga nizer is insta lled, you ca n load only onepla te t hrough the sa mple mana ger front door. You must loa d t he pla te on theright-ha nd t ra y. In this case, the r ight-ha nd t ra y becomes the num ber oneposition.

To load a sample plate:

1. Open the ACQUITY UP LC sample mana ger door .

2. Squeeze the t r ay but ton while you pull the t r ay toward you .

3. Load t he pla te onto the tra y so tha t w ell posit ion A,1 is a t t he r ight-rearcorner an d th e forw a rd edge of the plat e is behind the spring inside the

front of the carr ier.

4. Sl ide the tray into the sample ma nager unt i l i t cl icks into place .

Sliding tray into sample manager




5. Close the sample compartment door . A mechanism on the door ensuresth e plat es a re positioned correctly w hen t he door closes.

Selecting the optimum sample injection mode

The sa mple ma na ger support s t hree injection modes – P a rt ial L oop NeedleOverfill , Pa rt ia l Loop, an d Full Loop.

• P a r t ia l Loop Needle Overf il l Mode – the best genera l purpose mode forpar tia l loop injection. It provides the best pa rt ial loop a ccuracy,precision a nd linearit y for a w ide ra nge of sam ples, including str ong andw eak a cids a nd ba ses, hydrophilic, a nd hy drophobic compounds. It isrecommended a s t he first choice mode, except w here th ere a re clearindica tions for t he oth ers.

• P a r t i a l Loop Mode – should be reserved for those s itua t ions wherea na lysis time ta kes precedence over an y other concern, w here thesa mple volume is very limit ed, or w here th e inject ion volume is verylarge.

Caution: The plat es must be posit ioned corr ect ly t o a voiddam a ging the sa mple needle.

TP02389

Sample plate

Button

Plate tray

A-1 wellposition

• Ful l Loop Mode – should be chosen whenever accuracy and precision areth e primar y concerns. I t is t he recommended mode w hen using 1.0 mmI.D . columns.

Partial Loop Needle Overfill mode guidelines

• P rov ides opt imum per formance when in ject ion volumes a re ma in ta inedw ith in a ra nge of 10 t o 75% nomina l loop volume.

• The genera l ly accepted pract ice for par t ia l loop inject ion l inear i tyrest ricts t he inject ion volume select ed to ≤ 50%nom ina l loop volume




rest ricts t he inject ion volume select ed to ≤ 50% nom ina l loop volume.H owever, you can increa se th e usa ble loop volume to ≤ 75% nom in a l loopvolume by selectin g t he Needle Overfill techniq ue.

• These recommendat ions a re based on a chieving a n inject ion-to-inject ionvariability of ≤ 1% a cross t he specified volume ra nge. In a ddit ion, thecorrela tion betw een specified injection volume a nd pea k a rea must beR 2>0.999 a nd th e Area % RS D is < ±1.0.

Load Ahead mode guidelines

• The fi rst inject ion of a sample set w i ll not ut i l ize Load Ahead mode.• In ject ions sets w i th d i fferen t methods wi ll not u t i li ze Load Ahead mode.

• The min imum cycle t ime i s the lesser of ei ther the two run t imes orsample prepara t ion a nd w ash.

The follow ing f igures show a sta nda rd ACQUI TY U P LC system cycle t ime andload-ahead cycle time.

Standard ACQUITY UPLC system cycle time definition

Standard Injection mode and first injection in Load Ahead mode

Total ACQUITY UPLC system cycle time




ACQUITY UPLC system load-ahead cycle time definition

SM wash time

Start End

Chromatographic run time

SM sample preparationand positioning

Sampleinjection

System

setup

EndStart

Sampleinjection

Total ACQUITY UPLC system cycle time

Load Ahead mode after the first injection

Chromatographic run time

SM sample preparationSM wash time

Buffer delaySystem

setup andsample

positioning

into loop

The follow ing t a ble provides th e minimum a nd m a ximum injection volumesfor each of th e commonly used loops.

Selecting the injection mode and loop volume

Sample injectionvolume (µL)

Loop volume2 µL 5 µL 10 µL 20 µL 50 µL

a

P a rtia l loopneedle overfill

d i j ti

0.2 t o 1.5 0.5 t o 3.8 1.0 t o 7.5 2.0 t o 15.0 5.0 t o 37.0




a. The 50 µL loop must be used wit h a 250 µL sa mple syringe.

mode in jectionra ng e (µL)

P a rt ia l loop modeinjection ra nge(µL)

Notrecom-mended

Notrecom-mended

1.0 t o 5.0 2.0 t o 10.0 5.0 t o 25.0

Full loop modeinjection ra nge(µL)

2 5 10 20 50

Installing the sample manager shade

If your sa mples are light -sensitive, you should insta ll th e sam ple ma na gerfoa m sha de over the sa mple compar tment w indow.

Required material

• S ample ma na ger foa m sha de

To install the sample manager foam shade:




1. Inser t the foa m shade over the outside of the sample compartmentwindow.

Inserting the foam shade

Foam shade

Sample compartmentwindow

2. P ress the foam sha de into place over the window.

Foam shade pressed into place




Foam shade

Preparing the sample organizer

If your sy stem includes a sa mple orga nizer, prepar e it for opera tion a ccordingto t he procedures in t his section.

Initiating communications

To initiate communications between the sample manager and sampleorganizer:



Preparing the sample organizer 3-29

1. Open the sam ple ma nager door , load a pla te onto the r ight-hand tra y,a nd t hen close the sam ple ma na ger door.

Tip: When the system ha s both a sam ple mana ger and a sam pleorgan izer, th e right-ha nd t ra y of th e sample ma na ger becomes positionnum ber one, and t he left-ha nd t ra y is not a va ilable. The bott om shelf inth e sa mple orga nizer becomes position number t w o.

Exception: If t he syst em does not include a sa mple orga nizer, theleft-ha nd t ra y in t he sa mple ma na ger is designat ed position num ber

one, the right-ha nd tr a y, position number tw o.2. In the ACQU ITY UP LC Console, select Sa mple Mana ger from the

system t ree.

3. In the sample man ager informat ion window, cl ick Conf igure > Sa mpleOrga nizer. The Sa mple Orga nizer Configura tion dialog box appears.

4. Select the sample organizer f rom the l is t of ser ia l numbers in the

drop-down list, a nd t hen click OK.

5. The sam ple organizer a utomatica l ly detects w hich shelves conta inpla tes a nd i l luminat es their corresponding LE Ds.

Loading sample plates

The sa mple organ izer holds up to 21 ANSI plat es tha t y ou load t hrough t hefront door. How ever, th e actua l number of plat es you can load depends ontheir height. When the system ha s both a sam ple mana ger and sa mple

orga nizer , the r ight-ha nd shelf in t he sam ple ma na ger is referred to asposition 1 a nd t he bott om shelf in th e sa mple orga nizer a s position 2.

To load sample plates:

1. Open the sample organ izer door .




2. P u ll t h e s helf t ow a r d you .

3. Load t he pla te onto the shel f so tha t posit ion A,1 is a t the r ight-rearcorner an d the forw a rd edge of th e plat e is behind the stop a t t he leftfront corner.

Loading the sample plate

4. Ensure tha t the pla te does not extend beyond the pla t e s top a t the backof the shelf.

Caution: To prevent s pillag e, use Wa t ers-a pproved ca p ma ts ,sealing caps, or hea t sea l film on t he sam ples. Consult th e currentACQU ITY U P LC syst em relea se notes for a list of approvedsample covers.

TP02504

Shelf label

5. Sl ide the shel f in to the sample organizer unt i l i t s tops.

Shelf/plate combination and restrictors

Caution: To ensure the tra nsfer shut tle moves freely a nd w ithoutdam a ging the sa mple orga nizer , you must be able to slide ashelf/pla t e/via l combina t ion in or out w ith out int erfering w ith t he

restrictors directly a bove an d below it.




6. Repea t s teps 2 through 5 for the remain ing pla tes.

7. Close the sam ple orga nizer door. A mecha nism on the door ensures theshelves a re positioned corr ect ly w hen t he door closes.

8. Cl ick Conf igure > Scan an d store shel f layout to updat e and save thenew shelf configura tion.

To remove and replace the same plate on the same shelf:

1. Open the sam ple organizer door , and then remove pla t es tha t ha ve

finished processin g.2. P ull a shel f towa rd you , and then inser t a pla te of the same type and s ize

on th e shelf.

3. Load a pla t e onto a shelf so tha t posi t ion A,1 is a t the r ight-rear cornera nd th e forwa rd edge of th e plat e is behind the stop a t t he left-front

Caution: To avoid ja rring th e plat es from th eir shelves, do notslam th e sam ple organ izer door closed.

Shelf

Sampleplate

Restrictors

corner. Ens ure tha t the plat e does not extend beyond t he pla te stop atth e back of the shelf.


Caution: To prevent spillage, use Waters-approved cap mats, sealingcaps, or hea t sea l film on the sa mples. Consult t he current ACQU ITY

U P LC syst em relea se notes for a list of approved sa mple covers.

Caution: To ensure the tr a nsfer shut tle moves freely a nd w ith outdam a ging the sa mple orga nizer , you must be able to slide a




5. Repeat s teps 3 and 4 unti l a l l pla t es and holders are placed correct ly onth e shelves.

6. Close the sam ple organizer door. A mecha nism on the door ensures theshelves a re positioned corr ect ly w hen t he door closes.

7. Cl ick Verify . The sample organizer a utoma tica l ly scans t he pla tes andshelves, senses wh ich shelves cont a in plat es, compa res th em to th esa ved layout, verif ies that they ma tch, and i l lumina tes thecorresponding LE Ds inside the sa mple orga nizer door.

8. Configure the pla tes and shelves in the Empower or MassLynx da taapplication.

Tip: You can loa d plat es a nd sh elves eith er before or a fter configuringth em in the da ta a pplica tion, but you must configure them beforerunning sa mples.

To rearrange the shelves for a different plate configuration:

1. Open the sam ple organizer door , and then remove pla t es tha t ha vefinished processin g.

2. Add, move, or remove shelves f rom t he sample organizer so tha t theshelf configurat ion suits t he pla tes you intend to run. St a nda rdmicrot iter plat es need one slot. Int ermediat e pla tes need tw o slots, soa llow empty slots a bove the plat e or holder. Deep-w ell plat es an d a ll via l

dam a ging the sa mple orga nizer , you must be able to sl ide ashelf/pla t e/via l combina tion in or out w ith out int erfering w ith t he

restrictors directly a bove an d below it.

Caution: To avoid ja rring t he plat es from t heir shelves, do not slamth e sa mple orga nizer door closed.

holders need th ree slots, so a llow tw o empt y slots a bove the plat e orholder.

3. Load a pla t e onto a shelf so tha t posi t ion A,1 is a t the r ight-rear cornera nd th e forwa rd edge of th e pla te is behind t he stop, a t t he left-front

corner. Ens ure tha t the plat e does not extend beyond t he pla te stop atth e back of the shelf.

Caution: To prevent spillage, use Waters-approved cap mats, sealingcaps, or hea t sea l film on the sa mples. Consult t he current ACQU ITYU P LC syst em relea se notes for a list of approved sa mple covers.





5. Repeat s teps 3 and 4 unti l a l l pla t es and holders are placed correct ly on

shelves.

6. Close the sam ple orga nizer door. A mecha nism on the door ensures theshelves a re positioned corr ect ly w hen t he door closes.

7. Cl ick Configure > “Scan and s tore shel f l ayout ” .

The sa mple orga nizer initia lizes a nd sca ns t he shelves. When it detectsa new sh elf, it illumina tes th e LED to the left of the shelf, inside thesa mple orga nizer door.

Tip: In the ACQU ITY U P LC Console, a thin, grey bar a ppears for ea chempty shelf. After t he plat e or shelf is configured using E mpow er or

MassL ynx softw a re, plat e identif ica tion a ppears in a thicker ba r .8. Configure the pla tes and shelves in the Empower or MassLynx da ta

application.

Tip: You can loa d plat es a nd sh elves eith er before or a fter configuringth em in the da ta a pplica tion, but you must configure them beforerunning sa mples.

Caution: To ensure the tr a nsfer shut tle moves freely a nd w ith outdam a ging the sa mple orga nizer , you must be able to sl ide ashelf/pla t e/via l combina tion in or out w ith out int erfering w ith t herestrictors directly a bove an d below it.

Caution: To avoid ja rring th e plat es from th eir shelves, do notslam th e sam ple organ izer door closed.

Displaying sample plate information

To display sample plate information:

1. In the ACQUI TY UP LC Console, select Sa mple Organizer f rom the

system t ree.2. In t he sam ple organizer informat ion w indow, c lick Conf igure > Scan a nd

st ore shelf la yout t o upda te a nd sa ve th e configura tion of plat es onshelves.

The number designat ions of shelves th a t cont a in sam ple plat es aredisplayed beside informa tion about th e plat es




displayed beside informa tion about th e plat es.

Sample organizer information window

Tips:• The shel f and p la t e informat ion a re di sp layed on ly a f ter method

setup.

• Move the cursor over a shel f to d isplay the number of samplesremaining to be run from t ha t shelf .

Shelf number Plate information

Preparing the detector

If y our syst em includes a TU V detector, prepar e it for opera tion by follow ingth e procedures in t his section.

See also: If your syst em includes a P DA detector, see the ACQUI TY UPLCPhotodi ode Ar r ay Detector Gett i ng Star ted Gu i de for informa tion on prepar ingit .

See also: If your system includes a n EL S det ector, see th e ACQUI TY UPLC

Evaporat ive L igh t Scatter i ng Detector Gett i ng Star ted Gui de for informa tionon preparing it



Preparing the detector 3-35

on preparing it .

See also: If your syst em includes a ma ss spectr ometer, see the document a tiontha t a ccompanies the instr ument for informa tion on prepa ring i t .

Preparing the TUV detector

Starting the TUV detector

Tip: To prevent errors on st a rt up, be sure the flow cell conta ins dega ssed,tr a nspa rent s olvent (a cetonitrile or w a ter) a nd t he detector door is closedfirmly.

To start the TUV Detector:

1. Ensure the detector flow cell is f il led wi th degassed, t ra nsparent solvent(a cetonitrile or w a ter) and free of a ir bubbles. The detector might notinitia lize correctly if th e cell conta ins a ir.

2. Ensure the detector door is closed fi rmly.

3. P ress the pow er switch on th e door to pow er-on th e detector . Thedetector beeps t hree times a nd runs a series of sta rtup t ests w hile thelamp LE D blinks. The pow er LED shows st eady green.

Initia l iza tion usua lly requires approximat ely 2 minutes, a nd la mpw a rm-up requires approxima tely 3 minut es.

Caution: U se only th oroughly degassed HP LC-gra de solvents. G a s inth e mobile pha se ca n form bubbles in t he flow cell and ca use thedetector to fai l th e sta rtup dia gnostic tests.

4. When the lamp LED shows s teady green , sta r t the Empower orMa ssLy nx softw a re. You ca n monitor th e ACQU ITY UP LC C onsole formessages an d LE D indica tions. For best r esults, a l low a t least 30minut es to equilibrat e th e detector a nd sta bilize th e ba seline.

Tip: The absorba nce va lue appea rs in th e ACQU ITY UP LC Console a nda lso in Em power’s Run Sa mples w indow or Ma ssLynx’s In let Editorw indow . If t he detector is in dua l w a velength mode, tw o absorban cevalues appea r.

Absorba nce values ha ve a resolution of 0.0001 AU .

When the lam p is extin guished, “Lam p Off” appears in th e softw a re




inst ead of a bsorbance va lues.

5. Configure the detector according to the instruct ions in the Empower orMa ssLy nx online Help.

See also: “Configuring Empower” on pa ge 4-2 and “ConfiguringMassLynx software” on pa ge 4-12.

TUV Detector information window

Recording sample and reference energies

After you insta ll the detector or perform ma intena nce ta sks, like cha nging t helam p or flow cell, complete th e procedures in th is section t o verify t ha t th edetector optics and electronics work properly.

To record sample and reference energies:

1. Ensure tha t t he detector i s connected to the worksta t ion .

2. F lush the sys tem tub ing wi th f il tered , degassed HP LC-gradeacetonitrile.



Preparing the detector 3-37

3. P ump mobile phase for 15 minutes or more at 0.3 mL/min.

4. En sure the detector cell is f il led with solvent and free of air bubbles.

Tip: The detector ma y n ot initia lize correctly if a ir is present in t he cell.

5. When both LE Ds show st eady green, in it ia l iza t ion is complete .

6. S t a r t th e E mpow e r or Ma ssLyn x sof tw a r e.

7. La unch the ACQUI TY UP LC Console f rom the sa mple ma nager controlpanel.

See also: “St a rting t he ACQU ITY UP LC Console from E mpow er” onpage 4-11 a n d “St a rting t he ACQU ITY UP LC Console from Ma ssLynxsoftware” on pa ge 4-13.

8. Select t he TU V detector view in the ACQU ITY UP LC C onsole.

9. S et t he w a v elen gt h to 230 n m .

10. In the C onsole, select TU V Detector > Intera ctive Display from t he

system t ree.

11. Record t he sa mple a nd reference energies a t 230 nm.

Caution: The ma ximum a llow a ble pressure drop across the flowcell is 6895 kP a (69 bar , 1000 psi). I f t he solvent is viscous(meth a nol-w a ter, for example), you ma y n eed t o decrea se th ema ximum flow r a te to prevent brea king the cell.

Conditioning the column

Conditioning the column involves running a solvent gra dient th rough itw ithout injecting sa mples or run ning t he Event s ta ble. The run t ime forconditioning th e column should equa l the gra dient ta ble run t ime.

Caution: To prevent da ma ge to the detector flow cell, ensure th a t t hew a ste solvent does not flow t hrough t he detector during t hisprocedure. After inst a lling a new column, flush solvent th rough it a ndout t o wa ste before connecting t he column to t he detector (for exa mple,10 column volumes).




To condition the column:

1. Remove the column inlet l ine from t he detector , and place the end in asma ll w a ste conta iner.

2. I f Empower sof twa re controls the system, proceed as follows:

a . In th e Sa mples t a b le, a d d a r ow to th e meth od .

b. Select Eq uilibrat e/Condition Column (Isocra tic or Gr a dient) a s thefunction in th e new row.

c. Run the separa t ion method . The sys tem runs the grad ien t .

3. I f MassLynx sof twa re controls the system, proceed as fol lows:

a . Open the Sample Se t w indow, and select an in let method tha t

includes the chroma togra phic conditions you w a nt to use.b . In the Sa mples t ab le, add an in let prerun field.

c. In the Run Samples page , select S amples > Format > Customize.

d . In the Custom Field Display w indow, select In let prerun .

e . To save the column as par t of the window, save the sample set

format .f . Se lect method setup in the Sample Set w indow a s a pre-in let

method.

g . Select a method for the inlet f ile (these methods can be the same).

h . Run the sample set l ine . The system runs t he condit ion columnmethod and t hen runs the sepa ra tion method.

)

See also: For more informa tion a bout column conditioning, consult th eEm pow er online H elp.

Tip: The run t ime for conditioning a column should equa l the gra dient t a blerun time.

Shutting down the system

Caution: B uffers left in t he system can precipita te a nd da ma geinstrument components.



Shutting down the system 3-39

You might w a nt t o shut down th e system

• bet w een a na lyses

• overnight

• for a w eekend

• for 72 hours or m ore

Tips:

• I f you a re us ing Empower sof tware to control the system, set systemshutdown pa ra meters in the Inst rument Method Editor . Consult t heE mpower online Help for m ore informa tion.

• I f you a re us ing MassLynx sof tware to control your sys tem, set sys temshutdown par a meters in the Shutdown E ditor . Consult the MassLynxonline Help for more information.

Between analyses

To shut down the system between analyses:

1. B etween an alyses, cont inue to pump the init ia l mobile phase mixtureth rough the column. D oing so ma inta ins th e column equ ilibriumnecessar y for good r etention t ime reproducibility .

2. I f a few h ours wi ll pass before the next inject ion, s low the f low ra t e inth e interim t o a few tent hs of a mL/min t o conserve solvent.

Tip: E nsure tha t Auto-Shut down for your shut dow n method isdeactivated.

3. Keep the detector opera t ing and the column heat er a t opera t ingtemperature during this period.

Shutting down for fewer than 72 hours

To shut down the system for fewer than 72 hours:

1. Flush t he column w ith 100% a cetonitr i le. Doing so keeps th e column bedin an a ct ive, wet ted sta te .

See also: Wa ters ACQUI TY U PLC BEH Colum n Care and U se

Instruct ions .




Requirement: If you a re using buffers, you must first flush t he columnw ith a high -w a ter -cont ent mobile pha se (90% w a t er). Then st op t hepump flow .

2. Ext inguish the detector lamp to lengthen lamp l ife.

3. The column heat er can operat e overnight but should be shut down overa weekend.

Shutting down for more than 72 hours

To shut down the system for more than 72 hours:

1. Follow the steps for shutt ing dow n the system for fewer tha n 72 hours,above.

See also: Waters ACQU I TY U PLC B EH Colum n Care and U se

Instruct ions .

2. After f lushing the column and let t ing i t cool to ambient temperature ,disconnect the inlet a nd outlet tubes, and join th em w ith a union. Inst a l lend-plugs in th e column inlet a nd outlet fitt ings, an d th en return t hecolumn ca refully to its box for stora ge.

Warning: Risk of elect ric shock. The power sw itch on ea ch syst eminstrument cont rols the basic opera tiona l sta te of tha tinstr ument . Neverth eless, some instru ment circuits rema in livea fter t he instr ument is sw itched off. To completely interru ptpow er to a syst em instrument, set t he power switch to Off, an dth en unplug th e instru ment’s pow er cord from t he AC outlet.

3. P ump wa ter through the system (includ ing the b inary solvent ma nager ,sa mple ma na ger, a nd det ect ors) for 10 t o 20 min ut es at 0.5 mL /min ,followed by isopropyl a lcohol for a noth er 10 t o 20 minut es. Then t urn t hepump off, lea ving isopropyl alcohol in th e fluid lines.

Caution: B efore using an y syst em instruments for a nother type ofa na lysis, ensure tha t t he mobile pha se for t he new a na lysis ismiscible w ith meth a nol, w a ter, meth a nol/a cetonitrile, or isopropyla lcohol. Likewise, before resta rting the syst em, ensure tha t a nyresidua l ma teria l not miscible w ith t he init ial metha nol/w a termobile pha se ha s been flushed th oroughly from the syst em wit ha n a ppropriat e intermedia te solvent.



Shutting down the system 3-41




4 Configuring System Software

Contents:

Topic Page

C onfiguring E mpow er 4-2



4-1

S t ar t ing t he ACQU ITY U P LC Console from Empow er 4-11

C onfigur ing Ma ssLynx soft w a re 4-12

St a rting th e ACQU ITY UP LC C onsole from Ma ssLynx softw a re 4-13

Configuring Empower

P erform t hese ta sks to configure Em pow er:

• S ta r t the soft w a re a nd log in

• S elect sy st em in st rum en ts

• Na me t he system

Starting Empower and logging in

To start Empower and log in:



4-2 Configuring System Software

1. Select S ta r t > Programs (for Windows XP , All P rograms) > Empower >E mpower Login. The E mpower Login dia log box appears.

Alternative: You can a lso sta rt E mpow er through the E mpower desktopshortcut.

2. Type your user n a me a n d pa ssw or d.

3. C lick OK.

Selecting system instruments

To select system instruments:

1. In the Empower Pro window, cl ick Conf igure Sys tem.

2. In t he Conf igura t ion Mana ger window, c lick Acquisit ion Servers ,right -click t he node na me, a nd t hen select P roperties.

Tip: If you a re using Em pow er 1154 softw a re to control the syst em, thenode na me is referred to as t he a cquisit ion server nam e.

3. In th e Acquisit ion Server dia log box, cl ick the Inst ruments ta b. Thesystem instrum ents tha t a re successfully communicating w ith yoursystem a re show n w ith a Yes in the “OK?” column.

Instruments tab



Configuring Empower 4-3

4. Ensure tha t a b inary solvent ma nager (ACQ-BS M), sample mana ger(ACQ-SM), and detector (ACQ-TUV, ACQ-PDA, or ACQ-ELS) appear inthe instrum ent l ist , a nd t hen click OK.

5. Right-cl ick Systems, a nd then select New > Chromatographic System.

6. In the System Type area of the New Chromat ographic System Wizarddia log box, select C reat e New Sy stem, a nd t hen click Next.

System Selection dialog box

7. In the System Select ion dia log box, drag the na me of the instrument , orinstruments, you w a nt to include in t he new sy stem from the AvailableInst ruments pan e to the New Syst em Instru ments pane. Click Next.

8. When the Access Control dia log box appea rs, cl ick Next.

9. In t he Nam e Select ion dia log box, speci fy a nam e for your system. Entercomments, if a ny, a nd t hen click Finish. A confirma tion dia log boxappears.

10. Click Pr ojects, right-click a project, a nd t hen select Open.11. In the P roject window, cl ick Run Sa mples . The Run Sa mples window

appears.

Alternative: You can a lso a ccess the Run S a mples w indow viaEm power’s QuickSt a rt menu.

The Run S a mples window cont a ins cont rol pa nels for t he bina ry solvent




ma na ger, sa mple ma na ger, a nd detector .

Control panels

About the binary solvent manager control panel

If E mpower softwa re controls the system, th e bina ry solvent ma na ger’scontrol panel appea rs a t the bott om of the Run S a mples w indow . If Ma ssLynxsoftw a re controls th e system, the bina ry solvent ma na ger’s control pan el

a ppears a t t he bott om of the Inlet E ditor w indow .

Binary solvent manager control panel

The bina ry solvent ma na ger cont rol pan el displa ys flow st a tu s, systempressure, tota l flow ra te, a nd solvent composition para meters. You ma y editth ese when t he syst em is idle by clicking on th e underlined value. You can not

Status

Flow LED

System pressure

Total flow rateStop flow

Solvent composition

edit binary solvent ma na ger para meters wh ile the system is processingsamples.

The follow ing ta ble lists t he items in th e bina ry solvent ma na ger controlpanel.

Binary solvent manager control panel items

Control panel item Description

Flow LE D D ispla ys the a ctua l flow LE D on thefront panel of th e solvent ma na gerunless communications with thesolvent ma na ger are lost .




S ta t us D ispla ys t he st a tus of the currentoperation.

S yst em P ressure D ispla ys t he solvent ma na ger pres-sure, in kP a , bar , or psi . Pressureunit s ca n be cust omized th rough theACQU ITY UP LC C onsole.

Tot a l F low Ra t e D ispla ys t he tot a l flow ra t e of thesolvent ma na ger. Tota l flow r a teva lues ra nge fr om 0.000 to 2.000mL /min u nder norma l opera tion a nd0.000 to 8.000 mL/mi n w henpriming.

S olvent Composit ion D ispla ys t he percent a ge of solvent (1

a nd 2) to be dra w n from the pumps(A and B ). Composition va lues ra ngefr om 0.0 t o 100.0%.

(S t op Flow ) Immedia t ely stops a ll flow from t hesolvent m an a ger.

You can a ccess add itional functions by right -clicking a nyw here in the solventma na ger contr ol pan el:

Additional functions in the binary solvent manager control panel

Control panel function Description

Refresh syst em (S ys P rep) Refreshes t he fluid-ca rry ing linesa ccording to the current meth odconditions.

See also: “Refreshing the system”on pa ge 3-10.

S t a r t up B rings the syst em to opera t iona l




conditions a fter a n extended idleperiod or wh en swit ching t o differentsolvents.

See also: “Sta r t ing up the system”on pa ge 3-11.

P r ime A/B solvent s D ispla ys t he P r ime A/B S olvent sdia log box.

See also: “Pr iming the binarysolvent m an a ger” on pa ge 3-8.

P r ime sea l w a sh D ispla ys t he P rime S ea l Wa shdia log box.

See also: “Per forming a sea l w ashprime” on pa ge 3-5.

Reset B S M Reset s t he solvent ma na ger a ft er a nerror condit ion.

H elp D ispla ys t he ACQU ITY U P LCC onsole online H elp.

About the sample manager control panel

If Em pow er softw a re controls th e syst em, the sam ple man a ger’s control pan ela ppears a t t he bott om of the Run Sa mples window. If MassLy nx softw a recontrols the syst em, the sa mple mana ger’s control panel a ppears a t thebott om of the Inlet E ditor window.

Sample manager control panel

Run LED

Current column heater

temperature

Status

Current sample

compartment




The sam ple ma na ger control pan el displays current sa mple compart ment a ndcolumn h eat er temperat ures an d set points . These may be edited wh en thesyst em is idle by clicking on th e underlined value. You can not edit s a mplema na ger set points w hile th e system is processing sam ples.

The follow ing ta ble lists t he items in th e sam ple ma na ger control pa nel.

Sample manager control panel items


Run LE D D ispla ys t he a ct ua l run LE D on t hefront panel of the sa mple ma na gerunless communications with thesa mple ma na ger a re lost .

S t a t us D ispla ys t he st a tus of the currentoperation.

Current S ample CompartmentTemperature

Displa ys t he current sa mplecompartment t emperat ure to 0.1 ° Cresolution. When a ctive temperat urecont rol is disa bled, this fielddisplays “Off.”

Display ACQUITY UPLC

Console

Column heaterset point

p

temperature

Sample

compartment

set point

S a mple C om pa r tm en t Set P oin t D ispla y s t he cur ren t sa m plecompartment set point t o 0.1 ° C

resolution. When a ctive temperat urecont rol is disa bled, this fielddisplays “Off.”

Current Column H eaterTemperature

Displa ys t he current column hea tertempera ture to 0.1 ° C resolution,even w hen a ctive tempera turecontrol is disabled.

Sample manager control panel items (Continued)





You can a ccess add itional functions by right -clicking a nyw here in the sa mplema na ger contr ol pan el:

Column H ea ter S et P oin t D ispla ys t he current column hea terset point t o 0.1 ° C resolution. Whena ctive tempera tu re control isdisa bled, t his field display s “Off.”

(D ispla y C onsole) D ispla ys t he ACQU ITY U P LCConsole.

Additional functions in the sample manager control panel


Run Sys P rep P r imes t he sa mple ma na ger w it h

one wea k wa sh prime (w a sh an dsa mple syringes).

See also: “Refreshing the system”on pa ge 3-10.

P r ime syr inges D ispla ys t he P rime S yringes dia logbox.

See also: “P riming the samplemanager” on pa ge 3-16.

Wa sh needle D ispla ys t he Wa sh Needle dia logbox.

See also: “Wa shing th e samplema na ger needle” on pa ge 3-18.

Turn ligh t s on/off Turns t he sa mple ma na ger compa rt -ment a nd optiona l sam ple orga nizer

light s on or off.Reset S M Reset s t he sa mple ma na ger a ft er a n

error condit ion.


Additional functions in the sample manager control panel (Continued)





About the TUV detector control panelSee also: If your syst em includes a P DA detector, see the ACQUI TY UPLC

Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for informa tion on prepar ingit .

See also: If your system includes a n EL S det ector, see th e ACQUI TY UPLC

Evaporat ive L igh t Scatter i ng Detector Gett i ng Star ted Gui de for informa tionon preparing it .

If E mpower softw a re cont rols t he syst em, th e TU V detector’s cont rol pa nela ppears a t t he bott om of the Run Sa mples window. If MassLy nx softw a recontrols the syst em, the detector’s cont rol pa nel appears a t t he bott om of theInlet Editor window.

TUV Detector control panel

The TU V detector control pa nel display s a bsorba nce units a nd w a velengthva lues. You ma y edit t hese wh en th e system is idle by clicking on theunderlined value. You cann ot edit detector pa ra meters w hile th e system isprocessing samples.

Lamp On/Off LED

Turn detector

lamp On/Off

Status

Absorbance

units

Value of

wavelength A

The following table lists the items in the TUV detector control panel.

TUV detector control panel items


La mp On/Off LE D D ispla ys t he a ctua l la mp on/off LE Don the front pa nel of th e detectorunless communications with thedetector a re lost.

S t a t us D ispla ys t he st a tus of the currentoperation.

AU D ispla ys t he a bsorba nce unit s.




You can a ccess ad ditional fun ctions by r ight-clicking a nyw here in th e detectorcontrol panel:

nm D ispla ys t he va lue of w a velengt h A,in nm. I f the detector is in dua lw a velength mode, the va lue of w a ve-length B a lso a ppears.

(La mp On) Ignit es the detector la mp.

(La mp Off) E xt inguishes the detect or la mp.

Additional functions in the detector control panel


Aut ozero Reset s the a bsorba nce va lue to 0.

Reset TU V Reset s the detector , w hen present ,a fter a n error condition.


Starting the ACQUITY UPLC Console from Empower

To start the ACQUITY UPLC Console from Empower:

In th e Run sa mples w indow, click Displa y console in th e Sa mple Ma na ger

control panel.

ACQUITY UPLC Console window



Starting the ACQUITY UPLC Console from Empower 4-11

Configuring MassLynx software

P erform these ta sks to configure Ma ssLynx softw a re:

• S ta rt t he a pplica t ion

• S elect sy st em in st rum en ts

To start MassLynx software:

1. Select S t a r t > P r og r a ms > M a ssL yn x > M a ssL yn x.

Alternative: You ca n a lso sta rt Ma ssLynx softw a re through theMassLy nx desktop shortcut.

If MassL n Sec rit is not enabled MassL n soft a re sta rts a nd the




If MassLy nx Security is not enabled, MassL ynx softw a re sta rts, a nd theMassLy nx w indow a ppears. I f Ma ssLynx Security is ena bled, theMa ssLy nx Login dialog box appears.

2. Type your user na me a nd pa ssword, and select your doma in.

3. Cl ick OK. The MassLynx window a ppears .

To select system instruments:1. In the MassLynx window, cl ick In let Method .

2. In the Inlet Method window, select Instrument Configura t ion f rom theTools menu of the In let Meth od window .

3. In t he Inlet C onf igura t ion w indow, c lick Configure and then c lick Next .

4. In t he Select P ump dia log box, select Wat ers ACQUI TY as t he pumpingdevice, an d t hen click Next.

5. Select Wat ers ACQU ITY as the autosampler , an d then cl ick Next .

6. Select Wa ters ACQU ITY TU V, Wa ters ACQU ITY PD A, or Wa tersACQU ITY EL S a s t he detection device, and th en click Next.

7. C lick Next .

8. C lick F in ish .

9. Cl ick F inish , and then cl ick OK.

10. In t he Instrument Control Option P ack dia log box, ensure that “ Insta l lnew inst rum ent softw a re or upgra de existing inst a llat ion(s)” is selected,a nd t hen click Next.

11. Select ACQU ITY B inary S olvent Ma na ger, ACQU ITY Sam ple Ma na ger,a nd ACQ U ITY TU V Detector (or ACQ U ITY P D A Det ect or a nd/orACQU ITY EL S D etector), a nd t hen click Next. A progress bar a ppea rsa t t he bott om of the dia log box.

12. When the instr ument contr ol option pa ck insta l lation is f inished, the

Results screen of th e Inst rum ent C ont rol Option P a ck dia log boxappears.

13. Click Finish. The Inlet Method window a ppears.

Starting the ACQUITY UPLC Console from MassLynxsoftware



Starting the ACQUITY UPLC Console from MassLynx software 4-13

To start the ACQUITY UPLC Console from MassLynx software:

1. In t he MassLynx window, c lick Inlet Method. The Inlet Method windowappears.

2. Cl ick the ACQUI TY Addit iona l Sta tus ta b . The sample ma nager s ta t us

appears.3. Cl ick Display console . The ACQUI TY UP LC Console window

appears.




5 Verifying System Operation

Contents:

Topic Page

P repa ring the system 5-2

C rea t ing t he t est met hods 5-5

P erforming t he gra dient per forma nce test 5 9



5-1

This chapter explains how to run a gra dient performa nce test on a systemw ith a TU V detector t o verify th a t y our syst em is opera ting properly. Thesa mple you use to verify th e system is included in th e system st a rt up kit.

B efore you begin t his procedure, your sy stem must be set u p an d configured a sdescribed in C h a pt er 2, C h a pt er 3, and C ha pt er 4.

Tip: To ensure y ou ha ve th e lat est version of this procedure, visitht t p://w w w .w a t ers.com a nd click S upport C ent er.

Restrictions:

• This grad ien t per formance tes t i s not a pplicab le to ACQUITY UP LC

systems tha t ha ve a m a ss spectrometer.• There i s no grad ien t per formance test for ACQUITY UP LC sys tems tha t

ha ve a n EL S detector .

P erforming t he gra dient per forma nce t est 5-9

Preparing the system

P repa ra tion is the sa me wh ether the syst em is controlled by the E mpow er orMassLynx dat a system.

Requirements: Due to the increas ed sensit ivity of the ACQU ITY UP LCsystem a nd detectors:

• Al l solvents , includ ing wa t er and add i t ives , must be of the h ighes tchemical purit y (MS-gra de). Fa ilure to use MS -gra de solvents w ill resultin high ba ckground concent ra tion, low signa l-t o-noise, an d loss ofsensitivity.

• Al l MS-grade solvents used wi th the ACQUITY UP LC sys tem should be

properly filtered prior to their use. Wa ters r ecommends th a t t he solvent



5-2 Verifying System Operation

p p y pbe filtered thr ough an a ppropriat e membra ne filter using a solventclar ifica tion kit (47-mm a ll-gla ss filt er holder w ith 1-L fla sk)immedia tely before use. A genera l-purpose labora tory va cuum pump isa lso required for use with the a l l-glass solvent f i l tra tion a ppa ra tus.

• G lasswa re (such as solvent bot t les) should not be wa shed withdetergents or w a shed with other general glassw a re to prevent

conta mina tion. The glassw a re should be rinsed wit h th e high-puritysolvents to be used.

See also: Contr ol l ing Contam inat i on in L C/ M S Systems (par tnum ber 715001307) on the ACQU ITY UP LC Sy stem B ookshelf CD.

Wa ters recommends t ha t mobile phases a nd w a sh solvents be mixed a sfollows:

• Mobile phase A1 - 10:90 a cetonitr i le/w a ter (for A1, B2, strong and w eakneedle-w a sh, a nd t he plunger-seal w a sh lines)

• Mobile pha se A2 - 100% a cetonitr i le (for solvent l ine B1 an d A2)

• Mobile phase B1 - 100% aceton i t r ile

• Mobile phase B2 - 10:90 aceton i t r ile/wa ter

• Wea k w a sh - 10:90 a ceton it r ile/w a t er

• S t r on g w a sh - 10:90 a c eton it r il e/w a t er

• P lunger sea l wash - 10:90 aceton it r i le/wa ter

To prepare the system for verification:

1. P repare a 10:90 acetoni tr i le/wa ter mobi le phase:

a . Mea sure 100 mL of f il tered acetoni tr i le into a 100-mL gra duat edcylinder.

b . Ca refully t ransfer the a cetoni tr i le to a 1-L reservoir bot t le .

M 900 L f fi lt d HP LC d t i t 1000 L

Warning: Ch emica l exposure risk. Alwa ys observe sa fe la bora torypractices when you use this equipment a nd w hen you work withsolvents a nd t est solut ions. Know t he chemica l and phy sica l

propert ies of the solvent s a nd t est solutions you use. See th eMat eria l Sa fety D a ta Sheet for each solvent a nd test solution in use.



Preparing the system 5-3

c. Mea sure 900 mL of fi ltered HP LC-grade w at er into a 1000 mLgra dua ted cylinder.

d . Ca refully t ransfer the wat er to the same 1-L reservoir bot t le .

e. Cap the reservoi r bot t le and mix well .

f . Label the reservoir bot t le as 10:90 acetoni tr ile/wa ter .

g . Submerge l ines A1, B2, plunger sea l wa sh, s trong needle wa sh, andw eak n eedle wa sh in t he reservoir bott le conta ining th e 10:90a cet onitr ile/w a ter .

h . P lace the reservoi r bot t le in the solvent t r a y .

2. P repare a mobile phase of 100% acetonitr i le:

a . P our approxima tely 1L of fi ltered a cetonitr i le into a 1-L reservoirbottle.

b . Label the reservoi r bot t le a s aceton it r i le .

c. Submerge lines A2 a nd B 1 in the a cetoni tr i le reservoir bot t le .

d . P lace the reservoi r bot t le in the solvent t r a y .

3. Insta l l the ACQU ITY UP LC column in the column heat er , close thecolumn tr a y, a nd r eplace the column h eat er’s front cover. If you need

more informa tion a bout inst a lling t he column , see “Insta l ling thecolumn” on pa ge 2-27.

4. Access the ACQUI TY UP LC Console and per form these ta sks:

Caution: Never cha nge directly betw een im miscible eluents orbetw een buffered solut ions a nd orga nic eluents. I mm iscibleeluent s form emulsions in the flow pat h. B uffered solut ions a ndorganic eluents in combina tion ca n result in sa lt precipita tion inth e gra dient proport ioning va lves, pump hea ds, check va lves, orother par ts of th e system. Confirm tha t a l l f luids in the system a remiscible w ith a cetonitr ile. If you need a dditiona l informa tiona bout priming y our syst em, see “P riming the binary solventmanager” on pag e 3-8.




a . P r ime each solvent line of the binary solvent ma na ger for 10minut es. See “P riming a w etted binary solvent ma na ger” onpage 3-10.

b . P r ime the sea l wa sh of the b inary solvent ma nager for severa lminut es. See “P riming a w etted binary solvent ma na ger” on

page 3-10.

c. P r i me th e s a mple ma n a g er a t l ea s t 5 t imes . See “P r iming t hesample manager” on pa ge 3-16.

d . P erform the seal chara cter iza t ion procedure in the sample ma nager .See “Cha ra cterizing t he needle seal” on pa ge 3-20.

e. Ca l ibra te the system volume in the sample mana ger .

Alternative: U se the Refresh (Sy s P rep) function t o prime the binar ysolvent ma na ger. See “Refreshing the system” on pa ge 3-10.

5. P repa re the sa mple a s listed on the sam ple instructions, using 10/90acetonitr i le:water .

6. P lace the sample in the v ia l pla te , not ing the via l posi t ion, and put theplat e in position 2 of the S a mple Ma na ger.

Creating the test methods

The gra dient performa nce test method pa ra meters a re the sa me w hetherEm pow er or Ma ssLyn x softw a re controls the system. Follow th e steps below tocreat e the methods, sett ing the pa ra meter values to ma tch those pictured in

th e screen representa tions.Tip: Click on th e ta b pages to display online H elp.

Creating the instrument method

To create the instrument method:

1. Crea t e an inst rument method wi th the b inary solvent mana gert h i t h f ll i t ti



Creating the test methods 5-5

par a meters shown in t he follow ing screen represent a tion.

Binary solvent manager instrument parameters

Tip: The bina ry solvent ma na ger para meters ar e the sam e rega rdless ofw hether y our syst em includes th e TU V or P DA detector.

2. Se t ins t rument method para meters for the sample manager as shown inth e follow ing screen r epresenta tion.

Sample manager instrument parameters




Tip: The sam ple man a ger para meters ar e the sam e regar dless ofw hether y our syst em includes th e TU V or P DA detector.

3. Cl ick Advanced, on the G enera l tab , a nd set the para meters shown inth e follow ing screen r epresenta tion.

Sample manager instrument advanced settings



Creating the test methods 5-7

Tip: The sam ple man a ger para meters ar e the sam e regar dless of

w hether y our syst em includes th e TU V or P DA detector.4. I f your system includes the TUV detector , set instrument method

parameters as shown in the following screen representation.

See also:

• For in format ion on default va lues , consult the ACQUITY Consoleonline Help.

• For more informat ion on detector para meters , consult theACQU ITY U P LC Console online H elp, E mpower online Help, orMa ssLy nx online Help.

• I f your sys tem in cludes a P D A d etector , see th e ACQUI TY UPLC

Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for informa tion onverifying it .

• I f your sys tem in cludes a n E L S detector , see th e ACQUI TY UPLC

Evapora ti ve L i ght Scatt er i ng Detector Gett i ng Star ted Gui de forinforma tion on performing a run.

TUV detector instrument parameters




5. S a v e t h e in st r um en t m et h od .

Creating the sample set method

If E mpower softwa re controls the system, you must create a sam ple set

met hod. The sa mple set met hod pa ra met ers (In j Vol., # of In jections,Fun ction, Run Time, an d Next In jection Delay ) a re th e sam e whet her yoursyst em includes th e TU V or P DA detector. How ever, t he meth od set a ndreport methods vary. B efore a cquiring da ta , ensure tha t you chose theappropriate method set.

See also: For more informa tion on creat ing a sa mple set met hod, consult th eEm pow er online H elp.

To create the sample set method:

1. Set th e se sa mple set me thod pa r a me ter s:

• Con dit ion column on ce (r un t ime = 6.0 min u tes )

• I nject ion volum e = 5 µL (10 mm , 25 mm flow cell or E LS D)

• # of Inject ions = 3

• I f Empower sof tware controls the system, run t ime = 4.0 minutes(w ith Next Injection Delay = 2.5 minut es). If MassL ynx softw a recontrols the system, t ota l run t ime = 6.5 minutes

2. S a v e t h e s a mple s et m et h od .



Performing the gradient performance test 5-9

Performing the gradient performance test

Restriction: I f y our ACQU ITY U P LC system uses only a ma ss spectrometer,th e grad ient performa nce test is not a pplica ble.

When the syst em is prepared a nd t he test m ethods a re creat ed, you a re readyto perform the gra dient performa nce test . The steps for run ning th e test va ryslightly, depending on w heth er your syst em uses E mpower or Ma ssLy nxsoftw a re, but the desired results a re the sa me.

To perform the test:

1. S ta rt the run :• I f the sys tem is con trol led by Empower sof tware , open the project in

Run S a mples, select the gra dient performa nce test sam ple set , a ndth en select Run a nd Report .

• I f the system is con trol led by MassLynx sof tware , access theMassLy nx ma in pa ge, a nd select S ta rt from the Run menu.

2. When the sample set is complete, enter the appropria t e results in theta ble, below .

Retention time reproducibility (three replicates)

Peak

Peak

retention timemean value

Std devAcceptablestd dev

2-acetylfuran ≤1.5 secs

acetanil ide ≤1.5 secs

acetophenone ≤1.5 secs

propiophenone ≤1.5 secs

butylparaben ≤1.5 secs

benzophenone ≤1 5 secs




3. Review the gradient per forma nce repor t . The gradient per forman ce testresult is “passing” w hen th ese conditions a re realized:

• The peaks a re symmetr ica l , in tegra ted, and ident i fied cor rect ly .(Compa re the chroma togra m on the report t o th e sam plechroma togra m, below, to determ ine this.)

• The peak reten t ion t imes show a s ta ndard dev ia t ion of less than orequa l t o 2.0 seconds. (Consult th e t a ble you completed t o determinethis.)

benzophenone ≤1.5 secs

valerophenone ≤1.5 secs

Sample gradient performance test chromatogram

0.34

0.32

0.30

0.28

0.26

0.24

0.22

0.20

0.18

0.16

2 - a

c e t y l f u r a n

a c e t a n i l i d e

a c e t o p h e n o

n e

p r o p

l o p h e n o n e

b u t y l p

a r a b e n

b e n z o p

h e n o n e

e



Performing the gradient performance test 5-11

Note tha t t his is a representa tive chroma togra m. The results from yoursystem may vary s l ight ly .

0.16

0.14

0.12

0.10

0.08

0.06

0.04

0.02

0.00

-0.020.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00

Minutes

v a l e r o p h e n o n e




6 Maintaining the System

Contents:

Topic Page

Ma int ena nce schedule 6-2

C onta ct ing Wa ters t echnica l service 6-5

Ma int ena nce considera t ions 6-6

C onfigur ing ma int ena nce w a rnings 6-7



6-1

See also:

• I f your sys tem in cludes a P D A d etector , see th e ACQU ITY U PLC

Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for informa tion on

ma inta ining it .

• I f your system in cludes a n E L S dete ctor , see th e ACQUI TY UPLC

Evapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de forinforma tion on maint a ining it .

C onfigur ing ma int ena nce w a rnings 6 7

Ma int a ining the bina ry solvent ma na ger 6-8

Ma int a ining the sa mple ma na ger 6-29

Ma int a ining the column hea t er 6-61

Ma int a in ing the TU V det ect or 6-64Ma int a ining the sa mple orga nizer 6-77

Maintenance schedule

Wa ters recomm ends th a t y ou perform t he follow ing routine ma int enan ce onthe ACQUI TY U P LC system to ensure relia ble opera tion a nd a ccura te results.When using th e system t hroughout t he day (and on night s a nd w eekends), or

w hen using a ggressive solvents such as buffers, perform these ma intena nceta sks more frequently.

Recommended routine maintenance schedule

Maintenance procedure Frequency For information...

Repla ce th e mixer on t he

binary solvent ma na ger

Du ring scheduled

preventivemaintenance

See page 6-9

Replace th e check valve on th e Once every yea r or as See page 6-10



6-2 Maintaining the System

Replace th e check valve on th ebinary solvent ma na ger

Once every yea r or asneeded

See page 6 10

Repla ce solvent filt ers D uring scheduledpreventivemaintenance

See page 6-14

Clean t he air f i lters in th ebinary solvent ma na ger door

Du ring scheduledpreventivemaintenance

See page 6-14

Repla ce th e air filters in th ebinary solvent ma na ger door

Du ring scheduledpreventivemaintenance

See page 6-15

Repla ce th e bina ry solventmana ger head seals


See page 6-16

Repla ce th e bina ry solventma na ger plungers


See page 6-20

Repla ce th e vent va lvecartr idge

As needed S ee page 6-24

Defrost the sa mple

compartment


Repla ce t he sa mple needlea ssembly on t he sa mplemanager

Once every y ear or asneeded

See page 6-30

Ca librat e the needle Z-a xis onthe sample ma na ger

Whenever t he needle isreplaced

See page 6-40

Cha ra cterize the needle sea lon the sam ple mana ger

Whenever t he needleor needle sea l isreplaced

See page 6-41

Cha ra cterize th e needle andsa mple loop volumes on th esample mana ger

Whenever t he needleor sa mple loop isreplaced

See page 6-42

Repla ce the punctu re needle

holder on t he sa mple ma na ger


Repla ce t he sa mple loop on Once every yea r or as See page 6-45

Recommended routine maintenance schedule (Continued)




Maintenance schedule 6-3

the sample ma na ger needed

Repla ce th e sam ple syringe onthe sample ma na ger


See page 6-49

Repla ce th e w a sh syringes on

the sample ma na ger

Once every yea r or as

needed

See page 6-53

Modify th e sam ple syringeconfiguration pa ra meters onthe sample ma na ger


Replace th e inject ion valvecart ridge on th e sa mplemanager


See page 6-56

Test the backpressureregulator


Repla ce th e filter in t hecolumn in-line filter u nit onth e column hea ter

Du ring scheduledpreventive mainte-na nce or wh enever youexperience highpressure

See page 6-61

Replace t he colum n in -linefilter unit on t he columnheater

After y ou ha ve usedthe six fi l ters th a ta ccompanied th e unit

See page 2-27

Flu sh t he flow cell on t he TU Vdetector


P urge t he flow cell on theTUV detector


Reverse-flush t he flow cell ont he TU V detector As needed S ee page 6-68

Replace th e flow cell on t heTUV detector


Repla ce the la mp on t he TU Vdetector


See page 6-73

D efrost the sa mple orga nizer As needed S ee page 6-29

Clean t he air f i lter in thesample organizer


See page 6-77

Recommended routine maintenance schedule (Continued)





Repla ce th e fuses on t hesystem instruments

As needed S ee page 6-27,page 6-59,page 6-75, andpage 6-78

Clean th e system instrumentsw ith a soft, lint-free cloth , orpaper da mpened wi th w at er

As needed S ee page 6-28,page 6-61,page 6-76, andpage 6-79

Contacting Waters technical service

If you a re loca ted in t he US A or C an a da , report ma lfunctions or otherproblems to Wa t ers Technica l S ervice (800 252-4752). I f you a re locat eda nyw here else, phone Wa ters’ corpora te hea dqu a rt ers in Milford,

Ma ssa chuset ts (U SA), or conta ct your loca l Wa ters s ubsidiar y. Our Web siteincludes phone num bers a nd e-ma il a ddresses for Wa ters loca tions w orldwide.G o to ww w .wa ters.com, a nd click About Wa ters > Worldw ide Offices.

When y ou phone Wa t ers Technical S ervice, be prepar ed to provide th isinformation:

• E rror messa ge (if a ny)

• Na ture of t he sympt om• I nst r um en t ser ia l nu mber s (see “Loca ting sy stem serial numbers” on

page 7-5)



Contacting Waters technical service 6-5

page 7 5)

• F low ra te

• Opera t ing pressure

• Solvent (s)

• D etector set t i ng s (sen s it i vit y a n d w a v elen gth )

• Type a n d ser ia l number of column(s)

• Sa mple t ype

• E mpow e r or Ma ssL yn x softw a r e v er sion a n d ser ia l n umber

• ACQUITY UP L C sys tem w or ks t a t i on mod el a n d oper a t i ng sys tem

versionFor complete informa tion on report ing shipping da ma ges an d submitt ingclaims, see Wat er s L i censes, War r an ti es, and Su ppor t Ser vi ces .

Maintenance considerations

Safety and handling

Observe these wa rning a nd caut ion a dvisories w hen you perform ma intenanceon your syst em.

Warning: To prevent injury, a lwa ys observe good laborat orypra ctices when y ou han dle solvent s, cha nge tubing, or opera te t hesyst em. Know t he physica l an d chemical propert ies of th e solventsyou use. See the Mat eria l Sa fety Da ta Sheets for t he solvents in use.

Warning: To av oid electr ic shock, do not open t he power supplycover. The power su pply does not cont a in us er-serviceable pa rt s.




Proper operating proceduresTo ensure your sy st em run s efficient ly, follow th e opera ting procedures a ndguidelines in C h a pt er 3 and C h a pt er 5.

Spare parts

See Appendix C , for spa re pa rt s informa tion. You should not a tt empt to

replace a ny pa rts not l isted in Appendix C .

Caution:

• To a void da ma ging electr ical pa rts, never disconnect a n electr icala ssembly w hile power is a pplied to a n inst rum ent. To completelyinterrupt power t o an instr ument, set t he pow er switch to Off, an d

th en unplug t he pow er cord from the AC outlet. After power isremoved, w a it 10 seconds before you disconnect a n a ssembly.

• To prevent c ircui t dama ge due to s ta t ic cha rges, do not t ouchintegrat ed circuit chips or other syst em instruments tha t do notrequire manual adjustment.

Configuring maintenance warnings

Maint enance counters provide rea l-time usa ge sta tus informa tion tha t ca nhelp you det ermine w hen t o schedule rout ine ma int enan ce for s pecificcomponents. You ca n set usa ge thresholds and m aint enance w ar nings tha t

a lert you when a component r eaches th e designat ed threshold l imit . B ysett ing thr eshold limits a nd monitoring these usage counters regular ly, youca n minim ize unexpected failures an d unscheduled downt ime duringimporta nt work. Maintena nce wa rnings can be set for t hese conditions:

• Solvent ma nager pumped volume in l ite rs - The b inary solvent ma nagerrequires regular m a intena nce, the frequency of wh ich depends on thevolume of f luid i t pumps. You can set a ma intenan ce wa rning to a lert

you w hen t he pumped fluid rea ches a specified volume (pumped fluidvolume th reshold).

• Number of sample ma nager inject ion cycles - The sample ma nager



Configuring maintenance warnings 6-7

Number of sample ma nager inject ion cycles The sample ma nagerrequires regular m a intena nce, the frequency of wh ich depends on howma ny injections i t ma kes. You can set a ma intenan ce wa rning to alertyou w hen t he injection coun t r ea ches a s pecified count (injectionsthreshold).

• Detector lamp li fe in hours - The detector ’s lam p requires regularreplacement , th e frequency of w hich depends on its opera ting tim e. Youca n set a ma intenan ce w a rning to a lert you when the lamp has operat edfor a specified num ber of hours (t ime t hr eshold).

• Tota l number of inject ions on the column - Column l ife expecta ncy var iesw ith t he kinds of sam ples an d solvents you use an d on your usa gepat terns, including th e number of sam ple injections loaded onto thecolumn. You can set a ma intenance wa rning to alert you when th enum ber of injections on t he column rea ches a th reshold limit. Doing sohelps prevent chroma togra phic results from degra ding during a nimporta nt r un a nd protects t he system a ga inst sudden pressure cha ngesth a t sa mple build-up in the column can cause.

Maintaining the binary solvent manager

P erform t he procedures in th is section w hen you discover a problem wit h asolvent ma na ger component or during prevent ive ma intena nce. Forinforma tion about isolat ing problems in the binar y solvent ma na ger, see

“B inary solvent ma na ger troubleshooting” on pa ge 7-43.

Recommendation: Whenever you repla ce seals, a lwa ys w et t heir surfa ceswith methanol.

Warning: To prevent injury, a lwa ys observe good laborat orypra ctices when y ou han dle solvent s, cha nge tubes, or operat e thesolvent ma na ger. Know t he physica l and chemica l properties of thesolvents y ou use. Refer t o the Material S a fety Da ta Sheets for t hesolvents in use.




Binary solvent manager major components

TP02580

Primary

pump A

Primary

pump B

Accumulator

pump A

Accumulator

pump B

Degasser

Drain tube

(to waste)

PharMed tubing

(from degasser

motor to waste)Seal washleak tube

Solventselect valve A

Seal washVent valve

Solventselect valve B

Mixer

Replacing the mixer

Replace the mixer w henever t he int egra l filter in side of it becomes clogged.

Required materials



• AC QU I TY U P L C mixer (see Appendix C for t he part number)

Warning: To prevent injury, alw a ys w ear eye protection a ndpar t icle-free, pow der-free, non-la t ex gloves w hen ha ndlin g solvents .

Caution: To prevent contamination, wear particle-free, powder-free,non-lat ex gloves w hen r epla cing t he mixer.



Maintaining the binary solvent manager 6-9

To replace the mixer:

1. S t op t h e s olv en t flow .

2. U se the 5/8-inch open-end wr ench to hold the mixer in place.

3. U se the 1/4-inch open-end w rench to disconnect th e out let compressionfit t ing.

Disconnecting the outlet compression fitting

Mixer

Outlet

compression fitting

4. Aga in using t he 5/8-inch open-end w rench to hold the mixer, disconnectth e inlet compr essing fit t ings w ith t he 1/4-inch w rench.

Disconnecting the inlet compression fittings

Inlet compressionfittings

Mixer




5. Remove the old mixer f rom the bracket .

6. Ca refully remove the mixer f rom the packag ing .

7. In ser t th e new mi xer in to th e b ra cket .

8. Reat ta ch the compression f it t ings to the mixer , and t ighten thecompression fitt ings fing er-t ight plus a n a ddit iona l 1/4-t urn .

Replacing the check valve

Required materials


• 5/16-inch open-end wrench• C h eck va lve a s sem bly (see Appendix C for th e pa rt nu mber)

To replace the check valve:

1. P ow e r-off th e sol ven t ma n a g e r.

Caution: To prevent contamination, wear particle-free, powder-free,non-latex gloves when replacing the check valve.

2. Move the solvent bot t les to a loca t ion below the solvent ma nager hea d.

3. U se the 5/16-inch open-end w rench to hold the check valve in place.

4. U se the 1/4-inch open-end wr ench to disconnect the t ubing connectionfrom the check valve.

Disconnecting the tubing from the check valve

Warning: Solvent spill risk. Fa ilure to move the solvent bott les t o aloca tion below th e solvent m a na ger head ma y result in solvent spills.

5/16-inch

open-end wrench




5. U se the 1/2-inch open-end wr ench t o loosen t he check va lve, an d remove

th e check valve assembly from the solvent m a na ger head.

Check valve

Tubing connection

1/4-inchopen-end wrench

Disconnecting the check valve from the solvent manager head

Pressure transducer cable

Pressure transducer

Check valve

1/2-inchopen-end

Solvent manager head




6. Ca refully remove the new check valve a nd wa sher f rom the packaging.

7. Inser t the new P EEK wa sher into the new check va lve. Make sure thecha mfered edge of the w a sher faces aw a y from the check va lve.

Caution: When y ou remove t he check valve, ma ke sure you a lsoremove the P EE K w asher .

1/2 inch open endwrench

Inserting PEEK washer into check valve

8. Inser t the check valve assembly into the solvent ma nager head, andtight en t he check va lve nut finger tigh t plus a n a dditional 1/8 tu rn

PEEK washer

Check valve

Chamfered edge

facing away from

check valve




tight en t he check va lve nut finger-tigh t plus a n a dditional 1/8-tu rn.

9. Reat ta ch the f it t ing to the check valve , and t ighten the compressionscrew finger -t ight plus a n a ddit iona l 1/4-tu rn for exist ing st a inless st eeltu bing or finger-tight plus a n a dditiona l 1/2-tu rn for n ew s ta inless steeltubing.

Replacing solvent filters

To replace a solvent filter:

1. Remove the f il tered end of the solvent t ubing from the solvent bottle.

2. Remove the old solvent f i l ter from the short piece of P TFE t ubing, an dreplace i t w ith a new one.

Replacing the solvent filter

Caution: Wea r pa rt icle-free, pow der-free, non-la tex gloves w henha ndling t he solvent f i lter . Oil from your han ds can conta mina te th esolvent filter.




3. Inser t the f il tered end of the solvent tubing into the solvent bot t le .

Cleaning the air filters in the binary solvent manager door

To clean the air filters:

U se a va cuum to clean t he air filters loca ted inside the bina ry solventma na ger door.

TP02484

Solvent filter

PTFE tubing

Solvent

tubing

Replacing the air filters in the binary solvent manager door

If t he a ir f i lters cannot be clea ned by va cuuming, replace them w ith n ewfilters.

Required materials

• T10 TORX® driver

• B i na r y solven t m a na g er a ir filt er s (s ee Appendix C for t he part number)

To replace the air filters:

1. U sing the T10 TORX driver, remove the eight screws tha t secure the airf i lter fra mes an d a ir f i lters t o the inside of th e bina ry solvent ma na ger

door.

Air filters inside the binary solvent manager door




2. Remove the old a ir fi lters from the a ir f il ter f rames and discard t hem.

3. Al ign the new a i r fi lte rs w i th the a i r fi lte r fr ames.

4. U sing the T10 TORX dr iver, a t t ach the a ir fi lters and f rames to the

inside of th e binary solvent ma na ger door w ith t he eight screws.

TP02429

Air filter

Frame

Air filter

Frame

Removing and replacing the solvent manager head seals

See also: C ha pt er 7, to help determ ine wh ether y ou need t o replace th esolvent ma na ger pump head sea ls.

Required materials

• T27 TO RX d riv er (s t a rt u p kit )


• 5/16-inch open-end wrench

• H ea d sea l a n d sea l w a s h s pa cer (s ee Appendix C for t he part number)

• Met ha nol


non-lat ex gloves w hen removing a nd replacing the solvent ma na gerhead seals.




Met ha nol

• S ea l ext ra ct ion tool

To remove the solvent manager head:

1. Flush the binary solvent ma nager w i th a ppropr ia t e solvent .


3. In the binary solvent mana ger informa tion window, cl ick Mainta in >Heads.

4. In the Head Maintena nce dia log box, select the head (P r imary orAccumula tor) from the solvent ma na ger (A or B ) th a t y ou plan toperform ma intena nce on.

5. Cl ick Move B ackwa rd , and then w a i t for the plunger to s top.


7. Remove the sea l wash tubes secured to the sea l wa sh housing by tw obarbed f i t t ings.

Warning: Solvent spill risk. Fa ilure to move the solvent bott les t o aloca tion below t he solvent ma na ger head ca n result in solvent spills.

8. U se the 1/4-inch open-end wr ench to disconnect th e out let tube from t hetransducer.

9. U se the 5/16-inch open-end wr ench to hold the check valve ca rt ridge inplace.

10. U se th e 1/4-inch open-end w rench t o disconn ect t he t ubing connection

from the check valve.






11. Disconnect t he pressure tra nsducer ca ble from t he bulkhead.

Check valve

Tubing connection


12. U sing th e T27 TORX driver, remove the tw o hea d bolts, w hich area ccessible from th e front of the pressure tra nsducer.

Removing the head bolts securing the solvent manager head

TP02435




The bolts rema in ca ptive behind t he mounted tr a nsducer.

13. G ently pull the head off the a ssembly, making sure not t o t i l t the headduring t he extraction.

Removing the solvent manager head

Caution: To a void da ma ging th e plunger, support th e solvent ma na gerhea d from below a s you remove it.

To remove the solvent manager head seals:

1. S ta nd the solvent mana ger head upr igh t on a c lean sur face.

2. Remove the old sea l wa sh sea l and d iscard it .

Solvent manager head seals

Seal wash

housing

Seal wash

seal

Solvent manager head

PTFE O-ring

High pressure seal

Seal wash spacer




3. Inver t the solvent mana ger head so tha t the sea l wa sh housingsepa ra tes from t he hea d.

4. U sing the sea l extract ion t ool , carefully pul l the high pressure sea l out ofthe solvent ma na ger head. Ta ke ca re not to scra tch a ny surfa ces.

5. Ca refully remove the P TFE O-r ing .

6. P ress the new P TFE O-r ing in to i t s sea t w i th your thumbs .

7. Lubrica t e the new high pressure sea l w i th methanol, and place i t looselyin the sea t .

8. Center the sea l backup r ing over the high pressure sea l so tha t th ecross-side faces upward.

9. Orient t he sea l wa sh housing so tha t t he holes on the side of it l ine upw ith t he holes on th e side of the solvent ma na ger head.

10. Reinsta ll the seal wa sh housing into the solvent ma na ger head, pushingdown unt il you h ear (or feel) th e high pressure seal sn a p into place.

11. Lubrica t e the sea l w ash sea l wi th metha nol , and sl ide it onto theplunger. Slide th e sea l back until it conta cts t he support plat e.

sealSeal wash spacer

To reattach the solvent manager head:

1. Moisten the sea l s and p lunger w i th methanol.

2. Ca refully s lide the hea d assembly over the sapphire plunger .

3. Align the mounting holes in the support pla te wi th t he two 5-mm screws

tha t a re ca ptive in the head a ssembly.4. With the hea d assembly secure against t he support pla te , use the T27

TORX driver t o tighten t he head screw s evenly.

Replacing the solvent manager plungers

See also: C ha pt er 7, for help determining w heth er you need to repla ce the

plungers.

Caution: To prevent contamination, wear particle-free, powder-free,non-lat ex gloves wh en repla cing t he solvent m a na ger plungers.




Required materials

• 1/4-in ch open -en d w r en ch• 5/16-inch open-end wrench

• T27 TORX dr iver

• R epla cem en t plu ng er (s ee Appendix C for th e pa rt number)

• Met ha nol

To remove the solvent manager head:

1. Flush the binary solvent ma nager w i th a ppropr ia t e solvent .


3. In the binary solvent mana ger informa tion window, cl ick Mainta in >H eads. The Head Ma int enan ce dialog box appears.

4. Select t he head (P r imary or Accumulator) f rom the solvent m a nager (Aor B ) tha t you pla n t o perform m a intenan ce on.

5. Cl ick Move Forwa rd , and then wa i t for the plunger to s top.


7. Remove the sea l wash tubes secured to the sea l wa sh housing by tw o

barbed f i t t ings.

8. Remove the f it t ings a t ta ched to the check valve and pressure tra nsduceroutlet.

9. U se the 5/16-inch open-end w rench to hold the check valve in place.

10. U se th e 1/4-inch open-end w rench t o disconn ect t he t ubing connectionfrom the check valve.


Warning: Solvent spill risk. Fa ilure to move the solvent bott les t o aloca tion below t he solvent ma na ger head ma y result in solvent spills.




11. Disconnect t he pressure tra nsducer ca ble from t he bulkhead.

Check valve

Tubing connection


5/16-inch

open-end wrench

12. U sing t he T27 TORX driver, remove th e tw o support plat e bolts.

Removing the solvent manager head and support plate bolts

TP02436




13. G ently pull the solvent ma nager head a nd support pla te off the actuat orhousing, ma king sure not to t i l t t he head during t he extra ction.

Removing the solvent manager head and support plate

TP02436

To replace the plunger:

1. S ta nd the solvent ma nager head a ssembly uprigh t on a clean surface,a nd set i t a side i f you are not repla cing any seals as pa rt of thisprocedure. If you a re replacing seals, r efer t o “Removing a nd replacingthe solvent ma na ger hea d seals” on pa ge 6-16 for specific inst ruct ions.

Recommendation: Wa ters st rongly recommends t ha t y ou repla ce th esolvent ma na ger head sea ls when you replace th e plunger.

2. U se your f ingers, or th e T27 TORX driver, to apply pressure on th erelea se collar . This a ction disenga ges th e old plunger from t hemechanism.

R l i th l

Caution: To a void da ma ging th e plunger, support th e solvent ma na gerhea d from below a s you remove it.




Replacing the plunger

3. G rasp the new plunger wi th a clean, l in t-free cloth , and inser t theplunger w hile applying pressure to th e relea se collar .

T27 driver

Plunger

Piston

Spring-loadedrelease collar

To reattach the solvent manager head:

1. L ubr ica te th e plun ger w i th meth a n ol.

2. P lace the solvent mana ger head a ssembly in posit ion on the mechanism.In sert t he tw o T27 TORX screws into t he holes in t he support plat e, an dtight en the screw s evenly.

3. Connect the pressure tra nsducer cable to the bulkhead.

4. R ea t t a ch a ll fit t in gs .

Replacing the vent valve cartridge

Caution: To avoid brea king th e plunger, ensure t ha t t he solventma na ger hea d a ssembly is stra ight w hen you position i t on themechanism.





Required materials

• H ex w rench, 2-mm

• Ven t va lve ca r t ridg e (see Appendix C for th e pa rt n umber)

To replace the vent valve cartridge:


2. In the binary solvent mana ger informa tion window, cl ick Intera ct iveDisplay.

3. In t he binary solvent mana ger interact ive display d ia log box, cl ickControl .

p p pnon-lat ex gloves wh en repla cing t he vent va lve ca rt ridge.

4. E n sur e th e ven t v a lv e is set t o Ven t .

Tip: To cha nge t he sett ing t o Vent , click on Vent Valve ima ge a nd selectVent.

Interactive display showing vent valve setting

5. Remove the fit t ings a t t a ched to the vent va lve ca r t r idge.

Vent valve cartridge with fittings




6. U se the 2-mm hex wrench to remove the hex screw a t t he 10 o’clockposition on t he vent va lve ca rt ridge.

7. Remove the vent va lve car t r idge f rom the vent va lve a ssembly.

8. Ca refully remove the replacement vent va lve ca r tr idge from thepackaging.

Pump Bvent/waste

From pump A

(transducer,

accumulator)

Pump B

to systemPump A

vent/waste

Pump A

to system

From pump B

(transducer,accumulator)

Hex screw in10 o’clock position

9. Ensure that the groove in the car t r idge housing a l igns wi th the grooveon the drive cla mp. If it does not, t urn t he drive clam p unt il the groovesalign.

Aligning vent valve cartridge grooves

10. Insert th e new vent va lve ca rtr idge into the vent va lve ca rtr idgechamber.

Drive clamp

Aligned grooves




11. In sert t he 2-mm h ex screw a t t he 10 o’clock position on t he vent va lveca rt ridge. U se the 2-mm h ex w rench to tigh ten it.

12. Rea t t ach a l l f it t ings .

13. In th e ACQU ITY UP LC C onsole, select B inary S olvent Ma na ger fromthe syst em tr ee. The binary solvent ma na ger informa tion windowappears.

14. C lick Cont rol > P rime A/B S olvent s. The P rime A/B S olvent s dia log boxappears.

15. In t he Solvent Line(s) to P rime area , select the solvent l ine(s) you wa ntto prime.

16. Ch oose solvent A1 or A2 for line A a nd solvent B 1 or B 2 for line B .17. In t he Time (min) box, type the number of minutes tha t y ou w a nt to

prime the bina ry solvent ma na ger for .

18. Click St a rt . The vent va lve sw itches to th e Vent position, one or bothpumps begin priming at specified flow ra tes, an d th e bina ry solvent

Caution: The vent va lve cart ridge must slide fully into th e vent va lvea ssembly. If it does not, cont a ct your Wa ters service representa tive.

ma na ger sta tus displays “P ump P riming.” When priming is complete,the sta tus returns t o “Idle.”

Replacing the fuses

Sus pect a n open or otherw ise defective fuse wh en th e

• s olven t ma n a g er fa i ls t o pow er -on .

• solven t m a na g er s ta t us L E D s a r e u nlit .

• pow er su pply fa n does not oper a t e.

Required materials

Warning: To av oid electr ic shock, power-off an d un plug t he solventma na ger before examin ing t he fuses. For cont inued protectiona ga inst fire, replace fuses only w ith t hose of th e sam e type andrat ing.




• 5 A, 250 V, 5 × 20 mm, slow-blow, IEC type fuse (2) (see Appendix C forthe part num ber)

• F la t -bla de scr ew dr iver

To replace the fuses:

Requirement: Replace bot h fuses, even w hen only one is defect ive.

1. P ow e r-off th e sol ven t ma n a g e r.

2. Disconnect t he power cord f rom t he power entry module .

3. U se a f lat -blade screwdriver to open the fuse holder door, which islocat ed above th e pow er ent ry m odule on t he rear pa nel.

Removing the fuseholders

4. With minimum pressure, pull on each spring-loa ded fuse holder toremove it.

Fuse holders

Fuse holder door

Fuse

Power entry module




remove it.

5. D isca r d t h e fu ses.

6. Inser t the new fuses into the holders and t he holders into the powerentr y module.

7. Reconnect t he power cord to the power entry module .

Cleaning the instrument’s exterior

U se a soft cloth , dam pened wit h w a ter, to clean t he out side of th e solventmanager .

Warning: For cont inued protection a ga inst fire, replace fuses w ith t hea ppropriate type and ra ting.

Maintaining the sample manager

Defrosting the sample compartment

Defrost t he sample compart ment w hen the sa mple ma na ger is una ble to reach

a set tempera ture between 4 a nd 10 ° C (39.2 and 50 ° F) in a hot a nd humidenvironment.

Tip: To keep the sa mple compa rt ment from freezing, open it s door only w hennecessar y. (Opening t he door a dmit s humid a ir into the sam ple compa rt ment ,w hich ca uses condensa tion a nd freezing.)

To defrost the sample compartment:

1. Open the sam ple compartment door , and remove any sa mples.

Tip: The sa mple orga nizer (if present) aut omat ica lly defrosts a long w iththe sample mana ger .

Warning: To prevent hea vy condensa tion from forming inside t he



Maintaining the sample manager 6-29

2. I f your system conta ins a sample organizer , open the sample organizerdoor a nd remove a ny t empera tu re-sensitive sam ples.


4. In the sample ma nager informat ion window, cl ick Mainta in > Defrost .The D efrost dia log box a ppea rs.

5. In t he Defrost d ia log box, cl ick St ar t . The sample compartment defrosts

for 60 min ut es.

Warning: To prevent hea vy condensa tion from forming inside t hesam ple compart ment a nd da ma ging i t , the sa mple compar tment door

must rema in open th roughout th e defrost procedure.

Warning: To prevent hea vy condensa tion from forming inside t hesam ple orga nizer a nd da ma ging i t , the sa mple orga nizer door mustrema in open t hroughout t he defrost procedure.

Replacing the sample needle assembly

Repla ce the needle a ssembly wh en the

• s a mple ma n a g er is un a b le to r ea ch sa mple t r a n s fer pr essur e

• needle is ben t• needle t ip is da ma ged

• needle is plugged

Required material

Warning: To a void puncture w ounds keep ha nds or loose clothin g clea rof the needle as sembly mechan ism w hile it is moving. The sa mplema na ger beeps three times wh enever t he door is open a nd t he needlea ssembly mecha nism is about t o move.

Caution: To a void opera t iona l problems, ensu re th e sa mple needle isproperly inst a lled.




q

• Needle a ssem bly (see Appendix C for t he part number)

To remove the needle assembly:

1. In the ACQUI TY UP LC Console, cl ick Sam ple Mana ger , in the lef t pane.

2. Select Mainta in > Cha nge needle. A message request ing that youremove the r ight-ha nd sa mple pla te from t he sam ple ma na gercompa rtm ent a ppears.

3. Open th e s a mple ma n a g er door .

4. P ul l out the r ight-hand t ray, a nd remove the sample pla t e , i f one isloaded.

5. After removing the sample pla te f rom the tra y, cl ick OK on theACQU ITY U P LC Console message w indow.

6. U nscrew the needle assembly’s f it t ing f rom injector por t 3 .

Injection valve

Port 3fitting




7. Remove the fit t ing from the need le.

8. In t he sample compartment , loosen the red thumbscrew on the needlemounting bra cket.

Needle assembly

Red thumbscrew

Needle latch




9. P ush the needle la t ch ba ck to release the needle mounting cyl inder f romits mounting ca vity.

Releasing the needle cylinder from the mounting cavity

Latch

Mounting cavity

Mounting cylinder

Flag

Loose loop oftubing




10. Lift the needle t ip out of the white pla stic guide a t t he bott om of theXYZZ mechanism.

Removing the needle from the white plastic guide at the bottom of theXYZZ mechanism

Needle tip

White plastic guide




11. Remove the needle assembly from t he sample compart ment.

ACQUITY UPLC sample needle assembly

Warning: To avoid puncture w ounds, or da ma ge to the end of theneedle, do not t ouch or press th e end of the sa mple needle.

Needle tip

Fitting

ID sleeve

Clamping sleeve

Mounting cylinder

Protective cap




To install the sample needle assembly:

1. Inser t the needle t ip into the guide tube next to the injector .

Threading the needle assembly through the needle guide tube

Needle tip

Needleguide tube

2. G ently push the needle assembly into the sam ple compartment .

3. From inside the sample compartment , ta ke hold of the needle assemblytubing a s i t enters t he cha mber from a bove.

4. Remove the protect ive cap f rom t he needle t ip.

5. Hold the needle by the mounting cyl inder , and wi th t he needle t ip

pointing down w a rd, insert i t s t ip into the wh ite plast ic guide a t t hebott om of the XYZZ mecha nism.

Inserting the needle tip into the needle guide

Needle tip

Needle guide




Needle guide

6. Inser t the needle mounting cyl inder into the mounting cavi ty .

Inserting the needle cylinder into the mounting cavity

Needle latch

Mounting cavity

Mounting cylinder

Z-flag

Loose loop oftubing




7. P ul l the needle la tch forwa rd to secure the needle assembly.

8. Form a loop so tha t t he por t ion of the tubing wi th the black clampingsleeve ca n be secured by t he th umbscrew.

9. Route the tubing through the groove below the Z-f lag , a s shown in thefigure below.

Tubing routed through groove

Z-flag

Loose loop of

tubing

Groove




Incorrect placement of loose loop of tubing

Caution: To avoid dam a ging t he inst rument , do not place the looseloop of tubing a bove t he Z-fla g.

Z-flag

Loose loop of

tubing

10. Align t he tubing so tha t t he portion just t o the left of the clamping sleevefits into the notch behind the thum bscrew .

11. Tighten the th umbscrew so tha t i t secures the portion of the needle w ithth e black sleeve.

Needle latched with loop

Needle assembly

Thumbscrew

Loose loop of

tubing

Z-flag




12. En sure tha t t he needle tubing is fully inserted into port 3, on th e

injection va lve, a nd t hen th read t he f it t ing into the port , t ightening thefittin g securely.

13. Close the sample ma nager door .

14. On th e ACQU ITY UP LC Console, cl ick Control > Prime syringe.

15. In t he Prime Syringes dialog box, select S a mple syringe a nd wa shsyringes.

16. Type 1 in t he “Number of cycles” t ext box.

17. Click OK. The sample ma na ger sta tus display s “Pr iming.” Whenpriming is complete, the sta t us return s to “Idle.”

Calibrating the needle Z-axis

You n eed t o ca librat e th e needle Z-a xis w henever y ou repla ce th e sa mpleneedle.

To calibrate the needle Z-axis:

1. Cl ick Main ta in > Ca l ibra te needle Z ax is .2. In t he Cal ibra t e Needle Z Axis d ia log box, cl ick Sta r t , a nd then cl ick OK

in t he confirmat ion w indow .

3. U se t he +Z button to drive the sample needle down to with in 1 mm of thetray surface.

4. Sw itch the displacement increment to 0.1 mm, and lower the sample

needle unt il it is a lmost touching the surfa ce of th e sam ple tr a y holder.

Tip: To ea sily a nd efficient ly a chieve t he correct needle sett ing, slide abusiness card under t he needle. Then lower t he needle until it touchesth e ca rd lightly but does not restr ict t he ca rd’s free movement .

Adjusting the sample needle




5. C lick S ave.

Result: The confirma tion w indow a ppea rs.

6. C lick Yes.

Sample needle almost

touching surface of sample

tray holder or just touchingbusiness card

Characterizing the needle seal

The needle sea l cha ra cteriza tion function finds t he position a t w hich theneedle achieves a seal on the w a sh st a tion block. Cha ra cterizing th e needleseal is critical t o a ccepta ble sa mple ma na ger performa nce.

P erform th is procedure a fter y ou repla ce a nd/or a djust t hese items:

• The sample need le or any par t o f the need le assembly

• The needle (Z) or piercing needle (Zp) f lags (home and top-of-pla te)

• A h ome or t op-of-pla te sen sor

• The needle sea l

• Th e inject port sea l

• The w a sh st a tionRequirements:

• Make sure the sample mana ger is pr imed before chara cteriz ing theneedle seal.

• P er form this procedure before ca l ibra t ing the needle and sample loopvolumes




volumes.

To characterize the needle seal:

1. Cl ick Main ta in > Chara cteri ze > Needle sea l .

2. Cl ick S ta r t . The chara cter iza t ion beg ins , and the sample mana ger s ta tusdisplays “Cha ra cterizing seal .”

Result: When t he opera tion ends, the Results pa ne a ppea rs.

3. I f character iza t ion is unsuccessful , examine the needle to ensure youinsta lled i t properly. Make an y necessary a djustments, a nd th encha ra cterize th e needle sea l ag a in. If unsu ccessful, see C ha pt er 7,Dia gnostics a nd Troubleshooting.

4. I f the cha racter iza t ion is successful, cl ick Close.

Characterizing the needle and sample loop volumes

P erform t his procedure w henever you chan ge the

• mobile pha ses

• w a sh solvent s

• sa mple loop

• needle

• syr inges

Whenever y ou replace th e sa mple loop a nd/or sa mple needle, you mustcha ra cterize the volume of th e repla cement par ts t o accommodat e thedifferently sized pa rt s a nd to ensur e optim a l chroma togra phic results. Do thisrega rdless of whet her th e sizes of the repla cement pa rt s equa l those of th eorigina l part s or differ from th em.

Ch a nges in th e composition of the w eak w a sh solvent ca n a ffect viscosity ,surfa ce tension, an d/or polar ity. D uring sa mple injection, t he w eak w a shsolvent both precedes a nd follow s t he sa mple in th e fluid-carry ing lines, so thesa mple is directly a ffected by th e w eak w a sh solvent.

Requirements:




Requirements:

• Speci fy the s izes of the sam ple needle, loop, and syr inge in the Volumesdialog box before you characterize their volumes.

• P r ime the sample mana ger before chara cteri zing the volumes .

• P e r for m a meth od setup usin g E mpow e r or M a ssLyn x sof tw a r e w i th a n ymethod tha t ha s the same a ir gap a nd sample draw ra t e tha t you w i ll beusing.

To characterize the needle and sample loop volumes:

1. Cl ick Mainta in > Cha racter ize > Needle a nd loop volumes.

2. In t he Cha racterize Needle and Loop Volumes dia log box, cl ick St ar t ,a nd t hen a llow a pproxima tely 10 minut es for t he test t o run (see th e runtime bar gra ph).

• Cha racter iz ing the loop volume measures the volume of the loop (inmL).

• Chara cteri zing the need le volume measures the volume of theneedle (in mL ).

Tip: The syst em records this t est in t he log a s a dia gnostic event .

3. To v iew the da ta , cl ick Results .

Results: If t he loop test produces a fa iling result , suspect th a t t he loop isblocked or leaking. I f t he needle test produces a failing r esult, suspectth a t t he needle is bent , broken, or blocked. Also, ensu re th e needle,sa mple loop, an d sa mple syringe configura tions a re correct.

4. To print the result s , cl ick Pr in t .

5. C lick C lose.

Replacing the puncture needle holder

Repla ce th e puncture needle holder if it becomes da ma ged.

Required materials

Warning: For protection a ga inst injury , do not remove th e protectivecover from t he puncture needle unt il th e replacement procedure iscomplete. Touching or pressing t he end of th e needle can cau se apuncture wound.




• P u n ct u re n eed le ca r t r id ge k it (s ee Appendix C for th e pa rt n umber)• Needle-n ose plier s

• T6 TORX dr iver

• C lea n sheet of pa per

To replace the puncture needle:


2. Select Ma inta in > Replace > Needle. A message request ing tha t youremove the r ight-ha nd sa mple pla te from t he sam ple ma na gercompart ment a ppears.

3. Open the sample manager door . Pul l out the r igh t-hand t r a y , and

remove th e sam ple plat e, if one is loa ded. Use a sma ll, th in coin t o tu rnth e screw on the bott om of th e tra y 1/4 t urn count erclockwise, releas ingthe t r ay .

4. Remove the wel l p la te t r ay f rom the sample manager compar tment .

5. In t he ACQU ITY UP LC C onsole, cl ick OK to move the needle to themaintenance position.

6. P lace a sheet of paper on the floor of the sample ma nager compartment ,beneat h t he puncture needle. The pa per w ill ca tch t he needle if it fa llsduring replacement .

7. Remove the needle clean system tube from the barbed f it t ing on theexisting puncture needle holder.

8. Loosen the T6 set screw t ha t secures the puncture needle holder. Thepuncture needle holder should fall out of t he housing. If it does not,gra sp th e puncture needle holder w ith th e needle-nose pliers, a ndca refully pull downw a rd. I f th e punctu re needle holder does not comeout of th e needle block, loosen th e set screw un til it pulls free.

9. G ra sp the new puncture needle holder by the protective pla stic cover onth e needle tip.

10. Slide the puncture needle holder over t he sa mple needle and into thehousing. Orient th e needle so th a t t he barbed fitt ing enters th e slot onth e needle block.




Barbed fitting in slot on needle block

11. Hold the puncture needle a ssembly in position w hile you t ighten the setscrew w ith th e T6 TORX driver.

Needle block

Barbed fittingSlot

12. Reat ta ch the needle clea n system tube to the barbed fi t t ing.

13. Ca refully remove th e protective cover from the punctur e needle.

14. Remove the paper from the sample ma na ger compar tment .

15. Refit the tr a y, a nd t urn t he screw 1/4 tu rn clockw ise to secure it.

16. Select C ont rol > Reset S M to end the ma intenan ce procedure a nd resetthe sample mana ger .

Replacing the sample loop

Repla ce the sa mple loop wh en it clogs or w hen its ca pacity is ina ppropriat e foryour chroma togra phic need.

Caution:

• To prevent ba ndsprea ding or excessive ca rryover, be sure toreinsta ll a previously inst a lled sa mple loop in exa ctly t he sa meorienta tion tha t i t w a s insta l led before.

• The O-rings on the new sa mple loop are used to hold the sea ls inplace durin g sh ipping. To avoid leaks , remove th e O-ring s prior to




insta lling t he sa mple loop.

Tips:

• With a piece of tape, mark the ends of a previously insta l led sample loopto help ensure i t is insta l led in t he same orienta tion t ha t i t w a s insta l ledin before.

• A Wat ers field serv ice eng ineer must ca l ibra te the new a i r sensorw henever you replace th e sa mple ma na ger's air sensor assembly.

Required materials

• AC QU I TY U P L C sa m ple loop (s ee Appendix C for th e part number)

• Open -en d w r en ch , 1/4-i nch

To replace the sample loop:

1. S l ide out the sample mana ger flu id ics t r ay .

2. Hold the sample loop, a nd pul l the inject ion va lve towa rd you.

Caution: To av oid kinks, do not pull on th e volum e detection device(port 2) or sample needle (port 3) tubing.




3. U se the wrench t o remove the f it t ings on por ts 1 and 4 of the inject ionvalve.

Sample loop fittings on the injection valve

4. Remove the sample loop and it s f it t ings.

5. P lace the sample loop and i ts fi t t ings in a conta minat ion-proof bag .

Port 1fitting

Port 4

fitting

Sampleloop

Inlet frompump

Outlet to

column




6. Ca refully remove the replacement sample loop and f i t t ings f rom thepackaging.

Sample loop (assembled)

To ports 1 and 4 on

injection valve

7. Remove the O-r ings f rom the sample loop.

8. Slide a tw o-piece ferrule a nd compression screw onto one end of thesa mple loop, and th en seat th e end into injection va lve port 1.

9. Tighten the fi t t ing wi th the wrench unt il snug .

See also: “ Inst a l la tion recommenda tions for f i t t ings” on pa ge 2-13.

10. Remove the f it t ing an d inspect t he ferrule to ensure i t ha sn’t m oved.

Ferrule installation

Caution: The O-ring s on th e new sa mple loop a re used t o hold th esea ls in place during s hipping. To a void lea ks, remove the O-ring sprior t o insta lling t he sa mple loop.

Caution:

• To prevent dead volume in the system, ensure tha t t he ends of theloop a re fully insert ed into t he port s of the injection va lve.

• U se only Wat ers approved ACQU ITY UP LC loops wi th ACQU ITY

U P LC fitt ings. Ea ch loop should be fitt ed to its injector.




11. Repeat steps 7 through 9 for t he other end of the sam ple loop an d forinject ion va lve port 4.

12. Reinsta ll the sam ple loop f it t ing into va lve ports 1 a nd 4.

13. P ush th e injection va lve back to th e closed position.

Caution: To avoid kinks , do not pus h on t he volume detection device

(port 2) or sample needle (port 3) tubing.

TP02506 TP02507

TubingFerrule TubingFerrule

Correct Incorrect

14. Slide the f luidics tra y closed.

15. En sure the sam ple ma na ger compart ment door is closed.

16. In t he ACQU ITY UP LC C onsole, select S am ple Man a ger from thesystem t ree.

17. Click Maint a in > Ca libra te Syst em Volume. This cha ra cterizes the newsyst em volumes.

Replacing the sample syringe

Tip: Air bubbles in th e syringe a dversely a ffect syst em pressure, ba seline,volume, and peak a rea . Remove a ir bubbles by gent ly ta pping on the syringea s i ts plunger retracts.

Repla ce th e sam ple syringe w hen either of these conditions a rise:

• The syr inge p lunger t ip becomes worn or d i scolored

• You w a n t t o ch a n ge t o t h e ot h er sy r in ge s iz e

Caution: B e ca reful not t o crush t he volume detection device line.




• Th e sy rin g e lea ks or ca uses a ir bubb les

Required materials

• R epla cem en t sa m ple s yr in ge (s ee Appendix C for th e pa rt n umber)

• D ega ssed, w ea k w a sh solven t

Warning: To a void injury , ensure t ha t n o inject ion is in progress orpending before you remove the sa mple syringe.

Maximum syringe draw rates for the 30 µL needle

Aspiration (Rheodyne Sample Loops)

Solvent Type 2 µL 5 µL 10 µLa

20 µL 50 µLb

50:50met ha nol/w a t er

65 250 280 260 310

100% w a ter 120 450 500 480 550

100% a cet onit r ile 300 1130 1300 1250 1400

a. The 10 µL loop is the st a nda rd loop.b. The 50 µL loop must be used wit h a 250 µL sa mple syringe.

Maximum syringe draw rates for the 15 µL needle


Solvent Type 2 µL 5 µL 10 µL

a

a. The 10 µL loop is the st a nda rd loop.

20 µL 50 µL

b

b. The 50 µL loop must be used wit h a 250 µL sa mple syringe.

50:50met ha nol/w a t er

32.5 125 140 130 155

100% w a ter 60 225 250 240 225

100% a cet onit r ile 150 565 650 625 700

Maximum syringe draw rates for the 30 µL needle (Continued)


Solvent Type 2 µL 5 µL 10 µLa

20 µL 50 µLb




µ p µ p g

To replace the sample syringe:


2. Cl ick Mainta in > Replace > Sa mple syr inge. This invokes a w izard tha tmoves t he syringe t o the down position.

3. Remove the knurled screw t hat holds the syr inge barrel to the syr ingemounting bra cket.

Sample syringe assembly components

Samplesyringe valve

Sample syringe

Knurled collar




4. U nscrew the sample syr inge counterclockwise unt i l it separa tes f rom thesa mple syringe va lve.

Knurled screw

5. Depress the syr inge barrel to clear t he top mounting bracket , a ndremove th e syringe.

Removing the sample syringe

Caution: To a void breaking t he syringe, do not gra sp it by its gla ssba rrel. Alwa ys gra sp the syringe by its knu rled collar .

Sample syringe valve

Mounting bracket

Knurled collar




6. Ca refully remove the replacement sa mple syr inge from the packaging.

7. P ar t ia l ly f il l the new syr inge (by hand) w i th weak wa sh solvent (to helpremove a ir bubbles).

Requirement: Ma ke sure all air bubbles ar e removed.

8. Retract t he syr inge plunger so tha t t he plunger end slides over the

th rea ded post on the syringe guide mount ing bracket.9. Screw t he new sample syr inge par t ia l ly into the sam ple syr inge va lve.

10. Finger t ighten the sa mple syringe.

Threaded holder

Syringe plunger

11. Insta l l and f inger t ighten the knurled screw tha t holds the samplesyringe plunger to th e mount ing bra cket.

12. Run the P rime Sa mple Syr inge Only option until there a re no a irbubbles in t he sa mple syringe.

Replacing the wash syringes

The only wa sh sy ringe size ava ilable is 2.5 mL. To ensure a ccura te r esults,use only syr inges tha t a re approved by Wa ters for use in th e ACQUITY UPLC

system. S ee Appendix C for a list of Wa ters-a pproved syringes.

Repla ce the w a sh syr inges wh en th e syringe plunger’s t ip leaks or becomesw orn or discolored.

Required materials

• R epla cem en t w a s h sy rin ges (see Appendix C for th e pa rt number)

• D ega ssed wa sh solvent s

Caution: To a void injury , ensur e tha t n o inject ion is in progress orpending before you remove the syringe.




To replace the wash syringes:


2. Remove the knurled screws t hat secure the wa sh syr inge plungers to thew a sh syringe mount ing brackets.

Wash syringe assembly components

Weakwash

syringeStrong

wash

syringe

Mounting

bracket

Mounting

bracket




3. Looking dow n a t t he wa sh syr inges, unscrew each syr inge, clockwise,unti l i t sepa ra tes from t he mounting bra cket.

Knurled

screws

4. Depress the syr inge barrel to clear t he top mounting bracket , a ndremove th e syringe.

Removing wash syringes

Caution: To a void breaking t he syringe, do not gra sp it by its gla ssba rrel. Alwa ys gra sp the syringe by its knu rled collar .

Knurled collar Mounting bracket




5. Ca refully remove each replacement wa sh syr inge f rom the packaging.6. P ar t i a l ly f il l each new w ash syr inge (by hand) wi th weak or s t rong wash

solvent (t o help remove a ir bubbles).

7. P ul l each wa sh syr inge plunger down so that t he plunger end slides overth e threa ded post on th e syringe guide mount ing bra cket.

8. Screw each new wa sh syringe par t i a l ly in to the mount ing bracket . Do

not t ighten.9. F in ger t ig hten ea ch w a sh sy r in ge.

10. Insta l l and f inger t ighten the knurled screw tha t holds each wa shsyringe barrel to th e mounting bra cket.


12. Run the Syst em Prime option a t lea st six t imes to remove a ir from thesyringe.

Modifying sample syringe configuration parameters

To configure the system for a syringe size that is different from the onecurrently fitted:


2. Select Configure > Volumes. The Volume Configuration dialog boxappears.

3. In t he Loop Size text box, type the size of the loop that wi l l be used wi ththe syr inge.

4. Select t he appropria t e needle s ize from t he list , a nd then c lick OK.

5 Select t he appropria t e sample syr inge size from t he list a nd then cl ick





5. Select t he appropria t e sample syr inge size f rom t he list , a nd then cl ickOK.

Replacing the injection valve cartridge

Requirement: When y ou repla ce th e inject ion va lve, you mu st a lso repla ce t hesa mple loop.

Required materials

• H ex w rench, 2-mm

• I n ject ion va l ve ca r t r id ge (s ee Appendix C for t he part number)

• Sa mple loop

To replace the injection valve cartridge:


2. In the sample manager in format ion w indow, cl ick Main ta in > P arkIn ject Valve.

3. P ow e r-off th e s a mple ma n a g er .


5. Hold the sa mple loop, and pull the inject ion va lve about one inch towa rd

you.

6. Remove the f it t ings a t ta ched to the inject ion va lve car t r idge.

Injection valve cartridge with fittings

Caution: To av oid kinks, do not pull on th e volum e detection device(port 2) or sample needle (port 3) tubing.




7. U se the 2-mm hex wr ench t o remove the screw a t the 10 o’clock positionon th e injection va lve ca rt ridge.

8. Remove the injection valve ca rtr idge from the injection valve a ssembly.

9. Ca refully remove the replacement inject ion va lve car t r idge f rom t hepackaging.

FittingsFittings

10. En sure tha t t he groove in the ca rtr idge housing and t he groove on thedrive clamp a re a l igned. If they a re not, t urn t he drive clam p until thegrooves line up.

Aligning injection valve cartridge grooves

11. Insert the new injection valve ca rtr idge into the injection valveassembly.

I h h h ’ l k i i h i j i

Caution: If t he injection va lve ca rt ridge does not s lide fully int o theinjection va lve assembly, cont a ct your Wa ters service representa tive.

Drive clamp

Aligned grooves




12. In sert t he 2-mm hex screw a t th e 10 o’clock position on t he inject ionva lve cart ridge, a nd use the 2-mm h ex w rench to tight en it.

13. Reat ta ch all fi t t ings an d insta l l a new sa mple loop (see page 6-45).

14. P ush th e injection va lve back to th e closed position.


16. En sure the sam ple ma na ger compart ment door is closed.

17. P ower-on the sample ma nager .

18. In t he ACQU ITY UP LC C onsole, select S am ple Man a ger from thesystem t ree. The sam ple ma na ger informa tion window a ppears.

Caution: To avoid kinks , do not pus h on t he volume detection device(port 2) or sample needle (port 3) tubing.


19. Click Control > P rime syringes.

20. In the P r ime Syr inges dia log box

• s elect s a m ple s yr in ge a n d w a s h sy rin ges .

• t y pe 1 in t h e N um ber of C ycles t ext b ox.

• cl ick OK. The sample mana ger s ta tus d isplays “P r iming .” When

priming is complete, the sta tu s return s to “Idle”.

21. Click Ma inta in > Cha ra cterize > Needle and loop volumes.

22. In the C ha ra cterize Needle and Loop Volumes dia log box, cl ick St a rt .The test r esults a ppea r in t he Loop an d Needle area s.

Testing the sample manager backpressure regulator

To test the sample manager backpressure regulator:


2. Cl ick Main ta in > Test backpressure.

3. In the Ba ckpressure Test d ia log box, cl ick Sta r t .

Wh h i l h R l




4. When the test is complete, the Resul ts pan e appears .

Replacing the fuses

Sus pect a n open or otherw ise defective fuse wh en a ny of these conditionsapply:

• The sample mana ger or column hea t er fa i l s to power-on .

• Th e sa mple ma n a g er or column h ea te r st a tus L E D s a r e un lit .

• Th e pow e r supp ly fa n d oes not oper a te.

Warning: To avoid elect ric shock, pow er-off an d un plug t he sa mplema na ger before exa mining fuses. For continued protection a ga instfire, repla ce fuses only w ith t hose of th e sam e type and r a ting .

Required materials

• 10 A fuse (2) (see Appendix C for t he part number)

• F la t -bla de scr ew dr iver


Requirement: Replace bot h fuses, even w hen only one is open or defective.1. P ow e r-off th e s a mple ma n a g er .

Tip: The fan s inside th e sam ple ma na ger run cont inuously, even aft eryou power it off.


3. U se a f lat -blade screwdriver to open the fuse holder door, which islocat ed above th e pow er ent ry m odule on t he rear pa nel.


Fuse holder door




4. With minimum pressure, pull on each spring-loa ded fuse holder andremove it.

5. R emov e a n d d isca r d th e fuses .



Warning: For continued protection a ga inst fire, replace fuses w ith t he

a ppropria te type a nd ra ting.

Fuse holdersFuse

Power entry module


U se a soft cloth, da mpened with w a ter , to clea n t he outside of the sa mplemanager .

Maintaining the column heater

U nder norma l opera tin g conditions, th e column h eat er does not req uire anyroutine ma intena nce. If residue from solvent leaks a ccumula tes in th e columntra y, remove the column, an d w ipe the tra y clean w ith a soft cloth.

The column h eat er is powered by th e sam ple ma na ger. If the column h eat erloses pow er, the fuse in t he sa mple ma na ger ma y need to be repla ced. See“Repla cing t he fuses” on pa ge 6-59 for m ore informa tion.

Replacing the filter in the column in-line filter unit

If y our syst em ha s t he optiona l column in-line filter inst a lled, follow th einstr uctions below t o replace th e filter.


non-lat ex gloves or finger cots w hen r eplacing t he filter in t he columnin-line filter un it.



Maintaining the column heater 6-61

Tip: D ue to w ear on t he non-repla ceable compression fit ting s, Wa tersrecommends replacing t he column in -line filter un it a fter y ou use th e sixfil ters tha t a ccompan y th e unit .

Requirement: St op an y currently r unn ing mobile pha se before repla cing t he

column in-line filter un it.

Required materials

• Open-end wrench, 5/16-inch (2)

• P a r t icle-free, powder-free , non-la tex gloves or f inger cots

• R epla cem en t filt er (see Appendix C for th e pa rt n umber)

To replace the filter in the column in-line filter unit:

1. Open the column heat er door , li ft t he meta l la t ch , and t i l t the columnhea ter t r a y downw ard .

Opening the column heater tray

2. Remove the column components (column, in-line f il ter unit , andst a bilizer tu bing assembly).

Column components

Column heater trayMetal latch




3. Detach the column st abi l izer t ubing a ssembly f rom t he column in-l inefilter unit . Do not det a ch t he column in-line filter un it from t he columninlet.

4. Open t he column in-line filter unit by placing tw o 5/16-inch open-endw renches onto t he f la ts.

5. Remove the column in-line f il ter unit inlet nut , which contains th e usedfilter. Disca rd t he filter; it can not be reused.

TP02544

Flow

Column inlet In-line filter unit Column stabilizer tubing assembly

Reusable

finger-tight fitting

6. Inser t a new f il ter , and then reassemble the column in-l ine fi lter unitusing the wrenches.

Assembling and disassembling the in-line filter unit

7. Reat ta ch the column stabi lizer tubing a ssembly to the in-l ine fi lter unit .

8. St ar t the mobile phase and examine the assembled components for

leaks. Tight en fittings a s needed.

TP02543

Flow

Place wrenches on flats to

assemble and disassemble.

Outlet assembly Filter Inlet nut



Maintaining the column heater 6-63

9. Return the assembled components to the column tra y, push down t hemeta l lat ch to secure the tu bing, an d tilt t he tra y to the closed position.

Maintaining the TUV detector

Tip: Do not remove th e detector’s top cover. No user-servicea ble part s a reinside. If you open the det ect or door, close it firm ly before resum ing norm a loperation.

See also:


Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for informa tion onma inta ining it .


Evapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de forinforma tion on maint a ining it .

The detect or requires minima l routine ma intena nce. To achieve optima lperforma nce, follow th ese recommend a tions:

• Fi lter and degas solvents to prolong column li fe, reduce pressurefluctua tions, a nd decrea se baseline noise.

• To conserve lamp li fe , ext inguish the lam p whi le leaving the detectorrunn ing but idle. Note, however, tha t y ou should do so only w hen th e

lamp w ill rema in extinguished more tha n 4 hours.• Invoke the lamp opt imiza t ion sof tware rout ine a t leas t once a w eek by




Invoke the lamp opt imiza t ion sof tware rout ine a t leas t once a w eek bypow ering-off th e detector, w a iting 10 seconds, an d t hen powering-on t hedetector.

Alternative: I n t he ACQU ITY U P LC Console, select TU V Detector >Ma inta in > Ca librat e wa velengt h. This function w ill correct errors inw a velength a nd updat e and save the new calibrat ion values.

• I f you use buffered mobile phase, flush i t from the detector beforepowering -off t o prevent

– plugg ed solv en t l in es a n d th e flow cell

– d a m a ged in st r um en t com pon en t s

– microbia l grow th

Caution: To avoid da ma ging t he column, remove it before you flushth e system.

Flushing the flow cell

Flush t he flow cell wh en it becomes cont a mina ted w ith t he residues ofprevious runs a nd a lso a fter each detector shut down. A dirt y flow cell ca nca use ba seline noise, decrea sed sa mple energy levels, ca librat ion fa ilure, andother problems. Alwa ys flush a nd purge the flow cell as y our initia l at tempt tocorrect th ese problems. I f , a fter flushing, th e problems persist, reverse flush

th e flow cell. If reverse flushing a lso fa ils, replace the flow cell.If y ou use buffered mobile pha se, flush it fr om th e detector beforepowering-off.

The a ctua l flow ra te ra nge is a ffected by t he combinat ion of solvent viscosity

Caution:

• To prevent f low cell fai lure, do not connect a ny t ubing or deviceth a t ca n creat e backpressure exceeding t he flow cell’s ma ximum

ra tin g of 6895 kP a (69 ba r, 1000 psi).• The pressure through the f low cell must not exceed 6895 kP a (69

ba r, 1000 psi). Increasing th e flow ra te usua lly increa ses thepressure. High-viscosity fluids genera lly increase t he pressureth rough th e flow cell and t herefore require a lower flow ra te.Allow a ble flow r a tes a re based on the limit of pressure tha t ea chflow cell ca n w ithst a nd.



Maintaining the TUV detector 6-65

The a ctua l flow ra te ra nge is a ffected by t he combinat ion of solvent viscosityand backpressure exerted on the flow cell. Total pressure across the flow cellmust never exceed t he ma ximum pressure limit.

Ma inta in a minimum ba ckpressure of 1724 kP a (17 bar , 250 psi) to preventbubbles from forming in th e flow cell. You can a tt a ch a ba ckpressure regulat orfrom the flow cell out let to the w a ste cont a iner.

Tip: Alwa ys us e well-degassed eluents.

Required materials

• A wrench sui tab le for removing and replac ing the column

• P a r t icle-free, powder-free , non-la tex f inger cots or g loves

• S ta inless steel un ion (to replace the column dur ing tes t ing)

Warning: To a void spills, empty t he wa ste cont a iner at regularintervals.

• A solvent , l ike methanol, which is miscible in both the mobile phase andw a te r

• H P LC-qua lit y w a ter

• Strong cleaning solvent suita ble for your system (6N ni tr ic a cid or 15%phosphoric a cid a re commonly used if the m etha nol flush fa ils toa dequa tely clean t he cell)

• S epa r a t e con ta in er for a cid w a s t e

• A means for measur ing the pH of the acid ef fluent , if you use an a cid asyour clea ning solvent

To flush the flow cell:

1. E xt in gu is h t h e la m p.

2. Discontinue the eluent f low, a nd remove the column.

3. Using a s ta inless steel union , a t t a ch s ta in less s teel tubing withcompression screw s a nd ferrules from t he injector outlet (w here th ecolumn w a s removed) to t he detector’s sa mple inlet. If a notherinst rument is dow nst rea m of th e detector out let, brea k the connect ion a tthe other instrum ent a nd route the outlet tubing to wa ste w hile

flushing.

TUV detector flow cell inlet and outlet locations





Flow cell

assembly

Outlet tubing

Inlet tubing

Captive screw

4. Flush the mobile phase from the detector wi th a miscible solvent andwa ter .

5. F lush the detector w i th HP LC-qua l ity w a t er to remove contaminantsfrom t he f low pat h.

6. First , pump 100% a cetonitr i le through the f low cell to clea n th e interna lpat hw a ys. I f conta mina nt s persist, pump 6N nitric acid or 15%

phosphoric a cid thr ough t he flow cell.

7. F lush aga in wi th HP LC-qua l ity w a ter un t il the pH of the ef fluent i sneutra l .

8. Reat ta ch the column to resume f low of the mobile phase. Use aninterm ediary s olvent if you a re using a m obile pha se tha t is not misciblein wa ter .

Purging the flow cellAfter flushing t he flow cell assembly, purge th e flow cell wit h nit rogen, a nd

Warning: To prevent persona l exposure to st rong a cids or ba ses,a lwa ys w ear suita ble persona l protective equipment wh en ha ndlingthem. At a minimum, you must wea r splash g oggles a nd protectivegloves.




a llow it to dry.

To purge the flow cell:

1. Connect t he pure nitrogen supply l ine to the sample inlet . Route thesa mple outlet to wa ste.

2. P urge the cell for 25 to 30 minutes a t 172 to 207 kP a (1.7 to 2 ba r , 25 to50 psi).

3. Allow t he flow cel l to dry completely .

Reverse-flushing the flow cell

If d irectly flushing a nd purging t he flow cell does not improve th e cell’sperforma nce, reverse flush it .

To reverse-flush the flow cell:

1. I f insta l led, remove the backpressure regula t or .

2. Reverse the inlet a nd out let connect ions to the cell and f lush.

3. Continue running in this reverse conf igura t ion for a pproximat ely 15minutes.

Tip: Fa lling system pressure indica tes th a t a dirty flow cell is becomingclean.

4. If th e flow cell rema ins blocked, replace the f low cell by follow ing theRepla cing t he Flow C ell service procedure.

Replacing the flow cell

Caution:

• To a void conta mina ting the flow cell, wear part icle-free,pow der-free, non-la t ex finger cots or gloves w hen h a ndlin g,removing or r eplacing it




See also: Contr ol l ing Contami nat ion in L C/ M S Systems (par t

number 715001307) on the ACQU ITY U P LC Sy st em B ookshelf CD.

Required material

Fla t-blade screwdriver

To replace the flow cell:

1. P ow e r-off th e d etector .2. S t op t h e s olv en t flow .

removing, or r eplacing it .

• To avoid dama ging the f low cel l, handle i t wi th care . Do notdisa ssemble the flow cell.

3. Open the detector door , gent ly pull ing i ts r ight edge towa rd you.


4. Disconnect t he detector ’s in let an d out let t ubing f rom t he main column

connection.

Flow cell

assembly

Outlet tubing

Inlet tubing

Captive screw




5. R emov e th e f low cell:

• Using a 1/4-inch, f la t -blade screwdriver , loosen the t hree capt ivescrew s on th e flow cell assembly’s front plat e.

• G r a sp th e h a n d le a n d gen t ly pu ll th e a s sembly tow a r d you .

Removing the flow cell assembly

Front plate of

flow cell

assembly

Captive screw

Handle




TP02401

Captive screws

6. U npack and inspect t he new f low cel l. Ensure tha t t he flow cel l type(a na lytical or high sensitivity) suits y our applicat ion.

Top view of the analytical flow cell assembly

Caution: To a void da ma ging t he ca pillar y t ubing, do not touch it.

Capillary tubing

Flow cell body

Front of flowcell assembly




TP02583

Top view of the high sensitivity flow cell assembly

7. Inser t the new f low cel l assembly into the detector , and t hen t ighten theca ptive screws .

8. Connect t he inlet t ubing to the main column connect ion a nd f low cellinlet, a nd connect th e outlet t ubing t o the flow cell outlet.

TP02582

Capillary tubing

Flow cell body

Front of flowcell assembly




9. C l ose th e d etector d oor .

10. B efore you pow er-on t he detector, ensure th e flow cell is filled wit hdegas sed, tr a nspa rent solvent (a cetonitr ile or wa ter) a nd free of a irbubbles. The detector m a y n ot initia lize correctly if th e cell cont a ins a ir.

Replacing the lamp

Cha nge the la mp w hen it repea tedly fai ls to ignite or w hen th e detector fai lsto calibra te.

Tip: Record the new la mp’s serial n umber t o properly tr a ck la mp life. If you donot record a new la mp’s serial num ber in th e ACQU ITY UP LC Console, th eda te of th e previous la mp insta llat ion rema ins in the detector’s memory.

Required material

• P e rfor ma n ce m a in t en a n ce k it (s ee Appendix C for t he part number)

To remove the lamp:

1. P o w er -off t h e la m p.

Warning: To prevent burn injuries, a llow t he la mp t o cool for 30minut es before removing it. The la mp housing gets extremely hotduring opera tion.

Warning: To avoid eye injury from ultr a violet ra diat ion exposure

• power-of f the detector before changing the lamp.

• wear eye protect ion tha t f i lte rs u lt r av iolet l igh t .

• keep the lam p in the housing dur ing operat ion.




p

2. P ower-off the detector and disconnect th e power ca ble from the rea rpanel.

3. Allow t he la mp to cool for 30 minutes, and t hen open the door, gently

pulling its right edge towa rd you.

Warning: The lam p an d la mp housing ca n be hot. Wa it 30 minut esa fter pow ering off th e detector for t hese component s t o cool beforetouching them.

4. Detach the lamp’s power connector from the detector .

Removing the lamp

5. Loosen the two capt ive screws in the lam p base. P ull the lamp assemblyout of the lamp housing, and t hen gently w ithdra w t he la mp.

Lamp power

connector

Lamp base

Alignment pinCaptive screw

Captive screw




To install the lamp:

1. Unpack the new lamp from i t s packing mater ia l w i thout touching the

bulb.2. Inspect the new lamp and la mp housing for clea nl iness .

3. P osit ion t he lam p so tha t t he cut-out on i ts base pla te is a t the 1 o’clockposition, in line w ith t he alignm ent pin on th e la mp housing, and t hen

Warning: La mp ga s is under slight nega tive pressure. To preventsha tt ering t he glass, use ca re wh en disposing of the la mp.

Caution: Do not touch th e gla ss bulb of the new la mp. Dirt or

fingerprints on th e bulb adversely a ffect detector operat ion. If t hebulb needs clea ning, gently ru b it w ith eth a nol a nd lens tissue. Donot use a bra sive tissue. Do not a pply excessive pressure.

gently push the lam p forw a rd unt i l it bott oms into position. En sure tha ti t seat s f lush a ga inst t he optics bench.

4. Tighten the two ca ptive screws, and th en reconnect th e power connector .

5. P ower-on the detector , and then wa i t about 30 minutes for the lamp tow a rm before resuming opera tions.

Tip: C ycling pow er t o the detector (th a t is, pow ering-off a nd t henpow ering-on th e instr ument ) initia tes t he verificat ion procedures.

6. In the ACQU ITY UP LC Console, select Mainta in > Cha nge Lam p. TheCha nge Lam p dialog box appea rs.

7. Cl ick New Lamp. The New Lamp d ia log box appears .

8. Type the ser ia l number for the new lamp (see the label a t ta ched to the

la mp connector w ire), a nd t hen click OK.

Replacing the fuses

Required material

Warning: To a void electric sh ock, power-off a nd unplu g t he TU Vdetector before examining the fuses. For continued protectiona ga inst fire, replace fuses only w ith t hose of th e sam e type and

rat ing.




Required material

The TUV detector requires two 100 to 240 VAC, 50 to 60 Hz, F 3.15-A, 250-VFAST B LO, 5 × 20 mm (IE C) fuses. See Appendix C for th e part number.

Sus pect a fuse is open or otherw ise defective w hen

• t h e d et ect or fa i ls t o pow er -on .

• t h e pow er su pply fa n d oes not oper a t e.


Requirement: Replace bot h fuses, even w hen only one is open or defective.

1. P ower-off the detector and disconnect t he pow er cord from th e pow erentr y module.

2. P inch the sides of the spring-loa ded fuse holder, wh ich is above thepow er entry module on t he rea r pa nel of th e detector. With minimumpressure, with dra w t he spring-loa ded fuse holder.

Removing the fuseholder


4. Make sure tha t the new fuses are proper ly ra ted for your requirements ,a nd t hen insert them int o the holder an d t he holder into the pow er entrymodule, gently pushing un til t he a ssembly locks int o position.


Fuses

Fuse holder

Power entrymodule





U se a soft cloth , dam pened w ith w a ter, t o clean t he out side of the TU Vdetector.

Maintaining the sample organizer

Ma int a ining th e sam ple orga nizer consist s of defrosting, clea ning its a ir filter,

repla cing fuses, a nd cleanin g the outside of the instr ument .

Defrosting the sample organizer

For informa tion on defrosting t he sa mple orga nizer, refer t o “Defrosting thesam ple compart ment” on pa ge 6-29.

Cleaning the air filter

To clean the air filter:

1. P ower-of f the sample organ izer .

2. Remove the dra in tube f rom the dr ip t r ay .

3. P ul l the drip tra y towa rds you, and remove i t from the sample organizer .

Sample organizer drip tray and filter

Caution: Never lubricat e sa mple orga nizer component s. D oing s o willdama ge the instrument .



Maintaining the sample organizer 6-77

4. With the fi lter s t i ll a t t ached to the dr ip tray, r inse the f il ter in a s ink.

5. R efit t he dr ip t ra y .

Caution: Do not remove the air filter from t he drip tr a y.

Filter

Drip tray

Replacing the fuses

Sus pect a fuse is open or otherw ise defective w hen

• t h e s a m ple or ga n iz er fa i ls t o pow er on .

• t h e pow er su pply fa n d oes not oper a t e.

Required materials

•10 A

fuse (2) (see Appendix C for t he part number)• F la t -bla de scr ew dr iver


Requirement: Replace bot h fuses, even w hen only one is open or defective.

1. P ower-of f the sample organ izer .

Tip: The fa ns inside th e sam ple organ izer run cont inuously, even a fteryou power it off.

2. Disconnect t he power cord from t he power entry module .

Warning: To avoid elect ric shock, pow er-off an d un plug t he sa mpleorganizer before exa mining th e fuses. For cont inued protectiona ga inst fire, replace fuses only w ith t hose of th e sam e type andrat ing.





3. U se a f la t-blade screwdriver to open the fuse holder door, which is aboveth e power entry module on the rea r pa nel of the sa mple organ izer.


4. With minimum pressure, pull on each spring-loa ded fuse holder toremove it.


Warning: For cont inued protection a ga inst fire, repla ce fuses wit hthe a ppropria te type and ra ting.

Fuse

Fuse holder

Fuse holderdoor

Power entrymodule



Maintaining the sample organizer 6-79




U se a soft cloth, da mpened with w a ter , to clea n t he outside of the sa mpleorganizer.




7 Diagnostics and

Troubleshooting

This cha pter d escribes how to find the cause of problems t ha t ca n a ffectperforma nce of the ACQU ITY UP LC™ system a nd how t o correct t hem.

Contents:

Topic Page

Troubleshoot ing w ith Connect ions Insight 7-2

P roper opera t ing procedures 7-4

M on itor in g per for ma n ce fr om th e ACQUITY UP L C Con sole 7-8

Monitor ing column hist ory 7-33

Solving ba seline noise a nd dr ift problems 7-38

B ina ry solvent ma na ger t roubleshoot ing 7-43

Sa mple ma na ger t roubleshoot ing 7-60Sa mple orga nizer t roubleshoot ing 7-89

TU V det ect or t roubleshoot ing 7-102



7-1

P D A det ect or t roubleshoot ing 7-114

E LS det ect or t roubleshoot ing 7-128

C hroma togra phy t roubleshoot ing 7-141

Troubleshooting with Connections Insight

Connections Insigh t™ is an int elligent device ma na gement (ID M) w eb serviceth a t ena bles Wa ters t o provide proactive service a nd support for t heACQU ITY UP LC s yst em. To use Connections Ins ight, a Wa ters t echnicia nmust insta l l the Connections Insight service a gent softwa re on the syst em’sEm pow er or Ma ssLy nx w orksta tion. In a client/server syst em, th e servicea gent m ust a lso be inst a lled on the comput er from w hich you cont rol thesystem. The service a gent softwa re aut omat ica lly a nd securely captu res an dsends informa tion a bout t he support n eeds of your syst em directly to Wa ters.

If you encount er a performa nce issue as you a re using the ACQU ITY UP LCConsole, you ca n a lso ma nua lly submit a Connections Insight request t oWa ters cust omer support .

Tip: Consult t hese sources for more informa tion a bout C onnections In sight:• Connect i ons Insigh t Insta l l a ti on Gu ide

• Connect i ons Insigh t User Gu ide

• Connect ions In sigh t FAQ Document

• h t tp://w w w .w a t er s.com

• Your sa les represen ta t ive• You r loca l Wa t er s s ubs idia r y

• Waters Customer Suppor t , a t 800 252-4752 (In the US A or Ca nada )



7-2 Diagnostics and Troubleshooting

To submit a Connections Insight request:

1. In t he ACQU ITY UP LC C onsole, select Troubleshoot > SubmitConnections I nsight request . The Connections Ins ight R equest dia logbox appears .

Connections Insight request dialog box



Troubleshooting with Connections Insight 7-3

2. Type your na me, telephone number, email address, a nd a description ofthe problem.

3. Click Submit. Allow a pproxima tely 5 minutes to sa ve the service profi le.

Result: A .ZIP file conta ining y our Connections I nsight profile isforw a rded to Wa ters customer support for review . Sa ving a serviceprofile or plot file from th e ACQU ITY U P LC Console ca n r equire up t o150 MB of file spa ce.

Proper operating procedures

So tha t your system a lwa ys r uns optima lly, follow the operat ing proceduresa nd guidelines in C ha pt er 2, C ha pt er 3, C ha pt er 4, C ha pt er 5, and C h a pt er 6.

Contacting Waters technical service

If you a re loca ted in t he US A or C an a da , report ma lfunctions or otherproblems to Wa t ers Technica l S ervice (800 252-4752). I f you a re locat eda nyw here else, phone Wa ters’ corpora te hea dqu a rt ers in Milford,Ma ssa chuset ts (U SA), or conta ct your loca l Wa ters s ubsidiar y. Our Web siteincludes phone num bers a nd e-ma il a ddresses for Wa ters loca tions w orldwide.G o to ww w .wa ters.com, a nd click About Wa ters > Worldw ide Offices.

Support Center dialog box




Locating system serial numbersEa ch system inst rument’s unique serial num ber fa cil ita tes service an dsupport . Seria l numbers a lso provide a w a y t o crea te single log entries foreach instrum ent so tha t you ca n review t he usage history of only tha tinstrument.

Recommendation: B e prepar ed to provide the serial num bers of th e

instr ument s in your syst em wh en you conta ct Wa ters customer support.

To view instrument information:

1. In the ACQU ITY UP LC Console, select a n instrument from the systemtree.

2. Click Configure > View m odule informa tion. The Module Informat ion

dia log box displa ys t his informa tion:• Ser ia l Number

• F irmw a re Version

• F irmw are Checksum

• C om pon en t S oft w a re Version

Alternatives:

• From the main window, place the cursor over the v isua l representa t ionof the system inst rument y ou w a nt informa tion for .

• Obta in the ser ia l number f rom the pr in ted labe ls on the rear pane ls of



Proper operating procedures 7-5

p pinst rument s or inside their front doors.

Tip: Column pa rt num bers an d serial num bers appear on t he column pa ge. Tosee column informa tion, click Column on the na vigat ion t ree.

Generating a service profile report

Sa ving a service profile report crea tes a .ZIP file cont a ining deta iledinforma tion a bout y our syst em. Wa ters service personnel might n eed th isinforma tion t o diagn ose syst em problems or t o properly prepa re t he correctma terials a nd pa rts for a service visi t .

To save a service profile report:

1. In t he ACQU ITY UP LC C onsole, cl ick Troubleshoot > Sa ve serviceprofile. The Sa ve Service Pr ofile dialog box a ppear s.

2. Sa v e th e fil e w i th a n a me a n d loca t i on .

Resetting system instruments to their initial state

You ca n reset a system instrument to its default setting a nd return th esa mple needle and sy ringe to their home positions. If an inst rument isperforming a diagnostic test , resett ing i t stops the test a nd returns i t to i ts

init ia l s ta te .Rule: You cann ot reset the syst em or a ny inst rument if a complete loss ofcommunication occurs betw een the system a nd Em power™ or MassLy nx™ ,wh ich a solid red LE D on t he instrument indica tes.

Reset a syst em instr ument if an y of the follow ing conditions occur:

• I t displa y s a fla sh in g red L ED .

• The ACQUITY UP LC Console di sp lays the “communica t ions f a ilure”message.

• You res ta r t t he sys tem or one of it s inst ruments (power-of f and thenpower-on).

• An error ha s occurred.

To reset a system instrument:1. In the ACQU ITY UP LC Console, select a n instrument from the system

tree.

2 Cl ick Control Reset mod le




2. Cl ick Control > Reset module.

Alternative: In E mpower or Ma ssLyn x softw a re, right -click an inst rum ent'scontrol pan el, and th en click Reset.

Recommendation: If resett ing th e instrum ents fa ils to correct t he problem,inspect th e ca bles a nd connections, a nd t hen reboot th e system. I f rebootingdoes not correct t he problem, conta ct Wa ter s Techn ical S ervice.

Reviewing logs to identify problems

Logs record notes a bout th e syst em, a n inst rument , or a project. When

view ing logs, you can use selection filters to specify t he kind of informa tionyou seek. Some of the par a meters y ou ca n select include view ing t he logentries th a t you or a nother user crea ted for a part icular inst rument on aparticular day.

To view log information:

1. In t he ACQU ITY UP LC C onsole, cl ick Logs. The Logs dialog boxappears.

2. Select a da t e or t ime span: Al l or today , s ince yesterday, past w eek, pa stmonth, or past y ear .

3. Select t he content: All content , errors, user log entries, or diagnosticcontent.

Tip: P a y a tt ention to softw a re errors and diagnostics results, as t heyprovide key clues during troubleshooting.

4. Select the instrument or system, such a s the current system, a speci ficcomponent , or a ll.

5. From the records table, select a log , and then rea d the descr ipt ion tha ta ppears in t he details of the current record a rea a t the bottom of thepage.

To print a log:

1. S elect a log.

2. C lick P r int .

3. In t he Windows P r int d ia log box, review the pr int opt ions, and t hen cl ickPr in t .



Proper operating procedures 7-7

Monitoring performance from the ACQUITY UPLCConsole

The ACQU ITY UP LC Console is a s oftw a re applica tion tha t replaces thekeypads a nd sma ll-screen displa ys tr a ditionally found on the front s of syst eminstr ument s. It offers a convenient cent ra lized mea ns for configuring set tings,

monitoring performa nce, running diagn ostic test s, an d perform ing routinema intenan ce on th e system a nd i ts instruments.

Via t he ACQU ITY U P LC C onsole's Web-like int erfa ce, you ca n q uicklyna vigate t o pictorial representa tions of ea ch syst em instrument a nd i tscomponents. You can a lso na vigate t o interactive diagra ms, w hich showinstr ument interconnections a nd provide diagnostic tools for tr oubleshootingproblems.

The ACQUITY UPLC Console includes controls for configuring, monitoring,ma inta ining, and m a na ging system components. For exa mple, graphica lsta tu s indica tors monitor a nd r eport t he real-tim e use of component s like th ecolumn a nd det ector la mp. The sta tu s indica tors let you configure usa gethresholds tha t , wh en rea ched, display m essa ges and cha nge the stat usindica tor's color. These a lerts ca n help you plan a nd schedule routinema intena nce before problems t ha t ca n cause unscheduled dow nt ime occur.




G eneral cat egories ar e logica lly grouped on the ACQU ITY U P LC Console'smenu. Su bordina te to th ese, specific listings na me ta sks you perform on theindividual instru ments, int eractive displa y, or sy stem.

Main areas of the ACQUITY UPLC Console

System tree Main window Task buttons



Monitoring performance from the ACQUITY UPLC Console 7-9

Determining system status

On sta rtup, the ACQU ITY UP LC C onsole defa ults to the ACQU ITY UP LC

Syst em selection in the na vigat ion t ree. Corresponding syst em sta tusinforma tion appea rs in the ACQUI TY U P LC Console's ma in w indow,including a pictorial representa tion of the syst em ha rdwa re. Virtua l LE Ds(light-emitt ing diodes) on t he front pa nels of th e virt ua l instr ument scorrespond to those on t he a ctua l syst em inst rument s. The LE Ds cha nge color

Error messages

appear hereMove the mouse pointer over the detector shown here todisplay information for up to two detectors connected to

the system.

to indica te th e opera tional sta tus of the system, its inst ruments a nd th eircomponent s. You can display deta iled sta tu s informa tion for a pa rt icula rinstr ument a nd/or one of its component s by paus ing t he cursor on t he syst emsta tus LED.

Rule: You can not cont rol or m odify a ny syst em functions from t he ACQU ITYU P LC Console unti l the Em pow er or Ma ssLynx softw a re completes the runa nd releases cont rol of th e system. You ca n neverth eless monitor currentsystem set t ings and v iew their s ta tus .

Monitoring data with real-time plots

The plot s provide up t o 96 hours of rea l-t ime syst em usa ge a nd performa ncedat a . U se the da ta to monitor conditions such as tempera ture, pressure, flow

Console LED colors and their meaning

LED color Status is

No color Idle

G reen Rea dy for opera t ion

F la s hin g g reen I nit ia lizin g, s ta r t in g u p, or ca libr a t in g

Red E rror condit ion tha t prevent s fur ther opera t ion, sucha s a commu nicat ions problem

F la shing red H a rdw a re fa ilure t ha t preven ts furt her opera t ion a ndrequires a tt ention




ra te, a nd other functions w hen performing routine ma intenan ce, runningdia gnost ic t ests , an d/or troubleshooting problems. You can a lso monit or plotsduring sa mple processing so th a t y ou can q uickly identify problems a nd t a ke

appropriate action.

When t he syst em is operat iona l, it collects a nd displa ys t he most r ecent 96hours of da ta for a ll plots, w hether or not th e plots a re visible. As th e systemcollects n ew da ta , i t disca rds t he oldest dat a .

Tip: To bett er ident ify pat terns w hen tr oubleshooting a ll 96 hours of da ta ,enable the Ext reme in D at a P rocessing featur e. Right-click a plot, a nd t hen

click P roperties. On the B SM, S M, or TU V Ch a nnels ta b, select E xtreme inDa ta P rocessing, a nd t hen click OK.

Recommendation: When th e syst em is performing optim a lly, print t he plotsof runs w hose conditions, reagent s, a nd solvent s represent th ose you typica lly

use in your w ork. Sa ve these plots, w hich reflect t he syst em's ba selineperforma nce. If substa nda rd results in subsequent runs suggest syst em orinst rument fa ilure, print t he plots of th ose runs. Then compa re th e lat er plotsw ith th e baseline plots t o determ ine the problem's severity.

To print one or more plots:

1. In the ACQUITY UP LC Console’s ma in window, or any ins t rument

w indow, select P lots from th e system tr ee. The P lots w indow a ppea rs.

2. Scroll the P lots w indow to di sp lay the plots you wa nt t o pr in t .

Tip: The w indow can only displa y up t o ten plots a t a time. To print a lla va ilable plots , scroll to view a nd print th e first ten plots, a nd t hen scrollto view a nd print t he remaining plots.

3. From the Troubleshoot menu, cl ick P r int plots .

4. In t he Pr int d ia log box, conf irm tha t All is selected , and then cl ick OK.

Saving plots

You can sa ve all plots or ea ch plot individually.

To save a screen capture of the plots:

This process crea tes a sma ll file, usua lly 1 to 5 MB or less.

1. Right-cl ick in the plot a nd select C apture plot .

Result: In t he background, the syst em opens the P C’s defa ult screen




gca pture program a nd sa ves the plot. The defa ult file na me is “P lot.bmp”,a nd t he file is an exact copy of th e plots th a t a ppea r on th e console

screen.2. Sa ve the fi le in your screen capture program.

Tip: Rena me th e file before sa ving it.

3. Scroll to oth er plots, a nd repeat st eps 1 a nd 2 i f you need other plots.

To save the data in the plots:

Tip: Sa ving plot da ta usua lly crea tes a very lar ge file, 100 MB or more.

1. Right-cl ick in the plot , a nd select S ave plot f i le .

2. B y defa ul t , a l l channels are selected , so clear the cha nnels you do notwa nt to sa ve.

3. Se lect a t ime range from the oldes t da te and t ime to the newest da te andtime.

Rule: The ma ximum is four da ys of dat a (96 hours).

4. C lick B row se.

5. In the “Set Loca t ion of Plot Fi le” d ia log box, enter a f i le name.

6. I f you do not w ant to save the f ile in the *.plot format , then select * .csvfile forma t .

7. C lick S a ve > OK .




Binary solvent manager plots

Plot Use to monitor

S yst em P ressure Overa ll solven t pr essu re in t he sy st em , usua llyindica ting relat ive sta bili ty of the syst em.

Tip: A noticea ble increase in syst em pressuremight indica te a blocka ge at some point a long t hesolvent pa th . Conversely, a decrea se in pressuremight indica te a loss of fluid due to lea ks or empt yreservoirs.

F low Ra t e Set point of t he flow ra t e for fluids t hrough t hesystem.

Mea sured Flow Ra te Aand B

Actu a l meas urement s of flow ra tes for solvents Aa nd B th rough th e system. This plot only a ppliesto nanoACQUI TY UP LC™ systems.

A Composi t ion (%)

B Composit ion (%)

The per cent a ge of solven t A (A1 or A2) flow ingfrom t he left solvent m a na ger.

The percent a ge of solvent B (B 1 or B 2) flow ing

from t he r ight solvent ma na ger.

D ispla ys t he percent a ge of solvent (1 a nd 2) to bedra wn from t he solvent ma na gers (A and B ).C omposit ion of A plus B equa ls 100.0%. S how s




p p qeith er isocra tic or gra dient blends of tw o solvents,A and B .

Tip: When monitoring gra dients dur ing meth odprocessing , plots of A Composit ion (%) a nd BC omposit ion (%) sh ow t he progr ession of solvent sA a nd B thr ough the progra mmed gradient, as t hesyst em mixes the solvents from y our initia l condi-tions to your final condit ions. If you a re running

only isocra tic mixtur es, you ca n h ide these plots tosa ve screen spa ce.

P ist on pressures (psi):

• P r ima ry A

• Accum ula t or A

• P r ima ry B• Accum ula t or B

P ressures a t each of four pistons in t he binarysolvent m a na ger.

The bina ry solvent m a na ger incorporat es tw o

solvent ma na ger pumps, A on t he left a nd B on t heright. Ea ch ha s tw o heads, prima ry on the left a nda ccumulat or on the r ight .

Tip: When t roubleshootin g piston problems,monitor the primar y a nd a ccumulat or plots toisolat e a lea king piston or one you suspect is

blocked.Degasser P ressure(psia)

Solvent degas ser pressure, wh ich indica tesw hether t he degassing function is opera tingproperly.

Tips: D egassing is enabled by default , w hether ornot solvent a nd/or sa mples ar e flow ing t hrough t he

system.

Binary solvent manager plots (Continued)

Plot Use to monitor




Sample manager plots

Plot Use to monitor

Sa mple P ressure (psi ) Observed pressure a t the sample need le in thesample mana ger .

Tem per a t ur es (° C ) O bs er ved t em per a t ur es , m on it or ed a t sev er a lplaces:

• Sample Temp: tempera ture of the samplemanager compartment

• Column Temp: temperature of the columnheat er compar tment

• Room Temp: tempera ture a t t he a ir inta ke ofthe sample ma na ger

• Sample Organ izer Temp: tempera ture of thesample organizer compartment

Rule: When you cha nge the current t emperat uresett ing for the column a nd/or t he sa mple ma na ger,you ca nnot run sa mples unless th ey a re below theset t emperat ure and a larm band.

Tip: Monitoring th e room t empera tu re's effect onsa mple a nd column t empera t ures helps identifyconditions t ha t ca n cause sa mples to degra de. Thesyst em might not be a ble to compensa te for room




syst em might not be a ble to compensa te for roomtempera tures tha t a re signif ica ntly higher orlow er tha n th ose prescribed for the sam ple a ndcolumn.

In ject ion Marks (S ta r t ) Shows when the sample manager s ta r t s eachinjection. I f you see chan ges in oth er plots, you canrea d th e injection ma rk's time, an d then look atth e oth er plots t o determine wh ether th esecha nges correla te w ith t he sta rt of a n injectioncycle.

Tip: The sa mpled setting can cause injectionma rks t o be obscured. To see inject ion ma rksbett er , ena ble the E xtreme (ma x a nd min va luesplotted) fea ture in Da ta P rocessing.

TUV detector plots

Plot Use to monitor

TU V Cha nnels A a nd B(AU)

Wa velength tr a ces (in a bsorbance units) from t heTU V detector for t he w a velength s specified in t he

method.In the single wa velength m ode, TU V Cha nnel Bhas no da ta .

The data plot continues beyond the injectionruntime.

PDA detector plots

Plot Use to monitor

P D A C ha nnel A Ma ximum a bsorba nce va lue for a ny w a velengt hspecified in t he current method.

The da ta plot is suspended betw een in ject ions.

P D A C ha nnel B Absorba nce for cha nnel 1 w a velen gt h, a s specifiedin the current meth od.

The da ta plot is suspended betw een injections.




ELS detector plots

Plot Use to monitor

E LS S igna l Light sca t t ering unit s (LS U ) from t he E LSdetector.

E L S Nebu lizer (° C ) Act u a l t em per a t ur e of th e n ebu liz er .

E L S D rift Tu be (° C ) Act u a l t em per a t u re of t h e d rift t ube.

E LS G a s P r essure Act ua l ga s pressure rea dba ck in psi.

Examining smaller or larger segments of data plotsYou can na rrow t he view of a plot , restr icting i t t o a pa rticula r segment ortime period. Conversely, you ca n expan d t he view of a plot t o include la rgersegments.

Na rrowing th e view of a plot r educes the num ber of periods show n in t heon-screen segment but ma kes the periods a ppea r la rger, lett ing you more

closely exa mine th eir pea ks. Expa nding t he view of a plot in crea ses thenum ber of periods shown on-screen but ma kes the periods a ppea r sm a ller,lett ing you evaluat e a la rger plot segment a nd identify t rends in thechromatography.

Tip: You can s et t he default t ime period for a plot in t he Time Ra nge ta b of theP lot P roperties dialog box.

To view a smaller plot segment:

1. Cl ick and drag wi thin the plot , draw ing a box around a plot segment tozoom on it .

2. Cl ick Ful l View w hen you wa nt t o restore the plot t o i ts norma l view.

Alternative: C lick U nzoom repeat edly to restore the plots to th eirprevious view, one step a t a tim e.

To view a larger plot segment:

1. Right-cl ick a plot , an d then cl ick Plot P roper t ies .

2. In the Amount of Dat a S hown box, cl ick and drag the indica t or on thel i l h i i d h i h h d




sca le unti l the t ime period tha t y ou w a nt to see appea rs in the hours an dminut es fields.

Alternatives: En ter va lues int o th e hours a nd minu tes fields, or click th ea rrows next t o ea ch field to increa se or decrea se their va lues.

3. C lick OK.

Modifying plot displaysYou can enable or disa ble the displa y of real-tim e dat a in th e plots by clickingRea l-Time .

Recommendation: If y ou need to observe the plots more closely, a nd d o notwa nt them t o move across the screen as the syst em collects new da ta , disablereal-time dat a .

Rule: You ca nnot rea rra nge the order of th e plots displa yed in the P lotswindow, but you ca n show or hide them.

To show or hide plots:

1. Right-cl ick any plot , and t hen cl ick P lot P roper t ies .

2. In the BS M Chan nels ta b , select the checkboxes for the plots you wa nt

displayed, a nd clea r t he checkboxes of th e plots you w a nt to hide.

3. In t he SM Cha nnels ta b , select or clear the checkboxes for the plots youw a nt t o display or hide.

4. In the TU V, PD A, or ELS D etector Channels ta b , select or clea r thecheckboxes for t he plots you w a nt to display or hide.

Tip: If t he syst em does not include a detector, th e detector Cha nnel A

a nd B plots do not a ppear.

5. C lick OK .

Creating log entries




You can crea te log ent ries to record events a nd service th a t y ou or ot hersperform on th e syst em an d its instr ument s, using the log entries to tra ck yourma intena nce a nd t roubleshooting effort s. For exa mple, you ca n creat e a logentry ea ch t ime you perform t hese ta sks:

• Replace components , such as the sample need le and loop

• C ha nge solvent s

Tip: The syst em a utoma tically records individua l log entr ies for a ll messages,errors, and dia gnostic syst em event s. You ca n view a nd filter all logs by da tea nd inst rument to see w ha t kind of informa tion th e log records for ea ch event.Thus y ou ca n a dd your ow n comm ents w ith out duplicat ing th e system's logs.

To create a log entry:

1. In the ACQUITY UP LC Console’s ma in window, or any ins t rumentw indow , select Maint ain > Creat e log entry.

2. From t he “Applies to” l is t , select t he instrument to which the log entryapplies.

Tip: Select Sy stem t o record syst em events or an y informa tion not

rela ted to a part icular instrument. To save time, creat e a log entry in theinstrument page becaus e the instrument na me is already selected in th edrop-down list.

3. In th e Oper a tor b ox, t ype your n a me.

4. In t he Comment box, type any descr ipt ive text , up to 249 a lpha numericcharacters.

5. Cl ick OK . The system saves the log en try w i th a da te and t imesta mp. If you a re using E mpow er as t he dat a system, the ACQU ITYU P LC C onsole saves the log entry in the Empower da ta base.




Displaying the status of binary solvent manager components

To view binary solvent manager component settings:

In the ACQU ITY UP LC Console, select B inary Solvent Mana ger > Int eractiveDispla y from th e system t ree. The binary solvent ma na ger interactive displa ydialog box a ppea rs.

Binary solvent manager interactive display dialog box




Modifying binary solvent manager component settingsSet t hese condit ions in the binar y solvent ma na ger int era ctive display dia logbox:

• Th e flow r a t es for pum p A a n d B

• The system f low a nd composit ion for pump A and B

• Solvent select ion va lve A to solvent A1 or A2

• Sol ven t select i on va l ve B to sol ven t B 1 or B 2

• Th e v en t v a lv e to v en t , syste m, or closed

To modify binary solvent manager component settings:

1. In the ACQU ITY UP LC Console, select B inary Solvent Mana ger >Int eractive Displa y from th e system t ree.

2. Cl ick Control in the binary solvent ma nager interact ive displaydia log box to invoke contr ol mode. The modifia ble sett ings a reunderlined.

Tip: A green circle appea ring in th e upper left corn er of th e icon mea nscontrol mode is enabled. A red squ a re in th a t place mea ns cont rol modeis disa bled.

3. Cl ick the under lined set t ing you wa nt to modify.




Displaying the status of sample manager components

To view sample manager component settings:

In the ACQUI TY U P LC Console, select Sa mple Man a ger > Intera ctiveDispla y from the system t ree. The sam ple ma na ger intera ctive displa y dia logbox appear s.

Sample manager interactive display dialog box




Modifying sample manager component settingsSet t hese condit ions in th e sample ma na ger intera ctive display dia log box:

• The pressure assi st va lve to the “on” or “of f” pos it ion

• The weak wa sh fi ll va lve to the “on” or “of f” pos it ion

• The wa sh select va lve to the “on” or “of f” pos it ion

• The st rong wa sh fi ll va lve to the “on” or “of f” pos it ion• The sample syr inge va lve posi t ion to needle, fi ll , or bypass

• The in ject ion va lve pos it ion to load or in ject

• Th e s a m ple t em per a t u re s et poin t

• Th e colu m n t em per a t u re s et poin t

To modify sample manager component settings:

1. In the ACQU ITY UP LC Console, select Sa mple Mana ger > Interact iveDispla y from t he system tree.

2. Cl ick Control in the sample ma nager interact ive display d ia log box toinvoke control mode. The modifiable settings are underlined.

Tip: A green circle appea ring in th e upper left corn er of th e icon mea ns

control mode is enabled. A red squ a re in th a t place mea ns cont rol modeis disa bled.





Displaying the status of sample organizer components

To view sample organizer component settings:

In t he ACQU ITY UP LC C onsole, select Sa mple Orga nizer > Int eractiveDispla y from the syst em tr ee. The sam ple orga nizer intera ctive display dia logbox appear s.

Sample organizer interactive display dialog box




Tip: The P neuma tic pressure is the air pressure inside th e int erna lcompressor.

Modifying sample organizer component settingsYou ca n perform these ta sks via th e sa mple orga nizer intera ctive displaydia log box:

• Transfer sample p la tes between the sample organ izer and the samplemanager

• Shutt le the X-axis posi t ion home or aw ay (hor izonta l , lef t-to-r ight

direction)• Shutt le the Y-axis posi t ion home or aw ay (f ront-to-back direct ion)

• Al ter the Z-axis pos it ion (ver t i ca l d irect ion)

• M od ify t h e s a m ple com pa r t m en t t em per a t u re

To modify sample organizer component settings:

1. In the ACQU ITY UP LC C onsole, select S am ple Organizer > Interact iveDispla y from t he system tree.

2. Cl ick Control in the sample ma nager interact ive display d ia log box toinvoke control mode. The modifiable settings are underlined.

Tip: A green circle appea ring in th e upper left corn er of th e icon mea nscontrol mode is enabled. A red squ a re in th a t place mea ns cont rol mode

is disa bled.


Understanding plate transfer

The sam ple organ izer tr a nsfers plat es cont a ining sa mples or vials ba ck a nd




forth betw een the sa mple ma na ger and sa mple orga nizer . Pla te tra nsfer

occurs w henever t he Empower or MassLynx softwa re initia tes a request t oprocess a sa mple tha t is st ored in t he sa mple orga nizer. Three subas sembliesmove plates within the sample organizer:

• X-axis, or Shut t le X, which t r ave ls in r igh t and lef t d irect ions . Homeposition is all th e wa y t o the left (inside th e sam ple organ izer).

• Y-axis, or Shut t le Y, which t r avels between the fron t a nd back of the

organ izer. H ome position is a ll th e wa y to the ba ck.• Z-Dr ive, which t r a vels ver t i ca l ly . Home pos it ion i s a l l the way to the

bottom.

The Z-D rive moves the Y-a xis to th e ta rget s helf, wh ere th e Y-a xis picks th eplat e. Then t he Z-Dr ive moves th e Y-a xis to t he sa me eleva tion a s t he X-a xis.The Y-a xis shut tles th e pla te int o the X-a xis, which tr a nsfers th e pla te int oth e sample man a ger for processing. When the sa mple ma na ger finishes withth e plat e, the X-a xis pulls it ba ck into th e sa mple orga nizer. The process isreversed to return th e plat e to the shelf it cam e from.

To transfer sample plates from the sample organizer to the samplemanager:

1. In the ACQU ITY UP LC C onsole, select S am ple Organizer > Interact iveDispla y from t he system tree.

2. In t he “Sample organizer interact ive display” d ia log box, cl ick Controlt o invoke contr ol mode. The modifia ble sett ings a re underlin ed.

Tip: A green circle a ppea ring in t he upper left-ha nd corner of the iconmea ns cont rol mode is enabled. A red squa re in tha t pla ce mean s cont rolmode is disa bled.

3. The sam ple organizer detects w hich shelves conta in pla tes andillumina tes t he corresponding LE Ds t o th e left of the shelves, w hich a revisible wh en t he door is open. The ACQU ITY U P LC Console show s t hetra y num bers of shelves th a t conta in plat es as underlined.

4. Cl ick the shel f number tha t con ta ins the sample pla te you wa nt totra nsfer to the sample ma na ger.

5. Cl ick OK to conf irm your choice .




Displaying the status of TUV detector componentsTo view TUV detector component settings:

In t he ACQU ITY U P LC Console, select TU V Detector > Int era ctive Displayfrom th e syst em t ree. The TU V Detector In tera ctive Display d ia log boxappears.

TUV detector interactive display dialog box




Modifying TUV detector component settingsSpecify the detector wa velengt h via t he TU V Det ector Int eractive Displaydia log box.

To modify TUV detector component settings:

1. In t he ACQU ITY UP LC C onsole, select TU V Detector > Interact ive

Displa y from t he system tree.2. In the TU V Detector Int eractive Display dialog box, cl ick Control to

invoke control mode. The modifiable settings are underlined.


3. Cl ick the under lined set t ing you wa nt to modify .




Displaying the status of PDA detector componentsTo view PDA detector component settings:

In t he ACQU ITY UP LC Console, select P DA Detector > In tera ctive Displa yfrom the syst em tr ee. The P DA Detector In tera ctive Display dialog boxappears.

PDA detector interactive display dialog box




Modifying PDA detector component settingsSpecify the detector w a velength via t he P DA Detector Int eractive Displaydia log box.

To modify PDA detector component settings:

1. In the ACQU ITY UP LC C onsole, select P DA Detector > Interact ive

Displa y from t he system tree.2. In the P DA Detector Interact ive Display d ia log box, cl ick Control to

invoke control mode. The modifiable settings are underlined.






Displaying the status of ELS detector componentsTo view ELS detector component settings:

In t he ACQU ITY UP LC C onsole, select ELS Det ector > In tera ctive Displayfrom the syst em tr ee. The EL S D etector In tera ctive Displa y dia log boxappears.

ELS detector interactive display dialog box




Modifying ELS detector component settingsYou ca n perform t hese ta sks via t he ELS detector intera ctive display dia logbox:

• Ch a n g e th e nebu li zer h ea te r lev el a n d d rift tub e h ea te r t emper a tu r e

• C h eck t he s a m ple a n d refer en ce en er gy

• I gn it e or ext in guish th e la m p

• C heck t he P MT ga in

• I nit ia t e a n Au to Zer o offset

• S imula te a G a ussia n pea k

• R eq u es t Au t o G a in for a ch rom a t og ra p hy ru n

To modify ELS detector component settings:

1. In the ACQU ITY UP LC Console, select ELS Detector > Interact iveDispla y from t he system tree.

2. In the ELS Detector Interact ive Display d ia log box, cl ick Control toinvoke control mode. The modifiable settings are underlined.

Tip: A green circle appea ring in th e upper left corn er of th e icon mea ns

control mode is enabled. A red squ a re in th a t place mea ns cont rol modeis disa bled.





Monitoring column historyYou can predict a column 's reliability a nd life expecta ncy by r eviewing itsusa ge history. F or exam ple, knowing t he num ber of injections performed on acolumn can help you predict w hen its performa nce might begin t o diminish orw hen it w ill likely need replacing. ACQU ITY U P LC syst em columns includea n eCord™ informa tion chip wh ich records var ious t ypes of column

informa tion. The eCord column chip int era cts w ith t he ACQU ITY UP LCConsole, permitt ing you t o ea sily a ccess t he informa tion for up t o 50 sam plesets. This feat ure is part icula rly useful in regulat ed environments because itdocument s t he performa nce of th e column s used t o valida te m ethods. TheeCord column chip also enha nces Wa ters' service a nd support efforts byproviding th e mean s to qu ickly retrieve informa tion a bout th e column'smanufacturer .

Requirement: Att a ch the column 's eCord chip holder t o t he recepta cle on th eside of th e column hea ter. Once at ta ched, th e eCord column chipa utomat ica lly records informa tion a bout tha t column.

See also: Waters ACQUI TY U PLC B EH Colum n Car e and U se In st r uct ions for m ore deta ils on column a nd eCord column chip capa bilities a ndspecifications.



Monitoring column history 7-33

To display column history:In th e ACQU ITY U P LC Console, select C olumn from t he syst em tr ee. TheColumn H istory w indow a ppears.

Column history window

Rationale: If th e “eCord Not F ound” prompt a ppea rs, you ca nnot set ath reshold or tr a ck column u sa ge. To set a th reshold or tr a ck column use, thecolumn m ust include an a tt a ched eCord chip.

Rule: The eCord column chip sa ves the 50 most recent sa mple sets, a nd t hendeletes t he oldest entry a s each new ent ry is recorded.




Reviewing column history

The ta ble show s t he ty pes of column informa tion you can review .

Types of column information displayed

Column

information

Information available

G enera l identif ica -t ion (pre-set a t t hefactory)

• Column na me

• C olu mn pa r t n um ber

• C olu mn s er ia l nu mber

Finding column manufacturing information

The ACQU ITY UP LC Console provides th e ma nufa cturing hist ory of acolumn , w hich includes identifica tion informa tion, a Wa ters certifica te of

U sa ge counters • Tot a l in ject ions

• Tot a l sa m ples

• Tot a l sa m ple s et s

E nvironment • Ma ximum pressure t o w hich t he column ha sbeen s ubjected

• Maximum tempera ture to which the columnhas been subjected

Inject ion • D a t e of first inject ion

• Da te of most recent in ject ion

H istor ica l • D a t e st a r t ed, first inst a lla t ion• 50 mos t r ecen t s a mple set s

• Name of the users who used the column

• Na me of th e sys tems th e column ra n on

• Number of in ject ion s

• N um ber of s a mples

• M a xim um colu mn pr es su re• M a xim um colu mn t em per a t u re

Types of column information displayed (Continued)

Columninformation

Information available




a na lysis, and t he dat e quali ty control testing wa s performed. Columnma nufa cturing informa tion ca n be retr ieved from t hese sources:

• ACQUITY UP L C System t r ee pa n e > Column > Column QC

• AC QU I TY U P L C S y st em t r ee pa n e > C olu m n > B a t ch QC

• ACQUITY UP L C System t r ee pa n e > Column > Ca r e a n d Use. In th eWa ters Web site, log in, a nd t hen search for B EH Column Ca re a nd U se

Instructions.Requirement: You mus t h a ve int ernet a ccess t o retr ieve columnma nufa cturing informa tion from th e a bove source.

Determining column statusFactors to consider

You can expect th e best column life span under idea l conditions:

• C lea n sa mples.

• Columns exhib it t he longes t l if et ime when used below 68948 kPa

(689 ba r, 10000 psi), so check th e ma ximum pressur e to see if th ecolumn sust a ined pressures great er tha n 68948 kPa (689 bar,10000 psi).

• Tem per a t ur es n ot a t ext r em es.

Conversely, operat ion a t t he extr emes of pH a nd t empera tur e reduceslifetime. Additionally, t he higher t he pressure, the short er th e lifetime.Aggressive sample diluents such a s DMS O a lso shorten column lifetime.

P a y a tt ention to the ma ximum pressure an d temperature recorded in t heenvironment a rea. I t tel ls you w hether t he column ha s endured extremes,w hich t end to shorten t he life span of th e column.

You can a lso review each sa mple set in t he column h istory in grea t d eta il:number, da te, and t ime sta rted; sample set nam e; user name; system nam e;num ber of injections; number of sa mples; a nd ma ximum pressure a ndtempera tu re for each sa mple set.

Deciding when to clean the column

Caution: Opera ting a t t he extremes of pH, tempera tur e, and/orpressure will result in a shortened column life span.




Ch a nges in pea k sha pe, pea k splitt ing, shoulders on the pea k, shifts inretent ion, chan ge in resolution, or increasing ba ckpressure ma y indicat econta mina tion of th e column. F lushing t he column a ccording t o therecomm ended procedures in t he Waters ACQU IT Y UPL C BEH Column Care

and U se I nstruct ions ma nua l usually removes a conta minan t. I f f lushing fai ls,purge th e column , follow ing t he clean ing a nd r egenera tion procedure, below .

Consider the t ota l number of injections, sa mples, a nd sa mple sets t he columnha s been exposed to. Review t he logs to determine w hether a ny ent riesindica te previous column clea ning a nd regenerat ing, an d determine howma ny sa mple sets t he column processed since th e la st clea ning a nd

regenera t ing seq uence. You ca n expect a t lea st 1000 inject ions, or more, underidea l condit ions. Also cons ider t hese fa ctors:

• H ow a g gr essiv e a r e the solv en t s you h a ve been usin g ?

• What kinds of buffers do you run? Do you a lwa ys r inse thoroughly beforeshutt ing down for the da y?

• Ma xim um t em per a t ur e a n d pr es su re?

• How pure a re your samples? (Complex proteina ceous vs. simple)• Are you using the ACQU ITY UP LC column in-l ine fi lter unit? This fi lter

protects the column from microparticles, which can clog the column’sinlet frit. A significa nt chan ge in system ba ckpressure (usua lly > 10%)signals a need to change t he f il ter .

See also: Contr ol l ing Contami nat ion in L C/ M S Systems (par tnumber 715001307) on the ACQU ITY U P LC Sy st em B ookshelf CD.

To clean and regenerate the column:

See also: Waters ACQUI TY U PLC B EH Colum n Car e and U se In st r uct ions .

When y ou detect a loss of column efficiency or t he column becomesconta mina ted, clea n a nd r egenerat e the column.

1. Flush the column with a neat organic solvent , ta king care not toprecipita te buffers.

2. I f f lushing does not remove column conta minants , clean the columnusing t he recommended clea ning a nd regenera tion procedures in t heWaters ACQUI TY U PLC B EH Column Care and U se I nst ru ct ions manua l .

3. I f f lushing a nd cleaning does not remove conta minants , replace the




g g pcolumn.

To equilibrate the column:

Eq uilibra te t he column w ith a minimum of 10 column-volumes of the m obilephase you will use. I f running a n a utomat ed gradient method, ensuresufficient a nd r eproducible equilibrat ion times a re used betw een injections.

Tip: If using ion-pairing rea gents, ensure th a t t he first time you use thecolumn you provide a s ufficient t ime a nd volume of solvent t o adequ a telyequilibrat e the column .

Example: For a 2.1 × 100 mm ACQUI TY UP LC B EH column, run about 2.5mL of solution a t 0.5 mL/min for a bout 5 minu tes.

Solving baseline noise and drift problems

The follow ing t a ble list s possible ca uses for baseline noise, drift, a nd spikes inth e syst em a nd suggest s possible corrective a ctions.

Corrective actions for baseline noise and drift problems

Symptom Possible cause Corrective action

B a seline drif t duringgradient

Absorbance A a nd Bmobile phases a redifferent in a dditive

concent ra tion inwa ter and organicsolvents.

B a lance the absorbance of thetw o mobile pha ses by reducingth e concentra tion of the a ddi-

tive in t he higher-a bsorbingmobile phase.

Caution: This ma kes themobile phase slight ly differentw ith r espect to th e additive.




Flat baseline with nopeaks

No solvent flow C heck t he solven t reservoirsan d a djust th e flow ra t e .

Det ector lam p notignited

U se detector diagn ostics todetermine reference a nd

sa mple energy. Zero energyindica tes t he lamp isextinguished.

P ower-on the la mp. If t his doesnot r esolve th e problem,repla ce th e la mp.

Detector lamp

burned out

Repla ce th e la mp.

Incorrect detectorwavelength

Check w a velength sett ing.

L ea k i n solv en t pa th Ch eck fit t i ng s.

The m obile pha semight a bsorb too

much U V at thew a velength s selected

Do not monitor at such lowwavelengths.

Lower t he concentra tion of th eadditive.

U se a different a dditive orsolvent, one wit h less absor-ba nce a t t he chosenwavelength.

La rge, unexpected Air bubbles d ue t o Inst a ll the ba ckpressure

Corrective actions for baseline noise and drift problems (Continued)




Solving baseline noise and drift problems 7-39

spikes on baseline outgassing regulator .

Repa ir the dega sser.

Im proper or defec-tive Et hernet cable

Replace the ca ble with ashielded Et hernet cable.

Long-term ba selinenoise cycling

(approximately 10minut es to 1 hour)

Ambient tempera-ture f luctua tions

St a bilize a mbienttemperature.

Ra ndom baselinenoise

Air in det ector flowcell

1. Flush th e detector f low cellto remove a ir.

2. Check the ba ckpressureregulat or a nd ensure it is

properly insta lled.B ubble det ect ed P r ime t he solvent ma na ge-

ment sy stem. Remove andvacuum-filter solvents.

Solventscontaminated

U se fresh solvent .

Column

contaminated

Clea n or replace the column.

D ir t y flow cell C lea n t he flow cell.

System improperlygrounded

P lug into an outlet on adifferent electrical circuit.

U se a pow er conditioner.

U nit not cooling

properly

Opera te the system with a l l

covers in pla ce. Ch eck th e backpan els for proper clear a nce.

Ra dio frequencynoise

Eliminate interference.

The m obile pha semight a bsorb toomuch a t t he wave-

Do not monitor at such lowwavelengths.

L h f h






lengths selected Lower t he concentra tion of th eadditive.

U se a different a dditive orsolvent wit h less absorbance atthe chosen wavelength.

R a pid ba s elin e s hift C olu mn n otequilibrated

Eq uilibrat e th e column.

Det ector not a llow edto warm up

Allow t he detector to wa rm upunt il ba seline is s ta ble. Time

var ies wi th w a velength a ndsensitivity.

Solventcontaminated


Flow fluctuations(ra pid or slow d rift)

P rime th e pump, repla ce pumpseals, check va lves. See“Erra tic retention times” in the

ta ble t i t led “Chroma tographytroubleshooting” onpage 7-141.

Incorrect wave-length for solvent

E nsure solvent does not a bsorbat the w avelength used.

Short t erm baselinenoise cycling (30 to

60 seconds )

F low fluct ua t in g L ook for lea ks. S ee “E rr a ticretent ion times” in the table

ti t led “Chroma tography tr ou-bleshooting” on pa ge 7-141.

Ra dio frequencynoise (short - orlong-t erm cycling)

Eliminate interference.





Solving baseline noise and drift problems 7-41

S low ba selin e dr ift S olven tcontaminated


Ch eck or repair filters.

Decreased U V lampenergy

Determine la mp energy a t t heACQUITY UPLC Console.

Ambient tempera-ture f luctua tions

St a bilize opera ting environ-ment t emperat ure enough t oa llow full equilibra tion.

U V detector flow cellleaking

Ins pect flow cell, tight enconn ect ions. I f flow cell isleak ing, call Wa t ers Techn ica lS ervice (see page 7-4).

D ir t y flow cell C lea n flow cell.

Absorbance A a nd Bmobile phases a redifferent in a dditiveconcent ra tion inwa ter and organic

solvents

B a lance the absorbance of thetw o mobile pha ses by reducingth e concentra tion of the a ddi-tive in t he higher-a bsorbingmobile phase.

Caution: This ma kes themobile phase slight ly differentw ith r espect to th e additive.

Lea king connectionsa t flow cell inlet a ndoutlet

Tight en fitt ings.






Binary solvent manager troubleshooting

Power LED

The pow er LE D, on t he left-ha nd side of th e binar y solvent ma na ger’s frontpan el, indica tes w hen t he inst rum ent is pow ered-on or powered-off. It issteady green w hen the instrum ent is w orking properly.

Flow LED

The flow LE D, t o the r ight of the pow er LE D, indicat es the f low sta tus.

Binary solvent manager flow LED indications

LED mode and color DescriptionU nlit Indica t es the bina ry solvent ma na ger is idle.

S tea dy green Indica tes the bina ry solvent ma na ger is oper-a ting norma lly.

F la shing red Indica tes a ser ious error t ha t ha s stopped pumpopera tion. Informa tion about t he error ca n befound in t he ACQU ITY UP LC C onsole.

S tea dy red Indica t es a ser ious fa ilure w it hin t he bina rysolvent ma na ger tha t prevents further operat ion.C a ll Wa t ers Techn ical S ervice (see page 7-4).

Binary solvent manager troubleshooting



Binary solvent manager troubleshooting 7-43


B ot h LE D s unlit No pow er 1. Inspect line cordconnections.

2. Test electr ica l outletfor pow er.

Open (spent ) or defec-tive fuse Replace fuse (seepage 6-27).

Err at ic f low orpressures

Air in syst em P rime bina ry solventma na ger (see page 3-8).

Fa ult y check va lve 1. P rime bina ry solventma na ger (see

page 3-8).2. P er form the s ta t i c

deca y test t o identifyfa ult y check va lve (seepage 7-54).

3. Replace faulty checkvalve (see page 6-10).

H igh-pressure leak inth e system

Loca te a nd repa ir theleak (see page 7-56).

P lugged solvent reser-voir filter

Clea n or replace filter(see page 6-14).

High pressure sealleaking

Repla ce high pressureseal (see page 6-16).

Binary solvent manager troubleshooting (Continued)





Fa ns do not run Open (spent ) or defec-tive fuse

Replace fuse (seepage 6-27).

P ower supply fa nw iring or motor

problem

1. Reset the b inarysolvent m an a ger via

th e cont rol pa nel orACQU ITY U P LCConsole.

2. I f the problempersist s, pow er-off a ndpow er-on a ga in.

3. I f the problem

persists, call Wa tersTechnica l S ervice (seepage 7-4).

P ower supply notworking

1. Reset the b inarysolvent m an a ger viath e cont rol pa nel orACQU ITY U P LC

Console.2. I f the problem

persist s, pow er-off a ndpow er-on a ga in.

3. I f the problempersists, call Wa tersTechnica l S ervice (see

page 7-4).






page 7 4).P ump head leakssolvent

Worn high pr essureseal

Repla ce high pressureseal (see page 6-16).

Loose hea d Tight en hea d bolt sthrough pressuretransducer.

L oose check va lve Tight en t he check va lve.Seal wa sh tubes notproperly insta lled

Install tubes properly.

P riming problems S olvent in let tubes tothe solvent m a na gerhead ma y be leaking

Inspect solvent connecttubes.

P ump heads not

a dequat ely primed

P rime pump hea ds (see

page 3-8).Solvent t ube might bedefective

Repla ce solvent tu be.

Defect ive check va lve Replace check va lve (seepage 6-10).

Solvent leaks from sealw a sh leak t ube (see

page 6-8)

Wor n s ea l w a s h s ea l s R epla ce s ea l w a s h s ea ls(see page 6-16).

Startup diagnosticsfai lure

Int ernal problem w ithcontroller board,solvent ma na gementsyst em, or sa mplema nagement system

1. Reset the b inarysolvent m an a ger viath e cont rol panel orACQU ITY U P LCConsole.

2. I f the problempersist s, pow er-off a ndpow er-on a ga in.

3. I f the problempersists, call Wa tersTechnica l S ervice (seepage 7-4).






Alarm messages

To clear alarm messages:

1. Follow t he correct ive act ion in this table, and reset t he solvent mana gervia t he cont rol pa nel or ACQU ITY U P LC Console.

2. I f the a l a rm message st i ll appears on the ACQUITY UP LC Console a f te ryou reset the solvent ma na ger, reboot t he solvent ma na ger. I f th e ala rm

messa ge persists, cont a ct Wa ters Technical Service.

The following t a ble l ists t he solvent ma na ger ala rm m essa ges and theirresolutions.

Alarm message abbreviations

Abbreviation Explanation

H W H a rdw a re

S N S eria l number

Binary solvent manager alarm messages

Message Resolution

B a tt ery-backed m emoryHW fa ilure

Expla na tion: The bat tery ba cked memory gotcorrupt ed and reset to defa ult va lues.

P robable causes:

• B a t t ery fa ilure

• E lect r on ics fa i lu re

Resolution:

1. Reset t he binary solvent mana ger v ia t hecont rol pa nel or ACQU ITY U P LC Console




cont rol pa nel or ACQU ITY U P LC Console.

2. If t he a larm persists, power-off and power-onaga in .

3. If th e ala rm persists, call Wa ters Technica lS ervice (see page 7-4).

Cy cle pow er t o complet e

sof twa re updat e

This is a norma l messa ge in response to a firm-

wa re upgrade.

Degasser communicationsH W error

1. Reset t he binary solvent mana ger v ia t hecont rol pa nel or ACQU ITY U P LC Console.



Dega sser report ed HWfault

P robable ca use: Degass er wa ste line is blockedor kinked.

Resolution:

1. En sure the degasser wa ste l ine is not blockedor kinked.

2. I f the a larm persists , reset the bina ry solventma na ger via t he control pa nel or ACQU ITYU P LC Console.


4. If th e ala rm persists, call Wa ters Technica l

S ervice (see page 7-4).Degasser va cuum H Werror

P robable ca use: Degass er wa ste line is blockedor kinked.

Resolution:

1. En sure the degasser wa ste l ine is not blockedor kinked.

2. I f the a larm persists , reset t he binary solvent

ma na ger via t he control pa nel or ACQU ITYC C l

Binary solvent manager alarm messages (Continued)

Message Resolution




U P LC Console.

3. If t he a larm persists, pow er-off and pow er-onaga in .


G enera l fa ilure 1. Reset t he bina ry solvent ma na ger via thecont rol pa nel or ACQU ITY U P LC Console.



Int er pump communica-tions H W failure




S ervice (see page 7-4).Int er pump motion cont rolHW fa ilure




S ervice (see page 7-4).Liters pumped ma inte-na nce limit exceeded

Reset th e pumped volume ma intena nce count er,modify t he pumped volume ma int ena nceth reshold, or perform ma intena nce (seepage 6-8).

Modified S N D ispla yed for in forma t ion only . No a ct ion isrequired.

P ower su pply 15V out of 1. Reset t he binary solvent mana ger v ia t het l l ACQU ITY U P LC C l


Message Resolution




r ange cont rol pa nel or ACQU ITY U P LC Console.



P ower supply -15V out ofr ange




P ower su pply 24V out ofr ange




S ervice (see page 7-4).P ower supply 5V out ofr ange




S ervice (see page 7-4).P u mp hom in g er ror P r oba ble ca use: A blocka ge in th e bin ar y solven t

ma na ger plumbing ca used a r a pid spike inpressure.


2. If t he a larm persists, power-off and power-on

aga in .3 If th e ala rm persists call Wa ters Technica l


Message Resolution





P u m p H W ov er pr es su re P r oba b le ca u se:

• A b lockage in the b inary solvent ma nagerplumbing caused a ra pid spike in pressure.

• You set the flow w hile the vent va lve wa s in

t he blocked position.• You set a f low ra te tha t i s too h igh for the

system.

Resolution:

1. Verify method para meters .

2. Reposition the vent va lve.

3. Check for blocka ges.

P ump motor driver HWfault




P ump motor lostsynchronization




Softw a r e d ow n l oa d f a iled 1. R eset th e bi na r y sol ven t ma n a g er via th econt rol pa nel or ACQU ITY U P LC Console.


Message Resolution






St op Flow from remote

host

This is a norma l messa ge in response to st op-

ping t he f low during a sam ple run.

Stop Flow input a ct iva ted P robable cause: The pump received a s top flowcontact.

Resolution: Ch eck rea r-pa nel conn ect ions t odetermine wh a t is connected to the St op Flowinput a nd ca using t he cont a ct closure (seepage 7-57).

S y st em ov er pr es su re P r oba b le ca u se: Th e pr es su re w i th in t he b in a r ysolvent m a na ger exceeded t he limit th e user setin the bina ry solvent ma na ger method editor .

Resolution:

1. Verify method para meters .

2. Check to see if the f low ra te is set too high orth e tempera tur e is set too low .

3. Check for blocka ges.

Sys tem und er pr essure P r ob a bl e ca use: Th e pr essur e w i th in th e b in a r ysolvent ma na ger fell below t he limit th e user setin the bina ry solvent ma na ger method editor .

Resolution:1. Verify method para meters .

2. P er form a pr ime (see page 3-10).

3. Check solvent levels.

4. Check for lea ks (see page 7-56).

5. P er form t he sta t ic decay test (see page 7-54).


Message Resolution




Tr a nsdu cer ra n ge er ror P oss ible ca u ses :

• Transducer cable improper ly plugged in .

• In ter n a l flow r es t rict ion .

Resolution:


2. Check tra nsducer cables .



Vent va lve fail H Wdetected





Message Resolution




Performing the static decay test

The sta tic deca y t est performs a pressure ram p-up. It a lso monitors th epressure deca y in th e solvent ma na ger to determine w hether t he check valves,tubes, fi t t ings, or plunger seals a re faulty. P erform the st a tic deca y t estw henever y ou

• suspect leakage in the check va lves or plunger sea l s .

• per for m ma i n ten a n ce on th e b in a r y solv en t ma n a g er .

• r epla ce fit t in gs in t h e flu id pa t h .

Requirement: If you replaced the seals, run t he solvent m a na ger at 96527 kP a(965 bar , 14000 psi) for a h a lf-hour before performin g t he st a tic deca y t est.Neglecting to properly brea k-in th e seals ma y caus e testing fa ilure.

Tip: The sta tic decay t est is not recommended as a qu a lifica tion test . Youshould perform th is t est only if you a re experiencing performa nce problemswith t he binary solvent ma na ger.

To perform the static decay test:

1. Make sure tha t a l l the f it t ings on the solvent mana ger a re t igh t .


3. C l ick Ma i n t a in > S t a t i c d eca y t es t .

4. In the Sta t ic Decay Test d ia log box, in the Test P a rameters area , select amodule (pump) to test .

5. In th e Accumulat or text box, specify your typica l opera ting pressure:

Range: 34474 to 103421 kPa (345 to 1034 bar, 5000 to 15000 psi)

Default: 96527 kP a (965 ba r, 14000 psi)

Cl i k S Th i i h R Ti b h




6. Cl ick Sta r t . The test t ime appears in the Run Time bar graph.

7. When the tes t ends, the Results pane appears .

The ma in t est crit erion is th e deca y r a t e. A ra te of 3447 kP a /min (34 ba r,500 psi) for each a ctu a tor indica tes a successful, or “pa ssed” test. Ot her

a spects of the sta tic deca y t ha t determine pass /fail st a tu s include percent ofstr oke a nd fina l pressure. For exam ple, excessive prima ry pressure w hile thea ccumulat or is compressed triggers a n a ccumula tor fa ilure, th e result ofexcessive valve leakage.

Performing the dynamic leak test

The dyna mic leak t est performs a pressure ra mp-up, an d t hen employsfeedba ck control to ma inta in consta nt pressure wit hin th e solvent ma na ger.B y monitoring plunger tr a vel during t he consta nt pressure phase, i t calcula testhe lea k ra tes of the P rima ry a nd Accumulat or a ctua tors. These lea k ra tes ca nindica te w hether t he check valves, tubes, fitt ings, or plunger sea ls are fa ulty .P erform t he dyna mic leak t est w henever y ou

• suspect leakage in the check va lves or plunger sea l s .• per for m ma i n ten a n ce on th e b in a r y solv en t ma n a g er .

• r epla ce fit t in gs in t h e flu id pa t h .

Requirement: I f you replaced th e sea ls, run the binary solvent ma na ger at96527 kP a (965 ba r, 14000 psi) for 30 minut es before performin g t he dy na miclea k test . Neglecting t o properly brea k-in th e sea ls ma y cause t esting fa ilure.

Tip: The dyna mic leak t est is not recommended as a qua lifica tion test. Youshould perform th is t est only if you a re experiencing performa nce problemswith t he binary solvent ma na ger.

To perform the dynamic leak test:

1. Make sure tha t a l l the f it t ings on the solvent mana ger a re t igh t .


3. Cl ick Mainta in > Dyna mic leak t est . The Dynamic Leak Test d ia log boxappears.

4. In the Test P ara meters area , select a module (pump) to test .

5. In th e Accumulat or text box, specify your typica l opera ting pressure:

Range: 34474 to 103421 kPa (345 to 1034 bar, 5000 to 15000 psi)




Default: 96527 kP a (965 ba r, 14000 psi)

6. Click St a rt . The test t ime appears in t he Run Time bar gra ph.

7. When t he test ends, t he Results pan e appears.

The ma in t est crit erion is the lea k ra t e. A ra t e below 150 nL/min for ea cha ctua tor indica tes a successful , or “passed” test . Ot her a spects of the t est th a tdetermine pa ss/fa il include percent of stroke to a tt a in pressure a nd plung ertra vel l imits.

Example: I f the prima ry a ctua tor must inta ke solvent t o ma inta in consta nt

pressure wh ile the a ccumulat or is un der test , a n a ccumula tor fai lure istr iggered, t he result of excessive ba ck-leaka ge t hrough th e outlet check va lve.

Repairing leaks

To repair leaks:

1. B efore doing anything else, inspect a l l f it t ings. I f you can not ident i fy aleak a mong th em, inspect t he check va lves and pum p sea ls.

2. Rev iew the messages tha t appear in the Results pane of the test .

Tip: These messa ges ma y h elp identify t he source of th e problem.

3. Fa i lure of the accumulator test suggests t he fol lowing elements shouldbe exa mined:

• Out let check va lve

• Accumula to r plun ger a n d h ig h pr essur e pump sea l

• Tubing between the accumula tor and accumula t or pressuretransducer

• Tubing between the accumula t or pressure t r ansducer and ventvalve

• Vent va lve rot or

4. A fa i ling pr imary t est result suggests the fol lowing elements should beexamined:

• Inlet check va lve

• P r im a r y plu ng er a n d h ig h pr es su re pu mp s ea l

• Tubin g betw e en th e pr ima r y a n d p rima r y p ressur e t r a n sducer• Tubing between the primary pressure t r ansducer and out let check




gvalve

If you ar e una ble to resolve a dy na mic leak t est fa ilure, cont a ct Wa tersTechnical Service.

Displaying the binary solvent manager’s rear panel interfaceconnections

You ca n use the ACQU ITY UP LC C onsole to determine the sta tus ofinput/output signa l connections on t he rea r pa nel of t he bina ry solventma na ger. The ACQU ITY UP LC C onsole displa y gives you a rea l-time st a tus ofth e inst rum ent’s signa l connections. A green LED indica tes a signa l ca ble isconnected t o the term ina l. A red LE D indica tes n o signa l ca ble is connected to

the terminal .

To display rear panel interface connections:


2. In the bina ry solvent ma nager informa tion w indow, cl ick Troubleshoot >Rear P a nel. The Bina ry S olvent Ma na ger Rea r P a nel dialog boxappears.

Binary Solvent Manager Rear Panel dialog box




The follow ing t a ble describes th e bina ry solvent ma na ger I/O connections.

Analog-out/event-in connections


Auxilia ry 1 (In) Reserved for fut ure use.

Auxilia ry 2 (In) Reserved for fut ure use.

R un S topped (Ou t) I nd ica t es t he bin a ry solven t m a na g er ha s cea s edopera tion becau se of a n error condit ion or oper-ator request.

S wit ch 1 (Out ) U sed t o send t ime-ba sed cont a ct closure s igna lsto externa l devices.

0−2V An a log 1 (Ou t ) An a log ch a r t ou t pu t fu nct ion a lit y .

G ra dient (In) Init ia tes the pumps to begin gra dien t opera tion

by eith er cont a ct closure input or 0-volt input .S top F low (In) S tops the flow from t he bina ry solvent ma na ger

w hen a n error condition or ha rdwa re fai lureoccurs.

S wit ch2 (Out ) U sed t o send t ime-ba sed cont act closure signa lsto externa l devices.

S wit ch 3 (Out ) U sed t o send t ime-ba sed cont act closure s igna lsto externa l devices.

0−2V An a log 2 (Ou t ) Ou t pu t s t h e a n a log sig na l t o a d ev ice su ch a s a nint egra tor or st rip-char t recorder. You ca n selectone of th e follow ing signa ls a s th e cha rt outsignal :

• F low ra t e

• S yst em pressure• Composi t ion (%A, %B )




Changing the rear panel interface connections

Via th e rear pan el display , you ca n open a nd close certa in output connections.This can be useful when y ou need t o sta rt or stop an injection or tr oubleshootsyst em connectivity.

To change the solvent manager rear panel interface connections:


2. In the bina ry solvent ma nager informa tion w indow, cl ick Troubleshoot >Rear Pa nel .

3. In the B inary Solvent Mana ger Rear P ane l d ia log box , loca t e a s igna lconnection w ith a n Out la bel, a nd t hen click the red or green LE D. Theoutput sign a l for t he selected signal connection indicat es wh ether it isopened or closed .

Chart-out signal conditions

SignalParameter Setting at 0Volts (Minimum)

Parameter Setting at2.000 Volts (Maximum)

Flow Ra t e 0.000 mL/min 2 mL/min

S yst em P ressure –345 kP a (–3.45 ba r,

–50 psi)

103421 kP a (1034 bar ,

15000 psi )Composit ion 0.0% 100.0%




Sample manager troubleshooting

Power LED

The pow er LE D, on th e left side of th e sam ple ma na ger’s front pa nel, indicat esw hen t he instr ument is pow ered-on or pow ered-off. It is stea dy green w henth e inst rum ent is w orking properly.

Run LED

The run LE D, t o the r ight of the pow er LED , indica tes the run sta tus.

Sample manager run LED indications


U nlit Indica t es the sa mple ma na ger is curren t ly idle.

S tea dy green Indica t es the sa mple ma na ger is opera t ingnormally, a tt empting t o complete a nyoutst an ding sa mples or diagnostic requests.When sa mple and diagnostic requests a refinished, the LE D returns t o the unlit sta te.

F la sh ing green Indica tes the syst em is w a it ing for a n inst rumentto reach operational condition (for example,w a iting for t he column heat er to rea ch i ts setpoint).

F la shing red Indica tes a ser ious error t ha t prevent s fur t heropera tion. Informa tion about t he error ca n befound in t he ACQU ITY UP LC C onsole.

S tea dy red Indica t es a ser ious fa ilure w it hin t he sa mplema na ger tha t prevents furth er opera tion. Ca llWaters Technical Service (see page 7 4)




Waters Technical Service (see page 7-4).

Sample manager troubleshooting



2. Test electr ical outlet

for pow er.Open (spent ) or defec-tive fuse


Fa ns do not run S a mple ma na ger notplugged in

Ensure sa mplema na ger is plugged in.

P ower supply fan

w iring or motorproblem

1. Reset t he sample

ma na ger via cont rolpa nel or ACQU ITYU P LC Console.

2. I f the problempersist s, pow er-offa nd pow er-on a ga in.

3. I f the problem

persists, call Wa tersTechnical Service(see page 7-4).


1. Reset t he samplema nager v ia thecont rol pa nel orACQU ITY U P LC


persist s, pow er-off



Sample manager troubleshooting 7-61

p pa nd pow er-on a ga in.

3. I f the problempersists, call Wa tersTechnical Service

(see page 7-4).

Alarm messages


1. Follow t he correct ive act ion in this ta ble , and reset the sample ma nagervia t he cont rol pa nel or ACQU ITY U P LC Console.

2. I f the a l a rm message st i ll appears on the ACQUITY UP LC Console a f te ryou reset the sa mple ma na ger, reboot the sa mple ma na ger. I f the ala rmmessa ge persists, cont a ct Wa ters Technical Service.

Sa mple compart mentfreezes

Sa mple compar tmentdoor opened t oofrequently

1. Defros t samplema nager , thenchoose a sa mplecompartmenttempera ture 4 ° C

(39.2 ° F) or grea ter(see page 6-29).

2. Minimize sampledoor a ccess.



h/w H a rdw a re

S N S eria l number

sw S oft w a re

Sample manager troubleshooting (Continued)





The following ta ble l ists t he sam ple ma na ger ala rm messages an d t heir

resolutions.

Sample manager alarm messages

Message Resolution

Air s ensor ca libra t ion h/wfault

P ossible ca use: Air in sy stem.

Resolution: P rime sa mple ma na ger (see

page 3-16).P ossible ca use: Da ma ged volume detectiondevice tu be.

Resolution:

1. In spect volume detection device tube fordamage .

2. Replace i f necessary.P ossible ca use: Da ma ged volume detectiondevice elect rica l line.

Resolution:

1. In spect volume detection device ca ble fordamage .

2. Replace i f necessary.

Air sensor calibra tion outof ran ge (va lue)

1. Reset t he sample mana ger v ia t he controlpan el or ACQU ITY UP LC C onsole.



Air sens or volum e notcalibrated






2. If t he a larm persists, power off and power onaga in .


Ambient temperaturera nge error (va lue) Set room t empera ture t o opera tional t empera-ture ra nge. See the ta ble t i t led “Sa mplemanager environmental specifications” onpage B -7.

B a tt ery backed f ile systemreset (value)




B a tt ery ba cked memoryh/w fa ilur e


P robable causes:



Resolution:




S ervice (see page 7-4).Column hea ter overtempera tur e or h /w fa ult(value)




Column hea t er/cooler overtempera tur e or h/w fa ult(value)


2 If t he a larm persists power off and power on

Sample manager alarm messages (Continued)

Message Resolution




(value) 2. If t he a larm persists, power-off and power-onaga in .


Column tempera turera nge error (va lue)

Expla na tion: The instrum ent is una ble to reachth e set point tempera tu re or th e set point is outof ra nge.




Column tempera turesensor h /w fa ult (va lue)




Dia gnostic test inter-rupted by user

Displa yed for informa tion only.

Dra wing sample ra t eexcessive (value)

P ossible ca use: Restr iction in sa mple needle,sa mple loop, air sensor, or t he dra w ra te is toohigh.

Resolution: Set a low er draw ra te.

eCord connect ed D ispla yed for informa tion only .

eC ord da ta inva lid 1. D et ach eC ord colum n chip holder from therecepta cle on t he side of the column hea ter,

and t hen reat ta ch i t .2. Repla ce the column.

eC ord disconnect ed D ispla yed for inform at ion only .


Message Resolution




p y y

eC or d d river h /w f a ult 1. R eset t h e sa m ple m a n a ger via t h e con t rolpan el or ACQU ITY UP LC C onsole.


aga in .3. If th e ala rm persists, call Wa ters Technica l

S ervice (see page 7-4).

eCord h/w fa ult 1. E nsure t he eCord column chip holder isa tt a ched to th e recepta cle on t he side of th ecolumn hea ter.

2. I f the a larm persists , reset t he samplema na ger via t he control pa nel or ACQU ITY

U P LC Console.3. If t he a larm persists, power-off and power-on

aga in .


eC ord not present 1. At ta ch t he eC ord column chip holder t o t he

recepta cle on t he side of the column hea ter(see page 2-31).

2. Disa ble the eCord column chip requirementvia t he ACQU ITY U P LC Console.

eCord sw fa ult 1. Reset t he sa mple ma na ger via the cont rolpan el or ACQU ITY UP LC C onsole.





Message Resolution




Fa iled to ca libra te systemvolume


Resolution:

1. P r ime sample ma nager (see page 3-16).

2. Cha ra cterize sa mple needle a nd sam ple loop

volumes (see page 3-21).P ossible ca use: Da ma ged volume detectiondevice tu be.

Resolution:



3. Check a l l f it t ings.

Possible cause: Incorrect sample needle orsa mple syringe configured.

Resolution: Cha nge configura tion on t heACQU ITY UP LC C onsole.

P ossible ca use: Da ma ged sam ple needle or

sa mple loop.Resolution:

1. Inspect sam ple needle a nd sa mple loop fordamage .


P ossible ca use: Excessive dra w ra te.

Resolution: Set up a method with a low er drawra te. For deta ils, see Em power onl in e H elp orM assL ynx onl in e H elp .

F h f l R h l h l


Message Resolution




Fuse h/w fa ult 1. Reset the sa mple ma na ger via t he cont rolpan el or ACQU ITY UP LC C onsole.



G enera l fa ilure 1. Reset the sa mple ma na ger via the cont rolpan el or ACQU ITY UP LC C onsole.



H ome a nd/or plat e sensorsh/w fa ult




S ervice (see page 7-4).H ome a nd /or refer encesensors h/w fa ult




In jection coun t exceeded(value)

Explanation: Displayed for information only.

Resolution:

1. Ent er a higher injection count (see page 6-7).

2. If the a larm persists, perform scheduledmaintenance.



In ject ion va lve home h/wfault



Message Resolution







In ject ion va lve move h/wfault

Resolution:

1. Inspect t he injection va lve pod a nd ensure itis securely fa st ened to the va lve drive.


U P LC Console.




Message Resolution




Loop calibra t ion out ofra nge (va lue)


Resolution:

1. P r ime the sample mana ger (see page 3-16).

2. Cha ra cterize sa mple needle a nd sam ple loopvolumes (see page 3-21).

P ossible ca use: Clogged or kinked sa mpleneedle, injection va lve, or sa mple loop.

Resolution:



3. Check injector , tra nsducer, metering syringevalve, and sa mple syringe.

P ossible ca use: Da ma ged volume detectiondevice tu be.

Resolution:

1. In spect volume detection device tube for

damage .2. Replace i f necessary.

Possible cause: Incorrect sample loop, sampleneedle, or sample syringe configured.

Resolution: Cha nge configura tion on t heACQU ITY UP LC C onsole.

P ossible ca use: Excessive dra w ra te.Resolution: Set up a method with a low er drawra te. For deta ils, see Em power onl in e H elp orM assL ynx onl in e H elp .


Message Resolution




y p

Loop volume n otcalibrated

P ossible ca use: Ch a nge in configura tion a nd/orloop not char a cterized.

Resolution:


2. I f the a larm persists , reset t he samplema na ger via t he control pa nel or ACQU ITYU P LC Console.



S ervice (see page 7-4).Missing /w rong pla t e orvial

1. Ensur e tha t t he via l is in the correct position.

2. Check if correct pla te is selected a nd if theANS I version of the pla t e is select ed.




Modified S N D ispla yed for in forma t ion only . No a ct ion isrequired.


Message Resolution




Needle calibra tion out ofra nge (va lue)


Resolution:

1. P r ime the sample mana ger (see page 3-16).


P ossible ca use: Clogged or da ma ged sam pleneedle, injection va lve, or s a mple loop.

Resolution:



3. Check a l l f it t ings.

P ossible ca use: Da ma ged volume detectiondevice tu be.

Resolution:


2. Replace i f necessary.P ossible ca use: In correct sa mple needleconfigured.

Resolution: Cha nge sa mple needle configura tionon the ACQU ITY UP LC Console.

Possible cause: Incorrect sample loop, sample

needle, or sample syringe configured.Resolution: Cha nge configura tion on t heACQU ITY UP LC C onsole.

P ossible ca use: Excessive dra w ra te.


Message Resolution




Resolution: Set up a method with a low er drawra te. For deta ils, see Em power onl in e H elp orM assL ynx onl in e H elp .

Needle volume n otcalibrated

P ossible ca uses:

• C h a n ge in con fig ur a t ion .

• Needle and/or loop not chara cter ized.

Resolution:





P lat e present, no pla teexpected

Remove plate from sample manager.

P ower supply fan h/w faul t 1. Reset the sample ma nager v ia t he controlpan el or ACQU ITY UP LC C onsole.



P ower supply h/w fau lt(value)






Message Resolution




Sa mple fluidics high pres-sure limit (va lue)

P ossible ca use: C logged fluid line (volume detec-tion device, sa mple loop, sam ple needle,internal valve passa ges).

Resolution:

1. Ch eck for kinked or crushed tubing (volume

detection device, sample loop, and sampleneedle).

2. Replace dama ged tubing.




S a mple hea t er/cooler overtempera tur e or h/w fa ult(value)





Sa mple Man a ger doorclosed dur ing defrost

Open sa mple ma na ger door.

Sa mple Orga nizer door

closed dur ing defrost

Open sa mple orga nizer door.


Message Resolution




Sa mple Orga nizer t emper-a tu re ran ge error (va lue)

P ossible ca use: The sa mple orga nizer can ’treach the low set point t emperat ure.

Resolution:

1. Ensure the am bient temperature is s table.

2. Defrost t he sample compar tment (seepage 6-29).

P ossible ca use: The sa mple orga nizer can ’treach the high set point t emperat ure.

Resolution: C a ll Wa t ers Techn ical S ervice (seepage 7-4).

Sa mple pressure low

(value): possibly no washsolvent or needle da ma ged

P ossible ca use: Wa sh solvent s low or empty .

Resolution: Refresh wa sh solvents a nd primethe sample ma na ger.

P ossible ca use: Da ma ged sam ple needle.

Resolution: Run “C ha nge Needle” dia gnostica nd inspect sa mple needle for dam a ge. Replaceif dam a ged.

P ossible ca use: Loose plumbin g conn ect ion orleaking injection sea l.

Resolution: Verify fitt ings a nd ferrules a re tighta nd replace if necessar y.

P ossible ca use: Pr essure regulat or out of ran ge(low).

Resolution: C a ll Wa t ers Techn ical S ervice (see

page 7-4).


Message Resolution




Sa mple pressure sensorh/w fa ult (va lue)

P ossible ca use:

1. No pow er to tr a nsducer; loose electr ica lconnection.

2. Tra nsducer ma lfunction.

Resolution:




S a mple syrin ge h/w loststeps (value)




S a mple syr inge home h/wfault 1. Reset t he sample mana ger v ia t he controlpan el or ACQU ITY UP LC C onsole.



S a mple syr inge move h/w

fault

1. Reset t he sample mana ger v ia t he control

pan el or ACQU ITY UP LC C onsole.2. If t he a larm persists, power-off and power-on

aga in .



Message Resolution





Sa mple syringe move tosens or h/w fa ult




S ervice (see page 7-4).Sa mple tempera turera nge error (va lue)

P ossible ca use: The sa mple orga nizer can ’treach the low set point t emperat ure.

Resolution:


2. Defrost th e sa mple compar tment (see

page 6-29).P ossible ca use: The sa mple orga nizer can ’treach the high set point t emperat ure.


Sa mple tempera turesensor h /w fa ult (va lue)




S a mple va lve home h/wfault





Message Resolution




S a mple va lve move h/wfault




Tempera t ure cont rollerw a tchdog time-out




S ervice (see page 7-4).Wa sh syr inge h /w loststeps (value)

1. Open the fluidics draw er , examine the wa shsyringes for da ma ge, a nd replace if necessa ry(see page 6-53).




Wa sh syr inge h ome h/wfault

1. Open the fluidics draw er , examine the wa shsyringes for da ma ge, a nd replace if necessa ry

(see page 6-53).2. I f the a larm persists , reset t he sample

ma na ger via t he control pa nel or ACQU ITYU P LC Console.





Message Resolution




Wa sh s yr ing e move h/wfault




aga in .


Wa sh syringe move tosens or h/w fa ult





X & Y axes not ca l ibra ted Ca l ibra te the XYZ mecha nism (see page 2-47).

X-a xis h /w lost st eps(value)




X-a xis hom e h /w f a ult 1. R eset t h e sa m ple m a n a ger via t h e con t rolpan el or ACQU ITY UP LC C onsole.


Message Resolution






X-a xis move h /w fa u lt 1. R eset t h e sa m ple m a n a ger via t h e con t rolpan el or ACQU ITY UP LC C onsole.



S ervice (see page 7-4).Y-a xis h /w lost st eps(value)




S ervice (see page 7-4).Y-a xis hom e h /w f a ult 1. R eset t h e sa m ple m a n a ger via t h e con t rol

pan el or ACQU ITY UP LC C onsole.



Y-a xis move h /w fa u lt 1. R eset t h e sa m ple m a n a ger via t h e con t rolpan el or ACQU ITY UP LC C onsole.




Message Resolution




Z-a xis h /w lost st eps(value)

1. Ensu re the sam ple needle a ssembly isproperly inst a lled (see page 6-30).

2. Ensu re the pla te dimensions are definedcorrectly.

3. Ensure th e XYZ ca librat ions a re correct.

4. Insert t he correct pla te .

5. En sure the plate is inserted correctly (seepage 3-22).




Z-a xis hom e h /w f a ult 1. E n su re t h e s a mple n eed le a s sem bly isproperly inst a lled (see page 6-30).

2. Ensu re the pla te dimensions are defined

correctly.3. Ensure th e XYZ ca librat ions a re correct.



6. I f the a larm persists , reset t he sample





Message Resolution





Z-a xis move h /w fa u lt 1. E n su re t h e s a mple n eed le a s sem bly isproperly inst a lled (see page 6-30).








Z-a xis n ot ca libr a ted 1. C ha r a ct erize t he n eedle sea l (see page 6-41).







aga in .



Message Resolution




( p g )

Zp-a xis h/w needle loststeps (value)

1. Ensu re the sam ple needle a ssembly isproperly inst a lled (see page 6-30).








Zp-a x is h om e h /w f a u lt 1. E n su re t h e s a m ple need le a ss em bly isproperly inst a lled (see page 6-30).

2. Ensu re the pla te dimensions are defined

correctly.3. Ensure th e XYZ ca librat ions a re correct.



6. I f the a larm persists , reset t he sample





Message Resolution





Performing the sample syringe leak test

The sa mple syringe leak t est checks for leaks in t he pat h betw een t he sam plesyringe a nd t he injection va lve.

Tip: The sa mple syringe leak t est is not recommended as a qua lifica tion test.You sh ould perform th is t est only if you a re experiencing performa nceproblems w ith t he sample mana ger.

To perform the sample syringe leak test:

1. Make sure tha t a l l the f it t ings on the sample mana ger a re t igh t .

Recommendation: P rime the sa mple syringe three times before sta rtingthe sa mple syringe leak t est .

2. In the ACQU ITY UP LC Console, select Sa mple Mana ger from the

Zp-a x is mov e h /w f a ult 1. E n su re t h e s a m ple n eed le a s sem bly isproperly inst a lled (see page 6-30).









Message Resolution




system t ree.

3. Cl ick Mainta in > Leak Test > Sample syr inge (sta t ic).4. Cl ick Sta r t . The test t ime appears in the Run Time bar graph.


Performing the wash syringe leak test

The wa sh syringe lea k test checks for leaks in th e pa th betw een t he weak a ndstrong w a sh syr inges a nd t he injection valve.

Tip: The wa sh syr inge leak t est is not recomm ended as a qua lifica tion test.You sh ould perform th is t est only if you a re experiencing performa nceproblems w ith t he sample mana ger.

To perform the wash syringe leak test:


Recommendation: P erform a prime before sta rting t he wa sh syringelea k test .


3. Cl ick Mainta in > Leak Test > Wash syr inge (sta t ic).

4. Cl ick Sta r t . The test t ime appears in the Run Time bar graph.


Performing the needle seal leak test

The needle sea l lea k test det ermines wh ether sea l lea ks exist in t he pat hbetw een the wea k wa sh and sa mple syringes.

Tip: The needle sea l lea k test is not recommended as a qua lifica tion test . Youshould perform th is t est only if you a re experiencing performa nce problemswith t he sample mana ger .

To perform the needle seal leak test:


Recommendation: P erform a prime before sta rt ing th e needle seal leaktest .





Q , p gsystem t ree.

3. Cl ick Mainta in > Leak Test > Needle sea l (s ta t ic).4. Cl ick Sta r t . The test t ime appears in the Run Time bar graph.


Repairing leaks

To repair leaks:

1. When a leak test fa i ls in the sample ma na ger , inspect the samplema na ger for leaks a nd tight en f it t ings or syringes where lea ks a re seen.

2. I f the lea k test s t i l l fa i ls , conta ct Wat ers Technica l S ervice .

Displaying the sample manager’s rear panel interfaceconnections

You can use the ACQU ITY UP LC C onsole to determine the sta tus ofinput/output signa l connections on t he rea r pa nel of the sa mple ma na ger. The

ACQU ITY UP LC C onsole display gives a real-time sta tu s of the instrum ent’ssigna l connections. A green LE D indica tes a signa l ca ble is connected to th etermin a l. A red LE D indicat es no signa l ca ble is connected t o the termina l.

To display rear panel interface connections:





2. In t he sample mana ger informat ion w indow, c lick Troubleshoot > Rea r

P a nel. The Sa mple Man a ger Rea r P a nel dia log box a ppears.

Sample Manager Rear Panel dialog box

The following t a ble describes t he sa mple ma na ger I /O conn ections.

Analog-out/event-in connections


In ject S ta rt (Out ) Indica tes tha t a n inject ion ha s st a rt ed.

Auxilia ry (In) Reserved for future use.

In ject Hold (I n) I nput signa ls from ot her syst em inst rum ent s t odelay th e injection.

0−2V An a log 2 (Ou t) C ha r t ou tpu t fu nct ion a lit y.



S h (O ) U d d b d l l






Changing rear panel interface connections

Via th e rear pan el display , you ca n open a nd close certa in output connections.Opening or closing output connections ca n be useful w hen you need to st a rt orstop a n injection or t roubleshoot syst em connectivity.

To change sample manager rear panel interface connections:


system t ree.2. In t he sample mana ger informat ion w indow, c lick Troubleshoot > Rea r

Panel .

3. In the Sa mple Ma nager Rea r P anel dia log box, loca t e a s ignal connect ionw ith a n Out la bel, an d th en click the red or green LED . The out putsigna l for t he selected sign a l connection indicat es wh ether it is opened

or closed .

R un S topped (Ou t) I nd ica t es t h a t t h e sa m ple m a na g er st opped oper -a tion beca use of an error condition or a n opera torrequest (clicking Stop Flow button, for example).

Analog-out/event-in connections (Continued)





Sample organizer troubleshooting

Power LED

The pow er LE D, on t he left side of th e sa mple orga nizer’s front pa nel,indica tes w hen t he instr ument is pow ered-on or powered-off. It is stea dy gr eenw hen the instr ument is w orking properly.

Run LED

The run LE D, t o the r ight of the pow er LED , indica tes the run sta tus.

Sample organizer run LED indications:


U nlit Indica t es the sa mple orga nizer is curren t ly idle.S tea dy green Indica tes the sa mple orga nizer is opera ting

normally, a tt empting t o complete a nyoutst an ding sa mples or diagnostic requests.When sa mple and diagnostic requests a refinished, the LE D returns t o the unlit sta te.

F la sh ing green Indica tes the syst em is w a it ing for a n inst rumentto rea ch opera tiona l condition.

F la shing red Indica tes a ser ious error t ha t prevent s fur t heropera tion. Informa tion about t he error ca n befound in t he ACQU ITY UP LC C onsole.

S tea dy red Indica t es a ser ious fa ilure w it hin t he sa mpleorga nizer t ha t prevents further operat ion. Ca ll

Waters Technical Service (see page 7-4).



Sample organizer troubleshooting 7-89

Sample organizer troubleshooting



2. Test electr ica l outletfor pow er.

Open (spent ) or defec-tive fuse


Fa ns do not run S a mple orga nizer notplugged in

Ensure sam ple orga nizeris plugged in.

P ower supply fa nw iring or motor

problem

1. Reset t he sampleorga nizer via the

sample mana gercont rol pa nel orACQU ITY U P LCConsole.

2. I f the problempersist s, pow er-offa nd pow er-on a ga in.



1. Reset t he sampleorga nizer via thesample mana ger

cont rol pa nel orACQU ITY U P LCConsole.

2. I f the problempersist s, pow er-offa nd pow er on a ga in




a nd pow er-on a ga in.


S ample pla tes ja m Nonconforming via l ormicrotit er pla tes

Ensure that a l l pla t esconform t o ANS Is tandards .

B a d plate sensor f iberoptic

Call Waters TechnicalS ervice (see page 7-4).

Incorrect a lignment ofX an d Y stops

Call Waters TechnicalS ervice (see page 7-4).

Startup diagnosticsfai lure

Int ernal problem w ithcontroller board,solvent ma na gementsyst em, or sa mplema nagement system

1. Reset t he sampleorga nizer via thesample mana gercont rol pa nel orACQU ITY U P LC


persist s, pow er-offa nd pow er-on a ga in.

3. I f the problempersists, call Wa tersTechnica l S ervice (see

page 7-4).

Sample organizer troubleshooting (Continued)





Alarm messages

The follow ing ta ble lists th e sa mple orga nizer ala rm messa ges and t heirresolutions.


1. Follow the correct ive act ion in this ta ble , and reset the solvent organizervia t he cont rol pa nel or ACQU ITY U P LC Console.

2. I f the a l a rm message st i ll appears on the ACQUITY UP LC Console a f te ryou reset t he sa mple orga nizer, reboot th e sam ple orga nizer. If th ea lar m m essag e persists, cont a ct Wa ters Technical S ervice.

Sample organizer alarm messages

Message Resolution

Air pump t ime-out ha rd-wa re faul t Expla na tion: A time-out ha s occurred in the a irpump beca use it h a s been continuouslypumping for five minut es with out reaching th especified pressu re 758 kP a (7.6 ba r, 110 psi).

P robable causes:

• Broken or d isconnected p ipe


Resolution:

1. Reset the sam ple organizer v ia the samplema na ger control panel or ACQUI TY U P LCConsole.





B a tt ery ba cked memoryhardw are fa i lure


P robable causes:



Resolution:




Compart ment tempera-ture ra nge error

P robable ca use: The sa mple orga nizer door isbeing opened too oft en.

Resolution:


2. Minimize the number of t imes you open t hesa mple orga nizer door.

Compart ment tempera-ture sensor ha rdw a re fa ult

Explanation: The compartment temperaturesensor fa iled.

Resolution:

1. Reset the sam ple organizer v ia the samplema na ger control panel or ACQUI TY U P LC

Console.2. If t he a larm persists, power-off and power-on

aga in .


Et hernet cable Resolution: Ch eck t he E th ernet ca ble connec-

Sample organizer alarm messages (Continued)

Message Resolution




Et hernet cabledisconnected

Resolution: Ch eck t he E th ernet ca ble connection a t t he sample orga nizer a nd netw orksw itch. Reconnect th e cable.

F use h a r d w a r e fa u lt : n E xpl a n a t ion : Th e fuse n (n specif ies the f usena me) fa iled.

Resolution:



aga in .


H ea t er/cooler over t emper -at ure or hardw are faul t

Expla na tion: The over-tempera tu re sensor ha stripped.

Resolution:




Lower sample ma na gerport t oo high or t oo low

Expla na tion: The low er sam ple ma na ger portloca tion is not in t he specified ra nge a nd isdrooping t oo mu ch or t oo litt le.

Resolution: U nscrew a nd m a ke exactlyhorizontal.

Lower sample ma na gertra nsfer shutt le Home an dReference sensors bothON

Explana tion: The sample mana ger tra nsfershut tle home a nd r eference sensors failed.

P robable causes:


• Sensors f a ilure (sensors or cab le)


Message Resolution




Resolution: C a ll Wa t ers Techn ical S ervice (see

page 7-4).

Lower sample ma na gertra nsfer shuttle homingerror

Explanation: The sample organizer transfershuttle homing failed.

P robable causes:


• Sa mple organ izer t r ans fer shut t le home

sensor fa ilure• P n e uma t i cs pr essur e too l ow


Lower sample ma na gertra nsfer shuttle move

error

Explanation: The lower sample managertra nsfer sh utt le move fai led.

Resolution:1. Reset the sam ple organizer v ia the sample

ma na ger control panel or ACQUI TY U P LCConsole.



S ervice (see page 7-4).Organ izer genera l fa i lure 1. Reset the sample organ izer v ia the sample




P la te a lready beingprocessed in th e low ersample mana ger

Ca use: An a tt empt w a s made to bring a plateinto the lower sa mple ma na ger, when there isa lrea dy a pla te present there.

Resolution: C a ll Wa t ers Techn ical S ervice (seeg 7 4)


Message Resolution




page 7-4).

P lat e a lrea dy present onshelf <shelfNumber>

Ca use: An a tt empt wa s ma de to pla ce a pla te ona shelf wh ere a nother plat e is a lrea dy present.

Resolution: P lace the plat e on a different shelf.

P lat e incorrectly placed ona shelf

Ca use: A plat e is ha nging out of a shelf.

Resolution: En sure t he plat e is positionedcorrectly on t he shelf.

P lat e present on low ersample mana ger t ra nsfer

shutt le

Ca use: A plat e ha s been detected on t he low ersam ple ma na ger tra nsfer shutt le during a sca n.

Resolution: Remove the plat e from t he low ersample manager transfer shuttle.

P ow er supply fa n ha rd-wa re faul t

Expla na tion: The pow er supply fa n ha s failed.

Resolution:



aga in .


P ower supply hardw arefault

Explana tion: t he power supply ha s fa i led.

Resolution:






Message Resolution




Sa mple orga nizer Z-a xismove error

Expla na tion: The sa mple orga nizer Z-a xis failedt o move.

Resolution:



aga in .


Sa mple orga nizer dooropen

P robable causes:

• The door is open wh i le trying to invoke anoperation.

• An opera t ion wa s running while the door wa sopened.

Resolution: Close the s a mple orga nizer door.

Sa mple orga nizer tra nsfershut tle home an d refer-

ence sensors both ON

Explanation: The sample organizer transfershu t t le home an d/or reference sensors fa iled.

P robable causes:• E lect r on ics fa i lu re


Resolution:



aga in .



Message Resolution




Sa mple orga nizer t ra nsfershut tle homing error

Explanation: The sample organizer transfershuttle homing failed.

P robable causes:


• Sa mple organ izer t r ans fer shut t le home

sensor fa ilure• P n e uma t i cs pr essur e too l ow

Resolution:




Sa mple orga nizer t ra nsfershut tle move error

Explanation: The sample organizer transfershut tle move fa iled.

Resolution:





Message Resolution




Sa mple orga nizer Z-a xisHome a nd R eferencesensors both ON

Expla na tion: The sa mple orga nizer Z-a xis homea nd /or referen ce sens ors fa iled.

P robable causes:



Resolution:1. Reset the sam ple organizer v ia the sample




Sa mple orga nizer Z-a xishoming error

Expla na tion: The sa mple orga nizer Z-a xishoming fa iled.

P robable causes:


• Sa mple organizer Z-axis home sensor fa i lure

• Mot or fa ilure

Resolution:






Message Resolution




Sa mple orga nizer Z-a xismove to AWAY positionerror

E xplan a tion: The sa mple orga nizer Z-a xis couldnot complet e a move to t he AWAY sensor.

Resolution:



aga in .


Sa mple orga nizer Z-a xismove to low er sa mple

ma na ger port error

E xplan a tion: The sa mple orga nizer Z-a xis couldnot complete a move to the low er sa mple

mana ger por t .Resolution:





Sh elf numbers notconsecutive

Expla na tion: The shelf numbers assigned a t t heend of a sca n a re not consecut ive.

Resolution:

1. Check a l l shelves.

2. I f the a larm persists , reset t he sampleorga nizer via the sa mple ma na ger contr olpan el or ACQU ITY UP LC C onsole.




Message Resolution





Temperature controltime-out h a rdw a re fault

Expla na tion: A time-out occurred in th e temper-ature controller watchdog.

Resolution:



aga in .


Topm os t pla t e too t a ll C a u se: Th e pla t e on t h e t opm os t sh elf in t h esa mple organ izer is too ta ll , a nd its height goes

a bove the gua rd ra il locat ed at t ha t slot .Resolution: U se a sh orter plat e or move the topshelf dow n.


Message Resolution




TUV detector troubleshooting

Power LED

The pow er LE D, on t he left side of th e detector’s front pa nel, indica tes w henth e instru ment is pow ered-on or powered-off. It is stea dy gr een w hen t heinstr ument is working properly.

Lamp LED

The lamp LE D, t o the r ight of the pow er LED , indica tes the lam p sta tus.

Clearing bubbles from the flow cell

To clear bubbles from the flow cell:

1. Ensure the pressure regula tor a t the out let of the flow cell is in place. I fthe pressure regulat or ha s been bypassed, ensure that a device tha tdevelops a minimum of 1724 kP a (17 ba r, 250 psi), but not gr eat er t ha n6895 kP a (69 ba r, 1000 psi), for t he given flow r a te a nd m obile pha se isplaced downst rea m from the detect or.

TUV detector lamp LED indications

LED state Description

U nlit Indica t es the det ector la mp is ext inguished.S t ea d y g reen color I nd ica t es t h e det ect or is oper a t in g n or ma lly .

F la s h in g gr een color I n dica t e s t h e d et ect or is in it ia l iz in g orcalibrating.

S tea dy red color I ndica t es a ser ious error t ha t prevent s fur t heropera tion. Informa tion a bout t he error ca n be

found in t he ACQU ITY UP LC C onsole.F la s hin g r ed color I ndica t es a ser iou s fa ilu re w it h in th e det ect or

th a t prevents further operat ion. Ca ll Wa tersTechnica l S ervice (see page 7-4).




2. Esta blish l iquid flow t hrough the detector flow cel l wi th degasseda cetonitr i le or metha nol a t a f low ra te tha t is a nticipat ed to be used for

subsequent a na lysis.

TUV detector troubleshooting


An a l og ou tpu t in cor r ect AUF S set t i ng ch a n g ed R eset th e AUF Ssetting.


2. Test electr ical outletfor pow er.

Open or defect ive fuse Replace fuse (seepage 6-78).

Ca libra tion or energyerror on start up

No solvent flowingth rough flow cell

U V a bsorber in flow cell

Make sure tha t solventis flowing th rough th eflow cell.

En sure th e mobilephase is t ransparent(such as w a ter).

Flus h t he flow cell (see

page 6-65).P er form ma nualcalibration.

Communicationproblems

Con fig ur a t i on pr oblem Ch eck E th er n etconfiguration.

Im proper or defect ive

Et hernet cable

Repla ce th e ca ble w ith

a shielded Et hernetcable.



TUV detector troubleshooting 7-103

Deut erium lam p doesnot l ight; lam p LED is“off”, solid r ed, orf la shing red

F a ult y la mp Repla ce t he la mp (seepage 6-73).

L am p not plugged in P lug in the la mpconn ector (seepage 6-73).

B a d lamp power supplyboard 1. Reset the TU Vdetector via t hecont rol pa nel orACQU ITY U P LCConsole.

2. I f the problempersist s, pow er-off

a nd pow er-on a ga in.3. I f the problem

persists, call Wa tersTechnical Service(see page 7-4).

La mp input shorted in“off” position

Inspect the rea r pa nelconn ections (see

page 7-111) or look for atimed event w ithin amet hod (possiblyprogra mm ed to extin-guish t he lamp).

No sample an d refer-ence energy; la mp LE D

is off, solid red, orf la shing red

La mp burned out At t empt t o reign it e t helamp. If i t fa i ls to

ignite, replace thelamp.

L am p ext inguished E nsure t he la mp sw it chis in t he “on” position.

Run the Sa mple andReference En ergy dia g-nost ic (see page 3-37).

TUV detector troubleshooting (Continued)





Alarm messages

To clear alarm messages:1. Follow the correct ive act ion in this ta ble, and reset t he detector v ia the

control panel or ACQU ITY U P LC Console.

2. I f the a l a rm message st i ll appears on the ACQUITY UP LC Console a f te ryou reset t he detector, reboot the detector. If th e ala rm m essag e persists,cont a ct Wa t ers Technica l Service.

The follow ing t a ble lists t he TU V detector ala rm m essages a nd t heirresolutions.

P eak out of range errora t s t a r t up

Mobile pha se a bsor-ba nce too high; or a irbubbles in t he flow cell

En sure th e mobilepha se in t he flow celldoes not a bsorb a bove250 nm.

En sure no air bubblesa re in t he flow cell (seepage 7-102).

Reca libra te t hedetector.



A/D Ana log t o digit a l convert er

AD C Ana log t o digit a l convert er

Mux Mult iplexer

TUV detector alarm messages

Message Resolution

TUV detector troubleshooting (Continued)





Ca librat ion differs x.xx257 nm . F low cell bubblespossible. Try Ca libra te

Reca librat e the detector.





Ca libra tion not found.Defaults Set

1. Reset th e TU V detector via t he control panelor ACQU ITY UP LC Console.



Ca librat ion successful.Ma x nm deviat ion sincelast ca librat ion: x.xx

Displa yed for informa tion only. No action isrequired.

Ca librat ion unsuccessful.257 Pea k out of ran ge.Flowcell Bubbles?

1. Flush t he f low cell (see page 6-65).

2. P ow er the TU V detector off a nd on a ga in.


1. Flush t he f low cell (see page 6-65).2. P ow er the TU V detector off a nd on a ga in.




Communication failure.

Referen ce or S a mple A/D .

1. Reset th e TU V detector via t he control panel

or ACQU ITY UP LC Console.2. If t he a larm persists, power-off and power-on

aga in .


TUV detector alarm messages (Continued)

Message Resolution




Configura tion not found.Defaults Set




S ervice (see page 7-4).Da rk current. ReferenceDa rk too high

Close th e detector door.

Fil ter initial izat ionfa ilure. Erbium position

1. Clea r bubbles from th e flow cell (seepage 7-102).


Fil ter initial izat ionfa ilure. No filters found


2. P ow er the TU V detector off a nd on aga in.

Fil ter initial izat ionfa ilure. No referenceenergy



Fil ter initial izat ion

fa ilure. Order filterposition

1. Clea r bubbles from th e flow cell (see

page 7-102).


Fil ter initial izat ionfa ilure. Sh ut ter position



G rat ing ini t ia l iza t ion

failure. Ba cklash too high.Flowcell Bubbles? Try Cal.

1. Clea r bubbles from th e flow cell (see

page 7-102).2. P ow er the TU V detector off a nd on aga in.

G rat ing ini t ia l iza t ionfa ilure. Missing/B a dSensor

1. Reset the TU V detector via the control panelor ACQU ITY UP LC Console.



Message Resolution





La mp dat a not found. Life-Time Defa ults Set

P robable ca use: Defective ba tt ery on C P U .

Resolution:





La mp energy low . Repla ce t he la mp (see page 6-73).

La mp external inputconflict

Set t he input on the rear pa nel so tha t i t doesnot conflict w ith t he lam p comma nd (seepage 7-111).

La mp fa ilure 1. Try relight ing the la mp.

2. If the a larm persists, pow er the TU V detectoroff and on aga in.

3. If the a larm persists, replace the lamp (seepage 6-73).

La mp light ing fa ilure 1. Try relight ing t he la mp.



Light Level Error. Air inCell? Try t o Ca librat e



Light Level Er ror. Ref.Sa tura ted . Cycle Power

1. Flush t he f low cell (see page 6-65).2. P ow er the TU V detector off a nd on a ga in.

Lowered gain t o a voidsat ura tion. Ref. LightLevel Increased

Displa yed for informa tion only. Indica tes t ha tflow cell bubbles w ere clear ed. No act ion isrequired.

Lowered gain t o a void

i S l Li h

Displa yed for informa tion only. Indica tes t ha t

fl ll b bbl l d N i i


Message Resolution




sa t ura t ion. Sa mple LightLevel Increased flow cell bubbles w ere clear ed. No act ion isrequired.

Method not found.Defaults Set




S ervice (see page 7-4).Mux ADC Ca libra tioncomma nd fa i lure




Mux ADC default registererror




Mux ADC input selectionfailure




Mux ADC reset comma nd

failure

1. Reset th e TU V detector via t he control panel

or ACQU ITY UP LC Console.




Message Resolution




No interru pt from preAmpboa rd. Average Sa mples

1. Reset the TU V detector via the contr ol panelor ACQU ITY UP LC Console.



S ervice (see page 7-4).No interru pt from preAmpboard. Lock. CannotContinue




No interru pt from preAmpboa rd. Wa iting for D a ta inD ua l




P eak not found D 2 656nm . Flow cell B ubbles? TryCa l ibra te

1. Flush th e f low cell and ca librat e. (seepage 6-65).



P eak not found xxx.x.E rbium 257 nm. Flow cell

B ubbles? Try C a librat e

Flus h t he flow cell (see page 6-65).

P eak not found xxx.x.E rbium 379 nm. Flow cellB ubbles? Try C a librat e


P eak not found xxx.x.E rbium 521 nm. Flow cell




Message Resolution





Displaying TUV detector rear panel interface connections

You can use the ACQU ITY UP LC C onsole to determine the sta tus ofinput /output signa l connections on th e rea r pa nel of the TU V detect or This

Remote command receivewhile calibrating -- Waitfor calibration to complete


Scan not found. DefaultsS et

P robable ca use: B a tt ery is dea d.

Resolution:




Sys tem ca n n ot r espon d 1. R eset th e TUV d ete ctor vi a th e con t r ol pa n elor ACQU ITY UP LC Console.



Sy stem not ca librat ed. Try

to Cal ibra t e


This is n ot a n ACQU ITYTU V personalit y ca rd




Wavelengths span 370nm.Order Filter n ot in use fornm : XXX



Message Resolution




input /output signa l connections on th e rea r pa nel of the TU V detect or. Thisdispla y gives you a r eal-time st a tu s of the instr ument ’s signa l connections. A

green LED indica tes a signa l ca ble is connected to the term ina l. A red LEDindica tes no signa l ca ble is connected to th e termina l.

To display TUV detector rear panel interface connections:

1. In t he ACQU ITY UP LC C onsole, select TU V Detector from the systemtree.

2. Click Troubleshoot > Rea r P a nel. The TU V Detector Rea r P a nel dialogbox appears .

TUV Detector Rear Panel dialog box

The followin g t a ble describes t he TU V det ector I/O conn ections .

TUV detector analog-out/event-in connections

Signal connections DescriptionAna log 1 (Out ) Not used.




In ject S t a r t (In) Not used.

La mp Off (In) When t riggered it ext inguishes t he la mp




La mp Off (In) When t r iggered, it ext inguishes t he la mp.



To change TUV detector rear panel interface connections:

1. In t he ACQU ITY UP LC C onsole, select TU V Detector from the systemtree.

2. In t he TU V detector informa tion window , cl ick Troubleshoot > Rea rPanel .

3. In t he TUV Detector Rear P anel d ia log box, loca t e a s ignal connect ionw ith a n Out la bel, an d th en click the red or green LED . The out putsigna l for t he selected sign a l connection indicat es wh ether it is opened

or closed .

C ha rt Ma rk (In) When t riggered, ca uses bot h a na log out putcha nnels and the digi ta l da t a sent t o the dat asyst em to increa se their va lue for a period oftime.


TUV detector analog-out/event-in connections (Continued)





PDA detector troubleshooting

Power LED


Lamp LED

The lamp LE D, t o the r ight of the pow er LED , indica tes the lam p sta tus.

PDA detector lamp LED indications


U nlit Indica t es the det ector la mp is ext inguished.S t ea d y gr een color I nd ica t es t h e det ect or is oper a t in g n or ma lly .


S tea dy red color I ndica t es a ser ious error t ha t prevent s fur t heropera tion. Informa tion a bout t he error ca n befound in t he ACQU ITY UP LC C onsole.

F la s hin g r ed color I ndica t es a ser iou s fa ilu re w it h in th e det ect orth a t prevents further operat ion. Ca ll Wa tersTechnica l S ervice (see page 7-4).




PDA detector troubleshooting





Replace fuse (see t heACQUI TY UPLC

Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).

Ch a nge in referencespectrum

Mobile pha se cont a insga s or is conta mina ted

P repare fresh mobilephase and degasthoroughly.

Air bubbles tr a pped inflow cell

Resea t a nd check a lign-ment of flow cell (seethe ACQUI TY UPLC

Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).

Flus h t he flow cell (seethe ACQUI TY UPLC

Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ), ora pply slight ba ckpres-sure on th e detectorwaste outlet .

Ensure the ba ckpres-sure regulat or isconnected to thedetector w a ste outlet .



PDA detector troubleshooting 7-115

Det ector notresponding toACQU ITY U P LCConsole

B a d or disconnectedcable

In spect cable conn ec-tions, t ightenconn ect ors, or r eplacecable.

Configura t ion prob lem Check E thernet config-ura tion. For deta ils, seeEm power onl in e H elp .

S hut t er fa ilure messa ge S hut t er fa iled 1. C lea r bubbles fromthe flow cell (seepage 7-102).

2. P ow er th e P D Adetector off an d on

aga in .S olven t in dr a in lin e L ea k fr om flow cell

gasketReplace th e flow cell(see the ACQUITY

UPLC Photodi ode

Ar r ay Detector Gett in g

Star ted Gui de ).

Lea k from flow cell

inlet a nd outlet f it t ings

Ch eck fitt ings for over-

tightening orundertightening, a ndrepla ce fitting s ifnecessary.

Sta tus l ight b l inks andlam p light is off

The detector is runn ingconfidence test s

No correct ive a ctionrequired. Wa it un tiltests f inish.

PDA detector troubleshooting (Continued)





Sta tus l ight b l inks andlam p light on

Fa i led star t updiagnostics

Resea t a nd check a lign-ment of flow cell (seethe ACQUI TY UPLC

Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).


Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).

Dirt y flow cell ca usingshutt er diagnostic to

fai l


Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).

In sufficient energyreaching photodiodea rra y beca use of airbubble

To prevent a ir bubblesfrom formin g, checkth a t t here is a 1- to2-foot (30- to 60-cm)

leng th of 0.009-inch(0.23-mm) ID tubingconnected to thedetector w a ste outlet .

Wea k la mp Repla ce t he la mp (seethe ACQUI TY UPLC

Photodi ode Ar r ay

Detector Gett i ng

Star ted Gui de ).

PDA detector troubleshooting (Continued)





Alarm messages


1. Follow the correct ive act ion in this ta ble, and reset t he detector v ia thecontrol panel or ACQU ITY U P LC Console.


The follow ing ta ble lists th e PD A detector a la rm messa ges an d theirresolutions.



A/D Ana log t o digit a l convert er

AD C Ana log t o digit a l convert er

D sp D igit a l signa l processing

H W H a rdw a re

NV-RAM Non-vola tile ra ndom access memory (ba ckup up by abat tery)

Neg Nega t ive

P D A P hot odiode a rra y

P os P osit ive

Vcc P ow er supply volt a ge

PDA detector alarm messages

Message Resolution

Ala rm D sp Tra nsferFailed

1. Reset t he PD A detector via t he contr ol pa nelor ACQU ITY UP LC Console.







Aut o exposure t ime t oohigh




Ca l ib ra t i on successf ul D ispla ye d for i nf or ma t i on on ly . No a ct ion isrequired.

Ca l ibra t ion unsuccessful Flush the f low cell and repeat ca l ibra t ion (seethe ACQU I TY UPLC Photodi ode Ar r ay Detector

Gett i ng Start ed Gui de ).

Comma nd received while

initial izing. U na ble toprocess

1. Reset t he PD A detector via t he contr ol pa nel


aga in .


Configura tion not found,defa ults set




Delta betw een previousa nd current t imesta mp is

incorrect




PDA detector alarm messages (Continued)

Message Resolution




HW Commu nicat ionsFa ilure, PD A ReferenceADC

1. Check the Ethernet cable connection a t t heP DA detector and net w ork sw itch. Reconnectth e ca ble.

2. I f the a larm persists , reset t he PD A detectorvia t he control pan el or ACQU ITY UP LCConsole.



HW Commu nicat ionsFa ilure, P DA Sa mple ADC

1. Check the Ethernet cable connection a t t heP DA detector and net w ork sw itch. Reconnect

th e ca ble.2. I f the a larm persists , reset t he PD A detector

via t he control pan el or ACQU ITY UP LCConsole.



S ervice (see page 7-4).HW Commu nicat ionsFa ilure, P D A Temp ADC

1. Check the Ethernet cable connection a t t heP DA detector and net w ork sw itch. Reconnectth e ca ble.

2. I f the a larm persists , reset t he PD A detectorvia t he control pan el or ACQU ITY UP LCConsole.




Message Resolution




H W F a i lu r e. F ilt er F la g . 1. R eset th e P D A d etector v ia th e con t r ol pa n e lor ACQU ITY UP LC Console.



HW Fa ilure. Lea k Refer-ence A/D F a ilu re




HW Failure. Leak Sa mpleA/D F a ilure




Lam p data not found,defa ults set P robable ca use: B a tt ery is dea d.Resolution:





La mp energy low Repla ce t he la mp (see t he ACQUI TY UPLC

Photodi ode Arr ay Detector Gett i ng St ar ted

Gu ide ).


Message Resolution




La mp external inputconflict




La mp fa ilure 1. Try relight ing the la mp.


3. If the a larm persists, replace the lamp (seethe ACQUI TY U PLC Photod iode Arr ay

Detector Gett i ng Star ted Gu i de ).

La mp hours counterexceeded

Repla ce the la mp (see th e ACQUI TY UPLCPhotodi ode Arr ay Detector Gett i ng St ar ted

Gu ide ).

La mp light ing fa ilure 1. Try reignit ing t he la mp.


3. If the a larm persists, replace the lamp (seethe ACQUI TY U PLC Photod iode Arr ay


Method not found,defa ults set

P robable ca use: B a tt ery is dea d.

Resolution:





Message Resolution




NV-RAM B ad 1. Reset t he P DA det ect or via the cont rol pa nelor ACQU ITY UP LC Console.



P DA Da rk Current tooHigh


P DA Da rk Current tooLow




P ower Supply Fa ilure. 24Volts on Lam p B oa rd




P ower Supply Fa ilure. 24Volts on P ersona lityC ar d




S ervice (see page 7-4).P ow er Supply Fa ilure.Neg. 15 Volts onPersonali ty





Message Resolution




P ow er Supply Fa ilure.P os. 15 Volts onPersonali ty




P ower S upply F a ilure. Vccon Cha rge Amp B oa rd




Reference Dar k C urrenttoo High


Reference Dar k C urrenttoo Low




S hut t er fa iled to h om e Ru n t he sh ut t er dia gn ost ic.

S y st em ca n n ot r es pon d 1. R es et t h e P D A d et ect or via t h e con t rol pa n elor ACQU ITY UP LC Console.



This is n ot a n ACQU ITYP DA persona li ty card





Message Resolution




Displaying PDA detector rear panel interface connections

You can use the ACQU ITY UP LC C onsole to determine the sta tus ofinput /output signa l connections on th e rear pa nel of th e P DA detector. Thisdispla y gives you a r eal-time st a tu s of the instr ument ’s signa l connections. Agreen LED indica tes a signa l ca ble is connected to the term ina l. A red LED

indica tes no signa l ca ble is connected to th e termina l.

To display PDA detector rear panel interface connections:

1. In the ACQU ITY UP LC Console, select P DA Detector f rom the systemtree.

Wavelength VerificationFailure





Message Resolution




2. Click Troubleshoot > Rear P a nel. The P DA Detector Rea r P a nel dialogbox appears .

PDA Detector Rear Panel dialog box

The follow ing t a ble describes th e P DA det ect or I/O connect ions.

PDA detector analog-out/event-in connections


Ana log 1 (Out ) U sed for a na log cha rt out put funct iona lit y.

Ana log 2 (Out ) U sed for a na log cha rt out put funct iona lit y.




La mp Off (In) When t r iggered, it ext inguishes t he la mp.Cha rt Ma rk (In) Ma rks a ll da ta w ith a 0.1 AU tick ma rk.

Aut o Zero (In) Ca lcula tes a n offset va lue t ha t , w hen a dded t othe sa mple signal , ma kes the resulting ba selinesigna l zero for a ll w a velengt hs.






To change PDA detector rear panel interface connections:

1. In the ACQU ITY UP LC Console, select P DA Detector f rom the systemtree.

2. In t he PD A detector informa tion w indow , cl ick Troubleshoot > RearPanel .

3. In the P DA Detector Rear P a nel d ia log box, loca t e a s ignal connect ionw ith a n Out la bel, an d th en click the red or green LED . The out putsigna l for t he selected sign a l connection indicat es wh ether it is opened

or closed .




ELS detector troubleshooting

Power LED


Lamp LEDThe lamp LE D, t o the r ight of the pow er LED , indica tes the lam p sta tus.

ELS detector lamp LED indications


U nlit Indica t es the det ector la mp is ext inguished.

S t ea d y gr een color I nd ica t es t h e det ect or is oper a t in g n or ma lly .


S tea dy red color I ndica t es a ser ious error t ha t prevent s fur t heropera tion. Informa tion a bout t he error ca n befound in t he ACQU ITY UP LC C onsole.

F la s hin g r ed color I ndica t es a ser iou s fa ilu re w it h in th e det ect orth a t prevents further operat ion. Ca ll Wa tersTechnica l S ervice (see page 7-4).




Alarm messages


1. Follow the correct ive act ion in this ta ble, and reset t he detector v ia thecontrol panel or ACQU ITY U P LC Console.


ELS detector troubleshootingSymptom Possible cause Corrective action



Ca ble loose or defective Inspect l ine cordconnections.


Replace fuse (see t heACQU IT Y UPLC Evap-

or at ive Li ght Scatter i ng

Detector Gett i ng

Star ted Gui de ).



AD C Ana log t o digit a l convert erH W H a rdw a re

NV-RAM Non-vola tile ra ndom access memory (ba ckup up by abat tery)

P MT P hot omult iplier t ube



ELS detector troubleshooting 7-129

The following ta ble l ists the E LS detector a larm messages a nd t heirresolutions.

ELS detector alarm messages

Message Resolution

Configura t ion Not Found P robab le cause: Memory c leared, memoryfa ilure, or loss of bat tery ba ckup.

Resolution:

1. Reset t he ELS detector via the control pa nelor ACQU ITY UP LC Console.



Drif t tube hea ter is not atset temperat ure

P robable ca use: The detector bega n a cquiringda ta before the hea ter reached the set pointtemperature.

Resolution:

1. Wa it unti l the heaters reach their set pointtemperatures.

2. I f the a larm persists , reset t he ELS detector







Dr ift t ube or optics heat errose a bove its set tempera -ture (temperature)

P robable ca use: The hea ter’s out put is a ffectedby fluid flow or a nother hea ter, or the heat er isdefective.

Resolution:

1. Try increasing the a la rm band by a l lowinggreater t empera ture l imits.

2. I f the a larm persists , reset t he ELS detectorvia t he control pan el or ACQU ITY UP LCConsole.



S ervice (see page 7-4).Dr ift t ube or optics heat erunder tempera ture(temperature)

P robable ca use: The hea ter’s out put is a ffectedby fluid an d ga s flow , or th e heat er is defective.

Resolution:

1. Try increasing the a la rm band by a l lowinggreater t empera ture l imits.





ELS detector alarm messages (Continued)

Message Resolution




G a s pr es su re t oo h ig h P r oba b le ca u se: Th e g a s pr es su re h a s r is enmore tha n 14 kP a (.14 ba r, 2 psi) a bove the setpoint.

Resolution:




G a s pr es su re t oo low P r oba ble ca u se: Th e g a s pr ess ur e h a s fa llenbelow 103 kP a (1.03 bar , 15 psi) or more t ha n 14

kP a (.14 bar , 2 psi) below the set point.Resolution:

1. Check the gas source.



aga in .


HW Commu nicat ionFa ilure - Mult iplexed AD C

P robable cause: Detector initia lizat ion is notcompleted due t o a problem w ith t he persona lityboard.

Resolution:1. Reset t he ELS detector via the control pa nel





Message Resolution




HW Commu nicat ionFailure - Reference Photo-diode ADC

P robable cause: Detector initia lizat ion is notcompleted due to a problem w ith t he an a logfront -end cable or electronics.

Resolution:




HW Commu nicat ionFa ilure - Sa mple ADC

P robable cause: A problem w ith t he an a logfront -end cable or electronics.





In va lid operat ion ba sed oninstrument s ta t e

P robable ca use: An un expected error ha soccurred.

Resolution:






Message Resolution




L a mp da t a n ot foun d P r oba ble ca use: Th e la m p usa ge da t a could notbe retrieved from NV-RAM.

Resolution:

1. Reenter the informa tion through theACQU ITY U P LC C onsole.




La mp fa ilure P roba ble ca use: The la mp is turned off, or thelam p burned out a nd needs to be replaced.

Resolution: Try t o ignit e the la mp. If t he errorpersists , replace the la mp (see the ACQUITY

UPLC Evaporati ve L igh t Scatt er i ng Detector

Gett i ng Start ed Gui de ).

La mp hours thr eshold

exceeded

Probable cause: The user-configurable lamp

hours t hreshold ha s been exceeded.Resolution: Replace the lamp (see theACQUI TY U PLC E vaporat i ve L ight Scat ter i ng


Low Input Ga s Pressure P robab le cause: The gas is turned of f and/orsomeone request ed St op Flow.

Resolution:

1. Check the input ga s l ine connection a nd ga ssource.

2. I f the a larm persists , reset the detector v iath e cont rol pan el or ACQU ITY U P LCConsole.


Message Resolution




Missed da ta point P roba ble ca use: A da ta poin t wa s m issed w hilecollecting da ta .

Resolution:

1. The ACQU ITY UP LC C onsole sends awa rning a la rm to the data system.




Nebulizer, drift tu be, oroptics hea ter probe failure P robable ca use: Either t he nebulizer, drift t ube,or optics probe is defect ive, or not conn ect ed.

Resolution:




NV-RAM B ad P roba ble ca use: Memory clea red, memoryfa ilure, or loss of bat tery ba ckup.

Resolution:

1. Reset t he ELS detector via the control pa nel


aga in .



Message Resolution




P MT Da rk Curr ent tooHigh

P robable ca use: Light is entering t he opticsbench, or a problem w ith th e P MT or person-a lity boa rd.

Resolution:




P MT Da rk Curr ent tooLow

P robable ca use: A problem w ith th e P MT orpersonality board.





Remote comma nd notsupported

P robable cause: A comma nd sent t o th e inst ru-ment from t he data system w a s not supported.

Resolution: Reset th e ELS detector via t hecontrol panel or ACQU ITY U P LC Console.


Message Resolution




Signa l too high. Ba d P MTor light lea k in bench.

P robable ca use: Excessive a mount of lightentering th e optics bench. The norma lizat ionfactor rema ins unchanged.

Resolution:

1. Close the detector door.





Syst em Not Norma lized.Run Dia gnostic P robable cause: Memory a nd previous norma l-izat ion informa tion were reset.

Resolution:




User r eset of NV-R AM P r ob a ble ca use: Th e user h a s reset th e NV-R AMvalues thr ough t he ACQU ITY U P LC Console.

Resolution:

1. Reset t he ELS detector via the control pa nel


aga in .



Message Resolution




Displaying ELS detector rear panel interface connections

You can use the ACQU ITY UP LC C onsole to determine the sta tus ofinput/output signa l connections on t he rea r pa nel of the E LS detector. Thisdispla y gives you a r eal-time st a tu s of the instr ument ’s signa l connections. Agreen LED indica tes a signa l ca ble is connected to the term ina l. A red LEDindica tes no signa l ca ble is connected to th e termina l.

To display ELS detector rear panel interface connections:

1. In the ACQU ITY UP LC Console, select EL S Detector from the systemtree.

2. Cl ick Troubleshoot > Rear P a nel . The ELS Detector Rear P an el d ia logbox appears .

ELS Detector Rear Panel dialog box




The follow ing t a ble describes t he E LS detector I /O conn ect ions.

ELS detector analog-out/event-in connections


S ign al (Out put ) P r ovides a high resolut ion out put for t he sa mplesignal .

Out put volta ge ran ge: –0.1 to 2.1 VDC . User ma ya pply a n offset t o prevent nega tive-going signa ls

due to drift during a chromat ogra phic run.Au xilia r y (O ut pu t ) P r ovid es a h ig h r es olu t ion ou t pu t for mon it or in g

the nebulizer t emperat ure, drift tube t empera-tu re, or ga s pressure.

Out put volta ge ra nge: –0.1 to 2.1 VDC .

S t op-F low (Ou t pu t ) S t ops t h e flow t o t h e ch rom a t og ra p hic s ys t emw hen a potentia l safety problem such a s a low

input ga s flow condit ion, or tempera tur e cont rolproblem occurs.

S w it ch (Out put ) C on trols a t im ed even t or threshold level a nd is auser-programmable auxiliary output.


La mp On (In) When t riggered, ignit es or ext inguishes the la mp.

C ha rt Ma rk (I n) When t riggered, ca uses bot h a na log out put cha n-nels and t he digita l da ta sent t o the dat a systemto increase t heir va lue for a period of time.






Via th e rear pan el display , you ca n open a nd close certa in output connections.

Opening or closing output connections ca n be useful w hen you need to st a rt orstop a n injection or t roubleshoot syst em connectivity.

To change ELS detector rear panel interface connections:

1. In the ACQU ITY UP LC Console, select EL S Detector from the systemtree.

2. In t he ELS detector informat ion w indow, cl ick Troubleshoot > RearPanel .

3. In the ELS Detector Rear P anel d ia log box, loca t e a s ignal connect ionw ith a n Out la bel, an d th en click the red or green LED . The out putsigna l for t he selected sign a l connection indicat es wh ether it is opened

or closed .




Chromatography troubleshooting

The ta ble below lists chromat ographic sympt oms, possible causes, a nd

suggested corrective a ctions for the ACQU ITY UP LC syst em.B efore you use th e ta ble, rea d t he G eneral Troubleshooting section, a nd followth e basic troubleshootin g steps to isolat e the cause of the chroma togra phicsymptom.

Warning: To prevent injury, a lwa ys observe good laborat orypra ctices when y ou han dle solvent s, cha nge tubing, or opera te t heACQU ITY UP LC sy st em. Know t he physica l and chemica l propertiesof the solvents you use. Refer to the Mat eria l Sa fety Da ta Sheets forth e solvent s in use.

Chromatography troubleshooting

Symptom Possible cause Corrective actionBandspreading,poor peak s ha pe,or loss of columnresolution

Wrong fitt ings t oACQU ITY U P LCcolumn oranyw here in thesystem.

U se ACQU ITY UP LC fi t t ings(Parker-style).

One or more

connections isdefective.

Ch eck connections. All tubes mu st

be “bott omed out” int o fema leconnectors.

The lengt h ordia met er of one ormore connectiontubes is incorrect.

Connectin g tube betw een th e samplema na ger injection valve an d columnshould be no la rger t ha n 0.005” forU P LC systems. Most connectingtu bes a re pre-cut w ith specified

diameters and lengths.Incorrect wa shsolvents orvolumes.

Ch a nge w a sh solvent s a nd/orvolumes.

Defect ive column. Replace column.



Chromatography troubleshooting 7-141

Column is dir ty,defective, orcontaminated.

• F lu sh t he colu mn w it h a nea torgan ic solvent, t a king ca re not t oprecipita te buffers.

• I f flushing doesn't remove columnconta mina nts, clea n the columnusing the clea ning a nd

regenerat ion proceduresdescribed in Water s ACQU I TY

UPLC BEH Colum n Care and

U se I nstr uctions .

• I f flushing/cleaning doesn'tremove cont a mina nt , replace th ecolumn.

System not s ta bi-lized orchemicallyequilibrated.

Eq uilibrate column w ith a minimumof 10 column volumes of the mobilephase to be used. If running a n a uto-ma ted gra dient method, ensure youa re using sufficient a nd reproducibleequilibration times betweeninjections.

Example: For a 2.1 × 100 mmACQU ITY U P LC B EH column, runa bout 2.5 mL of solut ion a t 0.5mL /min for about 5 minut es.

See also: ACQUIT Y UPLC BEH

Column Car e and U se I nstruct ions .

Incorrect wa sh

solvents orvolumes.

Ch a nge w a sh solvents a nd/or

volumes.

Temperaturefluctuations.

Set a n a ppropria te tempera ture.

Chromatography troubleshooting (Continued)





Decreased reten-tion times

Incorrect flowra te .

Cha nge f low ra t e .

Incorrect solventcomposition.

Ch a nge composition.

High columntemperature.

Reduce column tempera t ure.

In correct mobilephase.

Use correct mobile phase.

Columncontaminated.

Clea n or r epla ce column.

Incorrect column. U se correct column.

Sa mple diluent

ma y be strongertha n the init ia lmobile phase.

Dilut e sample in a w eaker solut ion.

In ject less.

Err at ic retentiontimes

Air bu bble inpump hea d.

P rime binary solvent mana ger.

Malfunctioningcheck va lves.

Clean or replace inlet check valvecartr idges.

Lea king plungerseals.

Replace sea ls (see page 6-16).

Separa t ionchemistry.

Det ermine the suit a bility of mobilephase a nd column.

Clogged solventfilters.

Replace filters.

Equilibrationt ime a f ter agradient ma y beinsufficient.

Specify a longer equilibrat ion t ime.

Incorrect wa shsolvents orvolumes.

Ch a nge w a sh solvents a nd/orvolumes.








Flat-topped peaks Sa mple concen-tra tion orinject ion volumeexceeds volt a geoutput of detector.

Decrea se sa mple concentra tion orinject ion v olume.

Increased reten-

tion times

Incorrect flow

ra te .

Cha nge f low ra t e .

Incorrect solventcomposition.

Ch a nge solvent composition.

In correct mobilephase.

Use correct mobile phase.

Columncontaminated.


Incorrect column. U se correct column.

Fluid lea k (causeslow er flow ra te).

Ins pect fittings for lea ks. P erformsta tic deca y test .

Increase insystem pressure

Column isplugged.


• I f flushing doesn't remove columnconta mina nts, clea n the columnusing the clea ning a ndregenerat ion proceduresdescribed in Water s ACQU I TY




Tubing isplugged.

Syst emat ica lly check t ubing byma king an d breaking connections.






Loss of columnefficiency

Column is dir ty,defective, orcontaminated.


• I f flushing doesn't remove columnconta mina nts, clea n the columnusing the clea ning a nd

regenerat ion proceduresdescribed in Water s ACQU I TY




System not s ta bi-lized orchemicallyequilibrated.

Eq uilibrate column w ith a minimumof 10 column volumes of the mobilephase to be used. If running a n a uto-ma ted gra dient method, ensure youa re using sufficient a nd reproducibleequilibration times betweeninjections.

Example: For a 2.1 × 100 mmACQU ITY U P LC B EH column, runa bout 2.5 mL of solut ion a t 0.5mL /min for about 5 minut es.

See also: ACQUIT Y UPLC BEH

Column Car e and U se I nstruct ions .

Incorrect wa sh

solvents orvolumes.

Ch a nge w a sh solvents a nd/or

volumes.








Reproducibilityerrors

Incorrectchemis t ry /int e-gration.

Review chemistry a nd integra tion.

Sa mple mana ge-ment systemproblem.

Troubleshoot sa mple man a gementsystem.

Incorrect wa shsolvents orvolumes.

Ch a nge w a sh solvent s a nd/orvolumes.

In correct volumera nge for injectionmeth od or t ype.

Cha nge volume range.

S ensit ivit y loss Lea k in bina rysolvent m an a ger.

Troubleshoot solvent ma na gementsystem.

Degraded,conta mina ted, orimproperlyprepa red sam ple.

U se fresh sa mple.

Column

contaminated.


Loss of columnefficiency.







A 1

A Safety Advisories

Wa ters instrum ents display ha zard symbols designed to alert you to th ehidden da ngers of opera ting a nd ma inta ining the instruments. Theircorresponding user guides a lso include the ha za rd sym bols, wit ha ccompa nying text st a tements describing t he haza rds a nd t ell ing youhow t o a void th em. This a ppendix presents a ll th e safety sy mbols a ndsta tements t ha t a pply to th e entire l ine of Wa ters products.

Contents

Topic Page

Wa rning symbols A-2C a ut ion symbol A-6

Wa rnings t ha t a pply to a ll Wa ters inst ruments A-7

E lect rica l a nd ha ndling symbols A-14



A-1

A 2 Safety Advisories

Warning symbols

Wa rning sy mbols alert y ou to the risk of deat h, injury, or seriously a dversephysiological reactions a ssociat ed wit h a n inst rument ’s use or misuse. Heeda ll w a rnings wh en you insta l l, repa ir , and operat e Wa ters instruments.Wa ters a ssumes no liability for the failure of th ose w ho inst a ll, repair, oropera te i ts instruments to comply wit h a ny safety preca ution.

Task-specific hazard warnings

The follow ing w a rning symbols a lert you to r isks t ha t ca n a rise when youopera te or ma inta in a n instr ument or instrum ent component. S uch risksinclude burn injuries, electric shocks, ult ra violet r a diat ion exposures, an dothers.

When th e follow ing sym bols a ppea r in a ma nua l’s na rra tives or procedures,th eir accompany ing t ext identifies the specific risk a nd expla ins how t o avoid

it .

Warning: (G eneral risk of da nger. When t his symbol appears on a ninst rument , consult th e instr ument ’s user document a tion forimport a nt sa fety-relat ed informa tion before you use th e inst rument .)

Warning: (Risk of burn injury from cont a cting hot surfa ces.)

Warning: (Ris k of elect ric sh ock.)

Warning: (Risk of fire.)

Warning: (Risk of needle punctur e.)

Warning: (Risk of injury caused by moving ma chinery.)

Warning: (Risk of exposure t o ultra violet ra dia tion.)

Warning: (Risk of contacting corrosive substances.)



A-2 Safety Advisories

Warning symbols A-3

Warnings that apply to particular instruments, instrumentcomponents, and sample types

The following w a rnings can a ppear in t he user ma nua ls of part icularinstr ument s an d on la bels affixed to them or th eir component pa rt s.

Burst warning

This wa rning a pplies to Wa ters instr ument s fitt ed with nonmeta llic tubing.

Warning: (Risk of exposure to a toxic subst a nce.)

Warning: (Risk of persona l exposure to laser ra dia tion.)

Warning: (Risk of exposure t o biologica l a gents th a t can pose a seriousheal th threat . )

Warning: P ressurized nonmet a llic, or polymer, tubing can bur st .Observe th ese preca utions w hen w orking around such tubing:

• Wea r eye pr otect ion .

• E xt in gu is h a ll nea r by fla m es .

• Do not use tubing that is , or has been, s tressed or kinked.

• Do not expose nonmeta llic tubing to incompat ible compounds l iketet ra hydr ofura n (THF ) a nd nit ric or sulfuric acids.

• B e aw are tha t some compounds, l ike methylene chlor ide anddimethy l sulfoxide, ca n cause nonmet a llic tubing t o swell, w hichsignif ica ntly reduces the pressure at wh ich t he tubing can rupture.



Warning symbols A-3


Mass spectrometer flammable solvents warning

This warning applies to instruments operated with flammable solvents.

Mass spectrometer shock hazard

This w a rning a pplies to all Wa ters m a ss spectr ometers.

This w a rning a pplies to certa in instrument s w hen they a re in Operat e mode.

Warning: Where significa nt qua nt ities of fla mm a ble solvents a reinvolved, a continuous flow of nitrogen into the ion source is requiredto prevent possible ignition in th a t enclosed spa ce.

En sure tha t the nitrogen supply pressure never fa l ls below 400 kP a(4 bar, 58 psi) during an a na lysis in which flamm a ble solvents a reused. Also ensur e a ga s-fa il connection is connected t o the H P LC

syst em so th a t t he LC solvent flow st ops if th e nitrogen supply fails.

Warning: To avoid electr ic shock, do not r emove the m a ss

spect rometer ’s protective pan els. The component s th ey cover a re notuser-serviceable.

Warning: H igh volta ges ca n be present a t cert a in externa l surfaces ofth e ma ss spectr ometer wh en the inst rum ent is in Operat e mode. To

a void non-leth a l electr ic shock, ma ke sure th e instru ment is inSt a ndby mode before touching a reas ma rked w ith t his high volta gew a rning symbol.



A 4 Safety Advisories

Warning symbols A-5

Biohazard warning

This w a rning a pplies to Wa ters inst rum ents t ha t can be used to process

ma terial tha t m ight conta in bioha za rds: substa nces tha t conta in biologica la gents capa ble of producing ha rmful effects in huma ns.

Chemical hazard warning

This w a rning a pplies to Wa ters in str ument s t ha t can process corrosive, toxic,f la mma ble, or other ty pes of haza rdous ma terial .

Warning: Wa ters’ instruments a nd softw a re ca n be used to ana lyze orprocess potent ia lly infectious h uma n-sourced products, ina ctivat edmicroorgan isms, a nd oth er biologica l ma teria ls. To avoid infectionw ith t hese a gents, a ssume th a t a ll biologica l fluids are infectious,

observe good la bora tory pra ctices, an d consult your organ iza tion’sbioha za rd sa fety representa tive rega rding th eir proper use an dha ndling. Specific preca ut ions a ppea r in th e la test edition of the U SNational Institutes of Health (NIH) publication, Bi osafety i nM icrobio logical an d Bi omedi cal L abor ator ies(BMBL).

Warning: Wat ers instruments can be used to a na lyze orprocess potentia lly ha za rdous subst a nces. To avoid injuryw ith a ny of these ma terials, familia rize yourself with t hema terials a nd their haza rds, observe Good La bora toryP ra ctices (G LP ), a nd consult your orga nizat ion’s sa fetyrepresent a tive rega rding proper use a nd ha ndling.G uidelines ar e provided in the lat est edition of theNa tiona l Research C ouncil's publica tion, Pr uden t Pra cti cesin the L abor ator y: H and l i ng and Di sposal of Chem icals.



g y


Caution symbol

The ca ut ion sym bol signifies th a t a n inst rument ’s use or misuse ca n da ma geth e instr ument or compromise a sa mple’s int egrity. The follow ing sym bol a ndits a ssociat ed stat ement a re typica l of the kind tha t a lert you to the r isk ofdam a ging the instrum ent or sa mple.

Caution: To prevent st a tic char ges and resulta nt circuit da ma ge, donot remove the covers th a t protect integra ted circuit chips.



Warnings that apply to all Waters instruments A-7

Warnings that apply to all Waters instruments

When opera ting th is device, follow st a nda rd q ua lity cont rol procedures an dth e the equipment guidelines in th is section.

Attention: Cha nges or modificat ions t o th is unit n ot expressly a pproved byth e par ty responsible for compliance could void th e user’s a ut hority to opera tethe equipment.

Important: Toute modifica tion sur cette unit é n’a ya nt pas ét é expressémenta pprouvée pa r l’a ut orité responsa ble de la conformit é à la réglement a tionpeut a nnu ler le droit de l’utilisa teur à exploiter l ’équipement.

Achtung: J edwede Änderungen oder Modifika tionen an dem G erät ohne diea usdrückliche G enehmigung der für die ordnungsgemä ße Fun ktion-stüchtigkeit verant wortl ichen P ersonen kann zum E ntzug derB edienungsbefugnis des S ystems führen.

Avvertenza: eventu a li modifiche o al terazioni apporta te a questa unità enon espressam ente a pprova te da un ent e responsa bile per la conformitàannulleranno l’autorità dell’utente ad operare l’apparecchiatura.

Atencion: cualq uier ca mbio o modifica ción efectua do en est a unida d qu e noha ya sido expresa ment e aprobado por la pa rt e responsable del cumplimientopuede an ular la a utoriza ción del usua rio pa ra uti l iza r el equipo.




Warning: U se ca ution w hen working w ith a ny polymer tubing under

pressure:• Alwa ys wear eye protect ion wh en near pressur ized polymer tubing.

• E xt in gu is h a ll nea r by f la m es .

• Do not use tubing tha t ha s been severely s tressed or kinked.

• Do not use nonmeta l l ic tubing wi th te tra hydrofuran (THF) or concentra tednitric or sulfuric acids.

• B e aw are that m ethylene chlor ide and dimethyl sul foxide causenonmeta llic tu bing to swell, which grea tly r educes the rupt ure pressure ofthe tubing.

Attention: Manipulez les tubes en polymère sous pression avec precaution:

• P or tez systéma tiquement des lunet tes de protect ion lorsque vous voustr ouvez à proximité de t ubes en polymère pressurisés.

• Et eignez toute flamme se trouvant à proximité de l ’instrument .

• Evitez d'uti l iser des tubes sévèrement déformés ou endomma gés.• Evi tez d 'ut i liser des tubes non méta l l iques avec du té trahydrofurane

(TH F) ou de l'acide sulfur iqu e ou nit riq ue concentr é.

• Sa chez que le chlorure de méthylène et le diméthylesulfoxyde entra înent legonflement des tu ya ux non méta lliques, ce qui réduit considéra blementleur pression d e rupt ure.

Vorsicht: B ei der Arbeit mit P olymerschlä uchen unt er Druck ist besondereVorsicht angebracht:

• In der Nähe von unter Druck stehenden P olymerschläuchen stetsSchutzbrille tra gen.

• Al le of fenen Flammen in der Nähe löschen.

• Keine Schläuche verwenden, d ie s ta rk geknickt oder überbeanspruchtsind.

• Nichtm etall ische Schläuche nicht für Tetra hydrofura n (THF ) oderkonzent rierte S a lpeter- oder S chw efelsä ure verwenden.

• Durch Methylenchlorid und Dimethy lsulfoxid können nichtm eta llischeSchlä uche quellen; dadur ch wird der B erstdru ck des Schlau ches erheblichreduziert.




Attenzione: presta re a tt enzione dura nt e l’utilizzo dei tu bi di polimero

pressurizzati :• Indossare sempre occhiali da lavoro protett ivi nei pressi di tubi di polimero

pressurizzati .

• Est inguere ogni fonte d i ignizione circosta nte .

• Non ut i lizzare tubi soggetti che ha nno subito sollecita zioni eccessive o sonstati incurvati .

• Non uti l izzare tubi non meta ll ici con tetra idrofuran o (THF ) o a cidosolforico o nitr ico concentr a t o.

• Tenere presente che il cloruro di metilene e il dimetilsolfossido provoca norigonfia ment o nei tubi non meta llici, riducendo notevolment e la resistenzaa lla rott ura dei tubi stessi.

Advertencia: se recomienda precaución cuando se trabaje con tubos depolímero sometidos a presión:

• El usua rio deberá protegerse siempre los ojos cua ndo traba je cerca detu bos de polímero sometidos a presión.

• S i h ub ier a a lg un a l la ma la s p rox imid a d es .

• No se debe tra bajar con tubos que se haya n doblado o sometido a a l taspresiones.

• Es necesar io ut i liza r tubos de meta l cuando se tra baje contet ra hidr ofura no (TH F) o á cidos nít rico o sulfúr ico concentr a dos.

• Ha y que tener en cuenta que el cloruro de metileno y el sulfóxido dedimetilo dilat a n los tubos no metá licos, lo que reduce la presión de rupt urade los t ubos.




Warning: The user shall be ma de aw a re tha t i f th e equipment is used in a

ma nner not specified by the ma nufa ctu rer, the protection provided by t heequipment m a y be impaired.

Attention: L’utilisat eur doit être informé que si le ma tériel est utilisé d’unefaçon non spécifiée par le fabricant , la protection ass urée par le ma térielrisque d’être défectueuses.

Vorsicht: Der Benutzer wird da ra uf aufmerksam gema cht, da ss bei unsach-gemäßer Verw enddung des G erät es unter Umst ä nden nicht ordnungsgemäßfunktionieren.

Attenzione: l’utente deve essere al corrente del fatto che, se l’apparecchia-tur a viene ust a in un m odo specifica to da l produt tore, la protezione fornit adall ’a ppa recchiat ura potrà essere invalidata .

Advertencia: el usuar io deberá sa ber que si el equipo se utiliza de formadistint a a la especificada por el fabrican te, las medida s de protección delequipo podrían ser insuficientes.




Warning: To protect a ga inst fire ha za rd, repla ce fuses wit h t hose of the sa me

type and ra t ing .

Attention: Remplacez toujours les fusibles pa r d’aut res du mêm e type et dela même puissa nce afin d’éviter t out risq ue d’incendie.

Vorsicht: Zum S chutz gegen Feuergefa hr die Sicherungen nur m itSicherungen des gleichen Typs un d Nennw ertes ersetzen.

Attenzione: per una buona protezione contr o i rischi di incendio, sostit uire ifusibili con altri dello stesso tipo e amperaggio.

Advertencia: sust ituy a los fusibles por otros del mismo tipo y ca ra cterística spar a evita r el riesgo de incendio.




Warning: To a void possible elect rical shock, disconn ect t he pow er cord before

servicing the instrument.

Attention: Afin d’éviter t oute possibilité de comm otion élect riq ue,débran chez le cordon d’a limenta tion de la prise a va nt d’effectu er la ma inte-na nce de l’instr ument .

Vorsicht: Zur Vermeidung von St romschlä gen sollte das G erät vor der

Wa rtung vom Netz getrennt w erden.

Attenzione: per evit a re il rischio di scossa elet tr ica, scollega re il ca vo dia limenta zione prima di svolgere la m a nut enzione dello str ument o.

Precaución: para evita r desca rga s eléctrica s, desenchufe el ca ble de alimen-ta ción del instrumento a ntes de realizar cua lquier repara ción.




Electrical and handling symbols

Electrical symbols

These ca n a ppear in instr ument user ma nua ls an d on t he instrument’s frontor rea r pa nels.

Electrica l power on

Electrical power off

S tandby

Direct current

Alternating current

P rotective conductor termina l

Fra me, or chassis, termina l

Fuse

Recycle sym bol: Do not dispose in municipa l w a ste.



Electrical and handling symbols A-15

Handling symbols

These ha ndling symbols and t heir ass ociat ed text ca n a ppea r on la bels a ffixed

to the out er pa ckaging of Wa ters inst rum ent a nd component s hipments.

Keep upright !

Keep dry!

Fragile!

U se no hooks!



B-1

BSpecifications

Tip: The syst em specificat ions out lined in t his a ppendix depend on t heconditions in y our labora tory. Refer to the ACQUI TY U PLC Si t e Preparat ion

Gu ide or conta ct Wa ter s Techn ical S ervice for more informa t ion.

See also:


Evapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de forspecifications.


Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for specificat ions.

• I f your system includes a ma ss spect rometer, see the documenta t ion

included w ith it for specifica tions.

Contents:

Topic Page

B ina ry solvent ma na ger specifica t ions B -2

Sa mple ma na ger specifica t ions B -7

C olumn hea ter specifica t ions B -11

TU V det ect or specifica t ions B -13

Opt iona l sa mple orga n izer specifica t ions B -17

eC ord column ch ip specifica t ions B -19



B-2 Specifications

Binary solvent manager specifications

Binary solvent manager physical specifications

Attribute Specification

H eight 22.9 cm (9 inches)

Depth (w ithout ca bles connected atrear ; includes heads a nd tubes atfront)

66.0 cm (26 inches )

Widt h 34.3 cm (13.5 inches)

Weight 26.3 kg (58 pounds)

Binary solvent manager environmental specifications


Opera t ing t empera t ure 4 to 40 ° C (39.2 to 104 ° F)

Opera ting hum idity <90%, non conden sing

Shipping a nd storage tempera ture −30 t o 60 ° C (-22 to 140 ° F )

Shipping a nd storage humidity <90%, non conden sing

Acoust ic noise (inst rum entgenerated) <65 dB A

Int erna l cooling D C-pow ered fa ns provide in terna lcooling by pulling a ir t hrough t heunit , exha usting i t from t he rea r .

Air flow requirement s Air enters t he front of t he systema llow ing sid e-by-side pla cement of

systems.



Binary solvent manager specifications B-3

Binary solvent manager electrical specifications


P rotection classa

Class I

Overvolta ge ca tegoryb I I

P ollution degreec

2

Moisture protectiond

Normal (IPXO)

L ine volt a ges, nomina l G rounded ACVolt a ge ra nge 100 t o 240 VAC nomina l

Frequency 50 t o 60 H z

Fuse 5 A, 250 V, 5 × 20 mm, slow -blow , IE Ctype

P ow er consumpt ion 360 VA

Contact closure outputs (SW1 toSW3)

Ma ximum volta ge: 30 VDC

Ma ximum current : 0.5 A

Ma ximum VA ra ting: 10 W

Cont a ct resista nce (nomina l): 0.2 ohms

Screw t ermina l connector

Run st opped out put Ma ximum volt a ge: 30 VD C





B ehavior : I f a n error message exists,sw itch is closed an d th en opened wh en

error is clea redS top flow input TTL signa l or con t a ct closure:

Input volta ge range: ±30 VD C

Logic High: ≥3.0 VDC

Logic Low : ≤1.9 VDC

Minimum pulse width : 100 msec




B-4 Specifications

S ta r t gra dient input TTL signa l or cont a ct closure:






Auxilia ry input 1 TTL signa l or cont a ct closure:






Auxilia ry input 2 TTL signa l or cont a ct closure:





Screw t ermina l connectorCha rt output s (1 a nd 2) 0 t o 2 volt s full sca le, screw termina l

(D igita l to a na log convert er ran ge isa ctua lly -0.1 t o 2.1 to allow for offsets)

RS 232 (development a nd m a nufa c-turing use)

9-pin subminiature D connector(socket t ype)

E t hernet RJ 45 connect or

a . Protection Class I – The insula ting scheme used in th e instrum ent to protect from electri-cal shock. Cla ss I ident ifies a s ingle level of insula tion betw een live par ts (wires) a nd exposedconductive part s (meta l pan els), in w hich the exposed conductive part s a re connected to agrounding system. I n t urn, t his grounding syst em is connected to the t hird pin (ground pin)on th e electrical power cord plug.

b. Overvoltage Category II – P erta ins to instr ument s tha t receive their electrical pow er froma local level such as a n electrical w all outlet.

c. Pollution Degree 2 – A mea sure of pollution on electrical circuits, w hich ma y produce areduction of dielectr ic str engt h or surfa ce resistivit y. Degree 2 refers only to norma lly noncon-ductive pollution. Occasionally, however, expect a temporary conductivity caused by conden-sat ion.

d. Moisture Protection – Norma l (IP XO) – IPXO mean s tha t no Ingress P rotect ion aga instan y t ype of dripping or spra yed w at er exists. The X is a pla ceholder th at identifies protectionag ain st dust, i f applica ble.

Binary solvent manager electrical specifications (Continued)




Binary solvent manager specifications B-5

Binary solvent manager performance specifications


G enera l H igh pressure mixing, bina rygradient.

P r iming Able t o pump a nd pr ime bot t les a t thelevel of the pump or high er. Sy ringe(sta rt up kit) is ava ilable for dry

priming.Number of solvent s U p to four (4): A1, A2, B 1, a nd B 2.

U ser mu st choose A1 or A2 a nd B 1 orB 2 before the sta rt of the gra dient.

Solv en t a n d sea l w a sh con t a i ner s Customer select a b le, loca te d in sepa -ra t e bot t le t ray .

S olvent dega ssing B uilt -in solvent dega ssing module, 1chan nel per solvent a nd 2 cha nnels forsa mple ma na ger needle wa shdegassing.

P ump sea l w ash (1 pump) Int egra l, a ct ive, progra mma ble.

Opera t ing flow ra te ra nge 0.010 t o 2.000 mL/min, in 0.001 mLincrements.

E ffect iv e s ys t em dela y volu me L es s t h a n 140 µL , in depen den t ofsystem ba ckpressure (w ith sta nda rdmixer a nd 20 µL loop).

Flow accuracy a ± 1.0% (0.5000 - 2.00 m L/min ),degassed methanol at 41 bar (600 psi).

F low precision 0.075% RS D or ± 0.02 min S D , 6 repli-

cat es ba sed on r etent ion tim e (0.5000 -2.000 mL/min ), (dega ssed a cetonit rile:w a t er, 60:40 dia l-a -mix, 6 replica t es,a lkylphenone test mix, 254 nm ),ACQU ITY UP LC 2.1 × 50 column,35 ° C ± 0.1 ° C .

Composit ion ra nge 0.0 to 100.0% set t a ble in 0.1%

increments.



B-6 Specifications

Composit ion a ccura cy ± 0.5% a bsolut e (full sca le) from 5 t o95% fr om 0.50 t o 2.00 mL /min met h-a nol: meth a nol/propylpar a bens, 254nm.

Composit ion precision 0.15% RS D or ± 0.04 min SD , w hich-ever is great er, based on retent iontime (degassed acetonitrile:water,60:40 dia l-a -mix, 6 r eplica t es, a lky-lphenone test mix, 254 nm ), ACQ U ITYU P LC 2.1 × 50 column, 35 ° C ± 0.1 ° C.

G r a dien t com pos it ion cu rves E leven (11) g ra d ien t cu rves (in clu din glinea r, s t ep [2], conca ve [4], a nd conv ex[4]).

C om pr es sibilit y com pen sa t ion Au tom a t ic a n d con tin uou s.M a xim u m oper a t in g pr es su re 1034 ba r (15000 ps i) u p t o 1 m L /m in ,

621 ba r (9000 psi) up t o 2 mL /min .

P rima ry w et t ed ma ter ia ls 316 S t a in less S teel, U HMWP E,Sa pphire, Ruby, FEP , P TFE , ETFE ,Dia mond-like coa ting, P EE K a ndP EE K a lloys, zirconia ceramic.

a. F or high pressure gradient systems, compositional a ccura cy is driven by flow ra te perfor-ma nce of the individual pumps. HP LC pumps meter solvent a t syst em pressure with th e sol-vent pre-compressed to system pressure before delivery to the column; but most laboratorymeasu rements of flow ra te a re performed a t a tmospheric pressure. Consequently, th epre-compression of th e mobile phase is not a ccounted for in t he mea surement a nd calculat ionof the a pparent flow r a te. The positive bias of the appa rent flow ra te increa ses with sy stembackpressure and is dependa nt upon the specific mobile phase, t he pa rtia l pressure of dis-solved air in t he mobile phase, a nd t he operat ing tempera ture. Wa ters’ flow ra te specificat iondefines a solvent a nd syst em pressure which minimizes the positive bias of the mea surementimposed by t he solvent pre-compression. Wa ters r ecognizes t ha t pure metha nol pumped a t asystem pressure of 600 psi may not reflect t he ty pica l usa ge of solvent delivery syst ems a nd

ha s developed a n alt erna tive mea surement tha t reflects chromat ographic practice. TheWa ters Adva nced Qualificat ion Test (AQT) meth od measu res flow ra te ba sed on the mea sure-ment of retent ion t ime an d not volume. The method uses ura cil wh ich is not r eta ined (onreverse pha se columns) a nd elutes w hen th e void volume ha s been pumped th rough th e col-umn. A plot of the inverse of th e retention time of the ur a cil , at different flow ra tes is l inearan d a ny n on-zero X-intercept implies a systema tic flow ra te bias. This test verifies both thelinea rity a nd a ccura cy of flow ra te a t syst em pressure. This type of test is more suitable formeasurements of UP LC systems, at mult iple flow rat es and w here the f low ra tes measuredexceed 1 mL/min a nd t he sy st em ba ckpressure can exceed 10,000 psi.

Binary solvent manager performance specifications (Continued)




Sample manager specifications B-7

Sample manager specifications

Sample manager physical specifications


H eight 27.3 cm (10.75 inches)

D epth 71.1 cm (28 inches)



Sample manager environmental specifications


Opera t ing t empera t ure 4 to 40 ° C (39.2 to 104 ° F)Opera ting hum idity <90%, non conden sing



Acoust ic noise (inst rum entgenerated)

<65 dB A

Sample manager electrical specifications


P rotection classa

Class I

Overvolta ge ca tegory

b

I IP ollution degree

c2


Normal (IPXO)

L ine volt a ges, nomina l G rounded AC

Volt a ge ra nge 100 t o 240 VAC nomina l

Frequency 47 t o 63 H z

Fuse 10 A



B-8 Specifications


Cont a ct closur e out put s (S W 1 toSW4)

Ma ximum volta ge: 30 VDC





In ject S t a r t Out Ma ximum volt a ge: 30 VD C





Auxilia ry In TTL signa l or cont a ct closure:




Inject H old In TTL signa l or cont a ct closure:


Logic High: ≥3.0 VDCLogic Low : ≤1.9 VDC

0−2V Ana log 2 Out Ma ximum volt a ge: 30 VD C




Screw t ermina l connectorRun S t opped Out Ma ximum volt a ge: 30 VD C





Sample manager electrical specifications (Continued)




Sample manager specifications B-9

Cha rt output s (1 a nd 2) 0 t o 2 volt s full sca le, screw termina l(Digita l to a na log convert er ran ge isa ctua lly -0.1 t o 2.1 to allow for offsets)

a . Protection Class I – The insula ting scheme used in th e instrum ent to protect from electri-cal shock. Cla ss I ident ifies a s ingle level of insula tion betw een live par ts (wires) a nd exposedconductive part s (meta l pan els), in w hich the exposed conductive part s a re connected to agrounding system. I n t urn, t his grounding syst em is connected to the t hird pin (ground pin)on th e electrical power cord plug.

b. Overvoltage Category II – P erta ins to instr ument s tha t receive their electrical pow er froma local level such as a n electrical w all outlet.

c. Pollution Degree 2 – A mea sure of pollution on electrical circuits, w hich ma y produce areduction of dielectr ic str engt h or surfa ce resistivit y. Degree 2 refers only to norma lly noncon-ductive pollution. Occasionally, however, expect a temporary conductivity caused by conden-sat ion.

d. Moisture Protection – Norma l (IP XO) – IPXO mean s tha t no Ingress P rotect ion aga instan y t ype of dripping or spra yed w at er exists. The X is a pla ceholder th at identifies protectionag ain st dust, i f applica ble.

Sample manager performance specifications


In jection cycle time (5 µL pa rt ia lloop with a ir gaps)

~ 45 seconds (or less) w ith a single200 µL w eak wa sh – standa rd mode

~ 60 seconds (or less) w ith default dua l

w a sh (100 µL st rong a nd 500 µL w ea k)S ample delivery precision 0.5% RS D (pa rt ia l loop) 3 to 10 µL in

20 µL loop

2.0% RS D (pa rt ia l loop) 0.5 to 2.9 µL in20 µL loop

0.3% RS D (ful l loop) 2, 5, 20, a nd 50 µLw ith 3× overfi ll

In ject ion linea r it y 0.999 from 2 to 10 µL, pa r t ia l loop, in20 µL loop w ith a ir ga ps

In ject ion volume 0.5 to 50 µL, in 0.1 µL increments

Sample carryover >0.005% (or 1.25 nL) wit h du a l w a sha nd U V detection

Sample manager electrical specifications (Continued)




B-10 Specifications

H ea t er/cooler 4 t o 40 ° C (39.2 t o 104 ° F), ±3 ° C(±5.4 ° F) in 25 ° C (77 °F) ambientconditions

P rogra mma ble in 1 ° C (1.8 °F )increments

N um ber of AN S I sa m ple pla t es Tot a l of t w o (2) pla t es : 96- a n d384-w ell pla tes; via l plat e 2-mL via ls,(48); tube plates 0.65 mL micro-centri-fug e tu be, (48) or 1.5 mLmicro-centrifuge tube, (24)

ANS I-complia nt pla tes E xpa nda ble t o 22 pla t es w ith opt iona lsample organizer

Ma xim um sa mple ca pa cit y 768 in t wo (2) 384-w ell pla tes.

E xpanda ble to up to 8,448 sa mplesw ith optiona l sa mple orga nizer

S a mple probe XYZZ’ ba sed needle-in-needle design

Minim um sa mple req uired 5 µL, using Ma ximum Recovery 2-mLvials

Wa sh solvent s Tw o (2) dega ssed: st rong solvent a nd

sa mple compat ible solvent, progra m-ma ble to suit a pplica tion, ma dea vailable from the bina ry solventmanager

Sample manager performance specifications (Continued)




Column heater specifications B-11

Column heater specifications

Column heater physical specifications






Column heater environmental specifications







<65 dB A



B-12 Specifications

Column heater electrical specifications


P rotection classa

a . Protection Class I – The insula ting scheme used in t he instru ment to protect from elec-trical sh ock. Cla ss I identifies a single level of insula tion between live part s (wires) a ndexposed conductive parts (metal panels), in which the exposed conductive parts are con-

nected to a grounding system. In turn , this grounding system is connected to the th ird pin(ground pin) on th e electr ical power cord plug .

Class I

Overvolta ge ca tegoryb

b. Overvoltage Category I I – P erta ins to inst rument s tha t receive their electrica l powerfrom a local level such a s a n electrical w all outlet.

I I

P ollution degreec

c. Pollution Degree 2 – A mea sure of pollution on electrical circuits, wh ich ma y produce areduction of dielectric strength or surface resistivity. Degree 2 refers only to normallynonconductive pollution. Occasionally, however, expect a temporary conductivity causedby condensat ion.

2


d. Moisture Protection – Norma l (IP XO) – IPXO mean s tha t no Ingr ess Protectionag ainst an y ty pe of dripping or spray ed wa ter exists. The X is a placeholder tha t identifiesprotection a ga inst dust , i f applica ble.

Normal (IP XO)

Line volt a ges, nomina l G rounded ACVolt a ge ra nge 100 to 240 VAC nomina l

Frequency 50 to 60 H z

Ma ximum VA input 750 VA

Column heater performance specifications


H ea t ing ra nge 5 ° C (9 ° F) a bove a mbient t o 65 ° C (149 ° F).

Temperat ure a ccuracy ±1.0 ° C (±1.8 ° F).

Column hea t er Accommoda tes one column, up t o 150 m mlength × 4.6 mm ID , pivots out for use wi thoptiona l ma ss spectr ometer.

Column t r a cking U sa ge informa t ion t ra cked via the eCordcolumn chip.



TUV detector specifications B-13

TUV detector specifications

See also:


Photodi ode Ar r ay Detector Gett i ng Star ted Gu i de for specificat ions.


Evapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de forspecifications.

• I f your system includes a ma ss spect rometer, see the documenta t ionincluded w ith it for specifica tions.

TUV detector physical specifications



D epth 50.8 cm (20.0 inches)


Weight 9.3 kg (20.5 pounds)

TUV detector environmental specifications



Opera ting hum idity <90%, non conden sin g

Shipping a nd storage tempera ture −30 t o 60 ° C (−22 to 140 °F )

Shipping a nd storage humidity <90%, non conden sin g

Acoust ic noise (inst rum entgenerated) <65 dB A



B-14 Specifications

TUV detector electrical specifications


P rotection classa

a . Protection Class I – The insula ting scheme used in th e instrum ent to protect from electri-cal shock. Cla ss I ident ifies a s ingle level of insula tion betw een live pa rt s (w ires) a nd exposedconductive part s (meta l pan els), in w hich the exposed conductive part s a re connected to agrounding system. I n t urn, t his grounding syst em is connected to the t hird pin (ground pin)on th e electrical power cord plug.

Class I

Overvolta ge ca tegoryb

b. Overvoltage Category I I – P erta ins to inst rument s tha t receive their electrical power froma local level such as a n electrical w all outlet.

I I

P ollution degreec

c. Pollution Degree 2 – A mea sure of pollution on electrical circuits, w hich ma y produce areduction of dielectr ic str engt h or surfa ce resistivit y. Degree 2 refers only to norma lly noncon-

ductive pollution. Occasionally, however, expect a temporary conductivity caused by conden-sat ion.

2


d. Moisture Protection – Norma l (IP XO) – IPXO mean s tha t no Ingress P rotect ion aga instan y t ype of dripping or sprayed w at er exists. The X is a placeholder t ha t identifies protectionag ain st dust, i f applica ble.

Normal (IPXO)

L i ne vol t a ges , n omin a l G r oun ded ACVolt a ge ra nge 100 t o 240 VAC nomina l

Line freq uency 50 t o 60 H z

Fuse Tw o fuses, 100 to 240 VAC, 50 t o 60 H zF 3.15-A, 250-V FAS T B LO, 5 × 20 mm (IE C )


Tw o a tt enuat ed a na logoutput cha nnels:2 VFS /0 mVFS

Two event outputs

Four event input s

At t enua t ion ra nge: 0.0001 t o 4.000 AU2 V output r a nge: −0.1 to +2.1 V10 mV out put r a nge: −0.5 to 10.5 mV

Type: conta ct closureVolta ge: + 30VCurrent: 1 A

Input volta ge: + 30 V ma ximum; 100 ms

minimum period



TUV detector specifications B-15

TUV detector operational specificationsa

a. All specifica tions derive from inst rument performa nce aft er a 1-hour w a rm-up time.


Wa velength ra nge 190 to 700 nm

B a ndw idt h 5 nm

Wa velengt h a ccura cy ±1 nm

Wavelength

repeatabil i ty

±0.25 nm

B a seline noise, singlewavelength

b

b. ASTM S ta nda rd E1657-94.

6 × 10-6 AU , shu nt in pla ce, 230 nm, 1.0 secondfilter t ime constant , 2 Hz

B a seline noise, dua lwavelength b

< ±5.0 × 10-5

AU , shun t in pla ce, 230 nm a nd 280nm , 2.0 second filter time const a nt , 1 Hz

Linearityb

≤5% a t 2.5 AU , propyl pa ra ben, 257 nm

D rift 5.0 × 10-4 AU /hour, sh un t in place, a t 230 nm (a fter1 hour w a rm -up) ∆T= ±2 ° C/hour (±3.6 ° F/hou r)

M a xim u m d a t a r a t e 80 H z (poin t s/1.0 s econ d pea k )

Sensitivity settingrange

0.0001 to 4.0000 AUFS

F ilt er set t ing ra nge S in gle w a velengt h:

0.1 to 5.0 seconds, Hamming (default)

1 to 99 second s, RC

Dua l wa velength :

1 to 50 seconds, H a mming (default )

1 to 99 second s, RC

Optical Component Specifications

La mp source D eut erium a rc la mpLa mp w a rra nty 2000 hours

Flow cell 10 m ligh t -guiding flow cell (Ana ly t ica l)

Wet t ed ma t er ia ls 316 st a in less st eel, fused silica , P E E K , P TF EAF2400



B-16 Specifications

TUV detector optical specifications


Monochroma t or Fa st ie E bert configura t ion

G ra t ing P la ne hologra phic 1800 grooves/mm

Opt ica l ba ndw idt h 5 nm

La mp pow er 30 W

TUV detector flow cell specifications

TypeFlowRange

(mL/min)

Volume(nL)

PathLength

(mm)

Sample TubingInternal Diameter(µm)

PressureRating

(bar/psi)In Out

Ana lyt ica l 0 to 2 500 10 125 125 69/1000

High-sensitivity

0 to 2 2400 25 125 125 69/1000



Optional sample organizer specifications B-17

Optional sample organizer specifications

Sample organizer physical specifications




Widt h 25.4 cm (10 inches), 61.6 cm (24.25 inches) w it h sa mplemanager

Weigh t 58.1 kg (128 pounds), w it hout int erna l or ext erna l shelves

Sample organizer environmental specifications







<65 dB A

Sample organizer electrical specifications


P rotection class a Class I

Overvolta ge ca tegoryb I I

P ollution degreec 2

Moisture protection d Normal (IPXO)

L ine volt a ges, nomina l G rounded AC

Volt a ge ra nge 100 t o 240 VAC nomina l

Frequency 47 t o 63 H zFuse 10 A



B-18 Specifications

Ma x VA input 540 VAa . Protection Class I – The insula ting scheme used in t he inst rum ent t o protect from elec-

trical sh ock. Cla ss I identifies a single level of insula tion between live part s (wires) a ndexposed conductive parts (metal panels), in which the exposed conductive parts are con-nected to a grounding system. In t urn, th is grounding system is connected to th e third pin(ground pin) on th e electr ical power cord plug .

b. Overvoltage Category I I – P erta ins to inst rument s tha t receive their electrica l powerfrom a local level such a s a n electrical w all outlet.

c. Pollution Degree 2 – A mea sure of pollution on electrical circuits, w hich ma y produce areduction of dielectric strength or surface resistivity. Degree 2 refers only to normally

nonconductive pollution. Occasionally, however, expect a temporary conductivity causedby condensat ion.d. Moisture Protection – Norma l (IP XO) – IPXO mean s tha t no Ingr ess Protection

ag ainst an y ty pe of dripping or sprayed w at er exists. The X is a pla ceholder th at identi-fies protection a ga inst dust, i f a pplicable.

Sample organizer performance specifications


H ea t er /cooler 4 t o 40 ° C (39.2 to 104 ° F), ±3 ° C(±5.4 ° F) in 20 ° C (68 °F) ambientconditions.

P rogram ma ble in 1 ° C (1.8 ° F)increments.

Number of ANSI complian t p la t es Tota l o f up to:21 s tandard micro-tit er plat es, or 11 interm ediat eheight pla tes, or 7 deep-w ell (or2-mL vial) plat es, an d combina -tions t hereof.

Ma xim um sa mple ca pa cit y Tot a l of up t o 8,064 sa mples: intwenty one (21) 384-well plates.

Sample organizer electrical specifications (Continued)




eCord column chip specifications B-19

eCord column chip specifications

eCord™

column chip information

Parameter Generated By Maximum Bytes

P a r t number Wa t ers 10

Ma nufa ctur ing ID # Wa t ers 64

Chemist ry Wa t ers 64

P a r t icle S ize Wa t ers 4Column D imension Wa t ers 8

D a t e of Ma nufa ct urer Wa t ers 8

E fficiency Wa t ers 4

B a ckpressure (psi) Wa t ers 4

Ret en t ion fa ct or Wa t ers 4

U SP Ta iling Fa ctor Wa t ers 4D a t e Test ed Wa t ers 8

Test C ondit ions Wa t ers 100

Recomm ended Opera tingGuidelines

Wa t ers P D F

S tora ge S olvent Wa t ers 32

B ulk B a t ch Wa t ers 8

pH Opera t ing Ra nge Wa ters 8

S t a r t D a t e a nd Time U ser 8

Finish D a te a nd Time U ser 8

P roject Na me U ser 64

S a mple Set Na me U ser 64U ser Na me U ser 64

S yst em # U ser 64

# Inject ions U ser 2

# S a mples U ser 2

H ighest B a ckpressure U ser 8

Avera ge Inject ion Volume U ser 4



B-20 Specifications

Avera ge F low Ra t e U ser 4Avera ge Column Tempera t ure U ser 4

S tora ge S olvent U ser 32

S uit a bilit y Met? U ser 4

eCord™ column chip information (Continued)

Parameter Generated By Maximum Bytes



C-1

C Spare Parts

This a ppendix lists recommended spar e part s a nd options for t he ACQU ITYU P LC™ system. Un listed pa rts a re not recommended for customerreplacement.

See also:


Photodi ode Ar r ay Detector Gett i ng Star ted Gu id e for a l ist of spa re pa rts.

• I f your system in cludes a n E L S dete ctor , see th e ACQUI TY UPLCEvapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de for a list ofspare par t s .

• I f your system includes a ma ss spect rometer, see the documenta t ionincluded w ith i t for a l ist of spare part s.

Contents:

Topic Page

ACQU ITY U P LC syst em pa r t numbers C -2

B ina ry solvent ma na ger spa re pa r t s C-2

Sa mple ma na ger spa re pa rt s C-4

TU V det ect or spa re pa r t s C-5

Opt iona l sa mple orga nizer spa re pa rt s C-6



C-2 Spare Parts

ACQUITY UPLC system part numbers

Binary solvent manager spare parts

Part numbers for the ACQUITY UPLC system

Item Part Number

Syst em performa nce test for ACQU ITY U P LCsyst em wit h TU V or P DA detector

715001134

Syst em performa nce test for ACQU ITY U P LCsystem w ith Quat tro Micro Premier

715001119

Recommended spare parts for the binary solvent manager

Item Part Number

ACQU ITY dua l ba ll a nd sea t 700002968ACQU ITY prima ry check va lve (pkg 2) 700002596

ACQU ITY UP LC C olumn Mixer 4.6 mm × 50 mm 205000403

Air filt er 700002632

Air filt er , ha ndle 700002633

Compression screw , gold, long (10) 700002645

Compression screw , gold, shor t (10) 700002634

D ua l ba ll a nd sea l 700002968

Ferrule set , t w o-piece sw a gelock 700002635

Fus e, 5 A, 250 V, 5 × 20 mm, slo blo 700002604

H a stelloy bot t le filt ers (1) 700002850

H a stelloy bot t le filt ers (7) 700002824H ea d sea l a n d sea l w a sh spa cer , UP 30, .0787 ID (2) 700002599

Mixer , st a nda rd, 100 µL 205000272

Mixer , zirconia , 50 µL 700002911

P rima ry inlet check va lve filt er kit 700002869

P r im a ry in let ch eck va lv e filt er repla cem en ts (5) 700002870

P lunger (2) 700002600



Binary solvent manager spare parts C-3

P TF E O-ring WAT076152S olvent filt er WAT025531

Tube, “A” t ra nsducer t o vent va lve 430001200

Tube, “B ” t ra nsducer t o vent va lve 430001199

Tu be, dega s ser por t A1 t o s olven t s elect va lve A 430001115

Tu be, dega s ser por t A2 t o s olven t s elect va lve A 430001116

Tube, dega sser por t B 1 t o solven t select va lve B 430001114

Tube, dega sser por t B 2 t o solven t select va lve B 430001113

Tube, solvent inlet S D S 430001020

Tube, S S V to check va lve 430001172

Tube, system out let S D S 430001486

Tube, t ra nsducer in let 430001151Tube, t ra nsducer t o check va lve 430001121

Tube, vent va lve por t 1 t o w a ste 430001209

Tube, vent va lve por t 2 t o t ee/filt er 430001208

Tube, vent va lve por t 4 t o w a ste 430001210

Tube, vent va lve por t 5 t o t ee/filt er 430001207

Wa sh sea l (2) 700002598

Recommended spare parts for the binary solvent manager (Continued)

Item Part Number



C-4 Spare Parts

Sample manager spare parts

Recommended spare parts for the sample manager

Item Part Number

Fuse, 0.05 A, 250 V, SMD 4000 A INTE RR 242.05 700002575



Fu se, 10 A, 5 × 20 mm, slo blo (5) 700002577

Needle, FE P /met a l, 30 µL 205000369

Needle, P E E K, 15 µL 700002708

Needle, P E E K, 30 µL 700002644

Needle, st eel, 30 µL 205000362

P uncture needle car t r idge kit (for new Y-ca rr iage) 700002773

S a mple loop, 10 µL 430001326S a mple loop, 2 µL 430001264

S a mple loop, 20 µL 430001320



S yr inge, sa mple, 100 µL, H P 700002570

S yr inge, sa mple, 250 µL, H P 410001347

S yr inge, w a sh, 2.5 mL 700002569



TUV detector spare parts C-5

TUV detector spare parts

See also:


Photodi ode Ar r ay Detector Gett i ng Star ted Gu id e for a l ist of spa re pa rts.


Evapor ati ve L igh t Scatt er i ng Detector Gett i ng Star ted Gui de for a list ofspare par t s .

• I f your system includes a ma ss spect rometer, see the documenta t ion

included w ith i t for a l ist of spare part s.

Recommended spare parts for the TUV detector

Item Part Number

B a ckpressure regula t or 700002571

Flow cell, Ana ly t ica l 205015008

Flow cell, H igh S ensit ivit y 205015009

Fus e, 3.15 A, 250 V, 5 × 20 m m, fa st -a ct ing (5) WAT055634

Inlet t ube assembly (Ana lytica l a nd H ighSensitivity)

430001225

P er for ma n ce m a in ten a nce kit (in clu des la m p) 201000186



C-6 Spare Parts

Optional sample organizer spare parts

Recommended spare parts for the sample organizer

Item Part Number

Fu se, 10 A, 5 × 20 mm, slo blo (5) 700002577

S helf 700002730



D-1

D Solvent Considerations

The informa tion in t his a ppendix a pplies only t o the follow ing inst rum ents:

• AC QU I TY U P L C sy st em modules

• AC QU I TY P D A det ect or

• AC QU I TY TU V det ect or

• AC QU I TY U P L C E LS det ect or

Contents:

Topic Page

In t roduct ion D -2S olvent recommenda t ions D -4

Solvent miscibilit y D -9

Solvent st a bilizers D -10

Solvent viscosit y D -11

Wa velength select ion D -11

Warning: To a void chemica l ha za rds, a lwa ys observe safe laborat orypra ctices w hen opera ting y our syst em.



D-2 Solvent Considerations

Introduction

Preventing contamination

For informa tion on prevent ing conta mina tion, refer to Contro l l ing

Con tamina t ion in L C/ M S Systems (pa rt num ber 715001307) on th eACQU ITY UP LC Sy stem B ookshelf CD, or visit h tt p://w w w .wa ters.com a ndclick S upport Cent er.

Clean solventsClea n solvent s ensure reproducible results a nd permit y ou to opera te w ithminimal instrum ent maint enance.

Dirt y solvent s ca n cause baseline noise a nd drift, a nd t hey can clog solventreservoir filters, inlet filters, a nd ca pillar y lines.

Solvent quality

U se MS-gra de solvents for the best possible results; th e minimumrequirement is HP LC-gra de. Filter solvents t hrough a n a ppropria temembrane filter.

Tip: Alwa ys be sure your solvent choices ar e consist ent w ith th erecommendations of the filter manufacturer.

Solvent preparation

P roper solvent prepara tion, prima rily f il tra tion, ca n prevent ma ny pumpingproblems.

Tip: Alw ay s use brow n-tinted gla sswa re to inhibit microbial growth .



Introduction D-3

Water

U se wa ter only from a high-qua lity w a ter purif ica tion system. If the wa tersyst em does not deliver filtered wa ter, filter th e wa ter t hrough a 0.2-µmmembrane filter.

Using buffersAdjust t he pH of aq ueous buffers. Filter th em to remove insoluble ma teria l,a nd th en blend them w ith a ppropriat e orga nic modifiers. After you use abuffer, flush it from t he pump by run ning a w et-prime with a t leas t fivesyst em volumes of HP LC -gra de distilled or deionized wa ter.

For shut downs of more tha n a da y, flush the pump wit h a 20%meth a nol/w a ter solut ion t o prevent microbial gr owt h.

Tip: To a void sa lt precipita tion, nonvola tile buffer concent ra tions sh ould n otexceed 100 mM.

Buffered solvents

When using a buffer, choose good qua lity rea gents, filtering th em th rough a0.2-µm filt er.

Tip: To discoura ge microbia l grow t h, r eplace 100% mobile aq ueous pha sedaily.

See also: For informa tion on prevent ing cont a mina tion, refer to Contro l l ing

Con tamina t ion in L C/ M S Systems (pa rt num ber 715001307) on th eACQU ITY UP LC Sy stem B ookshelf CD, or visit h tt p://w w w .wa ters.com a ndclick S upport Cent er.

Caution: U sing 100% w a ter ca n cau se microbia l growt h. Wa tersrecommends cha nging 100% w a ter solut ions da ily. Adding a sma lla mount of an organ ic solvent (~ 10%) prevent s m icrobia l growt h.

Caution: Some buffers can be incompa tible with ma ss spectr ometers.Consult th e documenta tion tha t accompanies your instrument forcompatible buffers.




Solvent recommendations

General solvent guidelinesAlwa ys observe the follow ing genera l solvent guidelines:

• Use h igh-qua l ity , brown-t in ted g lasswa re to inh ib it microb ia l g rowth .

• F i lte r solvents . Sma l l par t icles can permanent ly b lock a sys tem’sca pillar y lines. F iltering solvent s a lso improves check va lveperformance.

Solvents to use

The ACQU ITY U P LC syst em w a s designed for reversed-pha sechromat ogra phy a nd ACQU ITY UP LC B EH column chemistr ies. Wa ters® evalua ted th e system’s reliability using t ra ditional reversed-pha se solvent s.

This section lists solvents recommended for t he ACQU ITY U P LC syst em.Cont a ct Wa ters C ustomer Service to determine whet her you ca n use solvent stha t do not a ppear in t he l ist w ithout a dversely a ffecting instr ument orsyst em performa nce.

See also: “Conta cting Wa ters t echnical service” on pa ge 6-5.

Recommended solvents

• Acet onit r ile

• Acet on it r ile/w a t e r mixt u res

• I sopropa nol

• Met ha nol

• Met ha nol/w a t er m ixt ur es

• Wa ter



Solvent recommendations D-5

Other solvents

You can use the follow ing solvents. Note, however, tha t t hese solvents canshorten instrument life.

• Tet ra h ydr ofur a n (TH F )

• H exa ne

See also: “ACQU ITY U P LC system recommendat ions” on pa ge D -7 fora dditiona l informa tion.

Consider solvent polar ity w hen you chan ge typica l reversed pha se solvents.

Flush t he system w ith a solvent of intermedia te polar ity like isopropa nolbefore int roducing nonpolar s olvent s like TH F or hexa ne.

Additives/Modifiers

• 0.1% ethylene diaminete t r aacet ic ac id (EDTA)

• 0.1% h exa fluor ob uter ic a cid

• 0.1% tr iet hy l a m in e (TE A)• 0.1% tr i fluoracet ic ac id (TFA)

• 0.2%formic a cid

• 10 m M a m mon ium bica rbon a te

• 10 mM ph ospha t e buffer

• 50 mM a mmonium a cet a t e• 50 m M a m mon ium h ydroxide




Sample diluents

• Acet onit r ile

• Acet on it r ile/w a t e r mixt u res• D im et hy lfor ma m ide (D MF )

• D im et h yl su lfoxid e (D MS O)

• Isopropa nol

• Met ha nol

• Met ha nol/w a t er m ixt ur es

• Wa ter

Recommendation: Do not use buffers a s needle wa sh.

Cleaners

Recommendation: S ee the clea ning procedures in Contr ol l in g Contam inat i on

in L C/ M S Systems (par t n umber 715001307) on the ACQU ITY UP LC Sy stemB ookshelf CD , or visit ht tp://w w w .wa t ers.com a nd click S upport Cent er.

• P hosphoric a cid (≤30%)

• Sodium hydroxide (≤1Μ)

Solvents to avoid

You sh ould a void t hese solvents:

• Solvents tha t con ta in ha logens : fluor ine, bromine, or iod ine.

• Strong acids . (Use them only in weak concentra t ion, <5%, unless ascleaning a gents. Avoid using a cids as m obile pha ses wh en their pH<1.0.)

• P eroxidizable compounds such as UV-grade ethers , non-sta bil ized

THF , dioxa ne, a nd diisopropylether. (If you m ust use peroxidizablecompounds, be sure t o filter th em th rough dry a luminium oxide toa dsorb formed peroxides.)

• Solut ions tha t con ta in s t rong concentra t ions of complexing agentslike ethylene diaminetetraacetic acid (EDTA).



Solvent recommendations D-7

ACQUITY UPLC system recommendations

Cont a ct Wa ters for recommended system cleaning a nd flushing procedures.See “Cont a cting Wa ters technica l service” on pa ge 6-5.

See also: Contr ol l ing Contami nat ion in L C/ M S Systems (par tnum ber 715001307) on the ACQU ITY U P LC Sy stem B ookshelf CD, or visitht t p‘://w w w .w a ter s.com a nd click Su pport C ent er.

• THF a nd hexane can be used as the mobile-phase in ACQUITY UP LCsystems. However, as w ith ma ny nona queous solvents, t hey ca n shortensystem a nd instrum ent l i fe compar ed with equipment run ning ty pica lreversed-phase solvents.

• When us ing unstab i li zed THF, ensure tha t your solvent i s fresh lyprepa red. P reviously opened bottles cont a in peroxide conta mina nt s,w hich cause ba seline drift.

• Chloroform, methylene chlor ide, ethyl aceta te , and toluene are genera l lynot recommended for use in ACQU ITY UP LC syst ems. Nevert heless,you can use t hese solvent s in w ea k dilut ions (<10%) a s a ddit ives, sa mplediluents, or modifiers.

• Contac t your Wat ers sa les representa t ive or loca l technica l suppor torgan iza tion to determine wh ether a specific meth od is suita ble to use

w ith the ACQU ITY UP LC syst em instrum ents an d components.• When us ing THF or hexane, ins ta l l st a in less steel tubing, and min imize

th e use of P EE K components.

• Aqueous solvents must not remain in a shut-down sys tem because theyserve as a substr a te for microbial colonies. Microbes can clog sy stemfilters a nd capillary lines. To prevent th eir proliferat ion, a dd a sma lla mount (~ 10%) of an organ ic solvent such a s a cetonitr ile or m etha nol.

Warning: Explosion ha zar d: P eroxide contam inant s in THF ca nspont a neously a nd destru ctively explode when you part ially orcompletely eva pora te t he TH F.

Warning: Hea lth ha zar d: Hexane is a neurotoxin, and THF ca nirri tat e eyes, skin an d mucous membra nes and ca use harm fulneurologic effects . If you use either or both of th ese vola t ile solvent s,loca te your ACQU ITY UP LC syst em inside a fume hood or wa lk-incha mber to minimize exposure to ha rmful solvent va pors.




Binary solvent manager recommendations

• Th e sea l w a sh system must n ev er r un d ry , pa r t i cu la r l y durin gsepa ra tions t ha t use a polar mobile pha se.

• Isopropyl a lcohol or mixtures of metha nol and w at er , l ike 20%meth a nol/w a ter, a re effective seal w a sh solvents for TH F solventmixtures.

• For reversed-phase appl ica t ions , use aqueous sea l wa sh solu t ions wi th aw ea k orga nic component (for exam ple 1:9 met ha nol/w a t er).

• Do not use 100% organ ic sea l wash solut ions .

Sample manager recommendations

• Insta l l the a l l-sta inless-steel needle (par t number 205000362) on t hesa mple ma na ger when using TH F or hexa ne.

• Do not use concentra t ions of THF or hexane grea ter than 10% asw eak-w a sh solvent.

• Typica l organic sample diluents such as d imethylsulfoxide (DMSO) anddimethy lforma mide (DMF ) a re support ed.

Detector recommendations

Observe th e follow ing guidelines wh en using solvents w ith ACQU ITY UP LCsystem detector flow cells:

• A f low cel l t r anspor ted whi le tempera tures a re below 5 ° C must be fi lledw ith a lcohol.

S l t i ibilit



Solvent miscibility D-9

Solvent miscibility

B efore you chan ge solvents, refer to th e ta ble below to determine solvent

miscibility . B e aw a re of th ese effects:

• Changes involving two miscible solvents can be made di rect ly . Changesinvolving t w o solvents t ha t a re not tota lly miscible (for exam ple, fromchloroform t o wa ter) require a n int ermediat e solvent like n -propanol.

• Tempera ture a f fects solvent miscibi li ty . I f you a re running ahigh-tempera tu re a pplica tion, consider t he effect of the h igher

tempera tu re on solvent solubility .• B uffers dissolved in wa ter can precipi ta te when mixed wi th organ ic

solvents.

When you sw itch from a str ong buffer t o an orga nic solvent , thoroughly flushth e system w ith dist illed wa ter before you a dd the orga nic solvent.

Solvent miscibility

PolarityIndex

Solvent

Viscosity

cP, 20 °C(@1 atm)

Boiling

Point °C(@1 atm)

Miscibility

Number(M)

λ Cutoff(nm)

0.0 N-hexa ne 0.313 68.7 29 ––

1.8 Triet hyla mine 0.38 89.5 26 ––

4.2 Tet ra hydrofura n (TH F ) 0.55 66.0 17 220

4.3 1-propa nol 2.30 97.2 15 210

4.3 2-propa nol 2.35 117.7 15 ––

5.2 E t ha nol 1.20 78.3 14 210

5.4 Acetone 0.32 56.3 15, 17 330

5.5 B enzyl a lcohol 5.80 205.5 13 ––

5.7 Met hoxyetha nol 1.72 124.6 13 –– 6.2 Acetonit r ile 0.37 81.6 11, 17 190

6.2 Acet ic a cid 1.26 117.9 14 ––

6.4 D imet hylforma mide 0.90 153.0 12 ––

6.5 D imet hylsulfoxide 2.24 189.0 9 ––

6.6 Met ha nol 0.60 64.7 12 210

9.0 Wa ter 1.00 100.0 –– ––

U i i ibilit b (M b )




Using miscibility numbers (M-numbers)

Use miscibility numbers (M-numbers) to predict the miscibility of a liquidwith a s ta ndard solvent .

To predict th e miscibility of tw o liquids, subt ra ct t he sma ller M-num ber va luefrom the la rger M-num ber value.

• I f the dif ference betw een the two M-numbers is 15 or less , the twoliquids a re miscible in a ll proportions a t 15 °C .

• A di fference of 16 indica t es a cr it ica l solut ion temperature f rom 25 to75 ° C , w i th 50 ° C a s the opt ima l tempera ture .

• I f the di fference is 17 or great er , the liquids are immiscible or theircrit ica l solution tempera tu re is above 75 ° C.

Some solvents prove immiscible with solvents a t both ends of the lipophilicityscale. These solvents r eceive a du a l M-num ber:

• The f ir s t number , a lwa ys lower than 16, ind ica t es the degree ofmiscibility w ith highly lipophilic solvent s.

• The second number a ppl ies to the opposite end of the sca le . A largedifference betw een t hese tw o numbers indica tes a limited ra nge ofmiscibility.

For exa mple, some fluoroca rbons ar e immiscible with a ll the sta nda rdsolvents a nd h a ve M-numbers of 0 an d 32. Tw o liquids w ith dua l M-num bersa re usua lly miscible w ith ea ch oth er.

A liquid is clas sified in th e M-num ber syst em by t esting for miscibility w ith asequence of sta nda rd solvent s. A correction t erm of 15 units is t hen eithera dded or su btra cted from t he cutoff point for m iscibility .

Solvent stabilizers

Do not leave solvents cont a ining sta bilizers, like THF w ith but yla tedhydr oxyt oluene (B H T), to dry in t he syst em’s flow pat h. A dry flow pat h,including t he detector flow cell, becomes cont a mina ted w ith residua lsta bil izer , and a substa ntia l clea ning effort is needed to restore the f low pat hto its in itia l condition.

Solvent viscosity



Solvent viscosity D-11

Solvent viscosity

G enerally, viscosity is not a considera tion wh en you opera te w ith a single

solvent or under low pressure. H ow ever, wit h gra dient chroma togra phy, theviscosity chan ges tha t occur as th e solvent s a re mixed in different proport ionsca n effect pressure cha nges during t he run. F or exam ple, a 1:1w a ter/meth a nol mixture produces tw ice the pressure of either w a ter ormetha nol alone.

If you do not know t he extent to wh ich pressure cha nges affect t he an a lysis,monitor the pressure during the run.

Wavelength selection

The t a bles in t his section provide UV cutoff values for th ese items:

• C ommon solvent s

• C om mon mixed m obile ph a ses• C hromophores

UV cutoffs for common solvents

The ta ble below shows t he U V cut off (th e wa velengt h a t w hich t he a bsorba nceof the solvent equals 1 AU) for some common chromatographic solvents.

Opera ting a t a w a velength n ear or below th e cut off increases bas eline noisedue to solvent a bsorbance.

UV cutoff wavelengths for common chromatographic solvents

SolventUVCutoff(nm)

Acetone 330

Acetonit r ile 190

D iet hyl a mine 275

E t ha nol 210

Isopropa nol 205

Isopropyl et her 220

UV cutoff wavelengths for common chromatographic solvents (Continued)




Mixed mobile phasesThe follow ing t a ble provides a pproxima te w a velength cutoffs for some oth ersolvents, buffers, det ergents, a nd mobile pha ses. The solvent concent ra tionsrepresented a re those most commonly used. If you w a nt to use a differentconcentra tion, you ca n determine a pproxima te a bsorba nce using B eer’s law ,beca use a bsorbance is proportiona l to concent ra tion.

Met ha nol 205

n -P ropa nol 210

Tetrahydrofuran (THF) 230

Wavelength cutoffs for different mobile phases

Mobile PhaseUVCutoff(nm)

Mobile PhaseUVCutoff(nm)

Acet ic a cid, 1% 230 S odium chlor ide, 1 M 207

Ammonium a cet a t e, 10 mM 205 S odium cit ra t e, 10 mM 225

Ammonium bica rbona te, 10 mM 190 Sodium dodecyl sulfa t e 190Polyoxyethylene (35) lauryl ether(B RI J 35), 0.1%

190 Sodium forma t e, 10 m M 200

3-[(3-cholamidopropyl)-dimethy-lammonio]-1-propanesulfonate)(C H AP S ) 0.1%

215 Triethyl a mine, 1% 235

D ia mmonium phospha te, 50 mM 205 Trifluora cet ic a cid, 0.1% 190(E th ylenedinitrilo) tet ra a cetica cid disodium sa lt (disodiumE DTA), 1 mM

190 TRI S H Cl, 20 m M, pH 7.0,pH 8.0

202,212

4-(2-hydroxyethyl)-1-pipera-zineeth a nesulfonic a cid (H EP E S),10 mM, pH 7.6

225 Triton-X™

100, 0.1% 240

UV cutoff wavelengths for common chromatographic solvents (Continued)

SolventUVCutoff

(nm)

Wavelength cutoffs for different mobile phases (Continued)



Wavelength selection D-13

Mobile phase absorbance

This section lists th e absorba nces a t severa l wa velengt hs for frequent ly usedmobile phases. Choose the mobile phase carefully to reduce baseline noise.

The best m obile pha se for your a pplica tion is one tha t is tr a nspa rent a t t hechosen detection w a velength s. With such a mobile pha se, ensure th a t a nya bsorba nce is due only to t he sa mple. Absorbance by t he mobile phase a lsoreduces the linea r dyna mic ra nge of the detector by th e amount of absorba nceth e au tozero function ca ncels, or “a ut ozeroes,” out. Wa velengt h, pH , a nd

concentr a tion of the mobile phase a ffects its a bsorba nce. Exa mples of severa lmobile pha ses a re given in t he ta ble below.

Tip: The a bsorba nces in t he ta ble below a re based on a 10-mm pat hlength .

H ydrochloric a cid, 0.1% 190 Wa t ers P IC®

Reag ent A,1 via l/liter

200

Morpholinoethanesulfonic acid(MES ), 10 mM, pH 6.0

215 Wa ters P I C Rea gent B -6,1 via l/liter

225

P ota ssium phospha te,

monobasic, 10 mMdiba sic, 10 mM

190190

Wa ters P IC Reagent B -6,

low U V, 1 via l/liter

190

Sodium a cet a te, 10 m M 205 Wa t ers P IC Rea gent D -4,1 via l/liter

190

Wavelength cutoffs for different mobile phases (Continued)

Mobile PhaseUVCutoff

(nm)

Mobile PhaseUVCutoff

(nm)




Mobile phase absorbance measured against air or water

Absorbance at Specified Wavelength (nm)

200 205 210 215 220 230 240 250 260 280

Solvents

Acetonit r ile 0.05 0.03 0.02 0.01 0.01 <0.01 — — — —

Methanol(notdegassed)

2.06 1.00 0.53 0.37 0.24 0.11 0.05 0.02 <0.01 —

Methanol(degassed)

1.91 0.76 0.35 0.21 0.15 0.06 0.02 <0.01 — —

Isopropa nol 1.80 0.68 0.34 0.24 0.19 0.08 0.04 0.03 0.02 0.02

Unstablizedtetrahydro-

fura n (TH F,fresh)

2.44 2.57 2.31 1.80 1.54 0.94 0.42 0.21 0.09 0.05

Unstablizedtetrahydro-fura n (TH F,old)

>2.5 >2.5 >2.5 >2.5 >2.5 >2.5 >2.5 >2.5 2.5 1.45

Acids and Bases

Acetic acid,1%

2.61 2.63 2.61 2.43 2.17 0.87 0.14 0.01 <0.01 —

Hydro-chloric a cid,0.1%

0.11 0.02 <0.01 — — — — — — —

Phosphoric

a cid, 0.1%

<0.01 — — — — — — — — —

Trifluoro-a cetic a cid

1.20 0.78 0.54 0.34 0.22 0.06 <0.02 <0.01 — —

Diammo-niumphosphate,

50 m M

1.85 0.67 0.15 0.02 <0.01 — — — — —

Mobile phase absorbance measured against air or water (Continued)




Triethy-la mine, 1%

2.33 2.42 2.50 2.45 2.37 1.96 0.50 0.12 0.04 <0.01

Buffers and Salts

Ammoniumacetate,

10 m M

1.88 0.94 0.53 0.29 0.15 0.02 <0.01 — — —

Ammoniumbicarbonate,10 m M

0.41 0.10 0.01 <0.01 — — — — — —

Ethylene-dinitrilo)tetraacetic

acid diso-dium sa l t(disodiumEDTA),1 mM

0.11 0.07 0.06 0.04 0.03 0.03 0.02 0.02 0.02 0.02

4-(2-hydrox-yethyl)-1-pi

pera-zineethanesulfonic acid(HEPES),10 mM,pH 7.6

2.45 2.50 2.37 2.08 1.50 0.29 0.03 <0.01 — —

Morpholino-

ethanesulfonic acid(MES),10 mM,pH 6.0

2.42 2.38 1.89 0.90 0.45 0.06 <0.01 — — —


200 205 210 215 220 230 240 250 260 280





Potassiumphosphate,monobasic(KH 2P O4),10 m M

0.03 <0.01 — — — — — — — —

Potassium

phosphate,dibasic,(K 2H P O4),10 m M

0.53 0.16 0.05 0.01 <0.01 — — — — —

Sodiumacetate,10 m M

1.85 0.96 0.52 0.30 0.15 0.03 <0.01 — — —

Sodiumchloride,1 M

2.00 1.67 0.40 0.10 <0.01 — — — — —

Sodiumcitrate,10 m M

2.48 2.84 2.31 2.02 1.49 0.54 0.12 0.03 0.02 0.01

Sodiumformate,10 m M

1.00 0.73 0.53 0.33 0.20 0.03 <0.01 — — —

Sodiumphosphate,100 mM, pH6.8

1.99 0.75 0.19 0.06 0.02 0.01 0.01 0.01 0.01 <0.01

Tris H Cl,20 mM, pH7.0

1.40 0.77 0.28 0.10 0.04 <0.01 — — — —

Tris H Cl,20 mM, pH8.0

1.80 1.90 1.11 0.43 0.13 <0.01 — — — —


200 205 210 215 220 230 240 250 260 280





Waters PIC ®

Reagents

P IC A,1 via l/L

0.67 0.29 0.13 0.05 0.03 0.02 0.02 0.02 0.02 <0.01

P IC B 6, 1

via l/L

2.46 2.50 2.42 2.25 1.83 0.63 0.07 <0.01 — —

P IC B 6, low

UV,1 v ia l/L

0.01 <0.01 — — — — — — — —

P IC D 4, 1via l/L

0.03 0.03 0.03 0.03 0.02 0.02 0.02 0.02 0.02 0.01

Detergents

B RI J 35, 1% 0.06 0.03 0.02 0.02 0.02 0.01 <0.01 — — —

3-[(3-chola -midopropyl)-dimethy-lammonio]-1-propane-

sulfonate)(CHAPS),0.1%

2.40 2.32 1.48 0.80 0.40 0.08 0.04 0.02 0.02 0.01

Sodiumdodecylsulfate(S D S), 0.1%

0.02 0.01 <0.01 — — — — — — —

4-octylphen-ol poly-ethoxylate(Triton

®

X-100), 0.1%

2.48 2.50 2.43 2.42 2.37 2.37 0.50 0.25 0.67 1.42


200 205 210 215 220 230 240 250 260 280





Polyoxyeth-ylenesorbitanmonolau-ra te(Tween™

20), 0.1%

0.21 0.14 0.11 0.10 0.09 0.06 0.05 0.04 0.04 0.03


200 205 210 215 220 230 240 250 260 280

Index



Index-1

Aa bsorbance values, display of 3-36, 4-9acids D-14ACQU ITY syst em

sta r t ing up 3-12ACQU ITY UP LC C olumns Ca lculat or

1-4

ACQU ITY U P LC Consolemonitoring system sta tus from 7-8sta r t ing

from E mpower softw a re 4-11from MassLyn x softw a re 4-13

ACQU ITY UP LC E LS detectorcontrol panel, using 4-4, 4-7, 4-9

ACQU ITY UP LC syst empar t numbers C -2plumbing 2-13refreshing 3-10, 4-6, 4-8sta r t ing up 3-11sta r tup 4-6

additives D -5

air filterbinary solvent man a gercleaning 6-14replacing 6-15

sa mple organ izer, clean ing 6-77ala rm messages

binary solvent man a ger 7-47ELS detector 7-129P DA detector 7-118sample mana ger 7-62sample organizer 7-92TUV detector 7-105

ANSI 3-22, 3-30a ut ozero cont rol 4-10

Bba ckpressure regula tor 2-17

explanation 2-15pictured 2-17

backpressure test , sa mple ma na ger 6-59

ban dsprea ding, preventing 2-13

baseline noise, troubleshooting 7-38baseline spikes 7-39bases D-14binary solvent man a ger

air filterscleaning 6-14replacing 6-15

ala rm messages 7-47check va lve, repla cing 6-9, 6-10checking for lea ks 7-54, 7-55cleaning

air filters 6-14component sett ings

modifying 7-21

viewing 7-20cont rol panel, using 4-4dry prime 3-8flow LE D 3-3, 4-5, 7-43fuses, replacing 6-27head seals

reat taching 6-20removing 6-19

I/O sig na l conn ectors 2-38install ing 2-10leaks, repa iring 7-56mainta ining 6-8overview 1-5plumbing 2-18plunger, replacing 6-23

l l



Index-2

power LE D 3-3, 7-43, 7-89preparing 3-5

pressur e flow envelope 1-5prime 3-8priming 3-8pumped volume warning 6-7rea r pa nel interfa ce connections

changing 7-59displaying 7-57

repairing leaks 7-56signa l connectors 2-38solvent filters, repla cing 6-14solvent ma na ger hea d

reat taching 6-24removing 6-16, 6-20

solvent recommenda tions D -8spare par t s C -2

specificationselectrical B -3environmental B -2performance B -5physical B -2

troubleshooting 7-43–7-59vent va lve ca rt ridge, replacing 6-24

w etted, prime 3-10biohaza rd w arning A-5brake, FlexCa rt 2-8buffered solvents D -3buffers D -3burst warning A-3

C

ca lcula tor, column s 1-4calibrating

needle Z-a xis 2-52, 6-40XYZ mechanism 2-47

calibrationguidelines v

cartr idge

inject ion va lve, repla cing 6-56vent va lve, replacing 6-24

caut ion symbol A-6characterizingneedle sea l 3-20, 3-21, 6-41needle volum e 3-21, 6-42sa mple loop volum e 3-21, 6-42

cha rt-out para meters 2-39, 7-58check va lve, repla cing 6-9, 6-10checksum, viewing 7-5chemical ha zard wa rning A-5chemistry technologies 1-12chromat ogra m, gra dient performa nce

test 5-11chromatography troubleshooting

7-141–7-146cleaners D -6

cleaningair filters

sample organizer 6-77clean ing, a ir filters

binary solvent ma na ger 6-14clea ring bu bbles, from flow cell 7-102column

calculator 1-4compatibility 1-12conditioning 3-38conn ectin g eCord chip 2-31history, monitoring 7-33information, see eCord column chipinstall ing

MS detection 2-32opt ical detection 2-27

ma nufacturing informa tion,finding 7-35

overview 1-12replacing

filter in colum n in-line unit 6-62usage threshold wa rning 6-7

column h eat er ELS detector 4-9



Index-3

I

column h eat ercurrent temperat ure 4-7mainta ining 6-61overview 1-8see eCord column chipset point 4-7specifications

electrical B-12environmental B -11performance B-12

physical B-11column sta bilizer

function 1-8install ing 2-27

comments iiicomposition, solvent 4-5compression screw ferrule assembly

2-13condit ioning colum n 3-38configuring

Empower software 4-2ma intena nce wa rnings 6-7MassLynx sof twa re 4-12syringe para meters 6-56

connectingeCord colum n chip 2-31, 2-34solvent supply 2-34

connectionselectr icit y s ource 2-45Et hernet , m aking 2-37signal , making 2-43

Connections Insight request,

submitting 7-3contacting Waters Technical Service

2-2, 6-5, 7-4contamination

preventing D -2control pa nel 4-4, 4-7, 4-9

binary solvent ma na ger 4-4

ELS detector 4-9P DA detector 4-9sample mana ger 4-7TUV detector 4-9

creatinginstrument method 5-5log entries 7-18sa mple set meth od 5-8system in E mpower 4-3test meth ods 5-5

cust omer comment s ii i

Dda ma ge, reporting 2-2, 6-5, 7-4da ta systems 1-11defrosting, sa mple compa rt ment 6-29degas ser vent line positioning 2-24

detectorEL S, overview 1-10lamp li fe thr eshold wa rning 6-7P DA, overview 1-10see ELS detector, P DA detector,

and TUV detectorsolvent recommenda tions D -8TU V, overview 1-10

detergents D-17diagnostics, start up 3-35diluents, sample D -6dirty flow cell 6-65dra in routing hole 2-12draw ra t es , maximum 6-49, 6-50drift problems, tr oubleshooting 7-38

drip ma na gement system, properplacement for 2-12

dry prime, performing 3-8dyna mic lea k test, performing 7-55

EeCord colum n chip

connecting 2-31, 2-34

overview 1 12 sta rting ACQU ITY UP LC Console



Index-4

overview 1-12specifications B-19

electrica l specifica tionsbinary solvent ma na ger B -3column hea ter B-12sample mana ger B -7sample organizer B-17TUV detector B-14

electrical symbols A-14electricity source, conn ect ions 2-45EL S detector

a la rm messages 7-129component sett ings

modifying 7-32viewing 7-31

cont rol panel, using 4-9I/O sig na l conn ectors 2-42

lamp LEDELS detector 7-128

mainta ining 6-1, 6-64overview 1-10plumbing 2-17power LE D 7-128preparing 3-35

rea r pa nel interfa ce connectionschanging 7-140displaying 7-138

signa l connectors 2-42specifications B -1, B -13, C -1, C -5troubleshooting 7-128–7-140

Empowersystem test mix

preparing 5-3running 5-9

Empower softwarelogging in 4-2overview 1-11sta r t ing 4-2

sta rting ACQU ITY UP LC Consolefrom 4-11

environment a l specificat ionsbinary solvent ma na ger B -2column h eat er B-11sample mana ger B -7sa mple organ izer B -17TUV detector B-13

equipment guidelines iv , 7Ethernet connections, making 2-37exhaust hood 2-23

Fferrule installation 6-48filter

binary solvent ma na gercleaning 6-14

replacing 6-15column in-linereplacing 6-62

sa mple orga nizer, cleaning 6-77firmw a re version, view ing 7-5fittings, tightening recommendations

2-13flammable solvents A-4FlexCart

adjusting height 2-6brake

locking 2-8releasing 2-8

components 2-4height, a djusting 2-6

moving 2-9overview 1-13pow er connections 2-46

flow celldir ty 6-65specifica t ions, TU V det ect or B -16TUV detector

clearing bubbles 7-102 full loop 3-24



Index-5

I

gpictured 2-16, 6-66, 6-69replacing 6-68

flow LED 4-5f low ra t e

during priming 3-8during shutdow n 3-40to ta l 4-5

flushin g, TU V detector flow cell 3-37,6-65

full loop mode 3-24fume hood 2-23fuses, replacing

binary solvent ma na ger 6-27sample mana ger 6-60sample organizer 6-78TUV detector 6-75

Ggradient performance test

chromatogram 5-1, 5-11guidelines

calibration vqua lity contr ol vi

Hha ndling symbols A-15hardware, preparing 3-1head seals, bina ry solvent m a na ger

reat taching 6-20removing 6-19

II/O sig na l conn ectors

binary solvent ma na ger 2-38EL S detector 2-42P DA detector 2-42sample mana ger 2-40TUV detector 2-41

injection mode

ppar tia l loop 3-23pa rt ia l loop needle overfill 3-23

injection valvecartr idge

connecting 2-22replacing 6-56

injections t hreshold w a rning 6-7install ing


columnMS detection 2-32opt ical detection 2-27

la mp, TU V detect or 6-74needle a ssembly 6-35P DA detector 2-12sample mana ger 2-11

sha de, sa mple ma na ger 3-27TUV detector 2-12

instrumentcleaning 6-4information, viewing 7-5method, creating 5-5resetting 7-6

selecting 4-2intended use vinterface connections

binary solvent ma na ger 7-57ELS detector 7-138P DA detector 7-125sample mana ger 7-86TUV detector 7-111

IVD a uthorized representa tive vii

Ll amp

life threshold w a rning 6-7tu rn on/tu rn off contr ol 4-9TUV detector

instal l ing 6-74 checking for lea ks 7-54, 7-55, 7-84,



Index-6

instal l ing 6 74removing 6-73

replacing 6-73lamp LED 4-10, 7-102, 7-114, 7-128leaks, repairing 7-56, 7-86LE D

binary solvent ma na gerflow 3-3, 7-43power 3-3

binary solvent ma na ger, flow 4-5EL S detector

lamp 7-128power 7-128

monitoring 3-3, 7-9P DA detector

lamp 7-114power 7-114

sample mana gerpower 7-60run 3-3, 4-7, 7-60, 7-89

TUV detectorlamp 3-4, 4-10, 7-102, 7-114,

7-128power 7-102

lights, tur ning off 4-9loa d ah ead modeguidelines 3-24

loading sample plates 3-22, 3-30log entries

printing 7-7viewing 7-7

log ent ries, creat ing 7-18logging in

Empower software 4-2MassLynx sof twa re 4-12

Mmainta ining


checking for lea ks 7 54, 7 55, 7 84,7-85

column h eat er 6-61ELS detector 6-1, 6-64P DA detector 6-1, 6-64sample mana ger 6-29system 6-2TUV detector 6-64

maintenancecolumn usa ge threshold w a rning

6-7considerations 6-6injections threshold wa rning 6-7lamp li fe thr eshold wa rning 6-7pumped volume wa rning 6-7safety considerations 6-6schedule 6-2

spare par t s 6-6warnings, configuring 6-7

manufacturing information, column

7-35mass spectrometer

preparing 3-35specifications B -1, B-13, C -1, C -5

ma ss spectrometer shock ha za rd A-4MassLynx sof twa relogging in 4-12overview 1-11sta r t ing 4-12sta rting ACQU ITY UP LC Console

from 4-13median ba seline filter 1-11miscibility

numbers D-10solvents D -9

mixed mobile pha ses, U V cutoff D-12mixer outlet, pictu red 2-21M-numbers D-10mobile pha se

for shut dow n 3-40 optical s pecifica t ions, TU V detector



Index-7

I

prepa ring for syst em verifica tion

5-3viscosity considera tions D-11wavelengths D-13

modifiers D -5modifying

component sett ingsbinary solvent ma na ger 7-21ELS detector 7-32

sample mana ger 7-23, 7-25TUV detector 7-28, 7-30

syringe configurat ion para meters 6-56

monitor a rm a ssembly 2-5monitoring

column history 7-33

d a t a 7-10performance 7-8system instrument LED s 3-3system sta tus 7-8

Nneedle

guide tube, pictured 6-35

holder, replacing 6-43install ing 6-35removing 6-30seal , char a cterizing 3-20, 3-21,

6-41volume, cha ra cterizing 3-21, 6-42w a sh, stopping 3-20

needle seal leak test , performing 7-85needle wa sh connections 2-21needle Z-axis, calibrating 2-52, 6-40

Oopera tiona l specificat ions

TUV detector B-15

B-16options

holding loop sizes C -4sa mple loop sizes C -4spare par t s C -1syringe s izes C -4

organ izer, see sam ple organ izeroverview


column heat er 1-8eCord column chip 1-12ELS detector 1-10Empower software 1-11MassLynx sof twa re 1-11P DA detector 1-10sample mana ger 1-7

sa mple organ izer 1-9solvent considera tions D -2TUV detector 1-10

Ppart numbers, ACQU ITY UP LC

system C -2par tia l loop mode 3-23

pa rt ia l loop needle overfill mode 3-23guidelines 3-24

P DA detectora larm messages 7-118component sett ings

viewing 7-29control pan el, using 4-9

I/O s igna l conn ectors 2-42install ing 2-12lamp LED

P DA detector 7-114mainta ining 6-1, 6-64overview 1-10plumbing 2-17

power LE D 7-114 pressure, system 4-5



Index-8

preparing 3-35

rea r pa nel interfa ce connectionschanging 7-127displayng 7-125

signa l connectors 2-42specifications B -1, B -13, C -1, C -5troubleshooting 7-114–7-127

performance specifications B -5binary solvent ma na ger B -5column hea ter B-12sample mana ger B -9sample organizer B-18

physical specificationsbinary solvent ma na ger B -2column hea ter B-11sample mana ger B -7

sample organizer B-17TUV detector B-13

platesANSI 3-22, 3-30information, displaying 3-34

plumbingbinary solvent ma na ger 2-18

EL S detector 2-17P DA detector 2-17sample mana ger 2-18TUV detector 2-15

plungerbinary solvent ma na ger, repla cing

6-23positioning sa mple plat e 3-30pow er, removing completely 3-40powering on 3-2precipita tion, preventing 3-39, 5-4preparing, system t est mix

Empower 5-3pressure flow envelope, binary solvent

manager 1-5

prime

w ett ed, performing 3-10prim e A/B solvent s cont rol 4-6prime sea l wa sh cont rol 4-6prime syr inges cont rol 4-8prime, performing 3-8priming

binary solvent ma na ger 3-8, 3-10sample mana ger 3-17seal wa sh 3-5

printinglog entries 7-7

printing plots 7-11pumped volume maint enance wa rning

6-7puncture needle holder, replacing 6-43

purgin g, TU V detector flow cell 6-67

Qqu a lity cont rol guidelines vi

Rreagents D-17rea l-tim e plots 7-10

rea r pa nel interfa ce connectionsbinary solvent ma na ger


ELS detectorchanging 7-140displaying 7-138

P DA detectorchanging 7-127displaying 7-125

sample mana gerchanging 7-88displaying 7-86

TUV detector

changing 7-113

displaying 7-111tt hi

lampTUV d t t 6 73



Index-9

I

rea t tachinghead seals, bina ry solvent ma na ger

6-20solvent ma na ger hea d 6-24

recommendationssystem D -7

recordingreference energies 3-37sa mple energies 3-37

reference energies, recordin g 3-37refresh syst em cont rol 4-6, 4-8refreshing

ACQU ITY UP LC syst em 3-10refreshing, ACQU ITY U P LC system

4-6, 4-8regulator, backpressure 2-17

removingflow cell, TUV detector 6-70head seals, bina ry solvent ma na ger

6-19la mp, TU V detect or 6-73needle assembly 6-30solvent ma na ger hea d 6-16, 6-20

repairing leaks 7-56, 7-86replacing

a ir f il ters, binary solvent ma na ger 6-15

check valve, binar y solventmanager 6-9, 6-10

filter in column in -line filter un it 6-62

flow cell, TUV detector 6-68fuses

binary solvent ma na ger 6-27sample mana ger 6-60sa mple organ izer 6-78TUV detector 6-75

injection va lve ca rt ridge 6-56

TUV detector 6-73plunger, bina ry solvent ma na ger

6-23punctu re needle holder 6-43sa mple loop 6-45sam ple syringe 6-49solvent filters 6-14vent valve ca rtr idge 6-24wa sh syr inges 6-53

reset cont rolB S M 4-6SM 4-9TUV detector 4-10

resett ing instrum ents 7-6restrictors 3-31reverse-flush ing, TU V detector flow

cell 6-68routine ma intenance 6-2run LED 4-7, 7-60, 7-89running

system t est mix, Empower 5-9

Ssafety advisories A-1

sa fety considera tions, ma intenan ce 6-6sa lts, prevent ing precipita tion 3-39,

5-4sample

compartment, defrosting 6-29displaying remaining 3-34energies, recordin g 3-37

loop, replacing 6-45plates, loading 3-22, 3-30

sample diluents D -6sa mple loop volume, cha ra cterizing

3-21, 6-42sample mana ger

a larm messages 7-62

backpressure repairing leaks 7-86



Index-10

testing 6-59

checking for lea ks 7-84, 7-85component sett ingsmodifying 7-23, 7-25viewing 7-22

cont rol panel, using 4-7flow expla na tion 1-7fuses, replacing 6-60I/O sig na l conn ectors 2-40

information window 3-17injection va lve ca rt ridge, repla cing

6-56injections t hreshold w a rning 6-7install ing 2-11leaks, repairing 7-86lights, tur ning off 4-9

loading sample plates 3-22mainta ining 6-29needle assembly

instal l ing 6-35removing 6-30

needle seal, char a cterizing 3-20,

3-21, 6-41

needle volume, cha ra cterizing 3-21,6-42needle Z-axis, calibrating 2-52,

6-40needle, w ash ing 3-19overview 1-7plumbing 2-18power LE D 7-60preparing 3-14priming 3-17puncture needle holder, replacing

6-43rea r pa nel interfa ce connections


resetting 4-9

run LED 3-3, 4-7, 7-60, 7-89sam ple compart mentcurrent temperat ure 4-7defrosting 6-29set point 4-7

sa mple loop volume, cha ra cterizing

3-21, 6-42sa mple loop, replacing 6-45

sa mple syringe, repla cing 6-49sha de, insta l ling 3-27signal connectors 2-40solvent recommenda tions D -8spare par t s C -4specifications

electrical B -7

environmental B -7performance B -9physical B -7

sta tus 4-7syringe configuration parameters,

modifying 6-56troubleshooting 7-60–7-88

tur ning lights off 4-9wa sh sy ringes, repla cing 6-53wa shing needle 3-18XYZ mechan ism, ca libra ting 2-47

sample organizera ir filters, clea ning 6-77ala rm messages 7-92clean ing a ir filt ers 6-77component sett ings, viewing 7-24displaying plate information 3-34drain 2-25fuses, replacing 6-78information window 3-34initia ting communications t o 3-29loading sample plates 3-30

overview 1-9preparing 3-29

solvent manager, see binary solventmanager



Index-11

I

preparing 3-29spare par t s C -6specifications

electrical B-17environmental B -17performance B-18physical B-17

troubleshooting 7-89–7-101unpacking 2-10

sam ple plat esloading 3-22, 3-30

sam ple set m ethod, crea ting 5-8sa mple syringe leak t est, performing

7-84sa mple syringe, repla cing 6-49sa mpling r a te recommendat ion 1-4

sa ving plots 7-11seal w a sh prime, performing 3-5select ing, syst em instr ument s 4-2, 4-12serial numbers

instrument 7-5service profile report, generating 7-5shelf spa cing 3-31shutting down

betw een ana lysis 3-39fewer t ha n 72 hours 3-40indefinitely 3-40long-term 3-40

signal ca ble, at ta ching 2-43signa l connections, ma king 2-43solvent

filters, r epla cing 6-14supply, conn ectin g 2-34t r a y 2-34

solvent ma na ger headreat taching 6-24removing 6-16, 6-20

managersolvents

buffered D -3buffers D -3changing 5-4clean D -2composition 4-5miscibility D -9other D -5

preparation D -2quality guidelines D -2recommendations

binary solvent ma na ger D -8detector D -8sample mana ger D -8

recommended D -4

stabilizers D-10strong wa sh 3-14tetrahydrofuran D -5, D -6, D -7, D -8,

D -9, D-10, D-12, D-14TH F D -5, D -6, D -7, D -8, D -9, D-10,

D-12, D-14to a void D -6UV cutoff D-11viscosity considera tions D-11w a te r D -3weak wash 3-14

spare par t sbinary solvent ma na ger C -2maintenance 6-6recommended C -1

sample mana ger C -4sa mple organ izer C -6TUV detector C -5

specificationseCord column chip B-19electrical B -3, B -7, B-12, B-14,

B -17

EL S detector B -1, B -13, C -1, C -5environmental B 2 B 7 B 11

configuration parameters,modifying 6 56



Index-12

environmental B -2, B -7, B-11,

B-13,

B-17ma ss spectrometer B -1, B-13, C -1,C -5

operational B-15optical B-16P DA detector B -1, B-13, C -1, C -5performance B -5, B -9, B-12, B-18physical B -2, B -7, B-11, B-13, B -17

stabilizerfunction 1-8install ing 2-27

sta bilizers, solvent D-10sta r t up 3-12sta r t up, ACQU ITY UP LC system 3-11sta r t ing

ACQU ITY U P LC Consolefrom E mpow er softw a re 4-11from MassLyn x softw a re 4-13

Empower software 4-2MassLynx sof twa re 4-12TUV detector 3-35

sta rting up, ACQU ITY syst em 3-12

sta rtup contr ol 4-6sta rtup diagnostics 3-35sta r tup tests 3-2sta rtup, ACQU ITY U P LC system 4-6static decay test, performing 7-54stop flow 4-5stopping, needle w a sh 3-20str ong needle w a sh connection 2-21symbols

caution A-6electrical A-14handling A-15warn ing A-2

syringe

modifying 6-56

draw ra t es, maximum 6-49

,6-50priming binary solvent ma na ger

wi th 3-9sa mple, replacing 6-49wa sh, repla cing 6-53

Sys P rep 4-6, 4-8system

pow ering on 3-2

recommendations D -7setup 2-2shutt ing down 3-39, 3-40test mix, Empower 5-9

system instrumentsselecting

from E mpow er softw a re 4-2

from MassLynx softw a re 4-12system pressure 4-5system sta tus

determining 7-9monitoring 7-8

syst em verifica tion 5-2

Tt ea ch block 2-47test methods, creat ing 5-5tests , s tar t up 3-2tetrahydrofuran D -5, D -6, D -7, D -8,

D -9, D-10, D-12, D-14TH F D -5, D -6, D -7, D -8, D -9, D-10,

D-12, D-14

tightening recommendations, fittings 2-13

tota l f low ra t e 4-5trademarks iitroubleshooting

baseline noise 7-38binary solvent ma na ger 7-43–7-59

chromatography 7-141–7-146drift problems 7-38

rear pan el interfa ce connectionschanging 7-113



Index-13

I

pEL S detector 7-128–7-140

P DA detector 7-114–7-127sample mana ger 7-60–7-88sample organizer 7-89–7-101TUV detector 7-102–7-113

tur n light s off control 4-9TUV detector

a la rm messages 7-105

component sett ingsmodifying 7-28, 7-30viewing 7-27

cont rol panel, using 4-9flow cell

clearing bubbles 7-102flushing 6-65

purging 6-67replacing 6-68reverse-flushing 6-68

fuses, replacing 6-75I/O sig na l conn ectors 2-41information window 3-36install ing 2-12lamp

cooling t ime 6-73install ing 6-74LE D 3-4, 4-10, 7-102LED indica tions 7-102, 7-114,

7-128life threshold w a rning 6-7removing 6-73

replacing 6-73tu rn on/tu rn off contr ol 4-9

mainta ining 6-64overview 1-10plumbing 2-15power LE D 7-102preparing 3-35

g gdisplayng 7-111

signal connectors 2-41spare par t s C -5specifications

electrical B-14environmental B-13flow cell B-16operational B -15

optical B-16physical B-13

sta r t ing 3-35troubleshooting 7-102–7-113verifying 3-37

Uunpa cking, sam ple organ izer 2-10U P LC syst em, guidelines 1-4U V cutoff values D-11, D-12

Vvent va lve cart ridge, replacing 6-24verifying

system 5-2

TUV detector 3-37version informa tion, softw a re a ndf irmware 7-5

viewingcomponent sett ings

binary solvent ma na ger 7-20EL S detector 7-31P DA detector 7-29sample mana ger 7-22sample organizer 7-24TUV detector 7-27

instrument informa tion 7-5log entries 7-7serial numbers 7-5

viscosity considera tions D-11

Wwa rning symbols A-2 A-7



Index-14

wa rning symbols A 2, A 7

warningscolumn usa ge 6-7configuring 6-7detector la mp life 6-7injections threshold 6-7pumped volume ma intena nce 6-7

w a sh needle control 4-8w a sh solvent s, selecting 3-14

wa sh syr ingesleak t est, performing 7-85replacing 6-53

w a shing, sa mple ma na ger needle 3-19w a ste line positioning 2-24wa ter , as solvent D -3Wa t ers Techn ica l Service, conta cting

2-2, 6-5, 7-4wavelength

cutoffs D-12display 4-9mobile phas e absorba nces D-13selection D-11

w eak needle wa sh connection 2-21

Windows operat ing syst ems 1-11XXYZ mecha nism, ca librat ing 2-47

Acquity Operators Manual

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