Waters Prep Series€¦ · Waters Prep series 1-3 Waters PrepLC system Delta-Prep system The Waters...

Waters Prep SeriesInstallation and Maintenance Guide

71500045204/Revision B

Copyright © Waters Corporation 2010All rights reserved

Copyright notice

© 2010 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA AND IN IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, its use.

Trademarks

Millennium, PrepPak, and Waters are registered trademarks of Waters Corporation, and AutoBlend, Delta-Prep, Empower, LAC/E, PowerLine, PrepLC, Radial-Pak, and “THE SCIENCE OF WHAT’S POSSIBLE.” are trademarks of Waters Corporation.Milli-Q is a registered trademark of Millipore Corporation.Other registered trademarks or trademarks are the sole property of their owners.

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Customer comments

Waters’ Technical Communications department invites you to tell us of any errors you encounter in this document or to suggest ideas for otherwise improving it. Please help us better understand what you expect from our documentation so that we can continuously improve its accuracy and usability.We seriously consider every customer comment we receive. You can reach us at [email protected].

Contacting Waters

Contact Waters® with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail.

Safety considerations

Some reagents and samples used with Waters instruments and devices can pose chemical, biological, and radiological hazards. You must know the potentially hazardous effects of all substances you work with. Always follow

Waters contact information

Contacting medium InformationInternet The Waters Web site includes contact

information for Waters locations worldwide. Visit www.waters.com.

Telephone and fax From the USA or Canada, phone 800 252-HPLC, or fax 508 872 1990.For other locations worldwide, phone and fax numbers appear in the Waters Web site.

Conventional mail Waters Corporation34 Maple StreetMilford, MA 01757USA

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Good Laboratory Practice, and consult your organization’s safety representative for guidance.

Safety advisoriesConsult Appendix A for a comprehensive list of warning and caution advisories.

Operating this instrument

When operating this instrument, follow standard quality-control (QC) procedures and the guidelines presented in this section.

Applicable symbols

Audience and purposeThis guide is intended for personnel who install and maintain the Waters® Prep Series HPLC system.

Symbol Definition

Manufacturer location

Authorized representative of the European Community

Confirms that a manufactured product complies with all applicable European Community directives

Australia C-Tick EMC Compliant

Confirms that a manufactured product complies with all applicable United States and Canadian safety requirements

Consult instructions for use

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Intended use of the Waters Prep Series HPLC systemUse the Waters Prep Series HPLC system to isolate and purify a wide variety of samples. The Waters Prep Series HPLC system is for research use only.

CalibratingTo calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards should include the entire range of QC samples, typical specimens, and atypical specimens.When calibrating mass spectrometers, consult the calibration section of the operator’s guide for the instrument you are calibrating. In cases where an overview and maintenance guide, not operator’s guide, accompanies the instrument, consult the instrument’s online Help system for calibration instructions.

Quality-controlRoutinely run three QC samples that represent subnormal, normal, and above-normal levels of a compound. Ensure that QC sample results fall within an acceptable range, and evaluate precision from day to day and run to run. Data collected when QC samples are out of range might not be valid. Do not report these data until you are certain that the instrument performs satisfactorily.

ISM classification

ISM Classification: ISM Group 1 Class BThis classification has been assigned in accordance with CISPR 11 Industrial Scientific and Medical (ISM) instruments requirements. Group 1 products apply to intentionally generated and/or used conductively coupled radio-frequency energy that is necessary for the internal functioning of the equipment. Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply network.

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EC authorized representative

Waters Corporation (Micromass UK Ltd.)Floats RoadWythenshaweManchester M23 9LZUnited Kingdom

Telephone: +44-161-946-2400Fax: +44-161-946-2480Contact: Quality manager

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Table of Contents

Copyright notice ................................................................................................... ii

Trademarks ............................................................................................................ ii

Customer comments ............................................................................................ iii

Contacting Waters ............................................................................................... iii

Safety considerations .......................................................................................... iii Safety advisories ................................................................................................. iv

Operating this instrument ................................................................................. iv Applicable symbols ............................................................................................. iv Audience and purpose......................................................................................... iv Intended use of the Waters Prep Series HPLC system ..................................... v Calibrating ........................................................................................................... v Quality-control ..................................................................................................... v

ISM classification .................................................................................................. v ISM Classification: ISM Group 1 Class B .......................................................... v

EC authorized representative ........................................................................... vi

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

Waters Prep series ............................................................................................ 1-2 PrepLC system................................................................................................. 1-2 Delta-Prep system............................................................................................ 1-3 Waters Prep series modules ............................................................................ 1-5 Accessories ..................................................................................................... 1-13

Getting started ................................................................................................. 1-14 Starting the system ....................................................................................... 1-14 Using the front panel..................................................................................... 1-16 Displaying help .............................................................................................. 1-19 Shutting down the system............................................................................. 1-19 Configuring the controller ............................................................................. 1-19

Table of Contents vii

2 Installing the System ............................................................................ 2-1

Setting up and inspecting ............................................................................... 2-2 Unpacking and installing the system............................................................. 2-2 Selecting the site.............................................................................................. 2-3

Electrical requirements .................................................................................. 2-5 Electrical conditions ........................................................................................ 2-5 Voltage selections............................................................................................. 2-5

Assembling the system ..................................................................................... 2-7 Connecting Rheodyne injectors ....................................................................... 2-7 Connecting the M-1000 PrepPak module (optional) ...................................... 2-9 Connecting additional devices......................................................................... 2-9 Attaching the interface cable and power cord.............................................. 2-13 Connecting IEEE-488 interfaces................................................................... 2-15 Connecting to the Waters 746 data module ................................................. 2-20 Liquid line connections.................................................................................. 2-21

Connecting a helium tank to the fluid handling unit ............................ 2-30 Helium specifications..................................................................................... 2-30 Sparging ......................................................................................................... 2-31 Connecting compression fittings ................................................................... 2-31

Setting up the plunger wash ........................................................................ 2-33 Considerations ............................................................................................... 2-34 Connecting the plunger wash tube ............................................................... 2-34 Connecting to the injection panel (PrepLC system)..................................... 2-35 Connecting to the column rack (Delta-Prep system) ................................... 2-37

Installing sample injection options ............................................................ 2-40 Installing the small-scale sample injection valve option (PrepLC system) 2-40 Using an autoinjector .................................................................................... 2-41 Making electrical connections ....................................................................... 2-41 Connecting liquid lines when using a Waters autoinjector......................... 2-41

Connecting a fraction collector ................................................................... 2-42 Making the remote start connection............................................................. 2-42 Making chart mark connections ................................................................... 2-42

viii Table of Contents

Installing analytical and preparative columns ....................................... 2-43 PrepLC system............................................................................................... 2-43 Delta-Prep system.......................................................................................... 2-45

Making detector connections ....................................................................... 2-48 PrepLC or Delta-Prep system ....................................................................... 2-48 Connecting detector semi-prep cells ............................................................. 2-48 Using the RI detector with the flow splitter option ..................................... 2-49

3 Troubleshooting, Testing, and Maintenance ................................... 3-1

Troubleshooting ................................................................................................ 3-2 Troubleshooting steps...................................................................................... 3-2 Troubleshooting flowcharts ............................................................................. 3-5 Chromatography troubleshooting ................................................................. 3-16 Hardware troubleshooting ............................................................................ 3-23 Software troubleshooting .............................................................................. 3-31

Performance tests ........................................................................................... 3-36 Self-diagnostics .............................................................................................. 3-36 Extended tests................................................................................................ 3-37

Maintenance procedures ............................................................................... 3-47 Column rack tubing configuration................................................................ 3-47 Fluid handling unit maintenance ................................................................. 3-48 Replacing fuses .............................................................................................. 3-67 Maintaining rheodyne manual injectors ...................................................... 3-71

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

Warning symbols ............................................................................................... A-2 Task-specific hazard warnings........................................................................ A-2 Specific warnings ............................................................................................. A-3

Caution symbol .................................................................................................. A-5

Warnings that apply to all Waters instruments ......................................... A-6

Electrical and handling symbols ................................................................. A-12 Electrical symbols .......................................................................................... A-12 Handling symbols .......................................................................................... A-13

Table of Contents ix

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

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

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

Clean solvent ..................................................................................................... D-2

Solvent miscibility ........................................................................................... D-3

How to use miscibility numbers (M-numbers) .......................................... D-5

Buffered solvents ............................................................................................. D-6

Solvent head height ......................................................................................... D-6

Solvent viscosity ............................................................................................... D-6

Solvent degassing ............................................................................................ D-7

Solvent degassing methods ............................................................................ D-8

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

x Table of Contents

1 Overview

Waters produces its Prep Series high performance liquid chromatography (HPLC) systems in two configurations:• Waters PrepLC™ System• Waters Delta-Prep™ SystemTip: Although the two systems share many features, certain aspects nevertheless differ. This guide addresses both differences and similarities.Contents

Topic PageWaters Prep series 1-2Getting started 1-14

1-1

Waters Prep series

The Waters Prep Series offers these features:• Two pump configurations:

– 150-mL flow rate with 500-µL pump heads at 4000 psi– 300-mL flow rate with 1000-µL pump heads at 2000 psiTip: Fitted with 1000-μL pump heads, the PrepLC or Delta-Prep system can operate at twice the flow rate of a 500-μL pump head system, permitting separation of larger sample loads.

• Four-solvent gradient capability (Auto•Blend™ method)• Pump purge valve to facilitate priming and purging of solvent lines• Plunger wash with integral pumping system• Single keypad control of PowerLine™ system and/or gradient conditions• Ability to configure the system to recall certain default conditions• Methods storage that lets you access previously created gradient, event,

and detector tables• Preprogrammed gradient profiles• System control of external PowerLine detectors and autoinjectors is

through an IEEE-488 interface• RS-232 serial communications interface for connecting to an optional

data workstation or integrator• Shelf unit to house system modules in an integrated assembly that

optimizes the volume of interconnecting tubing

PrepLC systemThe PrepLC system performs both analytical and preparative scale chromatography. Use its pump with either 500- or 1000-µL volume heads. The 500-µL pump heads allow flow rates of 1 to 150 mL/min and the 1000-µL heads a rate of 4 to 300 mL/min.The PrepLC system uses 0.040-inch ID tubing, which accommodates large- and small-scale operation. To optimize system performance, you can change critical tubing in the flow path to 0.009-inch ID or 0.040-inch ID, depending on flow rate and backpressure.

1-2 Overview

Waters PrepLC system

Delta-Prep systemThe Waters Delta-Prep system is an HPLC system capable of analytical and preparative scale chromatography.Use the system’s pump with either 500- or 1000-µL volume heads. The 500-µL pump heads permit flow rates of 1 to 150 mL/min, and the 1000-µL heads permit flow rates of 4 to 300 mL/min.The system allows easy scale up, so you need not replumb to change between small- and large-scale operations. Instead, select either operating mode with a column-switching valve.

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Injector panel

Controller

Fluid handling unit

Waters Prep series 1-3

Waters Delta-Prep system

Delta-Prep system components

PrepLC System controller – Houses system electronics and controls the solvent gradient, flow rate, sparging, and external events. It also provides connection terminals and communication ports for operating with external devices.Fluid handling unit – Contains components for delivering solvents from their reservoirs to the injector or column. It consists of the pump and plunger wash pump, solvent sparge valve assembly, and solvent proportioning valve assembly.Column rack – Holds these items:

• Rheodyne injectors• Gradient mixers• Pump purge valve• Scale switching valve• 0.009-inch ID and 0.040-inch ID stainless steel tubing arrangements

Controller

2487 detector

Fluid handling unit

Drip trayDrip tray

Column rack

1-4 Overview

• Space for three analytical or semi-prep columns• One 2-inch preparative column, and/or an M-1000 compression chamber

for PrepPak® cartridges.Rheodyne 7725i injector – For small scale injections, incorporates a 100-µL fixed sample loop, though you can install loops whose injection volumes vary. Optional loops, available from Waters, handle fixed volumes from 5 to 200-µL.Rheodyne 3725i injector – For large-scale injections, incorporates a 20-mL fixed sample loop, though you can install loops of varying injection volumes. Optional loops, available from Waters, handle fixed volumes from 2 to 20 mL.Purge valve – Lets you divert pump output to waste (e.g., when purging the system) or through the injector, columns, etc.Scale switching valve – Lets you choose large- or small-scale tubing systems. In the small-scale system, the injector is in a flow path plumbed with 0.009-inch ID tubing. In the large-scale system, solvent flows from the pump to the large-scale injector through 0.040-inch ID tubing.Inlet and outlet ports – Accepts pump outflow. The inlet port receives solvent for both small- and large-scale flow paths, which flow to separate outlet ports. The flow from these outlets goes to the detector flow cell.Purge outlet port – Flow goes from the inlet port to this outlet port when you set the purge valve to Purge.

Waters Prep series modulesThis section addresses these Prep Series major modules:

• Fluid handling unit• PrepLC controller• Rheodyne 3725i injector (PrepLC and Delta-Prep systems)• Rheodyne 7725i injector (Delta-Prep system only; optional for PrepLC

system)• Column rack (Delta-Prep system only)• Injector panel (PrepLC system only)

Fluid handling unit

The figure below shows the internal workings of the fluid handling unit, which delivers solvents from their reservoirs to the injector.


Waters Prep series fluid handling unit

Fluid meteringSolvent selection and proportioning take place on the low pressure (intake) side of the solvent delivery system.The fluid handling unit can proportion up to four solvents at a time with a high degree of reproducibility, even when those solvents compose extremely disproportionate mixtures. The solvent proportioning valve produces predictable gradient segments, regardless of solvent compressibility and system backpressure.Flow pathSolvents are helium-sparged in their reservoirs and proportioned at the solvent proportioning valve. They then flow to the pump inlet manifold, where the flow path branches at two inlet check valves. The pump heads draw solvent through the valves, moving 500 and 1000 μL with each stroke, with a maximum flow rate of 150 and 300 mL/min.From the pump heads, solvents converge at a tee fitting (150 mL) or common outlet check valve (300 mL) before flowing to a pressure transducer. Flow continues from the pressure transducer outlet to the system purge valve.Plunger washThe fluid handling unit contains an active plunger wash system. This helps lubricate the plunger, reducing wear, and flushes away any mobile phase or

Solventreservoirlines (4)

Plunger washinlet line

Connectorsand fittingsdrawer

Drip tray

Pump head

Outlet checkvalves (2)

Inlet checkvalves (2)

Manual sampleinlet valve

1-6 Overview

sample forced past the pump seal from the discharge side of the pump. Wash solvent flows from a reservoir to the wash pump and then to a cavity behind the main pump seal of one pump head. From there, it flows to the cavity behind the second head’s seal. It then returns to the reservoir or proceeds to waste, depending on your choice.The wash pump runs whenever the controller is on, circulating solvent by pushing a small volume along the flow path, one pulse per minute.

PrepLC controller

The figure below shows the Waters PrepLC controller, which controls solvent composition, flow rate, sparging, and external events, during a run.

PrepLC controller front panel

The controller’s rear panel, shown below, provides connection terminals and communication ports for operating with external devices.

Liquid crystaldisplay

Screen keys (4)Functionkeys (4)

HOME key Cursor movement keys

Number keys (11) CLEAR key

Contrastkeys (2)

ENTER key

On/offswitch


PrepLC controller rear panel

Rear panelThis section describes the electrical connections to the controller’s rear panel.

Event Switches (Connect to terminals S1, S2, S3, S4 and Ground) – Four switches on the rear panel control column-switching valves, fraction collectors, or similar external devices that require a contact closure at their input. These switches, whose maximum current capacity is 1 A each, are actuated directly or programatically, according to the mode you specify in the operating software. The controller’s OPERATE METHOD screen allows direct control of the switches. Program control is through the time-based PROGRAM EVENT screen, where you must specify each switch as ON, OFF, or PULSE.Tip: These terminals are always positive with respect to ground: +5 VAC = Off (open switch); 0 VAC = On (closed switch).The figure below depicts devices that require a TTL-compatible output connection.

Caution: Do not use cables longer than 9.8 feet (3 meters) when you connect to the screw-type barrier terminal strips. Also, attach the cable shield to the chassis ground of one instrument only

RS-232connector

IEEE-488connector

Pump interfaceconnector

Auxiliary fuse

Pump fuse

AC power inlet

Plug-interminalreceptacles

1-8 Overview

Tip: If the device you are connecting to has a Gnd (ground) terminal, you must connect it to the controller’s ground terminal.

Connection for devices to TTL-compatible outputs

The figure below depicts devices that require 12 Vac auxiliary power.Tip: The Gnd terminals are not used.

Connection for devices requiring 12-volt auxiliary power

Inject Switch (Connect to terminals Inject and Gnd) – For a system that includes a non-TTL injector, a run begins with the injector’s signal to the inject start terminal. When the controller acts as a PowerLine system device (see “Configuring the controller” on page 1-19), an autoinjector begins the run and sends its signal through the inject start terminal or the IEEE-488 communications interface.The inject start terminal becomes an output when you forgo using an injector in favor of starting the run with screen keys. Thus the inject start terminal synchronizes controller operation with that of fraction collectors, integrators, and other devices that require a start signal.Stop Flow (Connect Stop Flow and Gnd) – Accepts a contact closure signal to stop the pump when radial compression falls below 650 psi. The signal can originate with a programmable detector for Stop Flow scanning or with the pressure monitor on the M-1000 PrepPak module.Hold (Connect Hold and Gnd) – Produces an output signal that prevents further injections in cases of catastrophic events: power failure, pressure shutdown, or when abort conditions arise. Connect it with any compatible device that must be stopped when any of these events happen.

S1 to S4Gnd

Contact closure orTTL-to-gnd device

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Device requiring + 12V power

S1 to S4Gnd_


Chart (+, –) 10-mV full scale output – Plots output, which represents pump percentage composition (%A through %D) or flow rate, to a recording device you specify as a Chart OUT parameter on the SETUP screen.Tip: Because of the delay associated with system volume,“Pump percentage composition” refers to solvent composition at the proportioning valve and not the column or detector.Pressure (+, –) 10-mV full scale output – Directs pressure trace output from the pump transducer (0 to 4000 psi).Connector typesThe rear panel terminal strip contains these signal connections:Open Collector (S1, S2, S3, S4) – Output signals that control inputs requiring contact closures, TTL-level signals, or open collector outputs. They also trigger solenoid valves and other power devices that require current up to 1-A current. You can program the controller to turn these signals on and off or produce a 0.2-second pulse.Digital Output (Hold) – Useful for signaling the logic inputs of other devices. It is compatible with TTL inputs or inputs expecting contact closures. If the controlled input is polarized, connect the grounds together and the input terminal to the Hold output.Analog Output (Press +/–, Chart +/–) – Special output produces a DC voltage whose magnitude is proportional to the physical parameter monitored. The outputs’ full scale voltage range 10 mV. The Press terminals present only a voltage representation of the current system backpressure. The Chart terminals are configured through the controller to produce a voltage proportional to the composition of solvent reservoir A, B, C, or D, or to the current flow rate.Digital Input (INJ, Stop Flow) – Driven by several outputs, including TTL signals, open collector outputs, or contact closures. If the outputs driving these inputs are polarized, connect the ground terminals together and the signal terminals together.Tip: Stack multiple instruments (see the figure “Stacked IEEE-488 connections in a Waters workstation” on page 2-16) or connect them in series. Thus you might connect the 7725i injector, a Waters non-PowerLine autoinjector, and a Waters 746 integrator.Tip: An autoinjector connected to this terminal starts the run. (Waters autoinjectors start the run through PowerLine control.) The time required to

1-10 Overview

withdraw the samples means the autoinjector must generate the analysis start signal when the sample is ready for injection.The rear panel strip also contains these items:Signal ground – Terminals labeled Gnd should reference all controller digital input and output signals. Ground terminals should be connected together among system modules.Chassis ground – Internally connected to the controller’s chassis. These terminals connect the shield lead from an analog signal cable, like the one used with the Press or Chart analog output signals.+12 V supply – A terminal on the controller carries up to 1.2 A of current at +12 VDC. It is used with event output switches S1 to S4 for actuating solenoid valves and other automation accessories.AC power connector – Houses fuses and the inlet connection for the power supply cord. It also provides for selecting operating voltage.Auxiliary power fuse – Contains a 1.5-A fuse for the +12 VAC accessory power terminal.Pump power fuse – Contains a 4-A fuse (5 × 20 mm).Connecting cablesSystem interface – Thirty-seven pin connector that allows system communication between the controller and the fluid handling unit.IEEE-488 – Allows communication interface between system controller and PowerLine detectors and autoinjectors.RS-232 – Allows communication interface for Waters 746 integrator or Waters MassLynx™ or Empower™ workstations.

Injectors

Use the Rheodyne 3725i injector for large-scale separations and the 7725i model for small-scale ones. Both injectors let you load sample by syringe (injecting it into the injector) or suction (attaching the syringe to a vent tube and drawing it through the loop).Rheodyne 7725i injector (Delta-Prep system)The Rheodyne 7725i injector incorporates a 100-µL fixed sample loop. The loop and valve passages together form the 100-µL volume. This injector is for small-scale separations.


For large-scale operations, inject sample through the manual pump inlet line, the Rheodyne 3725i injector, or any available solvent delivery line.Rheodyne 3725i injectorThe Rheodyne 3725i injector incorporates a 20-mL fixed sample loop. The loop volume and its valve passages precisely determined sample volume.

Column rack (Delta-Prep system)

The column rack (see the figure “System column rack (Delta-Prep system)” on page 2-38) incorporates a small-scale (0.009-inch ID) and large-scale (0.040-inch ID) tubing system for the small- and large-scale injectors. The scale select valve lets you choose between the small-scale and large-scale flow paths.Small-scale flow pathWhen you select Small-Scale, solvent flows from the pump, through the purge valve, and continues to the mixer. From there it goes to the Rheodyne 7725i injector, the analytical column, the small-scale outlet port, and the detector.Large-scale flow pathWhen you select Large-Scale, solvent flows from the pump, through the purge valve, and continues to the mixer. From there it goes to the Rheodyne 3725i injector, the preparative column, the large-scale output port, and the detector.Purge valveWhen you set the purge valve to Pump Purge, the purge valve directs flow to the purge outlet. When set to System, it directs flow to the large-scale or small-scale flow path via the scale select valve.

Injector panel (PrepLC system)

Mounted on a bilevel shelf assembly, the injector panel houses the Rheodyne 3725i injector and pump purge valve. The figure “Injector panel (PrepLC system)” on page 1-13 shows the injector panel, indicating locations for the optional Rheodyne 7725i injector and scale select valves.

1-12 Overview

Injector panel (PrepLC system)

Solvent flows from the inlet port to the purge valve. When you set the purge valve to Pump Purge, solvent flows to the purge outlet. When you set the purge value to System, it flows through the injector to the column.

AccessoriesSeveral Waters Prep Series options let you to expand your system’s capability.

Waters fraction collector

You can program controller event options to integrate a Waters Fraction Collector as part of the Waters Prep Series. You can program the fraction collector itself for maximum flexibility and productivity.

SE 120 chart recorder

The SE™ 120 is a two-pen recorder that can simultaneously plot two analog signals whose full-scale range is from 0 to 10 mV to 0 to 10 VAC.

M-1000 PrepPak module

This product is the compression chamber module for Waters PrepPak cartridges. The pneumatically operated M-1000 PrepPak module applies radial compression to the flexible walls of a Waters PrepPak cartridge, creating a stable, uniform bed.

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Optional scale select valve site

Optional rheodyne 7725 injector site

Rheodyne 3725i injector

Purge valve


RCM modules

A range of radial compression modules (RCMs) accommodates Radial-Pak™ cartridges (8 × 10 mm, 25 × 10 cm, or 40 × 10 cm) for small-scale or large-scale operation. Radial-Pak cartridges are available in many and different packings.

Flow splitter option

The flow splitter option modifies the Prep Series for high flow rate use with a refractive index detector. It splits eluent flow so that 1% goes to the refractometer and 99% to the UV detector or fraction collector. This serves the necessary purpose of minimizing solvent pressure on the RI cell. The option includes an RI reference flush valve.Accessories, such as autoinjectors or detectors, are available from your Waters sales representative.

Getting started

This section describes using the Waters Prep Series, focusing specifically on these operations:

• Starting the system• Using the front panel of the PrepLC controller• Shutting down the system• Configuring the PrepLC controller

Starting the system

If this is the first time you are starting the system, make sure all components are properly installed.Press the On/Off switch on the front panel of the PrepLC controller to the ON position to start the system.

Caution: To ensure proper ventilation, operate the system with covers in place. Failure to do so shortens the life expectancy of some parts and invalidates the warranty.

1-14 Overview

During startup, the controller automatically runs self-diagnostic tests and then displays the Startup screen.

Startup screen

The Startup screen lists the diagnostic status, controller configuration, and software version number.

Diagnostic tests

The PrepLC controller is shipped as a PowerLine controller. Accordingly, the Startup screen appears in PowerLine mode.The self-diagnostics test each PowerLine module in the Waters Prep Series. These modules include:

• Controller and pump• Detectors (e.g., Waters 2487 or 2414)• Autoinjector (Waters 717plus)

If all tests run successfully, the Startup screen indicates the diagnostic status is “OK.”If any of the tests fail, the Startup screen displays a failure message.Controller or pump failureIf the controller or pump fails the diagnostics test, the Startup screen names the failed test. Turn the controller off, then on again. If the diagnostics fail a second time, call Waters Technical Service.

Getting started 1-15

Detector or autoinjector failureIf a detector or the autoinjector fails the diagnostics test, the Startup screen indicates so. Turn the controller off, then on again. If it fails again, call Waters Technical Service.

Leaving the Startup screen

To leave the Startup screen, press any function key. Function keys are explained in “Function keys” on page 1-17.

Using the front panelThe front panel (see the figure “PrepLC controller front panel” on page 1-7) consists of the display screen, the On/Off Switch, and three groups of keys.

Display screen

The display screen incorporates a screen for each operation and its entries as well as messages from the system. It also shows the cursor, a bright rectangle against a dark background.

Controller keys

The controller has three types of keys:• Keypad – Cursor movement keys, ENTER key, CLEAR key, and

number keys.• Function – A row of labeled keys, on the lower front panel, that switch

system operation modes.• Screen – A row of unlabeled keys, directly below the screen, that vary

with the screen you access.

Keypad

The Waters Prep Series controller’s main keypad includes these keys:• Cursor movement – Move the cursor in the direction each key’s arrow

indicates: up, down, left, and right. The cursor location on each screen shows as a bright rectangle. The width of the cursor generally indicates the size of the entry allowable for any given parameter field.

• HOME – Moves the cursor directly to the home, or first entry, position on that screen.

1-16 Overview

• ENTER – Registers the value you type for a parameter. The system evaluates each entry to determine whether it is within the proper limits. An error message appears when the value is not allowed.

• CLEAR – Erases the value typed immediately beforehand, if you did not press ENTER. The value that previously occupied the field reappears. This key also clears warning messages from the screen.

• Number – Numerals 0 to 9 plus the decimal point.Entering values

1. Use the arrow keys to move the cursor to the field in which you intend to enter a value.

2. Type a value. If you do not know the field’s allowable range, press the HELP screen key.

3. Press ENTER. The program evaluates the new entry for proper type and range.

4. An acceptable entry becomes the current parameter value, and the cursor moves to the next logical entry position.An unacceptable entry prompts an error message at the bottom of the screen, and the previous value remains the current parameter value. Press the CLEAR key to clear the error message and repeat steps 2 and 3.

Function keys

The five function keys are directly below the screen keys.

Function keys for a PowerLine or gradient controller

Key DescriptionSETUP Sets up the system hardware.DIRECT Programs the condition for the attended operation.OPERATE METHOD

Begins the method run.

PROGRAM METHOD

Sets up and stores Gradient, Event, and Detector tables.

PROGRAM TABLE Sets up and stores the Method table.


Screen keys

The five unlabeled screen keys are directly below the display screen. Labels appearing at the bottom of displayed screens, directly above each screen key, indicate the keys’ functions, which vary from screen to screen.Some screens, like the OPERATE METHOD and OPERATE GRADIENT screens, display the MORE screen key. Pressing this key displays a secondary set of screen keys that apply to those screens.Some screen keys act as toggles. When you press a toggle key, the screen label changes to display an alternate function. For example, when you press LOCK KEYBOARD, the screen label changes to UNLOCK KEYBOARD.The screen key labels might also indicate system status. For example, the leftmost key label on the OPERATE METHOD screen reflects the operating mode of the system as it changes from IDLE to Inject WAIT to RUNNING.The figure below shows the location of the label relative to the key.

Clear line screen key and its on-screen label

To initiate a function, press the screen key directly below its on-screen label.

Set Up Direct Operate Program Program Method Method Table

Help

Screenfunctionlabel

Clear linescreen key

Clearline

1-18 Overview

Displaying helpAll parameters in the Waters Prep Series have associated Help. A HELP screen key appears in the lower-right corner of every screen.

To display Help for a topic:

1. Move the cursor to the field for which you want help.

2. Press the HELP screen key. A Help screen topic appears that offers parameter specifications.

3. Press the RETURN screen key to quit Help.

Shutting down the systemIf you do not plan to use the Waters Prep Series system for overnight or longer, shut the system down, observing these precautions:

• Do not leave buffers in an unused system. First, flush the lines with Milli-Q® HPLC-grade water. After cleaning the lines of any salts, flush the system with Milli-Q® HPLC-grade water containing at least 10% methanol. (Remove the column if it is incompatible with this storage protocol.)

• Flush the plunger wash system with fresh solvent. Disconnect the plunger wash outlet line from the plunger wash pump (at the pump head). Using a syringe filled with fresh solvent, push the new solvent through the plunger wash lines. This ensures the cavity behind the pump seals is cleared of buffer while the pump remains idle.When you complete this operation, reattach the outlet line to the pump head.

Shut down the system with the On/Off switch on the front panel of the controller.

Configuring the controllerThis section helps determine which controller configuration to use.

Caution: Water or buffers that remain in the system can promote microbial growth.


The PrepLC controller, which Waters shipped in PowerLine configuration, allows maximum performance and programming flexibility. The Gradient configuration is usually used when an IEEE-based data system like MassLynx or Empower controls the Prep system. Nevertheless new users might find the Gradient controller configuration easier to operate initially.

PowerLine configuration

In its PowerLine configuration, the controller performs as follows:• Interfaces with PowerLine-compatible detectors (Waters 2487 and 2414

Detectors), PowerLine autoinjectors (Waters 717plus), data systems (MassLynx or Empower), and integrators (Waters 746 Data Module).

• Automatically sets up and controls all gradient, injection, external event, and detector parameters directly from the keyboard.

• Uses a Method table that links time-based control tables. These include a Gradient table, Event table, and Detector table.

The controller communicates with the PowerLine modules (autoinjector and detectors) through the IEEE-488 interface. It communicates with the Waters 746 integrator through the RS-232 serial interface.OperationBy specifying values for these parameters, you can set the PowerLine controller for product-specific operation with PowerLine detectors and autosamplers:

• Detectors– Detector filter constant– Autozero– Polarity– Absorbance units full scale (AUFS)– UV wavelength (λ)– Sensitivity

• Autosamplers– Syringe draw rate– Inject delay– Standards (how many and how often)

1-20 Overview

– Methods specifying vial numbers– Number of injections from each vial– Injection volume

The controller can contain 15 sets of Method tables. Each set of tables can contain these individual tables:

• Gradient – Includes up to 15 operational steps that define the varying solvent composition, flow rate, time range, and rate of change for each phase of the gradient.

• Event – Includes up to 15 operational steps to control output signals to external devices, internal alarms, sparge rate, and linking operation to other tables.

• Detector – Includes up to 15 operational steps to change wavelength and attenuation during a run.

You can create a Method table, a time-based program that can include up to 48 individual steps. Each step accesses a set of Gradient/Event/Detector tables. These are among the tasks the Method table can perform:

• Equilibrating the system for a specific period of time• Switching solvents• Purging the autoinjector• Purging the 2414 RI Detector flow cell• Performing a series of small-scale or large-scale injections• Cleaning up following buffer use• Lowering the system flow rate after an analysis

Having programmed the Method table, you can instruct the system to start at any step in the process.Screen layoutThe figure below shows the screens associated with the system’s PowerLine configuration.


t

mr

PowerLine controller screens

Gradient controller configuration

The controller allows these tasks in its gradient configuration:• Controls isocratic or gradient solvent flow• Controls output signals to external devices, internal alarms, sparge rate,

and linking operation with other tables while running a gradient• Programs and runs Gradient tables that define varying solvent

composition, flow rate, time range, and rate of change for each phase of the gradient

In gradient configuration, the controller does not communicate with any PowerLine modules. Nevertheless, it can operate with any Waters data system or integrator connected to the IEEE or RS-232 ports.OperationThe Gradient controller stores 15 sets of tables. Each set consists of a Gradient and an Event table. These run together during gradient operation. The Gradient and Event tables can each include up to 15 operational steps.

PowerLinecontroller

Setup Direct Opermeth

Progmeth

Progtable

Pumpsetup

screen

Directcontrolscreen

Initiateop methscreen

Programmethodsscreen

Programgradienscreen

Programeventscreen

Progradetectoscreen

Operatemethodsscreen

Autoinjsetupscreen

Systemconfigscreen

Pumpconfigscreen

Programeventscreen

1-22 Overview

You can program separate tables to perform these tasks:• Equilibrating the system• Switching solvent• Cleaning up following buffer use• Lowering the system flow rate after a run

You can program a table so that, once it has run, it initiates the running of another. You can also link a table back to itself to repeat a method.Screen layoutThe figure below illustrates the layout of screens for the system’s Gradient controller configuration.

Gradient controller screens

See the Waters Prep Series Operator’s Guide for details about configuring and operating the Gradient controller.

Gradientcontroller

Setup Direct Oper Meth Prog meth Prog table

Pumpsetup

screenIsocraticscreen

Initiateop gradscreen

Program gradientscreen

Program event

screen

Pumpconfigscreen

Operategradientscreen


1-24 Overview

2 Installing the System

Contents

Topic PageSetting up and inspecting 2-2Electrical requirements 2-5Assembling the system 2-7Connecting a helium tank to the fluid handling unit 2-30Setting up the plunger wash 2-33Installing sample injection options 2-40Connecting a fraction collector 2-42Installing analytical and preparative columns 2-43Making detector connections 2-48

2-1

Setting up and inspecting

PrepLC system

The main components of the PrepLC system arrive in three cartons, which together contain the following:

• PrepLC controller (electronics unit)• Fluid handling unit and Startup Kit• Shelf unit with injector panel• Rheodyne 7725i injector, when purchased separately• Gradient mixer

Accessories such as columns, integrators, detectors, etc. are packed separately.

Delta-Prep system

The main components of the Delta-Prep system arrive in four cartons, which together contain the following:

• PrepLC controller (electronics unit)• Fluid handling unit and Startup Kit• Shelf unit• Delta-Prep system rack with column clamps and injector valves

Accessories such as columns, integrators, detectors, etc. are packed separately.

Unpacking and installing the system1. Open the cartons, checking the contents of each against the packing slip

to confirm it contains its full complement of items.

2. Remove the system components from their respective cartons, and place them at the installation site.

3. Inspect all items for shipping damage. Immediately report any damage to both the shipping company and your Waters Technical Service representative.

2-2 Installing the System

Selecting the siteThis section describes the physical and environmental requirements of your Prep system.

PrepLC system

Install the system away from direct sunlight and heating and cooling vents. These additional conditions must apply:

• Temperature ranges from 4 to 38 °C (39 to 100 °F).• Relative humidity ranges from 20 to 90% noncondensing.• Adequate bench space and capacity to support 125 lbs (56.25 kg) plus

the detector’s weight.• Clearance between the unit’s rear panel and a wall or other structure is

at least 6 inches (15.24 cm). This allows adequate ventilation and access to cable connections.

• Vibration is negligible.• The shelf unit rests securely on the bench.• The mixer in the PrepLC system has to be mounted vertical with the

flow direction from bottom (inlet) to top (outlet).

Delta-Prep system

Install the system away from direct sunlight and heating and cooling vents. These additional conditions must apply:

• Temperature ranges from 4 to 38 °C (39 to 100 °F).• Relative humidity ranges from 20 to 90% noncondensing.• Adequate bench space and capacity to support at least 175 lbs (79.37 kg)

plus the detector’s weight.• Clearance between the unit’s rear panel and a wall or other structure is

at least 6 inches (15.24 cm). This allows adequate ventilation and access to cable connections.

• Vibration is negligible.• The shelf unit rests securely on the bench.

The figure below indicates system dimensions.

Setting up and inspecting 2-3

Waters Delta-Prep system dimensions

To install the shelf unit:

1. Bolt the column rack to the left side of the shelf unit with the 12 screws and nuts that accompany the column rack. Place the assembly in position on the bench.

2. Place the fluid handling unit in the bottom section of the shelf unit (see the figure “Waters Delta-Prep system dimensions” on page 2-4).Tip: Before installing the controller, determine whether you must reconfigure the operating frequency and/or operating voltage selections as outlined in “Electrical requirements” on page 2-5. Make any necessary changes, then proceed with step 3.

3. Slide the controller into the top slot, leaving the middle slot for the detector.

Caution: Do not stack fluid handling units and devices on top of electronic units without providing leak protection.

�� !�

21.5 in (54.61 cm)

23.7 in (60.2 cm)22.0 in (50.88 cm)


Electrical requirements

This section describes the Prep system’s electrical configurations.

Electrical conditionsThe system requires these electrical conditions:

• Grounded AC source, 100/120 or 220/240 voltage, 50/60 Hz, 500 VA• No nearby source of electronic noise such as that cause by electric

motors or arcing relay contacts• No abrupt load fluctuations• Proper fuses installed

Voltage selectionsYou can adapt the system to operate at the nominal voltages shown in the table below.

Changing the voltage

The system is preset for 120-VAC operation.

To change the voltage:

1. Ensure the front panel switch is to set to Off and the instrument disconnected from the electrical source.

Warning: To avoid electric shock, make sure the power cord is disconnected from the rear panel of the instrument before performing this section’s procedures.

Operating voltage ranges

Nominal voltage Voltage range Fuse (Amps) Power (VA)100 ±10% 4 400120 ±10% 4 400220 ±10% 2 400240 ±10% 2 400

Electrical requirements 2-5

2. Remove the power cord from its connector at the rear panel, and pry the power connector cover with a flat-blade screwdriver.

3. Remove the voltage selection barrel.

4. Select a suitable voltage setting.

5. Replace the voltage selection barrel so that the voltage you selected will show through the window when you close the cover.

Changing the voltage setting

6. Waters ships all controllers with 4-A fuses for 110/120-VAC operation. If you operate the unit with 220/240 VAC, change the fuse (see step 7). If you do not need to change the fuse, proceed to step 8.

7. To change the fuse, pull each fuse holder out, as though opening a drawer. (Spare fuses are included in the Startup Kit.)

8. Install the correct fuse in the holder, and slide it back in place. The arrow on each fuse holder points upward when it is in the correct position.

9. Close the power connector cover.

10. Plug the power cord into its connector at the rear panel.


Assembling the system

This section explains how to connect the Rheodyne injectors and the optional M-1000 PrepPak module to the controller and the controller to the fluid handling unit. It also describes additional, rear panel connections.

Ensure the voltage, frequency, and fuse selections match the power source as outlined in “Voltage selections” on page 2-5.The terminal strips, shipped with the controller, are attached to the terminal strip receptacles. The top and bottom terminal strips are marked for corresponding receptacles.

Connecting Rheodyne injectorsThe Rheodyne injector generates a signal output as it injects the sample. Connect the cable for this output to the Inject terminal on the controller’s rear panel.Tip: Cable polarity is irrelevant.

PrepLC system

To connect the Rheodyne injectors:

1. Plug the inject signal cable (Startup Kit) into the Rheodyne injector’s electrical connector. The figure “Injector electrical connector” on page 2-8 illustrates the location of the injector’s electrical connector. Route the cable along the top of the shelf and through the hole at the rear of the top tray.

2. Connect the two leads from the inject signal cable to the Inject and Gnd terminals.

Caution: Externally generated voltages can damage the instrument. Shorting the 12-VAC terminal to ground or and event output causes the +12 VAC fuse to blow.

Assembling the system 2-7

Injector electrical connector

Delta-Prep system

To electrically connect the Rheodyne 7725i and 3725i injectors to the controller, connect the leads from the inject signal cable, at the rear of the column rack, to the Inject and Gnd terminals.

Injector connections to the controller

Injector electricalconnector

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

Rheodyneinjector


Connecting the M-1000 PrepPak module (optional)The M-1000 PrepPak module generates an output that causes the controller to abort pump action when radial compression in its chamber falls below 650 psi.

To connect the optional M-1000 PrepPack:

1. Connect the signal output to the Stop Flow terminal on the controller’s rear panel.

2. Connect the M-1000 PrepPak module connector cable from the system interface terminals on the rear of the module to the Stop Flow input terminal on the controller’s rear panel.

If you are not using the M-1000 PrepPak module, see “Connecting additional devices” on page 2-9.

Stop flow connection from the optional M-1000 PrepPak module

Connecting additional devicesYou can connect devices to other terminals on the controller’s rear panel to access system features. This section describes connections to the Chart, Pressure, and S1 to S4 terminals.

Chart output

The Chart +/– terminals carry a 0- to 10-mV signal that represents a solvent’s flow rate or percent composition of a solvent. The second channel of an

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

PrepPAK Module

SYSTEMINTERFACE

CONNECTOR

M-1000


integrator or chart recorder plots the signal. The full-scale range of the chart outputs is +10 mV full scale.Tips:

• The integrator must invoke expanded memory to use the second channel on the 746 integrator to plot pressure or percent composition. It plots the second channel after the first channel finishes.

• The output signal reflects the solvent composition at the proportioning valve, which differs from its composition inside the column or detector cell.

Plotting

1. Connect the leads at one end of the event cable (Startup Kit) to the Chart+ and Chart– terminals.

2. Connect the leads at the other end of the cable to the corresponding +/– terminals for the second pen on the integrator or recorder.

Pressure and chart connections to the integrator or chart recorder

TP0

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

+

–

Integrator orchart recorder

Integrator orchart recorder second channel

Second channel


Pressure output

The Pressure output monitors pump pressure during ramp testing and other service procedures. It also monitors how different solvents affect pressure during various phases of gradient operation. The full scale range of the pressure output is +10 mV.

Plotting

1. Connect the leads at one end of the event cable (Startup Kit) to the Pressure+ and Pressure– terminals (see the figure “Pressure and chart connections to the integrator or chart recorder” on page 2-10).

2. Connect the leads at the other end of the cable to the corresponding +/– terminals for the second pen on the integrator or recorder.

External devices requiring contact closure or TTL switches

To connect external devices requiring contact closure or TTL switches:

1. Connect one lead from a two-conductor cable between the device and controller to a Gnd terminal on the controller. Connect the corresponding lead at the other end of that cable to a ground terminal on the device.

2. Connect the other leads on the cable to an available switch (S1 to S4) and the input terminal on the device.


External device connections

+12 VAC auxiliary power

To connect external devices requiring +12 VAC auxiliary power:

1. Connect one end of a two-conductor cable between the device and the controller to the appropriate +/– terminals on the device.

2. Connect the positive lead to the +12 VAC terminal on the controller and the negative lead to an available switch (S1 to S4) terminal.Tip: The maximum current available from the 12-VAC power source, divided among all the devices in use on S1 to S4, is 1.5 A. The maximum current capacity of any single event (S1 to S4) is 1.0 A.

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

+5 VAC switch closure or TTL

device


External device connections requiring auxiliary power

Connecting accessories

To connect a PowerLine autoinjector (Waters 717plus), see “IEEE-488 cables with external PowerLine devices” on page 2-17. To connect autoinjectors other than PowerLine devices, see “Using an autoinjector” on page 2-41.To connect fraction collectors, see “Connecting a fraction collector” on page 2-42. To connect PowerLine detectors (Waters 2487 and 2414), see “IEEE-488 cables with external PowerLine devices” on page 2-17. To connect detectors other than PowerLine devices, consult the documentation supplied with the detector.

Attaching the interface cable and power cordYou need to make the following cable connections for PrepLC controller use:

• Attaching the pump interface cable• Attaching the AC power cord

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

External device


Attaching the pump interface cable

To attach the pump interface cable:

1. Shut down the controller.

2. Connect the 37-pin interface cable to the pump interface connector on the controller rear panel (see the figure “Pump interface cable and power cord locations” on page 2-14).

3. Connect the other end of the 37-pin interface cable to the rear of the pump.

Pump interface cable and power cord locations

Caution: To avoid damaging the PrepLC controller and/or pump, make sure the controller power switch is turned off before performing this procedure.

AC powerconnection

Pump interfacecable

PrepLCcontroller

Fluidhandlingunit


Attaching the AC power cord

To attach the AC power cord:

1. Insert the D-shaped connector end of the power cord into the power receptacle on the PrepLC controller rear panel (see the figure “Pump interface cable and power cord locations” on page 2-14).

2. Insert the other end of the power cord into a wall outlet. For more information, see “Voltage selections” on page 2-5.

Connecting IEEE-488 interfacesThese are the different types of IEEE-488 interface connections you can make with the PrepLC controller:

• Making IEEE-488 connections with data systems• Making IEEE-488 connections with external PowerLine devices • Setting IEEE-488 addresses• Performing the IEEE-488 startup sequence

IEEE-488 cables with data systems

When you control the Prep system from a Waters data system (MassLynx or Empower workstations), use the IEEE-488 interface to receive information from the data system. Set up the controller for gradient operation. The data system operates as the system controller on the IEEE-488 interface.

To make IEEE-488 connections with data systems:

1. Connect the single receptacle end of the IEEE-488 cable (supplied with either the Prep system or the Waters data system) to your data system as follows:

• Empower workstation – Attach the cable to the busLAC/E™

(Laboratory Acquisition and Control/Environment) card (see the figure “Stacked IEEE-488 connections in a Waters workstation” on page 2-16).

• MassLynx workstation – Attach the cable to the GPIB (IEEE) card (see the figure “IEEE-488 connections in a Waters data network” on page 2-16).


Stacked IEEE-488 connections in a Waters workstation

IEEE-488 connections in a Waters data network

PrepLCcontroller

(as gradientcontroller)

717plusAutosampler

2487 Detector

busLAC/E card (Empower) or GPIB card (MassLynx)

IEEE-488connector

IEEE-488 cable

Data system

PrepLCcontroller

(as gradientcontroller)

PrepLC controller(as gradient controller)

LAC/Emodule

Data workstation

717plusAutosampler

2487 UVDetector

Thin wireethernet cable

IEEE-802.3ethernet connectorIEEE-488

cable

IEEE-488connector


2. Connect the other end of the cable, using a stackable connector for interconnecting additional instruments, to the IEEE-488 connector on the controller’s rear panel.

3. If you are using the controller as part of a networked multi-system, connect a second IEEE-488 cable to the stackable connector on the controller. Connect the other end of the cable to the IEEE-488 port on the next system component.

4. Repeat steps 2 and 3 for each additional component.Tip: The maximum total cable length between all IEEE-488 devices is 65 feet (20 meters). The maximum recommended cable length between any two IEEE-488 devices is 13 feet (4 meters). Longer total cable lengths can cause intermittent IEEE-488 communication failures.

5. Ensure all IEEE-488 cable screws are fastened to finger tightness.

6. Specify a unique IEEE-488 address, from 2 to 29, for each device connected on the IEEE-488 bus (see “Setting IEEE-488 addresses” on page 2-18).

Perform the correct IEEE-488 startup sequence for the data system.

IEEE-488 cables with external PowerLine devices

When controlling external PowerLine devices from the Prep system, use the IEEE-488 interface to communicate with the PowerLine devices (where the PowerLine controller is the controller on the IEEE-488 interface). Set up the PrepLC as a PowerLine controller.To ready the PrepLC controller as a PowerLine controller, see the Waters Prep Series Operator’s Guide.Supported PowerLine devices include the following:

• Waters autosamplers (for example, 717plus)• Waters detectors (for example, 2487 and 2414)

To make IEEE-488 connections with external PowerLine devices

1. Connect the single-receptacle end of the IEEE-488 cable (supplied with either the Prep system or the data system) to the IEEE-488 connector on the controller’s rear panel.


2. Connect the other end of the cable, using the stackable connector for interconnecting additional instruments, to the IEEE-488 connector on the next PowerLine instrument.

3. If using a multicomponent configuration, connect a second IEEE-488 cable to the stackable connector on the PowerLine instrument. Connect the other end of the cable to the IEEE-488 port on the next PowerLine component.

Setting IEEE-488 addresses

You must set a unique IEEE-488 address, between 2 and 29, for each device on the IEEE-488 bus so that the device is recognized on the IEEE-488 interface. When setting the address, note these instructions:

• When using a data system (MassLynx or Empower workstation), set the IEEE-488 address of the Gradient controller from the application’s Pump Configuration screen.

• When using the controller as a PowerLine controller, you do not set the IEEE-488 address of the Prep system pump. In the PowerLine system configuration, set the IEEE-488 address for each of the other PowerLine instruments through the instrument software or DIP switches. A previously set address has no effect in PowerLine configuration.

Recognizing IEEE-488 addressesAfter you set the address for an IEEE-488 device, cycle the device off and then on again. The new address is not recognized until the device undergoes its calibration or diagnostic routines (performed at startup).Startup sequence for IEEE-488 devicesThis section describes the correct IEEE-488 startup sequence when you use these devices:

• Prep system with a data system (PrepLC controller in Gradient mode)• PrepLC controller as a PowerLine controller

Caution: Perform the following steps to ensure that the data system or PowerLine controller recognizes other devices on the IEEE-488 interface.


Data system startup sequenceTip: This sequence requires you to start all devices in your data system operating before you start the computer. As you start each component, wait briefly to allow its internal diagnostic tests to run. This ensures each module functions and quickly identifies failures.

Perform the following startup sequence when you use the Prep system with a data system:

1. Start all equipment not controlled by the data system.

2. Start equipment controlled by the data system that is not under direct IEEE-488 control:• Empower workstation – All equipment controlled through the I/O

distribution box (busSAT/IN modules and connected analog detectors).

• Client/Server – All equipment controlled through the busLAC/E module (SAT/IN modules and connected analog detectors).

3. Start the pump, establishing eluent flow. Then start all other devices controlled through the IEEE-488 bus.Tip: You cannot use the Run Samples window, nor run methods or method sets, until all devices on an Empower system’s IEEE-488 bus are calibrated and operating. However, you need not start instruments not assigned to a system or assigned to one you do not intend to use.

4. Start the printer and monitor.

5. Start the computer.

PowerLine controller startup sequence

1. Start all equipment not controlled through the IEEE-488 interface.

2. Start all equipment controlled through the IEEE-488 bus.

3. Start the PrepLC controller.

Shutdown sequence for IEEE-488 devices

If you do not expect to use the Prep system overnight, or for longer periods, shut down the system observing this sequence:

1. Ensure the system is purged of salts.


2. Stop your chromatographic run (if necessary).

3. Press the On/Off switch on the controller’s front panel to shut down the system.

4. Shut down the other IEEE-488 devices.

Connecting to the Waters 746 data moduleYou can use the RS-232 interface to connect the controller to a Waters 746 Data Module. The interface transmits this data using:

• Control commands from the data module to the controller• Commands and report information from the controller to the data

moduleTip: The interface transmits no chromatographic data.

Connecting the RS-232

To connect the RS-232:

1. Connect the 25-pin connector (provided with the data module) to the rear panel of the data module.

RS-232 connections with a Waters 746 data module

2. Connect the other end of the cable to the RS-232 telephone jack connector on the controller’s rear panel.Tip: When connecting the 746 Data Module to the controller, you must also attach a cable from the Inject terminal to the 746 to coordinate the run start signals of the two systems.

746data module

PrepLCcontroller

(as PowerLinecontroller)

Run start signalfrom inject terminal

25-Pinconnector

RS-232 cable

Telephoneconnector


Liquid line connectionsThe Waters Prep system Startup Kit includes a variety of tubing and fittings for connecting to the fluid handling unit, injector, detector, and column. Refer to the list in the Startup Kit for information on all included tubing and fittings.This section describes how to select and cut tubing and how to install fittings.

Considerations

• When you install or change tubing observe safe laboratory practices regarding solvent handling. Refer to the Material Safety Data Sheets for the physical and chemical properties of the solvents you use.

• Always stop the flow and vent line pressure with the purge valve before loosening a fitting.

• Minimize dead volume by using appropriate tubing in all parts of the system. Cut tubing and install fittings as this section describes.

• Do not nick, kink, or sharply bend the tubing. This can restrict flow.Tip: Repeated bending of tubing causes tubing to fail.

• Subjecting the system to a wide temperature fluctuations can loosen fittings. Before operating the system at a new temperature, allow the system’s temperature to equilibrate with the room temperature. When you first start the system, check all connections for leaks, and retighten fittings as needed.

Tubing selection

Various tubing sizes for readying the system for large-scale operation accompany the Prep system. Use these guidelines when you select tubing to connect the Waters 2487 Detector:

• For flow rates up to about 10 mL/min in a small-scale operation, connect the column outlet to the reference side of the detector cell with 0.009-inch ID tubing.Tip: This varies with solvent viscosity.

Caution: Do not attempt to relieve pressure by loosening a fitting. This can cause tubing to separate from the fitting and spray solvent.


• For flow rates greater than ~10 mL/min and for large-scale operation, connect the column outlet to the sample side of the detector cell with 0.040-inch ID tubing.

The table below shows tubing sizes for the various flow cells used in Waters UV detectors.

Recycle and solvent inlet linesThe Startup Kit contains precut and prefitted lengths of PTFE tubing. These are solvent recycling and solvent inlet lines, and their respective sizes are 0.040-inch ID and 0.019-inch ID.Fluid pathwaysRefer to the following illustrations when plumbing your Prep system.The figure below shows tubing sizes for the standard PrepLC system’s fluid pathways. This system is used for both small-scale and large-scale injections.

Flow cell tubing sizes

DetectoreFlow cellpath length(millimeters)

ID inlet(inches)

ID outlet(inches)

2487AnalyticalSemi-prepVariable path lengthAutopurification:• Analytical• Preparative

1030.15 to 3

11

0.0090.040.04

0.0090.02

0.0090.040.04

0.040.04

2996AnalyticalSemi-prepVariable path lengthAutopurification:• Analytical• Preparative

1030.15 to 3

0.50.5

0.0090.040.04

0.0090.02

0.0090.040.04

0.040.04


PrepLC system fluid pathway

The figure “Delta-Prep system fluid pathway” on page 2-24 shows the fluid pathway in the Delta-Prep system.

Large-scale injector

Purgeoutlet

PrepLC pump

Plunger sealwash pump

Plunger sealwash solvent

B C

A D


Delta-Prep system fluid pathway

Cutting stainless steel tubing

When cutting tubing, avoid angled cuts. These create a bad fit between the tubing and connector port, which creates eddies (unswept volume) at the junction.

To cut stainless steel tubing:

1. Estimate or measure the length of tubing required to connect the components. Allow slack.

2. Use a cutting file to circumscribe the tube at the point you want to cut it.

3. Hold the tubing on either side of the scribe mark with two cloth-covered pliers, and gently work it back and forth until it separates.

Caution: Do not pull tubing tightly around sharp corners.

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Small-scale injector


Scale select

Purgeoutlet

Fractionvalve

Fractions/waste/recycle

Plunger sealwash pump

Solvents

Plunger sealwash solvent

Recycle to pump

PrepLC pumpB C

DA

valve

valve


Installing fittings

To install fittings:

1. Slide the fitting and then the ferrule (large end of the taper first) over the end of the tubing.

Compression screw assembly

2. Bottom the tubing and ferrule in the fitting seat.

3. As you press the tubing into the connector, tighten the compression screw 3/4 of a turn beyond finger tightness. Tightening the compression screw compresses the ferrule, fusing it onto the tubing in the correct position.Tip: Subsequent connections to this fitting will not require the extra 3/4-turn.

Cutting PTFE tubing

To cut PTFE tubing:

1. Estimate or measure the length of tubing required to connect the components. Allow slack.Tip: Do not pull tubing tightly around sharp corners.

2. Use a razor blade to cut the tubing squarely, taking care to avoid crushing it.

3. Inspect the cut for burrs or scratches. Make sure the tubing is not pinched closed.

Caution: Avoid angled cuts, which create eddies (unswept volumes) at the connection. Tubing cut at an angle fits poorly into the connector or port and can leak or separate.

FerruleTubing(Cut must be perpendicular to the tubing axis.)


Installing fittings

To install fittings:

1. Slide the polymeric compression screw and then the ferrule (small end of the taper first) over the end of the tubing.

Tefzel compression screw assembly

2. Bottom the tubing and ferrule into the fitting seat.

3. As you press the tubing into the fitting, hand-tighten the compression screw. Tightening the compression screw compresses the ferrule, fusing it onto the tubing in the proper position.

Fluid handling unit connections

This section describes these connections to the fluid handling unit:• Solvent reservoir setup• System sparge and helium setup• Seal wash setup• Rheodyne injector connections

The figure “Waters Prep series fluid handling unit” on page 1-6 shows the fluid handling unit.Solvent reservoir caps

Caution: Limit the solvent system to 5 psi pressure when you use pressurized solvent containers.

Compression screw1/4-28 x 1/8-inch tube

1/8-inch tubeReverse ferrule1/8-inch tube


Solvent reservoir caps help maintain sparged conditions with the reservoirs and ensure the delivery system’s optimal performance. Each cap has three feather-edged holes, which create a positive seal around the solvent, sparge, and vent tubes.

Solvent reservoir caps

The caps provided with the system fit 4-liter reservoirs, which Waters recommends. You can, however, obtain caps that fit 1-liter reservoirs.Tip: Choose solvent reservoirs onto which caps fit snugly.

To prepare the reservoir:

1. Position the reservoir in a convenient location, preferably at the same level as the fluid handling unit.

2. Remove protective wrapping from these nine PTFE tubes:• Solvent A, Solvent B, Solvent C, and Solvent D• Sparge A, Sparge B, Sparge C, and Sparge D• Wash Inlet

3. Uncoil the tubing, and train it to the right of the fluid handling unit.Solvent and sparge tube connections

The Startup Kit contains L-shaped tubing sections and 60-µm eluent filters. You can use either a section or filter on the ends of the four solvent inlet lines. The sections weigh the ends of the inlet tubing to keep it at the bottom of the larger solvent reservoirs but do not filter the solvent that goes to the pump.

Caution: Do not put reservoirs on top of the cabinet. Leakage can damage the system.


The filters press onto the ends of the solvent inlet lines. Their large-pore size and surface area help the pump operate effectively at preparative flow rates.

To connect a solvent reservoir to the fluid handling unit:

1. Push a labeled solvent tube through a hole in the cap of a correspondingly labeled reservoir.Tip: Push tubing far enough into the solvent reservoir to reach the bottom.

Solvent reservoir filter and diffuser placement

Fume hood

Vent tube

Diffuser

L-shaped tube

Eluent filter


2. Attach an eluent filter or L-shaped tube (both in Startup Kit) to the solvent line. The tubing must stop against a ridge inside the eluent filter.

3. Repeat step 1 for the sparge tube for that reservoir.

4. Attach a diffuser to the sparge line.

5. Repeat steps 1 through 4 for all solvent and sparge tubes.Vent tube connection

To connect the vent tube:

1. Cut a length of PTFE tubing (Startup Kit) to extend from a solvent reservoir to a fume hood. Push one end of the tubing about 1 inch into the remaining hole in the reservoir cap. This tubing serves as the reservoir’s vent.

Caution: Forcing tubing into the filter by pushing it beyond this ridge can create these problems:• Reduction of the filter area, which at higher flow rates can starve the

pump of solvent.• The inlet tube’s sharp edge can pare material from the inside the

filter, creating particles that can be sucked into the inlet line. These particles can foul the gradient proportioning valve, the pump check valves, the injector, or some other part of the system.

Warning: • Personal harm could result from improperly vented solvent/sparge

reservoirs. Always vent solvent vapors to a fume hood. Use particular care with a volatile solvent in a cold room, refrigerator, or other small, sealed environment.

• Sparging organic solvents like acetonitrile and methanol into the room air releases harmful vapors. If you use organic solvents, run the solvent reservoirs’ vent tubes to an exhaust hood to safely release vapors.

Caution: Make sure the end of the vent tubing inside the reservoir remains above the solvent level. Otherwise, solvent will be forced from the reservoir through this vent tube.


2. Connect the other end of the vent tube to an eluent filter (see the figure “Solvent reservoir filter and diffuser placement” on page 2-28).

3. Place the filter end of the vent tube in a fume hood.

4. Insert the solvent reservoir cap into the top of the reservoir.

5. Repeat steps 1 to 4 for each solvent reservoir.

Connecting a helium tank to the fluid handling unit

This section describes how to connect a helium tank to the fluid handling unit.Tip: The system does not include the high-pressure gas regulator required to connect the sparge inlet to a helium tank. The Startup Kit includes the shutoff valve, tubing, and other fittings for connecting the sparge inlet to a 50 to 90 psi helium supply.Helium sparging reduces the total dissolved gas in the solvent reservoirs and maintains that condition during operation. Use an ultra-pure carrier (UPC) grade of helium to prevent solvent contamination.

Helium specificationsRegulate the supply source to between 50 and 90 psi (3.4 and 6.1 atm) before connecting to the sparge inlet connection (see the figure “Sparge gas inlet connection” on page 2-33).

The required specifications for UPC-grade helium are as follows:• Less than 5.0 Mppm (molar parts per million) nitrogen• Less than 5.0 Mppm oxygen• Less than 1.0 Mppm total water• Less than 0.5 Mppm total hydrocarbon

Caution: Using an inferior grade of helium can void your system’s warranty.


SpargingSparging minimizes dissolved gases in the solvent. This decreases the potential for degassing when different mobile phases mix in the gradient proportioning valve. The helium flow through the system is as follows:

• The helium is introduced into the solvent through a diffuser, which disperses the gas into small bubbles. The bubbles increase sparge efficacy by increasing the surface area of solvent exposed to helium.

• A reduction in total dissolved gas occurs as the pure helium equilibrates at the gas-liquid interface.

• The displaced gases are carried to the surface and then expelled through the reservoir vent.

Tip: For more information on this process, see Appendix D.

Connecting compression fittingsUse 1/16-inch stainless steel tubing with an ID of at least 0.020 inches (preferably 0.040-inch ID) to connect the fluid handling unit with the helium supply. Connect each end of the tubing with compression fittings. See “Liquid line connections” on page 2-21 for details.

Connecting the helium supply

Shutoff valve assemblyAssemble the shutoff valve and sparge tubing as shown in the figure below. Use these Startup Kit components:

• 0.062-inch × 0.040-inch ID stainless steel tubing• Compression screws• Ferrules

Connecting a helium tank to the fluid handling unit 2-31

Shutoff valve assembly

Fluid handling unit connection

To connect the sparge tubing to the fluid handling unit:

1. Attach the shutoff valve to a helium tank regulator or regulated house supply.You might need fitting adapters at the regulator, depending on the type of source fittings.

2. Regulate the helium supply source to between 50 and 90 psi (3.5 and 6.3 atm).

3. Connect the supply line from the helium source to the sparge fitting on the fluid handling unit rear panel with the shutoff valve and tubing you assembled.

1/8 NPT × Z fitting 0.020 or 0.040-inch IDSS tubing

FerruleCompression

screw

Ferrule

To sparge Fitting

Shutoff valve1/8 NPT × 1/8 NPT

nipple

1/4 NPT M × 1/8 NPTFbushing (optional)

To regulator orhouse helium supply


Sparge gas inlet connection

Setting up the plunger wash

The fluid handling unit incorporates a plunger wash flow path, which carries wash solvent from its reservoir to a cavity behind the high pressure pump seals. The wash solvent is pumped along this pathway whenever the controller operates. Having rinsed the cavity, the wash solvent flows back to the reservoir or to waste. This lubricates the plunger, reducing its wear and flushing away any mobile phase forced past its seal from the high pressure side of the pump head.

Caution: • You must set up the plunger wash when you initially install the fluid

handling unit. Otherwise, the wash seals will fail, ultimately reducing the life of the high pressure seals in the pump heads.

• If the fluid handling unit has run without the plunger wash system installed, replace the wash seals before setting up the plunger wash, or leakage might result.

0.5A FUSE

CAUTIONDO NOT REMOVE CABLE

WHILE UNIT IS POWERED

CONNECT TO 4000

CONTROLLER ONLY

! !

! !

Spargeinlet connection

Setting up the plunger wash 2-33

ConsiderationsUse a solvent that is miscible with your mobile phase and which contains no dissolved salts that could crystallize as it evaporates. Also, empty and refill the plunger wash reservoir with new solvent about once a month (more often when used frequently) if you reuse it.

Connecting the plunger wash tube

To connect a solvent reservoir to the plunger wash:

1. Place the plunger wash pump’s inlet tubing into a reservoir containing a solvent that is compatible with your mobile phase.

Plunger wash connections

2. Connect outlet tubing to the wash fitting at the top of the left pump head. Put the other end of the tubing in a waste container or into the wash solvent reservoir to recirculate the wash solvent.

3. Force several milliliters of wash solvent into the inlet tube of the plunger wash pump, using a syringe. Keep pushing solvent into the tube until it comes out the left pump head and into the outlet tube.

Washer solvent

Inlet tubing

Pump head

Tubing fromwash pump Outlet tube Fluidics drawer


4. Put the inlet tube back into the wash solvent reservoir.

Connecting to the injection panel (PrepLC system)Tip: Refer to the figure “Injector panel (PrepLC system)” on page 1-13 when following the procedure in this section.

Connecting to the purge valve and Rheodyne 3725i injector

The pump outlet connects to the injection panel with 0.020-inch ID tubing. The PrepLC system uses the same size tubing to connect to the injector panel. The Rheodyne injector, designed for large-scale operation, has flow passages of 1.0-mm ID.

To connect the pump outlet to the injection panel:

1. Cut a length of tubing to extend from the pump outlet to the unused fitting on the injector panel’s purge valve, or that goes directly to the column inlet.

2. Install compression fittings on the pump end of the tubing as described in “Liquid line connections” on page 2-21.

3. Tighten the fitting at the pump outlet.

4. Remove the unused nut and ferrule from the purge valve, and install them on the other end of the pump outlet tubing. Attach the tubing to the purge valve and tighten (see the figure “Pump outlet connection to the column rack inlet (Delta-Prep system)” on page 2-39).

Put a waste beaker under the injector’s waste tubing (see the figure “Pump outlet connection to the purge valve (PrepLC system)” on page 2-36).


Pump outlet connection to the purge valve (PrepLC system)

To minimize bandspreading at flow rates less than ~10 mL/min, use 0.009-inch ID tubing between the injector and the small-scale column. Use 0.020-inch ID tubing throughout the system for flow rates above 10 mL/min to maintain adequate backpressure.Tip: Waters offers optional small-scale injector and scale select valves for the PrepLC system. Contact your local Waters Technical Service representative for details.

Connecting a prep column

You can introduce samples for large-scale chromatography through an unused solvent inlet line or the pump’s manual sample inlet. For large-scale operation, connect the pump outlet directly to the column inlet with 0.040-inch tubing.

Waste tubing

Inlet connector

From pump outlet(0.020-Inch ID

SS Tubing)

Purge valve (rear)


Pump outlet connection to the prep column (Large-scale use)

Connecting to the column rack (Delta-Prep system)The 0.040-inch ID and 0.009-inch ID tubing inside the Delta-Prep system’s column rack permit large- and small-scale operation. The scale switching valve isolates one tubing pathway from the other.

��

��

��

Column outlet

To detector

Column inlet

Injector panel

Pump outlet line


System column rack (Delta-Prep system)

Before you connect to the column rack, consult the table below for material types and sizes.

Column rack connections

Column rack connection

Type of tubing Tubing ID Connect to

Inlet from pump

Stainless 0.040 Transducer outlet on pump

Column outlet, small scale

Stainless 0.020 Detector inlet

Column outlet, large scale

Stainless 0.040 Detector inlet

Recycle to pump

PTFE 0.040 Recycle port on manual sample inlet valve

Inlet from detector

Stainless 0.040 Detector outlet

��"�

�� &�'��

Inlet (from pump)

Cabinet latchInlet (from detector)

Small-scale column outletLarge-scale column outlet

Recycle to pump

Fraction outlets selectorFraction outlets 1-8

Purge outlet

Vials

Fractions/waste/recycle valve

Purge/ system valve

Waste

Small/large-scaleswitching valve

Large-scaleinjector

Small-scaleinjector


The fluid handling unit connects to the column with a 0.040-inch ID tubing (Startup Kit) installed between the pump outlet and the Inlet bulkhead connector on the column rack’s front panel.

Pump outlet connection to the column rack inlet (Delta-Prep system)

To connect the pump and column rack:

1. Cut a length of 0.040-inch stainless steel tubing (Startup Kit) that reaches from the pump outlet to the Inlet connector on the column rack’s front panel.Tip: Use excess tubing, forming it into several coils, as shown in the figure “Pump outlet connection to the column rack inlet (Delta-Prep

Purge outlet Stainless 0.040 Waste containerFraction outlets

PTFE 0.062 Fraction containers

Waste PTFE 0.062 Waste container

Column rack connections (Continued)

Column rack connection

Type of tubing Tubing ID Connect to

Drip tray

Column rack

Controller

Drip tray

2487 Detector

Fluid handling unit


system)” on page 2-39. The coils act as a spring, letting the column rack slide open so that you need not disconnect the tubing from the Inlet connector. They also keep the tubing compact and manageable. The additional tubing nevertheless increases system volume, causing additional delay time. At preparative flow rates, however, this volume increase is insignificant and can even improve solvent mixing.

2. Install compression fittings on each end of the tubing (see “Liquid line connections” on page 2-21).

3. Tighten fittings at the pump outlet and column rack Inlet connector.

Installing sample injection options

Installing the small-scale sample injection valve option (PrepLC system)

If you expect to routinely inject sample volumes of less than 2-mL volume, Waters recommends you use the small-scale manual injector valve option kit. This kit includes these components:

• Rheodyne 7725i variable-volume injector valve assembly• 100-µL loop assembly with fittings• Inject start cable assembly• Small/large scale select valve with label• Fittings kit

To install the small-scale sample injection valve:

1. Remove the two blank-off plugs from the injector panel. Pressing them outward, from behind.

2. Place the self-adhesive “small / large scale” label over the opening below the existing “pump / purge” valve.

3. Fit the 2-way select valve in the “small / large scale” opening. The valve must rotate through both positions.

4. Remove the injector handle from the main valve assembly by loosening its two set screws.


5. Fasten the valve to the injector panel, to the right of the existing 3725i valve, using the two supplied mounting screws.

6. Replace the injector handle by tightening the two set screws on shafts flats.

7. Connect the two vent tubes (supplied with the valve) to ports 5 and 6 of the injector valve.

8. Remove the pump outlet tubing from the existing 3725i port 2 adapter.

9. Connect the pump outlet tubing to the two-way select valve inlet.

10. Connect the large scale select valve outlet to the 3725i injector port 2 adapter.

11. Connect the small scale select valve outlet to the new 7725i port 2.

12. Connect column(s) to either valve as appropriate.

Using an autoinjectorTip: Consult documentation provided with the autoinjector for complete information on this product.

Making electrical connectionsSee “Connecting IEEE-488 interfaces” on page 2-15 for information about connecting a PowerLine autoinjector (Waters 717plus).Tip: Position the autoinjector to the left of the system. This makes connecting to the column rack in the Delta-Prep system convenient.

Connecting liquid lines when using a Waters autoinjector

To bypass the Rheodyne injector connecting inlet and outlet lines:

1. Locate the autoinjector to the system’s left side, allowing space to access the solvent reservoirs.

2. Connect the blue line (inlet) from the pump outlet to the autoinjector.

3. Connect the red line (outlet) from the autoinjector to the column inlet.

Installing sample injection options 2-41

Connecting a fraction collector

Tip: See your fraction collector’s accompanying documentation for complete information on the product.This section explains how to make electrical connections when the Waters fraction collector is part of the Prep system.Tip: Position the fraction collector on the system’s left, close to the detector.

Making the remote start connectionFraction collectors need a signal from the injector to indicate when a separation method begins. A cable carries this signal between the remote start input on the fraction collector and the Inject and Gnd terminals on the controller. Use a standard signal cable (Startup Kit) to connect the remote start terminal on the fraction collector to the controller.

To make the remote start connection:

1. Connect the fraction collector’s pin 3 to any vacant Gnd terminal on the controller.

2. Connect the fraction collector’s pin 4 to the Inject terminal on the controller.

Making chart mark connectionsThe fraction collector outputs to a detector’s chart mark input to indicate tube changing during a separation.Connect the fraction collector to the detector’s chart mark input with a standard signal cable (Startup Kit). The fraction collector outputs a signal from terminals 9 and 10 of its I/O connector at every tube change.

To make chart mark connections:

1. Connect pins 9 and 10 on the collector to the appropriate input terminals on the detector. Here are two examples:• Connect to the Mark +/– terminals on the Waters 2487 or 2414

Detector.• Connect to any available Event In and Gnd terminals on the Waters

490 Detector.


2. Program the Mark function in the detector.

Installing analytical and preparative columns

This section describes how to install analytical and preparative columns in both the PrepLC and Delta-Prep systems.

PrepLC systemThe PrepLC system includes clamps and clamp holders for mounting a column vertically onto the shelf unit. The figure below illustrates how to attach a preparative column.

Attaching the column

Installing the column

To install the column:

1. Prepare a length of 0.040-inch ID tubing to extend between the pump outlet connector and the column inlet. Prepare another to reach between the column outlet and the detector.

Column outlet

Column inlet

To purge valve

Pump outlet

From injector outlet

Installing analytical and preparative columns 2-43

2. Install compression fittings on each end of the tubing as described in “Liquid line connections” on page 2-21.

3. Tighten fittings at all connectors.

Using an M-1000 PrepPak module

To use M-1000 PrepPak module:

1. Assemble the M-1000 PrepPak module as described in its operator’s guide.

2. Place the M-1000 PrepPak module beside the PrepLC system.

3. Prepare a length of 0.040-inch ID tubing to extend between the column outlet port (port 3 on the Rheodyne injector connector) and the column inlet on the front of the bottom end cap, and one that reaches between the column outlet on the top end cap (see the figure “Attaching the column” on page 2-43) and the detector.


5. Tighten the fittings at all connectors.

Connecting to a Radial Compression Module (RCM)

To install an 8 × 10 mm (small-scale) or 25 × 10 or 40 × 10 mm (large-scale) RCM:

1. Place the RCM to the left of the PrepLC system.

2. Prepare the inlet tubing.• For an 8 × 10 mm (small-scale) RCM, prepare a length of 0.020-inch ID

tubing that reaches between the column outlet port, port 3 on the Rheodyne injector, and the RCM inlet.

• For an 8 × 10 mm RCM, prepare a length of 0.020-inch ID tubing that reaches between the detector and the RCM outlet.

• For a 25 × 10 or 40 × 10 mm RCM, prepare the length of 0.040-inch ID tubing that reaches between the detector and the RCM outlet.



4. Tighten the fittings at all column connectors.

Column and cartridge preparation

See the Care and Use Manual that accompanies your column or cartridge for specific information about solvent selection and equilibration before use.

Delta-Prep systemThe Delta-Prep system column rack includes the necessary tubing, clamps, and clamp holders for mounting analytical and preparative columns. The figure below shows the tubing configuration in the column rack.

Tubing configuration in the column rack

Small-scale injector


Gradient mixer(small-scale)

Gradient mixer( large-scale)


Installing an analytical column

To install an analytical column:

1. Loosen the clamps at any available column location and slide the column holder apart.

2. Slide the column in place and tighten the clamps to secure it in position.

3. Prepare a length of 0.009-inch ID tubing to extend between the connector labeled Small Scale – To Column Inlet and the column inlet. Prepare another to extend between the connector labeled Small Scale – From Column Outlet and the column outlet.



Installing the prep column

To install the prep column:

1. Loosen the clamps at the prep column location and slide the column holders apart.

2. Slide the column into place, and tighten the clamps to secure it in position.

3. Prepare a length of 0.040-inch ID tubing to extend between the connector labeled Large Scale – To Column Inlet and the column inlet. Prepare another to extend between the Large Scale – From Column Outlet connector and the column outlet.

4. Install compression fittings on each end of the tubing (see “Liquid line connections” on page 2-21).


Using an M-1000 PrepPak module

To use M-1000 PrepPak module:

1. Assemble the PrepPak module as shown in its operator’s guide.


2. Place the PrepPak module inside the column rack, between the prep and analytical column holders.

3. Prepare a length of 0.040-inch ID tubing to extend between the connector labeled Large Scale – To Column Inlet and the column inlet (bottom end cap). Prepare another to extend between the connector labeled Large Scale – From Column Outlet and the column outlet (top end cap).



Installing a small-scale or large-scale Radial Compression Module (RCM)

To install a (small-scale) 8 × 10 mm, or (large-scale) 25 × 10 mm or 40 × 10 mm, RCM:

1. Place the RCM to the left side of the Delta-Prep system. You can, if you want, place it on the column rack floor.

2. Prepare the inlet tubing:• For an 8 × 10 mm (small-scale) RCM, prepare a length of 0.009-inch

ID tubing to extend between the Small Scale – To Column Inlet connector and the RCM inlet.

• For a 25 × 10 or 40 × 10 mm (large-scale) RCM, prepare a length of 0.040-inch ID tubing to extend between the Large Scale –To Column Inlet connector and the RCM inlet.

3. Install compression fittings on each end of the tubing as described in “Liquid line connections” on page 2-21, and tighten the fittings at all column connectors.

4. Prepare the outlet tubing:• For an 8 × 10 mm (small-scale) RCM, prepare a length of 0.009-inch

ID tubing to extend between the Small Scale – From Column Outlet connector and the RCM outlet.

• For a 25 × 10 or 40 × 10 mm (large-scale) RCM, prepare a length of 0.040-inch ID tubing to extend between the Large Scale – From Column Outlet connector and the RCM outlet.


5. Install compression fittings on each end of the tubing as described in “Liquid line connections” on page 2-21, and tighten the fittings at all column connectors.

Making detector connections

This section describes connecting the column outlet to the detector in the PrepLC and Delta-Prep systems for large-scale and small-scale operation.

PrepLC or Delta-Prep systemSmall-scale operation with a Waters 2487 Detector and semi-prep or prep flow cell requires these preparations:

• A 0.009- or 0.020-inch ID tubing and a union to connect the SMALL-SCALE OUTLET, on the front of the column rack, to the detector cell inlet

• An appropriate receptacle in which to put the waste line from the detector

Large-scale operation with a Waters 2487 Detector requires the appropriate flow cell (see the table titled “Flow cell tubing sizes” on page 2-22) and these preparations:

• A 0.020-inch ID or 0.040-inch ID tubing to connect the LARGE-SCALE OUTLET connector to the inlet side of the detector cell

• An appropriate receptacle in which to put the waste line from the detector

Tip: Minimize bandspreading. Keep the tubing length between the column outlet and detector as short as possible by placing the detector on the rack’s center level.

Connecting detector semi-prep cellsThe table titled “Flow cell tubing sizes” on page 2-22 lists the tubing sizes with which the detector’s semi-prep cells are plumbed. Consult it when you connect to the column outlet.The type of operation, small-scale or large-scale, can require the sample to flow on different sides of the detector. When replumbing the column outlet connection to a detector, make sure you reconfigure the signal source so that it


goes to the detector’s signal input. Your detector type determines whether you do this by interchanging two BNC signal cables or by keyboard input.

Using the RI detector with the flow splitter optionTip: When using a Waters 2414 Detector with flow rates above 5 mL/min, use the flow splitter option.The flow splitter eliminates excessive pressure on the RI cell. It diverts about 1% of the flow to the RI detector and, ultimately, to waste. The remaining flow goes to the function valve and fraction outlets or gets divided at the fraction collector.Tip: Consult the flow splitter option’s accompanying instructions for installation details.

Making detector connections 2-49

3 Troubleshooting, Testing, and Maintenance

Caution: To avoid instrument damage, wait about 3 minutes after shutting down your system before you disconnect any interconnecting cables.

Contents

Topic PageTroubleshooting 3-2Performance tests 3-36Maintenance procedures 3-47

3-1

Troubleshooting

You can easily correct many problems with your Prep Series system. If you cannot correct a condition, contact Waters Technical Service.

Contacting Waters technical service

North American customers who experience problems should contact Waters Technical Service at 800 252-4752. All others should call their local Waters subsidiary or Waters corporate headquarters in Milford, Massachusetts (USA).

When you call technical service

To expedite service, provide the following information when you call:• Symptom• Operating wavelength• Detector sensitivity setting (0.001 to 2.00 AUFS)• Flow rate• Type of column• Operating pressure• Sample type• Mobile phase(s)• Waters Prep system configuration• Data system

Troubleshooting steps1. Regard the system from a distance to observe whether something

obvious is causing the problem.

2. Compare current system operation with the performance before the problem started.Tip: Troubleshooting is easier when you know the system’s operating conditions when it works correctly. This step emphasizes the importance of tracking system parameters and performance during normal operation.

3-2 Troubleshooting, Testing, and Maintenance

3. Evaluate parameters in this sequence to rule out simple problem causes:

a. System pressure

b. Baseline

c. Peak retention time

d. Peak resolution

e. Qualitative and quantitative chromatography resultsIsolate the parameter whose behavior varies from its usual performance during normal system operation.

4. Run the performance test for a specific module to quickly determine whether a problem exists.

5. Use the troubleshooting flowcharts in “Troubleshooting flowcharts” on page 3-5 to deduce a symptom’s possible cause.

6. See the table below to determine corrective actions.

General PrepLC troubleshooting

Symptom Possible cause Corrective actionGrinding or shuddering noise from pump - high pressure shutdown not exceeded

Blockage in system before transducer

Clean pump head outlet tubing, outlet check valve and tubing to transducer.

Mechanical problem Call Waters Technical Service.

Pump shutting down, high pressure limit exceeded

High flow rate during injection valve actuation caused pressure spike

Decrease flow rate during injection.

Blockage in system after transducer

Remove components between pump and end of system to determine source of blockage. Eliminate blockage.

Column plugged Clean or replace column.Pump shutting down, low pressure limit exceeded

Leaks, empty reservoirs, or bubbles in the pump head

Eliminate leaks, fill reservoirs, prime pump.

Troubleshooting 3-3

Pump cavitating Solvent reservoirs positioned below solvent proportioning valve

Place solvents on bench beside fluid handling unit.

Leak in tubing connections or loose fittings prior to inlet check valve, causing aspiration of air (fluid leak may not be visible)

Eliminate leak, tighten fittings, replace ferrules.

Solvent inlet filter restricting flow

Clean or replace filters.

RI detector leaking internally or from purge port

Cell damaged – maximum flow rate or pressure of RI cell exceeded

Do not exceed 5 mL/min (2414, 2410, and 410 Detectors). Do not exceed 100 psi for any Waters RI detector.Replace detector flow cell if necessary (see detector operator’s guide).

Peak retention times halved

Incorrect pump head volume entered on PUMP CONFIGURATION screen

For PrepLC operation with 1000-µL pump heads, enter 1000 for pump head volume on the PUMP CONFIGURATION screen.

Unable to enter a flow rate above 150 mL/min

Incorrect pump head volume entered on PUMP CONFIGURATION screen

For operation with 1000-µL pump heads, enter 1000 for pump head volume on the PUMP CONFIGURATION screen.

Pulsing flow from waste line

Dirty check valve(s) Clean check valve(s).

General PrepLC troubleshooting (Continued)

Symptom Possible cause Corrective action


Troubleshooting flowchartsThe troubleshooting flowcharts in this section help isolate possible causes of these problems:

• High system pressure• Baseline noise• Erratic or incorrect retention times• Poor peak resolution• Column

If the troubleshooting flowcharts fail to address your problem, see “Chromatography troubleshooting” on page 3-16.

High system pressure

System pressure reference pointChoice of mobile phase, column, flow rate, and temperature can influence system pressure, and it can vary significantly from one method to another.You must therefore identify a pressure reference point. Each time you start a new method, record the system pressure. Thereafter, to diagnose a problem with system pressure, compare current system pressure to the reference pressure you established.

System controller screen is blank

Screen saver feature active

Press any function key to display the screen.

Increased system backpressure during injection using Rheodyne injector

Normal condition. When the injector is in the Inject position, the sample loop generates additional system backpressure.

To decrease system backpressure, after loading the sample, return the injector to the LOAD position.

General PrepLC troubleshooting (Continued)


Troubleshooting 3-5

ConsiderationsWhen troubleshooting high system pressure, consider these:

• Origin of the high pressure• Whether the pressure increase was gradual or sudden

Location of pressure buildupTo correct the high pressure condition, first isolate the part of the system in which the high pressure originates.Gradual pressure increaseSample or mobile phase particulates can increase pressure gradually over several injections.Sudden pressure increaseThese problems can cause pressure to rise suddenly:

• Particulates in one sample• Hardware malfunction

See the following figure to determine what can cause high system pressure.


High system pressure troubleshooting

No

Yes

Yes

No

Yes

No

Yes

No

High systempressure

Set flow to 0.0mL/min andvent system

Pressurereduced?

Pressurereduced?

Pressurereduced?

Pressurereduced?

Pressurereduced?

Adjust pressure

transducer

Replacepressure

transducer

Remove column.replace w/union.

run system.

Troubleshootcolumn

Releaseinjector outlet

fitting

Troubleshootdetector

Releaseinjector inlet

fitting

Troubleshootinjector

Call Waterstechnicalservice

Yes

Troubleshooting 3-7

Baseline noise

If the baseline is abnormal, determine whether the variation is short or long term. You can eliminate many causes of baseline abnormality by identifying the rate at which the baseline changes.See the following figure to determine what can cause an abnormal baseline.


Abnormal baseline troubleshooting

Baselineabnormal

Noisy?Pump

pressureconstant?

Troubleshootpump

Seechromatographytroubleshooting

table

Cycling?Solvent

not mixingproperly

Make surecorrect

pressure filteris installed

Long-term drift?

Wrong columnequilibration?

Allowcolumn toequilibrate

Detector notwarmed up

Allow detectorto warm up

Seechromatographytroubleshooting

table

Yes No

No Yes

Yes

Yes

No

No

Yes

No

No

Troubleshooting 3-9

Erratic or incorrect retention times

Retention time changesDetermine whether the retention times behave like this:

• Change from run to run• Remain constant from run to run but extend beyond the allowable assay

rangeExamining pressure fluctuations and changesWhen troubleshooting retention time problems, examine system pressure first. Look for these behaviors:

• Pressure fluctuations – Are they short term (with each pump cycle) or long term (over several minutes)?

• Absolute pressure change – Is the pressure higher or lower than the normal operating pressure?

Also, it is important to observe whether the retention time changes act like this:

• They appear suddenly at the end of a series of runs, indicating air is dissolving in the mobile phase, the mobile phase is degrading, or the column is contaminated.

• They occur early in a run and tend to become constant or within range after 3 or 4 minutes, indicating the column is not equilibrated, or that the solvent is improperly degassed and/or sparged.

See the figure “Erratic retention time troubleshooting” on page 3-11 and the figure “Incorrect retention time troubleshooting” on page 3-12 to determine what can cause a retention time problem.


Erratic retention time troubleshooting

Retention timeproblem

No

Yes

Retentiontime erratic?

Ret timeincorrect, but

reproducible?

Yes

No

Are pressurefluctuations sameas during normal

operation?

Chemistry problem?

Yes Yes

No No

Mobile phasedegrading?

Use freshmobile phase

Prime pumpto remove

air bubbles

No

Problemcorrected?

Mobile phaseseparating?

Yes

No

Stir mobilephaseProblem Is

operation

Wrongcolumn

equilibration?Equilibrate

column

No

Yes

Tempfluctuations

Stabilizeenvironment

No

Sparge conditionscausing solventcomponents toevaporate at

different rates

Yes

Suggested actions:

- Decrease sparge

- Use separatereservoir for puresolvents

rate

- Allow pump to make mixture.Mixing does notapply to RI detectors.

Yes

No

YesColumncontaminated?

Suggested actions:-Troubleshoot

column- Improve sample

prep

Yes

No

No

System leaking?

Isolate andtroubleshoot

leak

Troubleshoot pump:- Seals- Check valves- Plunger

Call Waterstechnical service

See next page

Troubleshooting 3-11

Incorrect retention time troubleshooting

Retention timeproblem

Retentiontime

erratic?

Ret. time incorrect, butreproducible?

Are pressurefluctuations sameas during normal

operation?

Possible causes:- Wrong mobile

- Wrong/degraded column

- Incorrect wavelength

phase

Solventproportioningvalve broken?

Troubleshootpump

See previous page

Is pressureconstant, butdifferent?

Is pump setto correct flow

rate?

Change flowrate

Large pressuredrop associatedwith one pump

head?

Is systemleaking?

Isolate/troubleshoot

leak

Purge pump.pressure Still

erratic?

Pump broken;troubleshoot

pump


Yes

No

Yes No

Yes Yes

Yes

Yes

No

No

Yes

YesNo

Yes

NoYes


Poor peak resolution

Before addressing problems with peak resolution, ensure that peaks are eluting at the correct retention time. The most common causes of poor peak resolution can also cause retention time problems.If peak retention time is correct, determine if the poor resolution occurs:

• Throughout the chromatogram• At a single peak pair

In a gradient method, if resolution of early peaks is poor but improves later in the chromatogram, precolumn band broadening might be the problem. If peak resolution is poor throughout the chromatogram, post-column band broadening, or loss of column efficiency, might be the cause.If one peak in the chromatogram is badly misshapen, it may imply that it interacts with the column by way of a different chemical mechanism.Refer to the figure “Peak resolution troubleshooting” on page 3-14 to identify a possible cause of your peak resolution problem.


Peak resolution troubleshooting

Are peakseluting at correctretention time?

Resolutionproblem

Troubleshootretention time

problem

Are peaksbroadening?

(column)

Are peaksbroadening?

(not column)

Check column bydetermining plate

count. low?

Replace columnand check plate

count. low?

Inline filter orguard column

clogged?

Clean/replace filteror guard column

Continued on next page Injector/columntubing diameter

too large?

Use correctdiameter tubing

Detector/columntubing diameter

too large?

Use correctdiameter tubing

Problem withinjector?

Troubleshootinjector


No

Yes

Yes

No

YesYes

No

Yes Yes

No

No

No

No

Yes

Yes

Yes

No


Peak resolution troubleshooting (Continued)

Column

Before using this troubleshooting tree, use those that appear earlier in the chapter to isolate the column as problem’s cause.Refer to the figure below to determine the possible cause of your column problem.

Continued fromprevious page Extra

column bandbroadening?

Detectorfilter constant

too high?

Adjust filterconstant

Coelutingpeaks?

If retention timecorrect, is samplecontaminated or

degraded?

Use freshsample andstandard


No

Yes

No

NoNo

Yes Yes


Column troubleshooting

Chromatography troubleshootingThis section lists symptoms of chromatography problems, possible causes, and suggested corrective actions.

Column problem

Replacecolumn inlet

frit

Pressurereduced?

Determine platecount to verifyperformance

Determineplate count

Replacecolumn outlet

frit

Retention time ofmarker compound

changed?

Clean/replacecolumn and

determine platecount

Pressurereduced?



Has platecount changed?



Call WatersTechnical Service

No

Yes

No

No

Yes

Yes

No

No

Yes

Yes

Has pressurechanged?


Before using the table below, see “Troubleshooting steps” on page 3-2 to isolate the cause of the chromatography symptom.

Chromatography troubleshooting

Symptom Possible cause Corrective actionErratic retention time Air in pump head Degas all solvents,

prime pump, or check sparge rate.

Malfunctioning pump check valves

Clean or replace pump check valves.

Leaking pump seals Replace pump seals (see “Replacing the pump seal” on page 3-58).

Separation chemistry Check mobile phase and column.

Clogged solvent filter Replace filters.Increased retention times

Incorrect flow rate or solvent composition.

Change flow rate or solvent composition.

Doubled retention times Air bubble in pump head

Prime pump to remove bubble.

Malfunctioning pump check valves

Clean or replace check valves.

Broken pump plunger Replace the plunger.Reduced retention time Incorrect flow rate Change flow rate.

Incorrect solvent composition

Change composition.

Incorrect mobile phase Use correct mobile phase.

Column contaminated Clean or replace column.

Incorrect column Use correct column.


Reproducibility errors Solvent not properly degassed or sparged

Degas or sparge solvent.

Incorrect chemistry Check chemistry.Incorrect integration Incorrect integration.Injector problem Troubleshoot injector.

Baseline drift, rapid Column not equilibrated

Equilibrate column.

Detector not allowed to warm up

Allow detector to warm up until baseline is stable (time varies based on wavelength and sensitivity).

Solvent contaminated Use fresh solvent.Solvent not properly degassed (rapid or slow drift)

Degas solvent.

Flow fluctuations (rapid or slow drift

Troubleshoot fluid handling unit.

Incorrect wavelength for solvent

Use correct wavelength and solvent (make sure solvent does not have a UV absorbance at the wavelength used).

Chromatography troubleshooting (Continued)



Baseline drift, slow Solvent contaminated Use fresh solvents.Decreased UV lamp energy

Check lamp energy(detector documentation).

Ambient temperature fluctuations

Stabilize operating environment temperature enough to allow full equilibration.

UV detector flow cell leaking (internal, cross-port)

Check flow cell; tighten connections.

Dirty flow cell Clean flow cell(detector documentation).

Baseline noise, short-term (30 sec to 1 min) cycling

Inadequate solvent blending in pump

Connect high flow mixer.

Flow fluctuating Stabilize flow.Solvent not mixed (short or long term cycling)

Stir solvent.

AC power source (short or long term cycling)

Disconnect other instruments on the power line; try a different wall outlet; have line voltage checked; use power conditioner.

Radio frequency noise (short or long term cycling)

Eliminate interference.




Baseline noise, long-term (approximately 1 hour) cycling

Ambient temperature fluctuations

Stabilize operating environment temperature.

Integrator or recorder faulty

Check integrator or recorder for excessive baseline noise.

Straight baseline, no peaks

No pump flow Set pump flow rate.Lamp not on Troubleshoot detector.Detector not zeroed Zero detector baseline.Improper connection between Prep system and recorder

Check cabling between Prep system unit and recorder.

Incorrect wavelength Check wavelength setting.

Leak in solvent path Check fittings.




Baseline noise, random Air in detector Purge UV detector to remove air.

Solvents not properly degassed

Sparge solvents.

Flow erratic, pump not primed

Prime the pump; check for air in the pump, failing seals.

Solvents contaminated Use fresh solvent.Column contaminated Clean or replace

column.Dirty flow cell Clean flow cell (see

detector documentation).

Analog output cable not properly connected between the Prep system and data system, recorder, or integrator

Properly connect cable.

System grounded improperly

Plug all system components into a single power strip.

Recorder voltage incorrect

Set recorder to proper voltage.

Unit not cooling properly

Operate unit with covers installed.

Radio frequency noise Eliminate interference.Defective detector Troubleshoot detector.




Flat-topped peaks Detector not zeroed Zero detector baseline.Incorrect recorder input voltage

Adjust recorder input voltage, or adjust detector output cable to proper position.

Sensitivity too high Select a less sensitive detector range.

Sample concentration or injection volume exceeds voltage output of detector

Decrease sample concentration or injection volume.

Sensitivity loss Leak in injector Troubleshoot injector.Degraded, contaminated, or improperly prepared sample

Use fresh sample.

Column contaminated Clean/replace column.Loss of column efficiency

Clean/replace column.

Peaks wider than expected

Troubleshoot injector.

Incorrect Filter Constant

Set correct Filter Constant value on DETECTOR SETUP screen.

Change in mobile phase composition

Correct mobile phase pH or ionic composition.

Leak in flow cell Tighten cell seal.Incorrect flow rate Change flow rate.




Hardware troubleshooting

System troubleshooting

The table below provides troubleshooting suggestions for overall system problems unrelated to a particular system module.

Sample energy decreased, reference energy not decreased

Contaminated mobile phase

Use fresh mobile phase.

Dirty flow cell Flush with water.Remove column, and flush system with stronger solvent to remove particulate or filmy matter from flow cell windows.Clean outside of flow cell windows.

General system troubleshooting

Symptom Possible cause Corrective actionUnit will not start Power cord not

connectedCheck power cord.

No power at outlet Check line voltage.Power supply fuse open or missing

Replace power supply fuse.

Fan(s) not running Unit not turned on Turn unit on.Fan wiring or motor problem

Call Waters Technical Service.

Screen is blank Unit not turned on Turn unit on.Startup diagnostic test failure

Power down and up again. If failure not corrected, call Waters Technical Service.




Fluid system troubleshooting

The table below addresses potential pump unit symptoms that can arise during operation.

No response from keypad

Keypad is locked Unlock keypad on PUMP SETUP screen.

Keypad is broken Check keypad.CPU in Hold mode Call Waters Technical

Service.

Pump troubleshooting

Symptom Possible cause Corrective actionPump will not run Loose control cable Reattach cable to rear

of fluid handling unit.High pressure limit set to 0

Set higher pressure limit on PUMP SETUP screen.

Broken plunger jamming the pump

Replace damaged plunger.

Defective pump motor Call Waters Technical Service.

Defective circuit board Call Waters Technical Service.

Defective pump transducer

Replace pump pressure transducer.

General system troubleshooting (Continued)



Pump leaks solvent Loose fitting Tighten leaking fitting.Salt crystals or debris in fitting

Stop Flow; remove and disassemble fitting; rinse fitting components with water; reassemble fitting.

Damaged fitting assembly

Stop Flow; remove and disassemble fitting; verify fitting components; replace ferrule and other damaged parts.

Worn pump plunger seals

Replace seals.

Loose pump head Tighten loose pump head(s) with a 5/32-inch hex-head wrench.

Loose pump inlet or outlet check valve

Tighten loose check valve(s).

Loose compression screw(s) on the pump head

Tighten loose compression screw(s) with a 5/16-inch open-end wrench.

Pump troubleshooting (Continued)



Injector troubleshooting

Before using the troubleshooting tables in this section, use those that appear earlier in the chapter to identify the part of the system in which the problem originates.

Erratic flow/pump pulsations

Gas dissolved in mobile phase

Sparge solvents.

Air bubble in pump head

Prime pump to remove bubble.

Dirty check valve(s) Clean check valve(s). Faulty check valve(s) Replace check valve(s).Plugged solvent reservoir filter

Clean or replace the filter.

Kinked or bent tubing Remove bend from tubing. If symptoms continue, replace tubing.

Leaky plunger seals Replace seals.Broken plunger Replace damaged

plunger.Defective solvent proportioning valve

Call WatersTechnical Service.

Squeak Pump seals not wetted Prime pump.Binding plunger seal Replace plunger seal

assembly.

Pump troubleshooting (Continued)



Rheodyne injector valveThe table below addresses Rheodyne injector symptoms that can occur during operation.

Waters autoinjectorThe table below lists autoinjector symptoms that can arise while operating a Waters autoinjector. Refer to the documentation that accompanies your autoinjector for additional troubleshooting information and service procedures.Tip: Replace both the needle and seal pack at the same time, because needle seal-pack sets are individually matched for optimal performance.

Rheodyne injector troubleshooting

Symptom Possible cause Corrective actionLeakage Stator screws need

tighteningTighten stator screws; replace rotor seal.

Scratches on rotor seal Replace rotor seal.Needle seal leakage Syringe needle smaller

than needle sealTry another syringe needle; gently push on plastic needle guide to deform PTFE sleeve to provide a better needle fit.

Plugged valve passages Unfiltered solvent or sample

Remove the stator and clean the passages with a small wire (0.015-inch maximum diameter).

Autoinjector troubleshooting

Symptom Possible cause Corrective actionLeak at seal pack Damaged needle, seal

packReplace needle and seal pack.

Cannot maintain high or low pressure

Leak at needle, seal pack

Replace needle and seal pack.


Reproducibility errors Syringe contains an air bubble (usually seen at the top of the syringe)

Remove bubble.

Leaks at the fluid tubing connections

Check for leakage at the connections with the corner of a tissue, and tighten connection slightly if the tissue is moistened.

Seal pack failure Replace seal pack.Compression check failed

Syringe contains air bubble (usually seen at the top of the syringe)

Remove bubble.

Mobile phase not properly degassed

Sparge mobile phase.

Leaks at the fluid tubing connections


High pressure limit on PUMP SETUP screen must be at least two times compression pressure

Change pressure limit on the PUMP SETUP page; change pump flow to adjust pressure.

Parameters used for Compression Check unacceptable for solvent

Repeat the test using default parameters on Purge and COMPressION test screen. If the test fails again, perform an autocalibration and repeat the test.

Seal pack failure Replace seal pack.

Autoinjector troubleshooting (Continued)



Detector troubleshooting

Before using the troubleshooting tables in this section, use those that appear earlier in the chapter to identify the part of the system in which the problem originates.

Autocalibration failed System not purged before autocalibration

Purge system and repeat autocalibration.

Solvent not properly degassed

Sparge solvent.

System pressure fluctuating more than 50 psi

Allow pressure to stabilize, then repeat autocalibration.

System pressure not at 400 +/− 50 psi

Set pump flow to achieve 400 +/– 50 psi.

Syringe contains a gas bubble (usually seen at the top of the syringe)

Remove bubble.

Leaks at the fluid tubing connection


Seal pack failure Disassemble ferrule connection, rinse, reassemble, check for leak.Replace ferrule and screw.Replace seal pack.

Carousel engages and turns, but does not find position 1

Dirty or defective carousel

Try another carousel; clean carousel.

Defective carousel reader


Autoinjector troubleshooting (Continued)



Consult your detector’s accompanying documentation for additional troubleshooting information and service procedures.UV detectorThe table below lists potential UV detector symptoms that can occur during operation, along with possible causes and corrective actions.

UV detector troubleshooting

Symptom Possible cause Corrective action“Unable to setup detectors” message on DIRECT CONTROL screen

External detector not calibrated

Calibrate external detector.

External detector turned off after it has been identified on the bus

RESCAN using the SYSTEM CONFIGURE screen.

Deuterium source lamp does not light

Faulty fuse Replace fuse on lamp power supply board.

Lamp leads not connected

Connect lamp leads.Replace lamp.

Normal effects due to lamp agingBad lamp power supply board


Unit will not calibrate (due to low reference energy)

Lamp aging Replace lamp.Reference side of flow cell is dirty

Clean flow cell.

Bad photodiode Call Waters Technical Service.


RI detectorThe table below addresses potential RI detector symptoms that can occur during operation.

Software troubleshootingMessages appearing on the controller screens during programming or operation require a response from you. They can be of two types: warning or shutdown. Other messages might not fit the above categories and do not require any action.Error messages appear on the controller screens during programming when you make these errors:

• Enter an improper value• Use an unacceptable entry method

RI detector troubleshooting

Symptom Possible cause Corrective actionSpikes on chart; spikes are synchronous with pump pulses

Solvent not properly degassed

Degas solvent. Place solvent at the level or above the level of the solvent proportioning valve to maintain slight backpressure.

Loss of sensitivity Foreign material in cell Clean cell and make sure solvent and column effluent are clean.

Source LED needs replacement or is misaligned


Short term noise or drift Fluctuating ambient temperature

Stabilize environment.

Dirt in cell Clean cell.Leak in cell Repair or replace cell

assembly.


To resume programming after a warning message, do one of these:• Press CLEAR.• Wait briefly until the message disappears.

Operational warning messages

Operational warning messages remind you to enter acceptable values and review your work.You must heed all warnings and take the appropriate corrective action. The table below lists the warning messages displayed on the controller.

Operational warning messages

Message DescriptionValue Entered is Out of Range

Programming/isocratic/direct control operation message: Enter a valid value. This message is accompanied by an audible alarm.

Enter Table Number for New Table

Gradient operation: Appears when using the SAVE function. The table currently in memory and the table number you entered display. The table currently in memory will be replaced with the new table in the next step. This message is accompanied by an audible alarm.

Replace with New Table1 = Yes 0 = No

Gradient operation: Only appears when using the SAVE function for the Gradient, Event, and Detector tables. Shows the table to be deleted. Allows you to abort the save if the table associated with the number entered should not be erased.

Rear Panel Stop Flow Input Detected

Message can appear anytime following a signal at the Stop Flow terminal. Suspends system operation until the source of the signal is removed.

Can’t Save Table Currently Executing

Gradient operation: The table number you wanted to save for a new or modified table is the same as the one running. Abort the run or wait until finished to replace it with the modified table. This message is accompanied by an audible alarm.


Program Method Table Saved…

Programming: Keyboard is locked while the SAVE function is storing the PROGRAM METHOD screen. This message is accompanied by an audible alarm.

Check Corresponding Gradient table/Events table/Detector table

Programming: Since the Gradient, Event, and Detector tables always run together (dictated by the table number), modifications made to the Gradient table should be consistent with the event or detector table of the same table number. This message is accompanied by an audible alarm.

An Inject Occurred Operation: Informational message indicating that an inject signal was sent. This message is accompanied by an audible alarm.

Line Is Incorrect Or Incomplete

Programming: Finish the entries required before attempting to move the cursor to a new location. This message is accompanied by an audible alarm.

UV Detector 1 Not FoundUV Detector 2 Not FoundRI Detector Not Found

Operation: Device has disappeared from the IEEE-488 bus. Check if the PowerLine module was turned on or if the PowerLine module was turned on before the controller was turned on. Go to the SYSTEM CONFIGURATION screen and rescan IEEE-488 bus. This message is accompanied by an audible alarm.

Illegal Vial Number Vial number is greater than the maximum value for the selected carousel. This message is accompanied by an audible alarm.

Number Out Of Range

Check the range for the parameter using the HELP key.

Composition MUST sum to 100%

The composition values for %A, %B, %C, and %D do not total 100%. Check entered values for all solvent reservoirs. This message is accompanied by an audible alarm.

Step Does Not Exist A step number was entered into the initial OPERATE METHODS screen which does not exist in the method table. Check the PROGRAM METHOD screen. This message is accompanied by an audible alarm.

Operational warning messages (Continued)

Message Description


Shutdown messages

Shutdown messages appear on the controller screens when an operator-set limit is exceeded. These require operator intervention. Determine appropriate action by consulting the table titled “Shutdown messages” on page 3-35.

To resume programming:

1. Press CLEAR.

2. Check the PUMP SETUP parameters.

3. Make adjustments to the parameters.

4. If the problem continues to occur, notify Waters Technical Service.System shutdown responseWhen a shutdown occurs, the system responds as follows:

• A message appears as:Shutdown Occurred - Reason:.............

• The system resets the flow rate to 0.0 mL/min. Flow and composition appear on the display.

Invalid Injection Number

The Injection Number is not part of the Step indicated. Check the PROGRAM METHOD screen. This message is accompanied by an audible alarm.

Invalid Vial Number

A vial number was selected that is not included in the selected step of the program method. Check the PROGRAM METHOD screen. This message is accompanied by an audible alarm.

First Vial Greater Than Last Vial

Refer to the PROGRAM METHOD screen. The last vial must always be in a higher location than the first vial. This message is accompanied by an audible alarm.

Table Is Full Exceeded the maximum 48 steps for this table. This message is accompanied by an audible alarm.

Operational warning messages (Continued)

Message Description


• The Hold output (on the controller’s rear panel) activates, signaling the connected instrument (for instance, the autoinjector) that the system is not ready for operation.

• Gradient, Event, and Detector tables that were running abort.• The Autostart time and Table # fields reset to blank.• External output switches S1 to S4, sparge valves, and the Alarm remain

in their current state, on or off.• Sparging continues at the last entered sparge rate.• The shutdown message remains on the screen until you press CLEAR.

When the stop flow signal occursIf a signal is detected at the Stop Flow terminal during a shutdown, a message appears on the screen. An inject input is not recognized while the system is in shutdown.If the cause is an obviously a system component, remove the component from the system. Then continue as follows.Shutdown not caused by stop flow signalIf a Stop Flow signal does not cause the shutdown, or if you correct the problem, press CLEAR to resume operation. Also take these actions where applicable:

• For isocratic operation, enter the flow rate• For gradient operation, press the RESUME key to resume operation

where the failure stopped it.Another shutdown can occur if you do not correct the cause before restarting.Tip: The controller only recognizes the first shutdown cause. Subsequent shutdown conditions are ignored once shutdown occurs.

Shutdown messages

Message DescriptionLow Pressure Limit The value entered on the SETUP screen was not

maintained. If the low-pressure limit was set high, check for small leaks at fittings and unions. Shutdowns with low limit settings are caused by large noticeable failures or an empty solvent reservoir.


Performance tests

These tests help you further isolate the problem’s cause:• Extended Test Routine for the controller• Pressure Transducer Calibration test• Transducer test• Solvent Proportioning Valve Pair test

For problems you cannot resolve, contact Waters Technical Service for assistance.

Self-diagnosticsThe Waters controller performs these self-diagnostic tests on startup:

• The 68000 processor, its clock and reset circuitry, LEDs, and drive circuitry, ROM-containing code preceding first entry

• ROM, associated address decoding, bus latch, and drive circuits• Addresses at each RAM location, associated address decoding, bus latch,

and drive circuits• Video RAM• EEPROM location with interrupt, read/write, associated address

decoding, bus latch, and drive circuits• Real-time clock, oscillator, counters, and associated interrupt circuitry

High Pressure Limit

The value entered on the SETUP screen was exceeded. If the high-pressure limit was set relatively low, check for an increase in backpressure (such as an older column with accumulated debris at the inlet and blockage in the fluid path).

Caution: To avoid damaging any equipment, wait about 3 minutes after shutting down your system before disconnecting any interconnecting cables.

Shutdown messages (Continued)

Message Description


When the self-diagnostic series successfully ends, the Startup screen appears.

Startup screen

No screen display

If the screen is blank, the failure might have prevented the program from controlling the LCD display.

Restart

Restart the program by turning the power switch off and back on. If characters fail to appear a second time, contact Waters Technical Service.

Extended testsAfter the automatic self-diagnostic routine ends, select the extended test routine by pressing the decimal (.) key on the keyboard.Perform the extended test routines if you suspect your problem originates with the controller, and you want to pinpoint the malfunction.

PowerLine

Multisolvent Delivery System

REVISION x.xx DATE: xx/xx/xx

DIAGNOSTIC STATUS: OK

CONT. TYPE: W4000 SYSTEM CONTROLLER

TO CONTINUE, PRESS ANY SCREEN KEY

TO START DIAGNOSTICS, PRESS DECIMAL PT.

Software Copyright 1988-1997

Waters Corporation

Performance tests 3-37

The table below lists diagnostic tests available through the extended test routine.

Tip: If any tests fail during this procedure, contact Waters Technical Service.

Tools

The only tools required for the extended test routine are a jumper wire to short various controller rear panel terminals and a screwdriver.

Extended test routine

Test DescriptionKeyboard test An echo of the key being pressed appears on the CRT.LCD test Displays all characters in upper- and lowercase.Stop Flow Input test

Tests the Stop Flow input switch on the rear panel.

External Inject test Tests the INJ input terminal on the rear panel.External Switches and Hold Output tests

Tests each of the four switch terminals and the Hold output terminal on the rear panel.

Chart test Produces a 0- to 10-mV sawtooth waveform at the chart terminals on the rear panel.

Sparge valve test Turns each sparge valve on and off in sequence.Pump and solvent valve test

Sets the pump to 1.0 mL/min (500-µL) or 4.0 mL/min (100-µL) and switches the solvent valves periodically.

IEEE-488 test CPU reads and displays the setting of the IEEE-488 address switch on the rear panel.

RS-232 test Tests RS-232 communication.12 VAC Fail test Operates the valves at a fast rate continuously

applying 12 VAC to the valves continuously. If this condition is detected, the test passes.

Counter test Verifies that the valves cannot switch to a 50/50 condition.


Rear panel terminals

General instructions

The following are general instructions for running the extended test routine. Each test instructs the operator to do one or more of the following:

• Press a key.• Place a jumper across terminals.• Make an observation.

You must cycle through the tests, either performing or bypassing them, to exit the extended test sequence.Advance to the next test by pressing ENTER. Skip unwanted tests by pressing ENTER a second time.When the tests are complete, the Startup screen reappears. You can then press the decimal key to rerun the extended test routine, or press any function key to resume normal operation.Some of the extended tests require additional actions that require explaining. These appear on the next several pages.

Stop flow input test

The Stop Flow switch accepts a signal from an external device that stops the pump. This test verifies that the Stop Flow input switch is functioning.

System Interface Connector

AC Power Connector

Screw Terminals

RS-232 Connector IEEE-488 Connector

INJE

CT

STO

PFL

OW

HO

LD

S1

S2

S3

S4

SWITCHESCHART+PRESSURE+

PRESSURE_

CHART_ G

ND

GN

D

GN

D

GN

D

AU

X.

+12V

Auxiliary +12 V Power Fuse

Pump Power Fuse

RS232

IEEE 488

Screwterminals

AC power connector

Pump power fuse

Auxiliary +12 VAC power fuseSystem interfaceconnector

RS-232 connector IEEE-488 connector


To perform a stop flow input test:

1. Attach a jumper wire from the Stop Flow switch on the rear panel to the Gnd terminal, also on the rear panel.

2. Press ENTER.• If the test succeeds, the screen displays GOT STOP FLOW.• If the test fails, the screen displays DID NOT GET STOP FLOW.

3. Press ENTER.

4. Remove the jumper wire.

5. Press ENTER to proceed to the next test.

External inject test

The external inject (INJ) switch accepts a signal from the injector to start a run. This test verifies that the INJ switch is functioning.

To perform an external inject test:

1. Attach a jumper wire from INJ on the rear panel to a Gnd terminal on the rear panel (see the figure “Rear panel terminals” on page 3-39).

2. Press ENTER.• If the test succeeds, the screen displays GOT INJECT.• If the test fails, the screen displays DID NOT GET INJECT.

3. Press ENTER.



External switch (S1 to S4) output test

This test verifies that the external switches, S1 to S4, are functioning. This test sets up for the next test, the Hold Output test, by directing you to move the jumper wire before pressing ENTER.


To perform an external switch (S1 to S4) output test:

1. Attach a jumper wire from the S1 terminal to the INJ terminal on the rear panel.

2. Press ENTER.The screen displays TEST PASSED or TEST FAILED.

3. Press ENTER.

4. Repeat steps 1 to 3 for the remaining switch terminals (S2 to S4).

5. When you have tested all four external switches, the screen displays CONNECT HOLD TO INJECT.Tip: If you want to perform the Hold Output test, do not press ENTER until after you attach Hold to INJ. If you want to bypass the Hold Output test, press ENTER.

Hold output test

To prevent further injections in case of power failure, the Hold switch outputs a signal to devices that can recognize the signal, pressure shutdown, or an abort procedure.

To perform a hold output test:

1. Attach a jumper wire from Hold to Gnd on the rear panel.

2. Press ENTER. The screen displays TEST PASSED or TEST FAILED.

3. Press ENTER.


5. Press ENTER.

Chart test

During this test, the Chart output ramps from 0 to 100% and then returns suddenly to 0%.

To perform a chart test:

1. Connect the chart recorder to the Chart + and – terminals.

2. Set the recorder to accept a 10-mV full scale signal.


A triangular (sawtooth) trace appears on the chart recorder if the test succeeds.


IEEE-488 address test

This test verifies that the system is reading the current address. When the test begins, the current address appears on the screen.

To perform an IEEE-488 address test:

1. Change the position of any IEEE-488 address DIP switch on the back of the controller panel. The address shown on the screen will change.

2. Repeat step 1 for each DIP switch.


Pressure transducer calibration test

Accurate operation requires that the pressure transducer is correctly calibrated to your system. The calibration procedure shows you how to adjust transducer slightly. Do this if you experience high or low pressure shutdowns to determine that the pressure transducer is causing your problem.Tip: Turn the screw slowly, because the screen’s response to the adjustment lags. A transducer output that falls below 0 psi can cause the screen to freeze. If the screen seems frozen, turn the potentiometer to increase the pressure value above 0 psi. The ideal setting is 6 to 24 psi.

To perform a pressure transducer calibration test:

1. Disconnect the column from the flow path by turning the Purge valve to PUMP Purge.

2. Start the Prep system and go to the DIRECT screen.

3. Set the flow rate to 0.0 mL/min and observe the pressure reading on the screen. The pressure value should be 0 psi.Tip: The pressure value can read 0 psi even when the pressure transducer is not set properly, because the screen does not display negative pressure.

4. Adjust the pressure transducer:


a. Insert a jeweler’s screwdriver into the hole.Adjusting the transducer

b. Turn the screwdriver slowly clockwise until the pressure display value reaches 0.

c. Turn the screwdriver slowly counterclockwise until a small positive number appears.

Solvent proportioning valve pair test

This test lets you evaluate the system when you suspect a defective proportioning valve is causing erratic chromatographic results.

You can perform the test in less than 1 hour. Use pure methanol as the mobile phase with a UV-absorbing sample.The sample is used to “spike” one reservoir of methanol, and must be linear in absorbance throughout the test range.Preparing solvent

To prepare the solvent:

1. Fill reservoirs A and B with solvent 1 (100% methanol).

Warning: Always observe safe laboratory practices when handling solvents.


2. Fill reservoirs C and D with solvent 2 (100% methanol with UV absorbing sample added).

The concentration of UV absorbing sample used in solvent 2 should produce about 0.45 AUFS in this test. Flushing solvent linesBe sure to flush any residual solvents from the system.Adjusting the recorder

To adjust the recorder:

1. Set the detector to 0.5 AUFS.

2. Pump solvent 1 (reservoir A or B) isocratically at 5 mL/min and adjust the recorder to 0.

3. Pump solvent 2 (reservoir C or D) isocratically at 5 mL/min, and adjust the recorder to full scale using the recorder’s variable full scale input.

The recorder is now set to respond so that 0% composition (pure methanol in solvent 1) = 0% full scale, and 100% composition (methanol containing sample in solvent 2) = 100% full scale.Program the gradient tableEnter and save Gradient table values as shown below.

Gradient table values

Time Flow %A %B %C %D CurveInitial 1.00 100 0 0 0 *5.00 1.00 90 0 10 0 1110.00 1.00 100 0 0 0 1115.00 1.00 90 0 0 10 1120.00 1.00 0 100 0 0 1125.00 1.00 0 90 10 0 1130.00 1.00 0 100 0 0 1135.00 1.00 0 90 0 10 1140.00 1.00 0 100 0 0 11


To perform a solvent proportioning valve pair test:

1. Go to the OPERATE METHODS screen.

2. Press the START RUN screen key.

3. When the detector baseline is stable at 0%, do the following:

a. Change the detector sensitivity to 0.1 AUFS.

b. Set the baseline to 10% full scale by adjusting the recorder’s zero adjustment.

c. A 5% change on the recorder now equals a 1% change in composition.

Evaluate resultsThe figure “Test results with all valves working” on page 3-46 is typical of the results obtained from this test when all valves are working. Valve failureA valve failure shows as a difference in height between the sections common to that valve.For example, if Valve A is malfunctioning, Peaks 1 and 2 (the only two containing contributions from Valve A) are irregular when compared to Peaks 3 and 4. Similarly, if Valve D is bad, Peaks 2 and 4 are irregular.

45.00 1.00 0 100 0 0 11

Gradient table values (Continued)

Time Flow %A %B %C %D Curve


Test results with all valves working


Maintenance procedures

This section includes maintenance procedures for the following components:• Fluid handling system• Rheodyne 7725i and 3725i injectors• Fuses

Column rack tubing configurationThe column rack does not typically require maintenance. However, the figure below illustrates the tubing configuration inside the column rack, in case you need to replace tubing.

Caution: To avoid damaging equipment, wait about 3 minutes after shutting down your system before disconnecting any interconnecting cables.

Maintenance procedures 3-47

Tubing configuration in the column rack

If you change fittings or tubing in the column rack, check for leaks before operating the unit.

Fluid handling unit maintenance

This section describes the maintenance or repair procedures for the fluid handling unit. These procedures include:

• Pressure transducer calibration• Check valve cleaning and replacement

Warning: Observe safe laboratory practices when handling solvents. Refer to the Material Safety Data Sheet for the solvents in use.


• Pump head removal• Pump seal replacement• Pump plunger replacement• Wash seal replacement

Pressure transducer calibration test

You must calibrate the pressure transducer to the system.

To perform a pressure transducer calibration test:

1. Disconnect the column from the flow path by turning the Purge valve on the column rack to PUMP Purge.

2. To start the system, access the DIRECT CONTROL (PowerLine controller) or ISOCRATIC (Gradient controller) screen.

3. Set the flow rate to 0.0 mL/min and observe the pressure reading on the screen. It should be a positive value slightly greater than 0 psi.Tip: The pressure value can read 0 psi even when the pressure transducer is not set properly, because the screen does not display negative pressure.Tip: Turn the screw slowly, because the screen’s response to the adjustment lags. A transducer output that falls below 0 psi can cause the screen to freeze. If the screen seems frozen, turn the potentiometer to increase the pressure value above 0 psi. The ideal setting is 6 to 24 psi.

4. Adjust the pressure transducer.Tip: A 2.5-second delay intervenes between the potentiometer adjustment and the on-screen pressure value.

a. Remove the left screw, below the transducer to reveal the coarse adjustment.

b. Insert a jeweler’s screwdriver into the center hole (fine adjustment),


below the transducer. Turn it to the center position.Adjusting the transducer

c. Move the screwdriver to the left hole (course adjustment). Turn the screwdriver slowly until the Pressure display reads 0 to 100.

d. Move the screwdriver to the center hole. Turn the screwdriver slowly until a small positive value appears.

e. Replace the cover screw in the left hole.

Cleaning and replacing check valves

These are three types of check valves in the Waters Prep system pump:• Inlet valves, located under the pump head• Outlet valves, located on top of the pump head• Dual outlet valves, located above the pump head (1000-mL only)


Cleaning check valvesClean check valves by running a nonionic detergent solution or 6N HNO3 through the pump heads. Flush with copious amounts of water to remove residual detergent or acid.Tip: Disconnect the column from the flow path. If the pump outlet is connected directly to the column (for large-scale operation), disconnect the column at its inlet from the flow path.Tip: For the PrepLC system, if the front panel is connected in the flow path, turn the Purge valve on the front panel to Purge.Tip: For the Delta-Prep system, turn the Purge valve on the column to Purge.

To clean the check valves:

1. Fill the solvent reservoir with a compatible cleaning solution like methanol:water (1:1) with liquid detergent or 6N HNO3 added.

2. Set the flow rate to 3.0 to 4.0 mL/min, and purge for 10 minutes.

3. Replace the contents of the solvent reservoir with water.

4. Set the flow rate to 3.0 mL/min and purge for 10 minutes.Replacing check valvesIf the previous procedure does not sufficiently clean the check valves, remove the check valves from the pump as described next and check them for damage. If none is apparent, put them in methanol, in an ultrasonic bath, for 10 minutes. If a part is damaged, replace it as necessary.Disassembling inlet check valves

To disassemble the inlet check valves:

1. Shut down the controller, and disconnect the power cord. The proportioning valve stops all solvent lines.

2. Disconnect the inlet tubing from the inlet check valve, and loosen the inlet check valve with a 1/2-inch open-end wrench.

3. Remove the inlet check valve housing assembly from the pump head, and disassemble it by tipping the housing upside down.Check the retainer cup and the ball-and-seat assembly for damage or debris.


The figure below shows an exploded view of the inlet check valve assembly.

4. Clean and replace worn or damaged parts.

Inlet check valve assembly

Caution: To protect the parts from contamination, use forceps or tweezers to handle the parts after cleaning them and when reassembling the inlet check valve.

PUMPHEAD

HOUSING SEAL

RETAINER CUP

SLEEVE

BALL

SEAT

SEAL GUIDE

INLET CHECKVALVE HOUSING

Pump head

Housing seal

Retainer clip

Sleeve

Ball

Seat

Seal guide

Inlet check valve housing


To reassemble inlet check valves:

1. Position the retainer cup so that you can look down into it.

2. Insert the sleeve into the retainer cup.

3. Place the ball in the retainer cup.

4. Place the seat guide over the ball with the shiny side of the seat facing the ball.

5. Place the seal into the check valve housing and press into place.

6. Lower the housing carefully over the stack of parts and invert the assembly.

7. Place the housing seal into the threaded hole on the bottom of the pump head.

8. Screw the assembly up into the pump head being careful to not tip the housing.

9. Firmly tighten the inlet check valve and attach the tubing.

To disassemble outlet check valves:


2. Disconnect the tubing from the outlet check valve, and loosen the outlet check valve with a 1/2-inch open-end wrench.

3. Disassemble the check valve assembly with a forceps.

4. Check the retainer cup. Also check the ball and seat assembly for damage or debris. The figure “Outlet check valve assembly” on page 3-54 shows the outlet check valve assembly in an exploded view.

5. Clean and replace worn or damaged parts.


Outlet check valve assembly

To reassemble outlet check valves:

1. Position the housing so that you can look down into the pump head.

2. Insert the housing seal into the pump head.

3. Place the seat assembly into the pump head. Face the shiny side upward.

Caution: To protect the parts from contamination, use forceps or tweezers to handle the parts after cleaning them and when reassembling the outlet check valve.

OUTLET CHECKVALVE HOUSING

SEAL GUIDE

RETAINER CUP

SLEEVE

BALL

SEAT

SEAL HOUSING

Pump head

Seal housing

Seat

Ball

Sleeve

Retainer clip

Seal guide

Outlet check valve housing


4. Place the ball in the seat.

5. Lower the sleeve around the ball, placing it on the seat.

6. Place the retainer over the ball and seat assembly. Center it within the pump head.

7. Place the seal guide into the outlet check valve housing. Press it into place.

8. Place the outlet check valve housing over the retainer.

9. Hand tighten the assembly.

10. Tighten the outlet check valve, and attach the tubing.

To disassemble and reassemble dual outlet check valves:

Pumps fitted with 1000-µL heads require a dual outlet check valve, shown in exploded view in the figure “Dual check valve assembly” on page 3-56.

1. If the flushing procedure in “Cleaning check valves” on page 3-51 does not sufficiently clean the dual outlet check valve, disassemble the check valve assembly as described in “Disassembling inlet check valves” on page 3-51 and “To disassemble outlet check valves:” on page 3-53.

2. Check the valve parts for damage. If none is apparent, put the parts in methanol, in an ultrasonic bath, for 10 minutes. If a part is damaged, replace it as necessary.

3. Reassemble the check valve assembly as described in “To reassemble inlet check valves:” on page 3-53 and “To reassemble outlet check valves:” on page 3-54.


Dual check valve assembly

Removing the pump head

The figure below is an exploded diagram of a pump head.

Pump head disassembly

To remove the pump head:


2. Disconnect the inlet line from the inlet check valve. If necessary, use a 5/16-inch wrench to loosen the inlet line. Let solvent drain onto an absorbent wipe.

Ball

Seat

Seal guide

Seat

Seal guide

Outlet checkvalve housing

Outlet checkvalve housing

ALLEN SCREW

PUMP HEAD

PUMP SEAL

Plunger bearings

Pump seal

Pump head

Hex-head screws


3. Start the pump, setting the flow rate to 0.1 mL/min. Run it until the indicator rod retracts fully into the pump head.

4. Shut down the pump.

5. Loosen the nut that secures the outlet line from the outlet check valve, then disconnect the line.

6. Remove the two hex-head screws with a 5/32-inch hex wrench. Slide the pump head assembly straight off the pump.

Tip: If you are replacing the pump seal, continue with the procedure in “Replacing the pump seal” on page 3-58.Tip: If you are replacing the pump plunger or the wash seal, refer to the next procedure to remove the head support assembly.

Removing the head support assembly

The figure below is an exploded diagram of the head support assembly.

Head support disassembly

Caution: The indicator rod must be fully retracted into the pump head. Otherwise, the plunger can sustain damage.

Caution: Causing the pump head to bind against the plunger can break the plunger.

SPRING

TEFLON WASHER

HEAD SUPPORTBUSHING

S S

O - RING

RETAINER

ALLEN SCREWS (4)

Indicator rod

Spring

Teflon washer

Hex-head screw (4)

Retainer

O-ring

Wash seal

Head supportbushing


To remove the head support assembly:

1. Remove the four plunger support screws with a 9/64-inch hex-head wrench.

2. Slide the head support assembly and indicator rod off the plunger.

Removing the head support assembly

Tip: Continue with “Replacing the pump plunger” on page 3-61 or “Replacing the plunger wash seal” on page 3-65.

Replacing the pump seal

During routine maintenance, replace the seal in both pump heads at the same time.Tip: The seal insertion tool that this procedure refers to consists of the tool itself and the large guide (Startup Kit). The tool presses the pump seal in position for correct alignment in the pump head. The guide ensures the seal is properly aligned during installation.

To replace the pump seal:

1. Remove the pump head

2. Remove the three plunger bearings from the back of the pump head to expose the pump seal.


3. Pry the seal out of the pump head with a small screwdriver. Do not scratch the surface of the pump head.

Removing the pump seal

4. Use the seal insertion tool to insert the seal in the pump head:

a. Put the seal onto the tip of the insertion tool so that the open end of the seal (with the spring) faces out from the end of the tool.

b. Fit the guide into the recess at the rear of the pump head. Tip: The circular opening in the guide is tapered, with one side larger than the other. The larger side of the opening must face you.Tip: Apply methanol to the pump head area for lubrication.

Caution: Do not damage the seat or score surrounding surfaces when you remove the seal.


c. Slide the insertion tool with the seal into the guide and press to firmly seat it in the pump head. Remove the insertion tool and guide.

Replacing the pump head seal

Tip: Wet the seal with a small amount of solvent before continuing with step 5.

5. Place the three plunger bearings behind the seal in the pump head as follows:• Bearing 1’s grooved side faces the rear of the pump head.• Bearing 2’s grooved side faces the front of the pump head.• Bearing 3’s grooved side faces the front of the pump head.

6. Carefully slide the pump head assembly onto the pump head support assembly. Ensure the indicator rod goes into the correct hole.

Caution: Inspect the plunger for scoring that could impair the seal.

�� #�

Seal

Guide

Insertion tool

Pump head


7. Gently press the pump head assembly into place. Apply even pressure to the front of the head.

8. Replace the hex-head screws on both pump heads. With a 5/32-inch hex wrench, tighten them evenly by alternately tightening one then the other by half turns.

9. Start the pump and set the flow rate to 3.0 mL/min.

10. Be sure the indicator rod moves freely.

11. Push solvent through the two pump heads with a syringe. Solvent should exit the second pump head. This primes the plunger wash, and ensures the plunger is lubricated when you start the pump.

12. Reconnect the solvent lines, start the pump, and check for leaks.

Replacing the pump plunger

Tip: This procedure requires moderate strength and dexterity as well as a special plunger insertion tool. Contact your Waters Service Representative before performing it.The plunger may experience excessive wear if you do not use the plunger wash. If your pump has been running for some time without the plunger wash, replace the wash seals (see “Replacing the plunger wash seal” on page 3-65).If the plunger sustains excessive wear despite use of the plunger wash system, rub it with a fine pumice to remove any coating. Then clean it thoroughly to remove all traces of the abrasive. Replace the plunger seal, and install new wash and high pressure seals. If this does not extend plunger life, replace the plunger.

To replace the pump plunger:

1. Remove the pump head and head support assembly.

2. Set the flow rate to 3.0 ml/min. Run the pump unit until the plunger fully extends, then shut the pump down.

Caution: Do not exert any side forces or you could damage the plunger.


3. Pry the piston cap out of the piston, using a wire whose end is formed into a small hook. Insert the hook behind the cap and pull it toward you.

Exposed plunger and piston

4. Insert the plunger insertion tool into the piston cavity. With one hand, force the inner part of the plunger insertion tool against the snap ring. Using a pair of snap-ring pliers, squeeze the snap-ring to release the it from the piston. The snap ring, washer, and spring will hang freely on the plunger rod.


Using the plunger insertion tool and snap-ring pliers to remove the plunger

5. Pull the plunger straight out of the cavity.

Removing the plunger

Snap-ring pliers

Plunger insertion tool


Reassembling the plunger

To reassemble the plunger:

1. Place the two halves of the ball retainer around the ball on the plunger.

2. Slide the spring over the ball retainer. The spring holds the ball retainer together.

3. Lubricate the end of the ball with a lithium-based grease like Lubriplate™.

4. Insert the plunger into the piston.

5. Slide the washer, then the snap ring onto the plunger.

6. Position the plunger insertion tool over the snap ring and washer.

7. Press the plunger insertion tool, forcing the snap ring, washer, and spring inside the piston.

8. Squeeze the snap ring with snap-ring pliers, simultaneously pressing on the tool’s central area. This forces the snap ring into the groove near the end of the piston.Tip: Seat the entire snap ring in the groove in the piston. Otherwise, it can spring out of position when you release the pliers.

9. Remove the pliers and plunger insertion tool. The snap ring remains firmly in place when it is positioned correctly.

10. Start the pump, and set the flow rate to 3.0 mL/min to retract the plunger. Run the pump until the plunger is fully retracted, then stop the pump.

11. Replace the piston cap by pressing it into position on the end of the piston.

12. Replace the head support assembly and indicator rod.Tip: The support assembly’s orientation is correct when the indicator rod is to the right of the assembly, and the number is on top.

13. Alternately tighten the four screws. Do not overtighten. The support assembly need only be flush against the front panel.

14. Gently slide the pump head onto the plunger. Ensure the indicator rod is in the correct hole. Alternately tighten the two screws.


15. Insert the two hex-head screws.

16. Reconnect the plunger wash lines and, with a syringe, push solvent through the two pump heads until it exits the second pump head. This primes the plunger wash and lubricates the plunger before you run the pump.

17. Start the pump, and set the flow rate to 3.0 mL/min. Pull out and release the indicator rod. If it does not snap back, the head is misaligned. Loosen the head and rotate it slightly for correct alignment.

18. Reconnect the solvent lines, and examine fittings for leaks.

Replacing the plunger wash seal

The wash seal is located in the head support assembly.

To replace the plunger wash seal:

1. Remove the pump head and head support assembly.

2. Remove the indicator rod from the head support assembly.

3. Push on the wash seal from the back of the head support assembly with a small, blunt-end tool. The seal and retainer pop out the front of the head support assembly. The front of the head support assembly is imprinted with a number.

Installing a new wash seal and reassembling the pump head

Tip: Refer to the figure “Head support disassembly” on page 3-57 for this procedure.

To install a new wash seal and reassemble the pump head:

1. Insert the seal insertion guide into the front of the head support assembly.Tip: The circular opening in the guide is tapered, with one side larger than the other. The larger side of the opening must face you.


2. Mount the new wash seal on the end of the insertion tool with the smooth side of the seal flush as shown below.

Wash seal insertion tool

3. Push the seal into the head support assembly, making sure it firmly seats. Remove the insertion tool and guide.Tip: Wet the seal with a small amount of solvent before continuing with step 4.

4. Replace the retainer on the front of the head support assembly.

5. Insert the indicator rod into its hole in the back of the head support assembly. Make sure the spring and PTFE washer are on the rod.

6. With the front of the head support assembly facing you, use both hands to replace the head support assembly on the pump casting.

7. Replace the four hex-head screws that hold the head support assembly in place. Alternately tighten them, but avoid overtightening.

Caution: Do not push from side to side or wiggle the assembly back into position. This can damage the plunger.

�� #�


8. Push the O-ring into place on the retainer.

9. Gently slide the pump head onto the plunger, ensuring the indicator rod is in the correct hole.

10. Insert the two hex-head screws, holding the pump head assembly in place, and alternately tighten them.

11. Reconnect the plunger wash. Use a syringe to push solvent through the two pump heads until it exits the second pump head. This primes the plunger wash and ensures the plunger is lubricated when you run the pump.

12. Start the pump, and set the flow rate to 3.0 mL/min. Pull out and release the indicator rod. If it does not snap back, the pump head is misaligned. Loosen the pump head and rotate it slightly to correct the alignment.

13. Reconnect the solvent lines, and examine them for leaks.

Replacing fusesThis section includes procedures for replacing fuses on the rear panel of the controller and fluid handling unit. These procedures describe how to replace the following fuses:

• Operating voltage fuses• Auxiliary +12 VAC fuse• Pump power fuse• Proportioning valve fuse

Operating voltage fuses

All controllers are supplied with two 4-A fuses installed for 110/120 VAC operation. If you intend to operate the unit on 220/240 VAC power, change to the IEC-approved 2-A, 250-VAC fuses, as outlined next.The Prep system is protected from abnormal line power situations or component anomalies by fuses located at the power cord connector. If the LCD fails to display the Startup screen and the fans fail to run when the controller is turned on, examine the main fuses for opens.


The table below lists the correct fuses for your operating voltage.

To replace fuses:

1. Make sure the instrument is connected to the electrical power supply.

2. Pry open the power connector cover on the back of the unit, next to the power cord.

3. Pull each fuse holder out as though opening a drawer. Spare fuses are in the Startup Kit and/or in the fluid handling unit shipping box.

4. Install the correct fuses in the holder. The arrow on each fuse holder points up when in the correct position.

Installing the fuse

5. Make sure one end of the fuse is closest to the pins. After both fuses have been inserted into the holder, put the holder back into the power

Operating voltage ranges

Nominal AC voltage

Voltage range Fuse (Amp)

100 ±10% 4120 ±10% 4220 ±10% 2240 ±10% 2


connector, pin end first. You can insert the holder into the connector in either orientation.

Auxiliary +12 VAC fuse

The 12-VAC fuse protects the AUX +12 VAC terminal on the controller rear panel from excessive current loads. If this fuse is open, no power is available from this terminal. The proportioning valve in the fluid handling unit drawer also fails to start.The 12-VAC auxiliary power fuse is housed separately, on the rear panel of the controller. Replace this fuse with a 1.5-A fuse (Startup Kit).

Auxiliary +12 VAC and pump power fuses

To replace the auxiliary +12 VAC fuse:

1. Insert a small flat-blade screwdriver into the fuse holder for the AUX +12 VAC fuse.

2. Turn the screwdriver to release the fuse holder.

3. Slide the holder with the fuse out from the rear panel.

4. Replace the open fuse with a new 1.5-A fuse.

5. Insert the new fuse and holder back into its location on the rear panel.

6. Lock the fuse holder in place using the screwdriver.

RS232

IEEE 488

CHART +

PRESSURE +

CHART —

PRESSURE —

INJE

CT

ST

OP

FLO

W

HO

LD

S1

S2

S3

S4

SW

ITC

HE

S

GN

D

GN

D

GN

D

GN

D

AU

X+

12V

Pump power fuse

Auxiliary +12 VAC power fuse


Pump power fuse

The pump power fuse protects the pump from excessive current loads. If this fuse is open, no power is available at the pump.

To replace the power pump fuse:

1. Shut down the controller and remove the power cord from its rear-panel connector.

2. Insert a small flat-blade screwdriver into the slot on the fuse holder for the pump power fuse.


4. Slide the holder with the fuse out of the rear panel.

5. Replace the open fuse with a new fuse. Tip: Pump fuse operating voltages are 120/220 VAC 4 A and 125-VAC IEC (time delay) 5 το 20 mm.

6. Insert the new fuse and holder back into the fuse chamber on the rear panel.

7. Lock the fuse holder into place using the screwdriver.

Proportioning valve fuse

A special power supply provides proper proportioning valve performance. If you fail to hear the proportioning valve operating within the fluid handling unit drawer during gradient operation, examine the fuse on the rear panel of the fluid handling unit.The 0.5-A fuse is housed on the rear panel of the fluid handling unit as shown in the figure below. Replace this fuse with a 0.5-A fuse (Startup Kit).

Warning: To avoid shock, turn off the controller front panel power switch and unplug the power cord from the rear panel.


0.5-A fuse

To replace the 0.5-A fuse:

1. Insert a small flat-blade screwdriver into the fuse holder for the 0.5-A fuse.


3. Slide the holder with the fuse out from the rear panel.

4. Replace the open fuse with a new 0.5-A fuse.

5. Insert the new fuse and holder back into its location on the rear panel of the fluid handling unit.

6. Lock the fuse holder in place using the screwdriver.

Maintaining rheodyne manual injectorsRheodyne manual injectors require minimal maintenance. With normal use, the injector should perform thousands of cycles without fail. This section describes the following maintenance procedures:

• Tightening the needle seal• Replacing the position sensing switch

0.5A FUSE

CAUTIONDO NOT REMOVE CABLE

WHILE UNIT IS POWERED

CONNECT TO 4000

CONTROLLER ONLY

! !

! !

ATTENTION

0.5-A fuse


• Fixing rotor seal leakageRefer to the figure “Rheodyne 7725i injector: exploded view” on page 3-73 for each procedure.

Tightening the needle seal

The needle seal, a PTFE sleeve in the rotor seal, may not seal correctly around a needle that is smaller than average. A poor seal results in a loss of accuracy in sample loading.

To reform the PTFE sleeve to make a good seal:

1. Remove the needle from the needle port.

2. Push gently on the plastic needle guide with the eraser end of a pencil. Do not squash the PTFE sleeve. Repeat if necessary.

To check for a proper seal around the needle:

1. Fill the syringe with water. Place the injector in the Load position and slowly discharge the water in the syringe into the injector. Notice the lack of resistance to syringe discharge.

2. Repeat step 1 with the injector handle halfway between Load and Inject (the pump must be off). The resistance to discharging the syringe should now be much greater.

The needle seal holds only a few psi of pressure, and will not completely prevent syringe discharge with the handle in the halfway position.

Replacing the position sensing switch

The position sensing switch is a magnetic reed switch actuated by a magnet in the shaft. The switch sends the inject signal to the controller, signaling the controller to begin a gradient. If a gradient does not begin after an injection, replace the switch. The switch is rated for 100 VAC at 200 mA.

To replace the switch:

1. Remove the stator, stator ring, and 60° stop ring.

Caution: Always use the correct type and gauge needle to load the injector, or damage to the rotor seal can result.


2. Pull the position sensing switch out of the stop ring.

3. Insert a new position sensing switch.

4. Reassemble the injector according to the instructions in “Replacing the rotor seal” on page 3-74.

Rheodyne 7725i injector: exploded view

Handle screwHandle

Needle guide

Set screws (2)

Pressure adjusting screws

BodyThrust bearing

Spring washers (4)Rotor pinPosition sensing switchStop ringSeal pinsNeedle tube

Bearing ringIsolation sealPTFE sleeveRotor seal

Stator ringStator locating pinStator face assembly

Stator locating holeStator

Stator screws (3)

Needle seal

Knob


Fixing rotor seal leakage

If you see liquid leak between the stator and stator ring, tighten the pressure adjusting screw. If you see it leak from the needle port or vent tube, replace the rotor seal.

To tighten the pressure adjusting screw:

1. Locate the pressure adjusting screw on the injector shaft (see the figure “Rheodyne 7725i injector: exploded view” on page 3-73). The pressure adjustment is set so that spring force between the valve rotor and stator is enough to hold 4000 psi (40.8 MPa).

2. Remove the injector knob.

3. Slip the pressure adjusting nut onto the shaft so the tabs on the nut slip into the slots in the adjusting screw.

4. Use a wrench to tighten the adjusting screw approximately 1/20th of a turn. Use the 20 dial markings on the body and the painted spot on the adjusting screw to gauge how far to tighten the screw.

5. If the new setting fails to accomplish leak-free operation, repeat the procedure by an additional 1/20th of a turn. Avoid excessive tightening, which increases rotor seal wear.

6. If this fails to stop the leak, replace the rotor seal. Otherwise, finish by removing the adjusting nut and replacing the knob.

Replacing the rotor sealPremature rotor seal failure can be caused by any of these conditions:

• Abrasive particles in the sample or mobile phase that scratch the rotor seal surface.

• A wrong needle tip that chips the ceramic stator face, scoring of the rotor seal surface.

• Buffer or salt crystallization, caused by a failure to flush the flow passages and needle port with water after using aqueous buffers or salt solutions. The abrasive particles scratch the rotor seal surface, resulting in leakage.

Caution: After using buffer solutions, flush the valve with water.


When replacing the rotor seal, always check the ceramic stator face for chips or cracks. If you find damage, replace it. Refer to the figure “Rheodyne 7725i injector: exploded view” on page 3-73 for this procedure.

To replace the rotor seal:

1. Leave the injector attached to the front panel of the pump, and leave the knob on. If you need to remove the injector from the front panel, remove the knob.

2. Remove the three stator screws.

3. Remove these parts by pulling axially:

a. Stator and stator face assembly (remove together). If the stator face assembly is damaged, replace it.

b. Stator ring.

4. Pry the rotor seal off the four seal pins. Leave the isolation seal and bearing ring in place.

5. Install a new rotor seal on the four seal pins.

Reassembling the injector

To reassemble the injector:

1. Loosen the pressure adjusting screw a half-turn. Note the original position of the two red dots.


2. Orient the rotor seal as shown below, with rotor seal slots facing the stator.

Rotor seal orientation (viewed from the stator)

3. Replace the stator ring so the pin in the 60° stop ring enters the mating hole in the stator ring.

4. Install the stator face assembly on the stator. The three pins on the assembly fit into the mating holes in the stator only one way.

5. Install the stator and stator face assembly on the valve so that the pin in the stator ring enters the mating hole in the stator.

6. Alternately tighten each of the three stator screws to keep the stator surface parallel to the stator ring surface until all parts are firmly in place.

7. Retighten the pressure adjusting screw until the red dots align as noted in step 1.

8. Replace the knob, and tighten the two set screws against the two flat areas of the shaft.

Rotor PinNotch

Rotor pinNotch


A Safety Advisories

Waters instruments display hazard symbols designed to alert you to the hidden dangers of operating and maintaining the instruments. Their corresponding user guides also include the hazard symbols, with accompanying text statements describing the hazards and telling you how to avoid them. This appendix presents all the safety symbols and statements that apply to the entire line of Waters products.

Contents

Topic PageWarning symbols A-2Caution symbol A-5Warnings that apply to all Waters instruments A-6Electrical and handling symbols A-12

A-1

Warning symbols

Warning symbols alert you to the risk of death, injury, or seriously adverse physiological reactions associated with an instrument’s use or misuse. Heed all warnings when you install, repair, and operate Waters instruments. Waters assumes no liability for the failure of those who install, repair, or operate its instruments to comply with any safety precaution.

Task-specific hazard warningsThe following warning symbols alert you to risks that can arise when you operate or maintain an instrument or instrument component. Such risks include burn injuries, electric shocks, ultraviolet radiation exposures, and others. When the following symbols appear in a manual’s narratives or procedures, their accompanying text identifies the specific risk and explains how to avoid it.

Warning: (General risk of danger. When this symbol appears on an instrument, consult the instrument’s user documentation for important safety-related information before you use the instrument.)

Warning: (Risk of burn injury from contacting hot surfaces.)

Warning: (Risk of electric shock.)

Warning: (Risk of fire.)

Warning: (Risk of sharp-point puncture injury.)

Warning: (Risk of hand crush injury.)

Warning: (Risk of exposure to ultraviolet radiation.)

Warning: (Risk of contacting corrosive substances.)

Warning: (Risk of exposure to a toxic substance.)

Warning: (Risk of personal exposure to laser radiation.)

A-2 Safety Advisories

Specific warningsThe following warnings can appear in the user manuals of particular instruments and on labels affixed to them or their component parts.

Burst warning

This warning applies to Waters instruments fitted with nonmetallic tubing.

Warning: (Risk of exposure to biological agents that can pose a serious health threat.)

Warning: (Risk of tipping.)

Warning: (Risk of explosion.)

Warning: (Risk of eye injury.)

Warning: Pressurized nonmetallic, or polymer, tubing can burst. Observe these precautions when working around such tubing:• Wear eye protection.• Extinguish all nearby flames.• Do not use tubing that is, or has been, stressed or kinked.• Do not expose nonmetallic tubing to incompatible compounds like

tetrahydrofuran (THF) and nitric or sulfuric acids.• Be aware that some compounds, like methylene chloride and

dimethyl sulfoxide, can cause nonmetallic tubing to swell, which significantly reduces the pressure at which the 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 warning applies to all Waters mass spectrometers.

This warning applies to certain instruments when they are in Operate mode.

Warning: Where significant quantities of flammable solvents are involved, a continuous flow of nitrogen into the ion source is required to prevent possible ignition in that enclosed space. Ensure that the nitrogen supply pressure never falls below 690 kPa (6.9 bar, 100 psi) during an analysis in which flammable solvents are used. Also ensure a gas-fail connection is connected to the LC system so that the LC solvent flow stops if the nitrogen supply fails.

Warning: To avoid electric shock, do not remove the mass spectrometer’s protective panels. The components they cover are not user-serviceable.

Warning: High voltages can be present at certain external surfaces of the mass spectrometer when the instrument is in Operate mode. To avoid non-lethal electric shock, make sure the instrument is in Standby mode before touching areas marked with this high voltage warning symbol.


Biohazard warning

This warning applies to Waters instruments that can be used to process material that might contain biohazards: substances that contain biological agents capable of producing harmful effects in humans.

Chemical hazard warning

This warning applies to Waters instruments that can process corrosive, toxic, flammable, or other types of hazardous material.

Caution symbol

The caution symbol signifies that an instrument’s use or misuse can damage the instrument or compromise a sample’s integrity. The following symbol and its associated statement are typical of the kind that alert you to the risk of damaging the instrument or sample.

Warning: Waters instruments and software can be used to analyze or process potentially infectious human-sourced products, inactivated microorganisms, and other biological materials. To avoid infection with these agents, assume that all biological fluids are infectious, observe Good Laboratory Practices, and consult your organization’s biohazard safety representative regarding their proper use and handling. Specific precautions appear in the latest edition of the US National Institutes of Health (NIH) publication, Biosafety in Microbiological and Biomedical Laboratories (BMBL).

Warning: Waters instruments can be used to analyze or process potentially hazardous substances. To avoid injury with any of these materials, familiarize yourself with the materials and their hazards, observe Good Laboratory Practices (GLP), and consult your organization’s safety representative regarding proper use and handling. Guidelines are provided in the latest edition of the National Research Council's publication, Prudent Practices in the Laboratory: Handling and Disposal of Chemicals.

Caution: To avoid damage, do not use abrasives or solvents to clean the instrument’s case.

Caution symbol A-5

Warnings that apply to all Waters instruments

When operating this device, follow standard quality control procedures and the equipment guidelines in this section.

Attention: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user’s authority to operate the equipment.

Important: Toute modification sur cette unité n’ayant pas été expressément approuvée par l’autorité responsable de la conformité à la réglementation peut annuler le droit de l’utilisateur à exploiter l’équipement.

Achtung: Jedwede Änderungen oder Modifikationen an dem Gerät ohne die ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen.

Avvertenza: qualsiasi modifica o alterazione apportata a questa unità e non espressamente autorizzata dai responsabili per la conformità fa decadere il diritto all'utilizzo dell'apparecchiatura da parte dell'utente.

Atencion: cualquier cambio o modificación efectuado en esta unidad que no haya sido expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equipo.

注意：未經有關法規認證部門允許對本設備進行的改變或修改,可能會使使用者喪失操作該設

備的權利。

注意：未经有关法规认证部门明确允许对本设备进行的改变或改装，可能会使使用者丧失操作该设备的合法性。

주의: 규정 준수를 책임지는 당사자의 명백한 승인 없이 이 장치를 개조 또는 변경할 경우, 이 장치를 운용할 수 있는 사용자 권한의 효력을 상실할 수 있습니다.

注意：規制機関から明確な承認を受けずに本装置の変更や改造を行うと、本装置のユーザーとしての承認が無効になる可能性があります。


Warning: Use caution when working with any polymer tubing under pressure:• Always wear eye protection when near pressurized polymer tubing.• Extinguish all nearby flames.• Do not use tubing that has been severely stressed or kinked.• Do not use nonmetallic tubing with tetrahydrofuran (THF) or concentrated

nitric or sulfuric acids.• Be aware that methylene chloride and dimethyl sulfoxide cause nonmetallic

tubing to swell, which greatly reduces the rupture pressure of the tubing.Attention: Manipulez les tubes en polymère sous pression avec precaution:• Portez systématiquement des lunettes de protection lorsque vous vous

trouvez à proximité de tubes en polymère pressurisés.• Eteignez toute flamme se trouvant à proximité de l’instrument.• Evitez d'utiliser des tubes sévèrement déformés ou endommagés.• Evitez d'utiliser des tubes non métalliques avec du tétrahydrofurane (THF)

ou de l'acide sulfurique ou nitrique concentré.• Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le

gonflement des tuyaux non métalliques, ce qui réduit considérablement leur pression de rupture.

Vorsicht: Bei der Arbeit mit Polymerschläuchen unter Druck ist besondere Vorsicht angebracht:• In der Nähe von unter Druck stehenden Polymerschläuchen stets

Schutzbrille tragen.• Alle offenen Flammen in der Nähe löschen.• Keine Schläuche verwenden, die stark geknickt oder überbeansprucht sind.• Nichtmetallische Schläuche nicht für Tetrahydrofuran (THF) oder

konzentrierte Salpeter- oder Schwefelsäure verwenden.• Durch Methylenchlorid und Dimethylsulfoxid können nichtmetallische

Schläuche quellen; dadurch wird der Berstdruck des Schlauches erheblich reduziert.

Warnings that apply to all Waters instruments A-7

Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione:• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero

pressurizzati.• Spegnere tutte le fiamme vive nell'ambiente circostante.• Non utilizzare tubi eccessivamente logorati o piegati.• Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido solforico

o nitrico concentrati.• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano

rigonfiamenti nei tubi non metallici, riducendo notevolmente la pressione di rottura dei tubi stessi.

Advertencia: se recomienda precaución cuando se trabaje con tubos de polímero sometidos a presión:• El usuario deberá protegerse siempre los ojos cuando trabaje cerca de tubos

de polímero sometidos a presión.• Si hubiera alguna llama las proximidades.• No se debe trabajar con tubos que se hayan doblado o sometido a altas

presiones.• Es necesario utilizar tubos de metal cuando se trabaje con tetrahidrofurano

(THF) o ácidos nítrico o sulfúrico concentrados.• Hay que tener en cuenta que el cloruro de metileno y el sulfóxido de dimetilo

dilatan los tubos no metálicos, lo que reduce la presión de ruptura de los tubos.

警告：當在有壓力的情況下使用聚合物管線時，小心注意以下幾點。

• 當接近有壓力的聚合物管線時一定要戴防護眼鏡。

• 熄滅附近所有的火焰。

• 不要使用已經被壓癟或嚴重彎曲管線。

• 不要在非金屬管線中使用四氫呋喃或濃硝酸或濃硫酸。

• 要了解使用二氯甲烷及二甲基亞楓會導致非金屬管線膨脹，大大降低管線的耐壓能力。


警告：当有压力的情况下使用管线时，小心注意以下几点：

• 当接近有压力的聚合物管线时一定要戴防护眼镜。

• 熄灭附近所有的火焰。

• 不要使用已经被压瘪或严重弯曲的管线。

• 不要在非金属管线中使用四氢呋喃或浓硝酸或浓硫酸。

• 要了解使用二氯甲烷及二甲基亚枫会导致非金属管线膨胀，大大降低管线的耐压能力。

경고: 가압 폴리머 튜브로 작업할 경우에는 주의하십시오.• 가압 폴리머 튜브 근처에서는 항상 보호 안경을 착용하십시오.• 근처의 화기를 모두 끄십시오.• 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오.• 비금속(Nonmetallic) 튜브를 테트라히드로푸란(Tetrahydrofuran: THF) 또는 농축 질산 또는 황산과 함께 사용하지 마십시오.

• 염화 메틸렌(Methylene chloride) 및 디메틸술폭시드(Dimethyl sulfoxide)는 비금속 튜브를 부풀려 튜브의 파열 압력을 크게 감소시킬 수 있으므로 유의하십시오.

警告：圧力のかかったポリマーチューブを扱うときは、注意してください。

• 加圧されたポリマーチューブの付近では、必ず保護メガネを着用してください。

• 近くにある火を消してください。

• 著しく変形した、または折れ曲がったチューブは使用しないでください。

• 非金属チューブには、テトラヒドロフラン(THF)や高濃度の硝酸または硫酸などを流

さないでください。

• 塩化メチレンやジメチルスルホキシドは、非金属チューブの膨張を引き起こす場合があり、その場合、チューブは極めて低い圧力で破裂します。


Warning: The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired.

Attention: L’utilisateur doit être informé que si le matériel est utilisé d’une façon non spécifiée par le fabricant, la protection assurée par le matériel risque d’être défectueuses.

Vorsicht: Der Benutzer wird darauf aufmerksam gemacht, dass bei unsachgemäßer Verwenddung des Gerätes die eingebauten Sicherheitseinrichtungen unter Umständen nicht ordnungsgemäß funktionieren.

Attenzione: si rende noto all'utente che l'eventuale utilizzo dell'apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall'apparecchiatura.

Advertencia: el usuario deberá saber que si el equipo se utiliza de forma distinta a la especificada por el fabricante, las medidas de protección del equipo podrían ser insuficientes.

警告：使用者必須非常清楚如果設備不是按照製造廠商指定的方式使用，那麼該設備所提供的保護將被消弱。

警告：使用者必须非常清楚如果设备不是按照制造厂商指定的方式使用，那么该设备所提供的保护将被削弱。

경고: 제조업체가 명시하지 않은 방식으로 장비를 사용할 경우 장비가 제공하는 보호 수단이 제대로 작동하지 않을 수 있다는 점을 사용자에게 반드시 인식시켜야 합니다.

警告：ユーザーは、製造元により指定されていない方法で機器を使用すると、機器が提供している保証が無効になる可能性があることに注意して下さい。


Warning: To protect against fire, replace fuses with those of the type and rating printed on panels adjacent to instrument fuse covers.Attention: pour éviter tout risque d'incendie, remplacez toujours les fusibles par d'autres du type et de la puissance indiqués sur le panneau à proximité du couvercle de la boite à fusible de l'instrument.Vorsicht: Zum Schutz gegen Feuer die Sicherungen nur mit Sicherungen ersetzen, deren Typ und Nennwert auf den Tafeln neben den Sicherungsabdeckungen des Geräts gedruckt sind.Attenzione: per garantire protezione contro gli incendi, sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento.Advertencia: Para evitar incendios, sustituir los fusibles por aquellos del tipo y características impresos en los paneles adyacentes a las cubiertas de los fusibles del instrumento.

警告 : 為了避免火災，更換保險絲時，請使用與儀器保險絲蓋旁面板上所印刷之相同類型與規格的保險絲。

警告 : 为了避免火灾，应更换与仪器保险丝盖旁边面板上印刷的类型和规格相同的保险丝。

경고: 화재의 위험을 막으려면 기기 퓨즈 커버에 가까운 패널에 인쇄된 것과 동일한 타입 및 정격의 제품으로 퓨즈를 교체하십시오.

警告 : 火災予防のために、ヒューズ交換では機器ヒューズカバー脇のパネルに記載されているタイプおよび定格のヒューズをご使用ください。


Electrical and handling symbols

Electrical symbolsThese can appear in instrument user manuals and on the instrument’s front or rear panels.

Electrical power on

Electrical power off

Standby

Direct current

Alternating current

Protective conductor terminal

Frame, or chassis, terminal

Fuse

Recycle symbol: Do not dispose in municipal waste.


Handling symbolsThese handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments.

Keep upright!

Keep dry!

Fragile!

Use no hooks!

Electrical and handling symbols A-13

B Specifications

This appendix contains specifications for the following:• Operational• Electrical• Environmental

Operational specifications

Item SpecificationNumber of solvents One to fourModes of operation Gradient, isocratic, and flow programmingOperating flow range

Programmable from 1 to 150 mL/min in 0.01-increments (500-µL heads) and from 4 to 300 mL/min in 0.01-increments (1000-µL heads).

Composition range 0 to 100% programmable in 1%-increments for each of four reservoirs: A, B, C, and D. Total composition must sum to 100%.

Compositional accuracy

Better than 3.0%.

Automatic solvent Sparging

Standard, helium gas, input gas pressure range 50 to 150 psi (3.5 to 10.5 Kg/cm2). Helium flow rate range 0 to 100 mL/min, programmable in 1-mL/min increments.

Pressure maximum 2000 psi (140.6 kg/cm2) over entire flow rate range (1000-µL heads); 4000 psi (281.2 Kg/cm2) over entire flow range (1 to 150 mL/min) with 500-µL pump heads.

Programmable pressure limits

Upper: 51 to 2000 psi (3.6 to 281.2 kg/cm2)Lower: 0 to 3950 (0 to 277.7 kg/cm2)

B-1

Program storage Storage for sets of 15 tables (Gradient, Event, and Detector), with up to 15 steps per table.Storage for a time-based multimethod program with up to 48 individual steps, linking sets of tables.EEPROM stores setup parameters and tables and preserves memory indefinitely following power loss or extended shutdown.

Gradient profiles Eleven gradient curves (including linear, step, concave, and convex).

Keyboard Dedicated keys provide direct access to all display screens. Cursor movement keys simplify programming, editing, and parameter entering. All keys are solvent resistant.

Programmable timed events

Fifteen timed events can be programmed and stored for each table:• Set S1 to S4 for accessory control• Audible alarm• Set sparge rate• Return to initial conditions• Change to new Gradient table

Electrical specifications

Item SpecificationPower requirements

Maximum power 500 VA

Power consumption 350 wattsFrequency 50 or 60 HzFuse Nominal Voltage Fuse (Amp)

100, 120 4 Amps 220, 240 2 Amps

Pump power fuse 4 A, 125 VAC, IEC (Time Delay) 5 × 20 mm. Protects the pump from excessive current loads.

Operational specifications (Continued)

Item Specification

B-2 Specifications

Time or operator controllable switch closures

Four controllable switch closures to ground. Open collector terminals pulled up to +5 VAC through 10 K ohm resistors. Can be pulled up to 12 VAC.Maximum allowable current = 1.0 A total. Maximum allowable voltage = 12 VAC. Outputs are controlled directly from keyboard or events program.Two modes available from keyboard and events program:• ON = Switch closed to ground.• OFF = Switch open (+5 VAC to ground).Additional mode available from events program:• Pulse = Single contact closure to opposite state for

100 ms.Auxiliary power supply

12 VAC to ground fused supply. 1.3-A maximum current.

Hold output One open connector switch closure to ground. Maximum current sink 40 mA. Low output indicates pumps stopped due to:• Exceeding high pressure limit• Below low pressure limit• Abort input• Startup conditionUsed to communicate with Hold input on Waters autoinjectors or other compatible inputs to prevent further injections.

Chart output (analog signal)

Two terminals (+, –) for recording pump flow rate traces or monitoring solvent composition (10-mV output).

Press output (analog signal)

Two terminals (+, –) for recording pump pressure traces (10-mV output).

Ground terminals Connected to signal ground used as reference for outputs (Hold, INJect, and switches S1 to S4), inputs (Inject and Abort), or 12 VAC power supply (not for use with 10-mV outputs).

Electrical specifications (Continued)

Item Specification

B-3

Environmental specifications

Item SpecificationTemperature Operating range: 4 to 38 °C (39 to 100 °F)

Storage range: 0 to 50 °C (32 to 122 °F)Humidity Operating range: 20 to 90% relative humidity

Storage range: 0 to 90% relative humidity (noncondensing)

Solvent compatibility

Compatible with solvents consistent with materials of construction

Physical specifications

Item SpecificationWetted surface material

316 stainless steel, ruby, sapphire, fluorocarbon polymer

System controller Height: 7.19 inches (18.26 cm)Depth: 21.75 inches (55.25 cm)Width: 11.32 inches (28.75 cm)Weight: 42 pounds (19.1 kg)

Fluid handling unit Height: 9.625 inches (24.4 cm)Depth: 20.5 inches (52.1 cm)Width: 11.125 inches (28.3 cm)Weight: 60.2 pounds (27.3 kg)

Shelf unit (PrepLC

system)Height: 26 inches (66 cm)Depth: 22 inches (50.88 cm)Width: 14 inches (35.5 cm)Weight: 26.4 pounds (11.9 kg)

Shelf unit and column rack (Delta-Prep system)

Height: 21.5 inches (54.61 cm)Depth: 22 inches (50.88 cm)Width: 23.7 inches (60.2 cm)Weight: 25 pounds (11.3 kg)

B-4 Specifications

C Spare Parts

This appendix lists the spare parts that are recommended for customer installation. Use the number in the Recommended Stock column as a guide to the quantity of each item to keep on hand.

Common parts

Item Recommended stock Part numberValve, 3-way purge 0 700001309System interface cable 0 WAT088972

3725 injector

Item Recommended stock Part number3725 Rebuild Kit 1 201000116Stainless Steel Needle #16

1 600000111

Needle Port Plug 0 700000593Adapter, 1/8 in. – 1/16 in.

0 WAT073574

Sample loops

Item Recommended stock Part number5-mL sample loop 0 WAS07370310-mL sample loop 0 WAS07370420-mL sample loop 0 WAS073705

C-1

150-mL configuration

Item Recommended stock Part number150-mL Conversion Kit 0 205000135Inlet/Outlet Check Valve Rebuild Kit

1 WAT037495

Inlet Check Valve Assembly

1 WAT033325

Outlet Check Valve Assembly

1 WAT033326

Plunger Assembly 2 700002123Plunger Seal Kit 2 WAT037490Gradient Proportioning Valve Assembly

0 WAT082823

300-mL configuration

Item Recommended stock Part number300-mL Conversion Kit 0 205000136Inlet Check Valve Assembly

1 WAT052230

Outlet Check Valve Assembly

1 WAT052220

Inlet Manifold 0 WAT052201Plunger Assembly 2 WAT052111Plunger Seal Kit 4 WAT052223Gradient Proportioning Valve Assembly

0 WAT082823

C-2 Spare Parts

D Solvent Considerations

Contents

Topic PageClean solvent D-2Solvent miscibility D-3How to use miscibility numbers (M-numbers) D-5Buffered solvents D-6Solvent head height D-6Solvent viscosity D-6Solvent degassing D-7Solvent degassing methods D-8

D-1

Clean solvent

The use of clean, particle-free solvents is important not only for reproducible results but also for operation with minimal maintenance. An unclean solvent can cause baseline noise and drift. Recommended solvent preparation includes using Milli-Q® water and filtering through a 0.45-µm filter.When operating a system with water, clean the water just before use to eliminate the presence of bacteria. To clean water, distill it in a glass and pass it through a Norganic water purification system or prepare water with the Milli-Q system. Contact your Waters® representative for more information.Inlet solvent filters can be used at low flow rates. To avoid drawing foreign particles into the pump where they can accumulate and affect pump operation, keep the solvent reservoir filter clean. If the filter becomes plugged, clean or replace it.If the Waters Prep system is not used for several weeks, flush the system (outlet line, inlet lines, and check valves) with filtered, distilled water containing 10% methanol or acetonitrile. Remove the column if it is not compatible with this storage protocol.

Warning: Always follow safe laboratory practices when handling solvents. Know the chemical and physical properties of the solvents. Refer to the Material Safety Data Sheet for the solvents in use.

Caution: Do not allow the solvent system to exceed 5 psi pressure when you use pressurized solvent containers.

D-2 Solvent Considerations

Solvent miscibility

Before changing from one solvent to another, see the table below to determine the miscibility of the solvents being used. The following considerations apply when changing solvents:

• Changes involving two miscible solvents are made directly. Changes involving two solvents which are not totally miscible (for example, from hexane to water) require an intermediate solvent (such as methanol or THF).

• Solvent miscibility is affected by temperature. If operating at an elevated temperature, consider the effect of the higher temperature on solvent solubility.

• Buffers dissolved in water can precipitate salts when mixed with organic solvents. When switching from a strong buffer to an organic solvent, flush the buffer out of the system with distilled water before adding the organic solvent. Also, when using a strong buffer, flush all pathways with distilled water before shutting the system down and leave distilled water in the system (flush with 10% methanol in water for shutdowns scheduled to be more than one day).

Physical properties of solventsa

Polarity index Solvent Viscosity

CP, 20 °C

Boiling Point °C (1 atm)

Miscibility Number (M)

Wavelength Cutoffs (nm)

0.3 N-decane 0.92 174.1 29 —0.4 Isooctane 0.50 99.2 29 2100.0 N-hexane 0.313 68.7 29 —0.0 Cyclohexane 0.98 80.7 28 210C.7 Butyl ether 0.70 142.2 26 —C.8 Triethylamine 0.38 89.5 26 —2.2 Isopropyl ether 0.33 68.3 — 2202.3 Toluene 0.59 10C.6 23 2852.4 P-xylene 0.70 138.0 24 2903.0 Benzene 0.65 80.1 21 2803.3 Benzyl ether 5.33 288.3 — —

Solvent miscibility D-3

3.4 Methylene chloride

0.44 39.8 20 245

3.7 Ethylene chloride

0.79 83.5 20 —

3.9 Butyl alcohol 3.00 117.7 — —3.9 Butanol 3.01 177.7 15 —4.2 Tetrahydro-

furan0.55 66.0 17 220

4.3 Ethyl acetate 0.47 77.1 19 2604.3 1-propanol 2.30 97.2 15 2104.3 2-propanol 2.35 117.7 15 —4.4 Methyl acetate 0.45 56.3 15, 17 2604.5 Methyl ethyl

ketone0.43 80.0 17 330

4.5 Cyclohexanone 2.24 155.7 28 2104.5 Nitrobenzene 2.03 210.8 14, 20 —4.6 Benzonitrile C.22 19C.1 15, 19 —4.8 Dioxane C.54 10C.3 17 2205.2 Ethanol C.20 78.3 14 2105.3 Pyridine 0.94 115.3 16 3055.3 Nitroethane 0.68 114.0 — —5.4 Acetone 0.32 56.3 15, 17 3305.5 Benzyl alcohol 5.80 205.5 13 —5.7 Methoxyethanol C.72 124.6 13 —6.2 Acetonitrile 0.37 8C.6 11, 17 2106.2 Acetic acid C.26 117.9 14 —6.4 Dimethylforma-

mide0.90 153.0 12 —

Physical properties of solventsa (Continued)


CP, 20 °C





How to use miscibility numbers (M-numbers)

Miscibility numbers (M-numbers) are used to predict the miscibility of a liquid with one of the standard solvents.To predict the miscibility of the two liquids, subtract the smaller value from the larger.

• If the difference between the two M-numbers equals 15 or less, the two liquids are miscible in all proportions at 15 °C.

• A difference of 16 units shows that the two liquids possess a critical solution temperature between 25 and 75 °C, with 50 °C as the optimal temperature.

• If the difference equals 17 or more, the two liquids are immiscible or their critical solution temperature is above 75 °C.

Interaction between the molecules of the two liquids can sometimes change the expected degree of miscibility. For example, ethers or tertiary amines show unpredicted miscibility with hydroxylic solvents due to hydrogen bonding. Unusually strong hydrogen bonding is also responsible for the miscibility of long-chain alcohols or carboxylic acids with standard solvents of low M-number. Conversely, they show anomalous immiscibility with aprotic solvents of low M-number.Some solvents prove immiscible with solvents at both ends of the lipophilicity scale. These solvents receive a dual M-number. The first number, always lower than 16, indicates the degree of miscibility with highly lipophilic

6.5 Dimethylsul-foxide

2.24 189.0 9 —

6.6 Methanol 0.60 64.7 12 2107.3 Formamide 3.76 210.5 3 —9.0 Water C.00 100.0 — —

a. Adapted from Godfrey, Norman B., Solvent Selection via Miscibility Number, CHEMTECH, 359-363 (1972).

Physical properties of solventsa (Continued)


CP, 20 °C




How to use miscibility numbers (M-numbers) D-5

solvents. The second number applies to the opposite end of the scale. A large difference between these two numbers indicates a limited range of miscibility. For example, some fluorocarbons are immiscible with all the standard solvents and have M-numbers of 0, 32. Two liquids with dual M-numbers are usually miscible with each other.A liquid is classified in the M-number system by testing for miscibility with a sequence of standard solvents. A correction term of 15 units is then either added or subtracted from the cutoff point for miscibility.

Buffered solvents

When using a buffer, obtain a good quality reagent and filter it through a 0.45-µm syringe filter.Do not leave the buffer in the pump upon shutdown. Once the run is complete, remove the column and clean the buffer out completely by flushing the outlet line, inlet line, and filter with approximately 40 mL of Milli-Q water containing 0.2% sodium azide. Use a syringe to flush distilled water through the vent holes while running the pump.After flushing with water, flush the pump with 10% methanol before shutdown. Store the pump in 10% methanol/water during the shutdown.

Solvent head height

The Waters Prep system is flexible in allowing you where to position the solvent reservoirs. Ideally, the reservoirs should be positioned approximately level with the pump heads, particularly at flow rates of 2.0 mL/min or less. However, well maintained (good plunger seals) pump heads are able to self-prime and draw well sparged solvent from a level beneath the pump heads. When drawing solvent from containers on the floor beneath the bench, well sparged solvent is critical. Gas in the solvent can come out of solution as the pump draws and reduces pressure in the solvent tubing.

Solvent viscosity

It is important to keep in mind that when running a gradient, the viscosity changes which occur as the solvents are mixed in different proportions can result in Pressure changes during the run. For example, a 1:1 mixture of


water and methanol produces twice the pressure of either water or methanol alone. If the extent to which the pressure changes will affect the analysis is not known, monitor the pressure during the run using the controller’s Chart output provided for this purpose (select %A or %B).

Solvent degassing

Mobile phase difficulties account for 70% or more of all problems in liquid chromatography. Degassing solvents used in the mobile phase is one of the best measures to eliminate these problems. The benefits are:

• Stability in the baseline and enhanced sensitivity in some types of chromatographic detectors

• Reproducible retention times for eluting peaks• Reproducible injection volumes• Stable pump operation

Only a finite amount of gas is dissolved in a given volume of liquid under specific conditions. This amount depends on the temperature of the liquid, pressure on the liquid, and the chemical affinity of the gas for the liquid.Generally, a gas is most soluble in a solvent where the intermolecular attractive forces between molecules of the solvent are similar to those of the gas (“like dissolves like”). If the main attractive forces are Van der Waals forces, the gas is more soluble in this solvent than in one with dipole forces or hydrogen bonding. Thus, a larger amount of helium, nitrogen, oxygen, or hydrogen dissolves in alkanes and benzene than in water.Temperature affects the solubility of gases in two ways. First, the higher the boiling point of a gas, the more soluble the gas is in a given solvent. Second, increasing the temperature of the gas/liquid solution affects the percentage of gas in solution. If the heat of solution results in an exothermic reaction, the percentage of gas in solution reduces. If the reaction is endothermic, the percentage increases. For example, solubility of helium in water decreases with an increase in temperature, but the solubility of helium in benzene increases in direct proportion to the temperature.The mass of gas dissolved in a given volume of solvent is proportional to the partial pressure of the gas in the vapor phase of the solvent. If the gas pressure decreases, the amount of that gas in solution also decreases.

Solvent degassing D-7

Dissolved oxygen affects UV-VIS detector performance in several ways1. Oxygen dissolved in solvents can form a UV absorbing complex, the amount of which is considerably different in different solvents. The effect is particularly strong with wavelengths below 260 nm. Therefore, any change in dissolved oxygen content can affect a UV baseline considerably. This phenomenon is particularly evident in the solvent tetrahydrofuran (THF). Dissolved oxygen does not seem to affect the absolute sensitivity of a UV system, but primarily causes baseline drift. This effect is especially noticeable during gradient operation where the dissolved oxygen content varies between the different solvents and, as the composition changes, causes erratic baselines or even peak-shaped artifacts on the baseline.Dissolved oxygen in a fluorescence detector has quite a different effect. It causes a tremendous loss of sensitivity. Bowen and Williams2 have discussed the quenching of aromatic hydrocarbons by dissolved oxygen in fluorescence detectors. Parker and Barnes3 have reported a 95% reduction in sensitivity of the fluorescence of borate-benzoin complex in air equilibrated ethanol. The oxygen quenching varies with different types of compounds, and aromatic hydrocarbons, aliphatic aldehydes, and ketones are especially susceptible.If for any reason the characteristics of the solvent change, the precision and accuracy of the solvent delivery system can be adversely affected. This can cause variations in both peak retention and, to some extent, peak height or area.

Solvent degassing methods

Sparging, or bubbling a gas through solvent, reduces the partial pressure of the unwanted gas at the surface of the solvent. This removes unwanted gas from solution and saturates the solvent with the second gas. Sparging with helium removes background absorbance on a UV detector and the quenching phenomenon caused by dissolved oxygen on a fluorescence detector.Helium sparging combines the convenience of short initial degassing time required, ease of maintaining the solvent condition during operation, and complete control within the framework of the Waters Prep systems.

1. S. R. Bakalyar, M. B. T. Bradley, R. Hoganen, Journal of Chromatography, 158 (1978) 277.2. E. J. Bowen and A. H. Williams, Trans. Faraday Soc., 35 (1939) 65.3. C. A. Parker and W. J. Barnes, Trans. Faraday Soc., 82 (1957) 606.


The degassing operation should be as efficient as possible. To remove the gas as quickly as possible, be aware of the following considerations:

• Helium sparging gives stable baselines and better sensitivity in a fluorescence detector, and prevents the reabsorption of atmospheric gases. Solvents sparged with helium generally are well degassed by the time they reach the pump because the helium will diffuse out of the solvent into the atmosphere through the PTFE tubing connecting the reservoirs to the pump. The solvent arrives at the pump with less than 1 atmosphere partial pressure of helium.

• Vacuum alone is too slow as an acceptable means of degassing solvent.

In the techniques listed above, except for helium sparging, the solvent reequilibrates to air saturation in 12 to 24 hours (depending on the solvent).Degassing by vacuum or sonification is often performed for improved pump performance in high-pressure applications but might not yield the required baseline stability for high-sensitivity absorbance detection. Therefore, the most practical solution for most applications is helium sparging since, due to the low solubility of the gases used, it does not impair pump performance in most solvents.

Warning: It is dangerous to apply vacuum to the brown gallon bottles in which solvent is shipped. They stand a high risk of implosion under these conditions.

Solvent degassing methods D-9

Index

Symbols+12 V supply terminal 1-11

AAbsolute pressure change 3-10AC power, cabling 2-15Accessories 1-13

flow splitter 2-49M-1000 PrepPAK module 1-13,

2-44, 2-46SE 120 chart recorder 1-13

Analog output signal 1-10audience and purpose ivAutoinjector troubleshooting 3-27

reproducibility errors 3-28seal pack leak 3-27

Autosamplers, IEEE-488 interface connections 2-15

Auxiliary power fuse 3-69

BBaseline drift, slow 3-19Baseline noise 3-8–3-9

long-term cycling 3-20random 3-21

biohazard warning A-5Blank screen 3-23, 3-37Buffered solvents D-6burst warning A-3

CCabling

AC power 2-15IEEE-488 2-15PowerLine 2-17

Can’t Save Table Currently Executing message 3-32

caution symbol A-5Chart and Press switches 1-10Chart test 3-41Chassis ground terminal 1-11Check Corresponding Table message

3-33Check valves

cleaning 3-50, 3-51replacing 3-50

inlet check valves 3-51–3-53outlet check valves 3-50

chemical hazard warning A-5Chromatography troubleshooting 3-16

baseline noiselong-term cycling 3-20random 3-21

doubled retention times 3-17flat-topped peaks 3-22rapid baseline drift 3-18reduced retention time 3-17sensitivity loss 3-22slow baseline drift 3-19straight baseline 3-20

CLEAR key 1-17Column rack 1-12

large-scale 1-12tubing configuration 3-48

Column switching valve 1-5Column troubleshooting 3-15Composition Must Sum to 100%

message 3-33Connection ports 1-13Connector types 1-10Contacting Waters Technical Service

3-2Controller

Index-1

choosing configuration 1-19failure on powerup 1-15Gradient configuration 1-22Inject terminal 1-9PowerLine configuration 1-20

Controller keys 1-18–1-19Help 1-19keypad 1-16, 1-17screen keys 1-18

Cursor movement keys 1-16

DData system 2-15

Empower IEEE-488 cabling 2-15IEEE-488 addresses 2-18powerup/power down sequence

2-18Waters 746 RS-232 cabling 2-20Waters 845/860 IEEE-488 cabling

2-15Degassing solvents D-7Delta-Prep system 1-3, 1-5

fluid handling unit 1-4inlet and outlet ports 1-5purge outlet port 1-5purge valve 1-5Rheodyne injector 1-5scale switching valve 1-5

Detector failure 1-16Detector table 1-21Detectors

IEEE-488 interface connections 2-15

lamp not lit 3-30troubleshooting 3-30

RI 3-31UV 3-30

Digital input signal 1-10Display screen 1-16

Doubled retention times 3-17

EEC Authorized Representative viElectrical connections

AC power 2-15IEEE-488 interface 2-15RS-232 interface 2-20

electrical symbols A-12Empower

IEEE-488 addresses 2-18IEEE-488 cabling 2-15powerup/power down sequence

2-18ENTER key 1-17Entering values 1-17equipment guidelines iv, A-6Erratic retention times 3-10Error messages

clearing 1-17reproducibility 3-18

Event table 1-21Extended test routines 3-36–3-42

Chart 3-41External Inject 3-40External Switch Output 3-40general instructions 3-39Hold Output 3-41IEEE-488 address 3-42required tools 3-38self-diagnostics 3-36–3-37starting up 3-37Stop Flow Input 3-39–3-40

External Inject test 3-40External Switch Output test 3-40

FFirst Vial Greater Than Last Vial

message 3-34flammable solvents A-4

Index-2

Flat-topped peaks 3-22Flow path 1-6

large scale 1-12small scale 1-12

Flow splitter option 1-14, 2-49Fluid handling unit 1-5–1-7

Delta-Prep system 1-4maintenance 3-48–3-67plunger wash 1-6

Fluid metering 1-6Fluidic system troubleshooting 3-24

pump leaks 3-25pump not running 3-24pump squeak 3-26

Fraction collectors, Waters 1-13Front panel 1-16–1-19

controller keys 1-18–1-19display screen 1-16

Fuses+5 A 3-70–3-71auxiliary power 3-69operating voltage 3-67pump power 3-70replacing 3-67–3-71

GGradient controller 2-15

description 1-22screen layout 1-23

Gradient table 1-21, 3-44

Hhandling symbols A-13Hardware troubleshooting 3-23

detectors 3-30fluidic system 3-24injector 3-26system 3-23–3-24

Head support, removing 3-57Help 3-2

I

online 1-19High system pressure 3-5

considerations 3-6Hold Output test 3-41Hold signal 1-10HOME key 1-16

IIEEE-488 address test 3-42IEEE-488 interface 1-20

cable length 2-17communication 2-15, 2-17data system powerup sequence

2-19Empower cabling 2-15IEEE-488 address, setting 2-18PowerLine controller cabling 2-17shut down sequence 2-19Waters 845/860 cabling 2-15

Illegal Vial Number message 3-33Incorrect retention times 3-10Inject start terminal 1-9Injector panel 1-12–1-13

connection ports 1-13purge valve 1-13

Injector troubleshooting 3-26Rheodyne injector 3-27

needle seal leakage 3-27plugged valve passages 3-27

Waters autoinjectors 3-27reproducibility errors 3-28seal pack leak 3-27

InjectorsRheodyne 3725i 1-12Rheodyne 7725i 1-11

Inlet and outlet ports 1-5Delta-Prep system 1-5

Inlet check valvesdisassembly 3-51

Index-3

replacing 3-51–3-53retainer cup 3-51

Inlet port connection 2-38Installing

plastic tubing fittings 2-25Rheodyne injectors with Delta-Prep

2-40intended use vInvalid Injection Number message

3-34Invalid Vial Number message 3-34ISM classification v

KKeyboard, dead 3-24Keypad 1-16Keys

controller 1-18cursor movement 1-16function 1-16screen 1-16

LLamp not lit 3-30Large-scale flow path 1-12Leaking solvent 3-25

MMaintenance

+5 A fuse replacement 3-70auxiliary power fuse replacement

3-69check valve cleaning 3-50fluid handling unit 3-48–3-67fuse replacement 3-67–3-71plunger wash seal replacement

3-65–3-67pump head removal 3-56pump plunger replacement

3-61–3-67

pump seal replacement 3-58replacing +5 A fuse 3-70–3-71replacing auxiliary power fuse 3-69replacing operating voltage fuse

3-67mass spectrometer shock hazard A-4Messages

clearing 3-32shutdown 3-34warning 3-32–3-34

Metering, fluid 1-6Method table 1-21Miscibility numbers D-5

NNeedle port 3-74Needle seal 3-72

leakage 3-27No key response 3-24Number keys 1-17Number Out of Range message 3-33

OOnline Help 1-19Open collector signal 1-10Outlet check valves

assembly 3-54Outlet port 1-5

PPeak resolution troubleshooting 3-13Peaks, flat-topped 3-22Performance tests

extended test routines 3-36–3-42pressure transducer calibration

3-42–3-43, 3-49solvent proportioning valve pair

test 3-43Physical specifications B-4Plugged valve passages 3-27

Index-4

Plunger wash 1-6fluid handling unit 1-6seal replacement 3-65–3-67

Potentiometer, adjusting 3-49Power Up screen, leaving 1-16Powering down

sequence 2-19system 1-19

Powering updata system sequence 2-19system 1-14–1-16

PowerLine controllerdescription 1-20screen layout 1-21

PrepLC system 1-2accessories 1-13, 1-13–1-14choosing controller type 1-19injector panel 1-12rear panel connections 2-7

PrepPAK module 1-13Pressure fluctuations 3-10Pressure transducer, calibration test

3-42–3-43, 3-49–3-50Program Method Table Saved message

3-33Pump

does not run 3-24leaks solvent 3-25power fuse 3-70squeak 3-26

Pump head, removing 3-56Pump plunger, replacing 3-61–3-65Pump seal, replacing 3-58Purge outlet port, Delta-Prep system

1-5Purge valve 1-5, 1-12

Delta-Prep system 1-5purpose and audience iv

I

RRack, column 1-12Radial-Pak cartridges 1-14Random baseline noise 3-21Rapid baseline drift 3-18Rear panel connections, terminal strip

3-39Recycle to pump port, connection 2-38Reduced retention time 3-17Removing

head support 3-57pump head 3-56pump seal 3-59

Replace with new Table message 3-32Replacing

inlet check valves 3-51plunger wash seal 3-65pump plunger 3-61

Replacing fuses 3-67–3-71+5 A fuse 3-70–3-71auxiliary power fuse 3-69operating voltage fuse 3-67pump power fuse 3-70

Reproducibility errors 3-28Required tools 3-38Restarting program 3-37Retainer cup 3-51Retention time troubleshooting 3-4Retention times

changes 3-10doubled 3-17erratic 3-10incorrect 3-10reduced 3-17

Rheodyne injectors 1-53725i injector 1-127725i injector 1-11Delta-Prep system 1-5installing with Delta-Prep 2-40

Index-5

needle seal leakage 3-27plugged valve passages 3-27sample loading methods 1-11troubleshooting 3-27

RI detectors, troubleshooting 3-4, 3-31Rotor seal, replacement 3-74RS-232 2-20

cabling to Waters 746 2-20communication 2-20

Ssafety advisories A-1Sample loading methods, Rheodyne

injector 1-11Scale switching valve, Delta-Prep

system 1-5Screen keys 1-18Screens

blank 3-23, 3-37display 1-16layout of PowerLine controller 1-21

SE 120 chart recorder 1-13Seal insertion tool 3-58Seal pack leaks 3-27Self-diagnostics 1-15, 3-36–3-37Sensitivity loss 3-22Setting IEEE-488 address 2-18Shutdown messages 3-34Signal

analog output 1-10digital input 1-10ground terminal 1-11hold 1-10open collector 1-10

Slow baseline drift 3-19Small-scale flow path 1-12Small-scale sample injection valve

option 2-40Software troubleshooting 3-31

shutdown messages 3-34warning messages 3-32–3-34

Solvent considerationsclean D-2degassing D-7miscibility D-3viscosity D-6

Solvent proportioning valve pair test 3-43–3-46

Gradient table 3-44solvent preparation 3-43valve failure 3-45

Solvent, leaking 3-25Specifications B-1Startup sequence, IEEE-488 devices

2-18Stator 3-74Stator ring 3-74Step Does Not Exist message 3-33Stop Flow and Hold switches 1-9Stop Flow Input test 3-39–3-40Stop Flow terminal, shutdown 3-35Straight baseline 3-20Switch outputs, extended test 3-40Switches

Chart and Press 1-10Start Flow and Hold 1-9

symbolscaution A-5electrical A-12handling A-13warning A-2

System modules, Waters Prep Series 1-5

System pressure, reference point 3-5System shutdown response 3-34System troubleshooting 3-23–3-24

blank screen 3-23no key response 3-24

Index-6

SystemsDelta-Prep 1-3–1-5PrepLC 1-2

TTables

Detector 1-21Event 1-21Gradient 1-21, 3-44Method 1-21

Technical assistance 3-2Terminal

+12 V supply 1-11chassis ground 1-11Inject start 1-9signal ground 1-11

Tests, performance 3-36These 3-50Tools

required 3-38seal insertion 3-60

Troubleshooting 3-2chromatography 3-16hardware 3-23software 3-31

Troubleshooting flowchartsbaseline noise 3-8–3-9column 3-15erratic retention times 3-10high system pressure 3-5incorrect retention times 3-10poor peak resolution 3-13

Tubing, installing plastic fittings 2-26

UUV detectors

lamp not lit 3-30troubleshooting 3-30

I

VValues, entering 1-17Viscosity D-6

WWarning messages 3-32–3-34

Can’t Save Table Currently Executing 3-32

Check Corresponding Table 3-33First Vial Greater Than Last Vial

3-34Illegal Vial Number 3-33Invalid Injection Number 3-34Invalid Vial Number 3-34Number Must Sum to 100% 3-33Number Out of Range 3-33Program Method Table Saved 3-33Replace with new Table 3-32Step Does Not Exist 3-33

warning symbols A-2, A-6Waters 746 Data Module

communication information 2-20RS-232 cabling 2-20

Waters 746 integrator 1-20Waters 845/860, powerup/power down

sequence 2-18Waters autoinjectors, troubleshooting

3-27Waters data system 2-15Waters Delta-Prep system. See

Delta-Prep systemWaters Prep Series

choosing controller 1-22powering down 1-19powering up 1-14system modules 1-5, 1-13

Waters PrepLC system. See PrepLC system

Index-7

Waters Technical Service, contacting 3-2

WorkstationsEmpower 2-15MassLynx 2-15

Index-8

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