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Waters 1525µ and 1525EF HPLC Pump

Installation and Maintenance Guide

34 Maple StreetMilford, MA 01757

71500152504, Revision A

NOTICE

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, the use of this document.

© 2003 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

Micromass and Waters are registered trademarks, and FractionLynx and MassLynx are trademarks of Waters Corporation.

Microsoft is a registered trademark, and Windows is a trademark of Microsoft Corporation.

National Instruments is trademark of National Instruments Corporation.

Tefzel is a registered trademark of E. I. du Pont de Nemours and Company.

All other trademarks or registered trademarks are the sole property of their respective owners.

Note: When you use the instrument, follow generally accepted procedures for quality control and methods development.

If you observe a change in the retention of a particular compound, in the resolution between two compounds, or in peak shape, immediately determine the reason for the changes. Until you determine the cause of a change, do not rely on the separation results.

Note: The Installation Category (Overvoltage Category) for this instrument is Level II. The Level II Category pertains to equipment that receives its electrical power from a local level, such as an electrical wall outlet.

STOPAtención: 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: eventuali modifiche o alterazioni apportate a questa unità e non espressamente approvate da un ente responsabile per la conformità annulleranno l’autorità dell’utente ad operare l’apparecchiatura.

Atención: 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.

Caution: 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 Tefzel tubing that has been severely stressed or kinked.

• Do not use Tefzel tubing with tetrahydrofuran (THF) or concentrated nitric or sulfuric acids.

• Be aware that methylene chloride and dimethyl sulfoxide cause Tefzel tubing to swell, which greatly reduces the rupture pressure of the tubing.

Attention : Soyez très prudent en travaillant avec des tuyaux de polymères sous pression :

• Portez toujours des lunettes de protection quand vous vous trouvez à proximité de tuyaux de polymères.

• Eteignez toutes les flammes se trouvant à proximité.

• N'utilisez pas de tuyau de Tefzel fortement abîmé ou déformé.

• N'utilisez pas de tuyau de Tefzel avec de l'acide sulfurique ou nitrique, ou du tétrahydrofurane (THF).

• Sachez que le chlorure de méthylène et le sulfoxyde de diméthyle peuvent provoquer le gonflement des tuyaux de Tefzel, diminuant ainsi fortement 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 Tefzel-Schläuche verwenden, die stark geknickt oder überbeansprucht sind.

• Tefzel-Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte Salpeter- oder Schwefelsäure verwenden.

• Durch Methylenchlorid und Dimethylsulfoxid können Tefzel-Schläuche quellen; dadurch wird der Berstdruck des Schlauches erheblich reduziert.

Precauzione: prestare attenzione durante le operazioni con i tubi di polimero sotto pressione:

• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati.

• Estinguere ogni fonte di ignizione circostante.

• Non utilizzare tubi Tefzel soggetti a sollecitazioni eccessive o incurvati.

• Non utilizzare tubi Tefzel contenenti tetraidrofurano (THF) o acido solforico o nitrico concentrato.

• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano rigonfiamento nei tubi Tefzel, che riducono notevolmente il limite di pressione di rottura dei tubi stessi.

Advertencia: manipular con precaución los tubos de polímero bajo presión:

• Protegerse siempre los ojos en las proximidades de tubos de polímero bajo presión.

• Apagar todas las llamas que estén a proximidad.

• No utilizar tubos Tefzel que hayan sufrido tensiones extremas o hayan sido doblados.

• No utilizar tubos Tefzel con tetrahidrofurano (THF) o ácidos nítrico o sulfúrico concentrados.

• No olvidar que el cloruro de metileno y el óxido de azufre dimetilo dilatan los tubos Tefzel, lo que reduce en gran medida la presión de ruptura de los tubos.

Caution: 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 unter Umständen nicht ordnungsgemäß funktionieren.

Precauzione: l’utente deve essere al corrente del fatto che, se l’apparecchiatura viene usta in un modo specificato dal produttore, la protezione fornita dall’apparecchiatura potrà essere invalidata.

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.

Caution: To protect against fire hazard, replace fuses with those of the same type and rating.

Attention : Remplacez toujours les fusibles par d’autres du même type et de la même puissance afin d’éviter tout risque d’incendie.

Vorsicht: Zum Schutz gegen Feuergefahr die Sicherungen nur mit Sicherungen des gleichen Typs und Nennwertes ersetzen.

Precauzione: per una buona protezione contro i rischi di incendio, sostituire i fusibili con altri dello stesso tipo e amperaggio.

Advertencia: sustituya los fusibles por otros del mismo tipo y características para evitar el riesgo de incendio.

Caution: To avoid possible electrical shock, disconnect the power cord before servicing the instrument.

Attention : Afin d’éviter toute possibilité de commotion électrique, débranchez le cordon d’alimentation de la prise avant d’effectuer la maintenance de l’instrument.

Vorsicht: Zur Vermeidung von Stromschlägen sollte das Gerät vor der Wartung vom Netz getrennt werden.

Precauzione: per evitare il rischio di scossa elettrica, scollegare il cavo di alimentazione prima di svolgere la manutenzione dello strumento.

Precaución: para evitar descargas eléctricas, desenchufe el cable de alimentación del instrumento antes de realizar cualquier reparación.

Commonly Used Symbols

Direct currentCourant continuGleichstromCorrente continuaCorriente continua

Alternating currentCourant alternatifWechselstromCorrente alternataCorriente alterna

Protective conductor terminalBorne du conducteur de protectionSchutzleiteranschlussTerminale di conduttore con protezioneBorne del conductor de tierra

Frame or chassis terminalBorne du cadre ou du châssisRahmen- oder ChassisanschlussTerminale di struttura o telaioBorne de la estructura o del chasis

Caution or refer to manualAttention ou reportez-vous au guideVorsicht, oder lesen Sie das HandbuchPrestare attenzione o fare riferimento alla guidaActúe con precaución o consulte la guía

Caution, hot surface or high temperatureAttention, surface chaude ou température élevéeVorsicht, heiße Oberfläche oder hohe TemperaturPrecauzione, superficie calda o elevata temperaturaPrecaución, superficie caliente o temperatura elevada

Commonly Used Symbols (Continued)

Caution, risk of electric shock (high voltage)Attention, risque de commotion électrique (haute tension)Vorsicht, Elektroschockgefahr (Hochspannung)Precauzione, rischio di scossa elettrica (alta tensione)Precaución, peligro de descarga eléctrica (alta tensión)

Caution, risk of needle-stick punctureAttention, risques de perforation de la taille d’une aiguilleVorsicht, Gefahr einer SpritzenpunktierungPrecauzione, rischio di puntura con agoPrecaución, riesgo de punción con aguja

Caution, ultraviolet lightAttention, rayonnement ultrvioletVorsicht, Ultraviolettes LichtPrecauzione, luce ultraviolettaPrecaución, emisiones de luz ultravioleta


UV

FuseFusibleSicherungFusibileFusible

Electrical power onSous tensionNetzschalter einAlimentazione elettrica attivataAlimentación eléctrica conectada

Electrical power offHors tensionNetzschalter ausAlimentazione elettrica disattivataAlimentación eléctrica desconectada


1

0

1525µ and1525EF HPLC Pump Information

Intended Use

The Waters® 1525µ and 1525EF HPLC Pumps can be used for in-vitro diagnostic testing to analyze many compounds, including diagnostic indicators and therapeutically monitored compounds. When you develop methods, follow the “Protocol for the Adoption of Analytical Methods in the Clinical Chemistry Laboratory,” American Journal of Medical Technology, 44, 1, pages 30–37 (1978). This protocol covers good operating procedures and techniques necessary to validate system and method performance.

Biological Hazard

When you analyze physiological fluids, take all necessary precautions and treat all specimens as potentially infectious. Precautions are outlined in “CDC Guidelines on Specimen Handling,” CDC – NIH Manual, 1984.

Calibration

Follow acceptable methods of calibration with pure standards to calibrate methods. Use a minimum of five standards to generate a standard curve. The concentration range should cover the entire range of quality-control samples, typical specimens, and atypical specimens.

Quality Control

Routinely run three quality-control samples. Quality-control samples should represent subnormal, normal, and above-normal levels of a compound. Ensure that quality-control sample results are within an acceptable range, and evaluate precision from day to day and run to run. Data collected when quality-control samples are out of range may not be valid. Do not report this data until you ensure that chromatographic system performance is acceptable.

Table of Contents

Preface ....................................................................................... 21

Chapter 1 Overview .......................................................................................... 24

1.1 Fluid-Handling Components ................................................. 26

1.2 Electronic Components......................................................... 29

1.3 1525µ and1525EF Pump Control ......................................... 30

Chapter 2 Installing the 1525 Pump ................................................................. 31

2.1 Selecting the Site .................................................................. 31

2.2 Unpacking the Pump............................................................. 33

2.3 Making Electrical Connections.............................................. 34

2.3.1 Connecting the Power Supply.................................... 34

2.3.2 Setting the IEEE-488 Address ................................... 34

2.3.3 Making IEEE-488 Connections.................................. 36

2.4 Making Fluidic Connections .................................................. 37

2.4.1 Making Eluent Supply Connections ........................... 37

2.4.2 Installing Different Mixer Configurations .................... 39

2.4.3 Installing an Optional 1500 Series Manual Injector....................................................................... 42

2.4.4 Installing an Optional 1500 Series Column Heater........................................................................ 42

2.4.5 Connecting the Pump Outlet...................................... 43

2.4.6 Connecting Fluid Waste Lines ................................... 44

Table of Contents 15

Chapter 3 Preparing for Operation ................................................................... 46

3.1 Powering On and Selecting the Pump .................................. 46

3.2 Preparing the Pump.............................................................. 46

3.2.1 Priming the Pump ...................................................... 48

3.2.2 Equilibrating the Pump............................................... 49

3.3 Powering Off the Pump ......................................................... 50

Chapter 4 Maintaining the 1525 Pump ............................................................. 52

4.1 Maintenance Considerations ................................................ 52

4.2 Diagnostic Tests .................................................................... 53

4.2.1 Retention Time Stability Test ..................................... 53

4.2.2 Ramp-and-Decay Test ............................................... 53

4.3 Replacing 1525µ Plunger Seals and Plungers ..................... 55

4.3.1 Preparing for Plunger Seal Replacement .................. 55

4.3.2 Removing and Replacing the 1525µ Plunger Seal .... 57

4.3.3 Cleaning and Replacing 1525µ Plungers .................. 58

4.4 Replacing 1525µ Check Valves ............................................ 61

4.5 Replacing 1525EF Plunger Seals and Plungers................... 64

4.5.1 Preparing for Plunger Seal Replacement .................. 65

4.5.2 Removing and Replacing the 1525EF Plunger Seal ........................................................................... 66

4.5.3 Cleaning and Replacing 1525EF Plungers................ 67

4.6 Replacing 1525EF Check Valves.......................................... 71

4.7 Replacing a Draw-Off Valve .................................................. 73

4.8 Replacing Fuses ................................................................... 75


Chapter 5 Troubleshooting ............................................................................... 78

5.1 Troubleshooting Pump Problems .......................................... 78

5.2 Troubleshooting Noises......................................................... 84

5.3 Troubleshooting Chromatographic Problems ........................ 85

Appendix A Specifications ................................................................................... 88

Appendix B Spare Parts and Accessories ........................................................... 92

Appendix C Solvent Considerations..................................................................... 96

C.1 Introduction .......................................................................... 96

C.2 Solvent Compatibility ........................................................... 97

C.3 Solvent Miscibility ................................................................ 99

C.4 Buffered Solvents .............................................................. 102

C.5 Head Height ...................................................................... 102

C.6 Solvent Viscosity ............................................................... 102

C.7 Mobile Phase Solvent Degassing ...................................... 102

C.7.1 Gas Solubility ......................................................... 103

C.7.2 Eluent Degassing Methods ..................................... 103

C.8 Wavelength Selection ........................................................ 105

Index ..................................................................................... 108


List of Figures 18

1-1 Waters 1525µ Pump ...................................................................... 241-2 Waters 1525EF Pump with an Optional Column Heater................ 251-3 Fluid-Handling Components in the Waters 1525µ Pump............... 261-4 Fluid-Handling Components in the Waters 1525EF Pump ............ 271-5 Rear Panel Electronic Components in the Waters 1525 Pump ..... 29

2-1 Pump Dimensions.......................................................................... 322-2 IEEE-488 Connection Example ..................................................... 372-3 Reverse Ferrule and Compression Screw Assembly..................... 392-4 1525EF Pump Mixer Configurations .............................................. 412-5 Standard Ferrule and Compression Screw Assembly ................... 44

3-1 Priming the Pump .......................................................................... 49

4-1 Unscrewing Pump Head Mounting Screws (1525µ Pump)............ 574-2 Exposed Head Support Assembly (1525µ Pump) ......................... 594-3 Disassembled Plunger (1525µ Pump) ........................................... 604-4 Disconnecting the Inlet and Outlet Tubing (1525µ Pump) ............. 624-5 Unscrewing Pump Head Mounting Screws (1525EF Pump) ......... 664-6 Exposed Head Support Assembly (1525EF Pump) ...................... 684-7 Assembled Plunger (1525EF Pump) ............................................. 694-8 Inserting the Plunger (1525EF Pump) ........................................... 704-9 Disconnecting the Outlet Tubing (1525EF Pump).......................... 724-10 Draw-Off Valve Inlet Manifold Assembly ........................................ 744-11 Rear Panel Fuse Holder ................................................................ 76

List of Figures

List of Tables

1-1 Fluid-Handling Components in the Waters 1525 Pump ................ 271-2 Electronic Components in the Waters 1525 Pump ................... 30

2-1 Installation Site Requirements ...................................................... 312-2 IEEE-488 DIP Switch Settings................................................. 35

3-1 Pump Preparation Recommendations .......................................... 47

5-1 Correcting Pump Problems ........................................................... 795-2 Correcting Noises ................................................................... 845-3 Correcting Chromatographic Problems.................................... 85

A-1 Physical Specifications for 1525 Pumps ....................................... 88A-2 Physical Specifications for the Column Heater............................. 88A-3 Environmental Specifications........................................................ 89A-4 Electrical Specifications ................................................................ 89A-5 Performance Specifications for the 1525µ Pump ......................... 90A-6 Performance Specifications for the 1525EF Pump....................... 90

B-1 Spare Parts for the 1525µ Pump .................................................. 92B-2 Spare Parts for the 1525EF Pump................................................ 93B-3 Accessories for the 1525µ Pump.................................................. 94B-4 Accessories for the 1525EF Pump ............................................... 94B-5 Maintenance and Operation Kits and Tools.................................. 95

C-1 Aqueous Buffers for Use with the 1525 HPLC Pump ................... 98C-2 Acids for Use with the 1525 HPLC Pump..................................... 98C-3 Bases for Use with the 1525 HPLC Pump.................................... 98C-4 Organic Solvents for Use with the 1525 HPLC Pump .............. 99C-5 Solvent Miscibility ....................................................................... 100

List of Tables 19

C-6 UV Cutoff Wavelengths for Common Chromatographic Solvents ...................................................................................... 105

C-7 Wavelength Cutoffs for Different Mobile Phases .................... 106C-8 Refractive Indices for Common Chromatographic Solvents ....... 107

List of Tables 20

Preface

The Waters 1525µ and 1525EF HPLC Pump Installation and Maintenance Guide describes the procedures for unpacking, installing, operating, maintaining, and troubleshooting the Waters® 1525µ and 1525EF (Extended Flow) HPLC Pumps. It also includes appendixes for specifications, spare parts and accessories, and eluent considerations.

This guide is intended for use by individuals who need to install, maintain, and/or troubleshoot the Waters 1525µ and 1525EF HPLC Pumps. You should be familiar with HPLC terms, practices, and basic HPLC system operations such as making fluidic connections.

Organization

This guide contains the following:

Chapter 1 describes the Waters 1525 HPLC Pumps, including features and options.

Chapter 2 explains how to unpack and install Waters 1525 HPLC Pumps.

Chapter 3 describes how to power on the pump, purge the pump, and power off the pump.

Chapter 4 covers routine maintenance, diagnostics, and troubleshooting procedures.

Chapter 5 explains how to diagnose and resolve problems with the pump.

Appendix A provides specifications of the Waters 1525 HPLC Pumps.

Appendix B lists recommended spare parts and accessories.

Appendix C provides information for proper preparation and use of solvents.

Related Documentation

Printed Documentation

Waters Licenses, Warranties, and Support: Provides software license and warranty information, describes training and extended support, and explains how Waters handles shipments, damages, claims, and returns.

Documentation on the Web

Related product information and documentation can be found on the World Wide Web. Our address is http://www.waters.com.

21

http://www.waters.com

Related Adobe Acrobat Reader Documentation

For detailed information about using Adobe® Acrobat® Reader, see the Adobe Acrobat Reader Online Guide. This guide covers procedures such as viewing, navigating, and printing electronic documentation from Adobe Acrobat Reader.

Printing This Electronic Document

Adobe Acrobat Reader lets you easily print pages, page ranges, or the entire document by selecting File > Print. For optimum print quantity, Waters recommends that you specify a PostScript® printer driver for your printer. Ideally, use a printer that supports 600 dpi print resolution.

Documentation Conventions

The following conventions can be used in this guide:

Convention Usage

Purple Purple text indicates user action such as keys to press, menu selec-tions, and commands. For example, “Click Next to go to the next page.”

Italic Italic indicates information that you supply such as variables. It also indicates emphasis and document titles. For example, “Replace file_name with the actual name of your file.”

Courier Courier indicates examples of source code and system output. For example, “The SVRMGR> prompt appears.”

Courier Bold Courier bold indicates characters that you type or keys you press in examples of source code. For example, “At the LSNRCTL> prompt, enter set password oracle to access Oracle.”

Underlined Blue Indicates hypertext cross-references to a specific chapter, section, subsection, or sidehead. Clicking this topic using the hand symbol brings you to this topic within the document. Right-clicking and selecting Go Back from the shortcut menu returns you to the origi-nating topic. For example, “Electrical components are described in Section 1.2, Electronic Components."

Keys The word key refers to a computer key on the keypad or keyboard. Screen keys refer to the keys on the instrument located immediately below the screen. For example, “The A/B screen key on the 2414 Detector displays the selected channel.”

… Three periods indicate that more of the same type of item can optionally follow. For example, “You can store filename1, filename2, … in each folder.”

22

Notes

Notes call out information that is helpful to the operator. For example:

Note: Record your result before you proceed to the next step.

Attentions

Attentions provide information about preventing damage to the system or equipment. For example:

Cautions

Cautions provide information essential to the safety of the operator. For example:

> A right arrow between menu options indicates you should choose each option in sequence. For example, “Select File > Exit” means you should select File from the menu bar, then select Exit from the File menu.

STOPAttention: To avoid damaging the detector flow cell, do not touch the flow cell window.

Caution: To avoid burns, turn off the lamp at least 30 minutes before removing it for replacement or adjustment.

Caution: To avoid electrical shock and injury, unplug the power cord before performing maintenance procedures.

Caution: To avoid chemical or electrical hazards, observe safe laboratory practices when operating the system.

Convention Usage

23

1
Chapter 1Overview
This chapter describes the Waters®1525µ and 1525EF (Extended Flow) HPLC Pumps, discusses fluid-handling and electronic components, and explains control of the pumps.

The pumps combine the most important aspects of eluent delivery for HPLC: high precision, reliability, smooth eluent flow, and proven performance.

Both pumps perform their intended functions equally well:

• The 1525µ Binary HPLC Pump is designed for precise, reproducible gradient delivery at low flow rates up to 5 mL/minute (Figure 1-1).

• The 1525EF Binary HPLC Pump is designed for increased flow rates of up to 22.5 mL/minute (Figure 1-2).

Both pumps are controlled over the IEEE-488 interface by Waters MassLynx™ 4.0 SP1 and Breeze 3.3. The microprocessor-controlled stepper motor and noncircular gears of each pump ensure smooth and precise flow regardless of backpressure, flow-rate setting, or eluent compressibility.

Figure 1-1 Waters 1525µ Pump

24

1

Figure 1-2 Waters 1525EF Pump with an Optional Column Heater

��

Bottle Holder

Column Heater

Overview 25

1

1.1 Fluid-Handling Components

Before installing a Waters 1525 Pump, become familiar with its fluid-handling components, as illustrated in Figure 1-3 and Figure 1-4.

Figure 1-3 Fluid-Handling Components in the Waters 1525µ Pump

GradientMixer

InletManifold

RestrictorTee(Patent Pending)

PressureTransducer

Pulse Dampeners(C overed by Shroud)

Pump HeadAssembly

Draw-Off Valve

Drip Tray Waste Exit

Drip Tray

Waste LineVent Valve

Pump Outlet

Vent Valve

Inlet Check Valve Assembly

Outlet Check Valve Assembly

Indicator Rod

Fluid-Handling Components 26

1

Figure 1-4 Fluid-Handling Components in the Waters 1525EF Pump

Table 1-1 describes the functions of the fluid-handling components in the 1525 Pump.

Table 1-1 Fluid-Handling Components in the Waters 1525 Pump

Component Description

Column heater (optional for 1525EF Pump – not shown)

Maintains elevated column temperature to facilitate method reproducibility.

Draw-off valve Enables attachment of a syringe for drawing eluent through the eluent reservoir line into the pump for priming.

Reference Valve

Outlet Check

Pump Head

Valve Assembly

AssemblyDraw-Off

Gradient

Manual Injector

Pressure

Tee

Pump Outlet

Inlet Check Valve Assembly

Pump Module B

Pump Module A

Reference Valve

Drip TrayDrip Tray Waste Exit

(Optional)

Valve

Manual Injector

Mixer

ManifoldInlet

Transducer

Pulse Dampeners(Covered by Shroud)

Waste Line

Valve Waste Lines

Indicator Rod

Overview 27

1
Drip tray Catches fluid leaks.
Drip tray waste exit Drains accumulated fluids to the waste container.

Gradient mixer Increases homogeneity. Also adds volume to the system.

Inlet and outlet check valve assemblies

Maintain flow direction and pressure by opening in one direction only.

Inlet manifold Provides the connection for eluent inlet tubing and routes eluent to the inlet check valve on each pump head.

Manual injector Enables a manual sample injection. Generates an inject start signal to the detector.

Manual injector waste line (1525EF Pump)

Routes flow from the manual injector to the waste container.

Plunger indicator rods Show the position of each pump head plunger.

Pressure transducer Senses operating pressure and converts values to elec-tronic signals for monitoring.

Pulse dampeners Dampen operating pressure fluctuations. Located on the left side of the unit under the mixer and behind the shroud.

Pump head assembly Draws in and delivers eluent. Defines pump capacity.

Pump outlet Routes eluent to the injector, column, and detector.

Reference (or vent) valve Directs flow from the pump to waste for purging, or through the injector, the column, and the rest of the system.

Reference (or vent) valve waste line

Routes flow from the reference (or vent) valve to the waste container.

Restrictor tee (1525µ Pump)

Blends two solvents together (with backflow prevention).

Seal wash holes(not visible in Figure 1-3 or Figure 1-4)

Allow manual flushing of plunger seals.

Tee (1525EF Pump) Blends two solvents together.

Table 1-1 Fluid-Handling Components in the Waters 1525 Pump (Continued)


Caution: To prevent eluent from leaking out of the pump outlet, position the eluent reservoir below the pump inlet manifold before disconnecting the pump outlet fitting.

Fluid-Handling Components 28

1

1.2 Electronic Components

Before installing the Waters 1525 Pump, become familiar with its electronic components as illustrated in Figure 1-5.

Figure 1-5 Rear Panel Electronic Components in the Waters 1525 Pump

Ethernet PortPower Cord

Power Switch

Fuse Holder

(Side of Unit)

I/O Terminal Block

IEEE-488 Port

IEEE-488 Address Switches

Cooling Fan Vent

RS-232 Port

(reserved for future use)

(for service use only)

Power Module

Connection

Overview 29

1

Table 1-2 describes the functions of the electronic components in the Waters 1525 Pump.

1.3 1525µ and1525EF Pump Control

Waters software allows monitoring and control of Waters 1525µ and 1525EF Pumps in binary applications. Using Waters software, you can do the following:

• Set all pump control parameters and operating ranges (see the appropriate software documentation for details).

• Define binary gradient conditions for a run (see Table A-5 or Table A-6 for more information, or refer to the software documentation).

• Prime the pump (see Section 3.2.1 on page 48).

Table 1-2 Electronic Components in the Waters 1525 Pump


Cooling fan vent Exhausts air for cooling internal electronics.

Ethernet port Provides an Ethernet network connection for remote opera-tion (reserved for future use).

Fuse holder Contains power fuses.

I/O terminal block Provides input and output contact closures for connection to external devices.

IEEE-488 address switches

Sets the IEEE-488 address for the pump.

IEEE-488 port Provides a National Instruments™ GPIB interface to the MassLynx workstation.

Power module Provides receptacles for power cord connection and the fuse holder.

Power switch Powers the pump on and off.

RS-232 port Provides a digital signal connection for remote operation (for service use only).

1525µ and1525EF Pump Control 30

2

Chapter 2Installing the 1525 Pump

This chapter lists the site requirements for a Waters 1525 Pump, and describes how to unpack the pump, make electrical connections, and make fluidic connections.

2.1 Selecting the Site

Install the pump at a site that meets the specifications indicated in Table 2-1.

Figure 2-1 shows the dimensions of a 1525 Pump.

Table 2-1 Installation Site Requirements

Factor Requirement

Temperature 4 to 40 °C (39 to 104 °F)

Relative humidity 20 to 80%, noncondensing

Bench space Width: 17 in. (43 cm) including bottle holder[24 in. (61 cm) with column heater]Depth: 24 in. (61 cm)Height: 17 in. (43 cm)

Vibration Negligible

Clearance Minimum 6 in. (15 cm) at rear for ventilation and cable connections

Static electricity Negligible

Input voltage and frequency

Grounded AC, 120/240 VAC, 50/60 Hz, single phase

Electromagnetic fields No nearby source of electromagnetic noise, such as arcing relays or electric motors

Power requirements 200 VA for pump150 VA for column heater

STOPAttention: To avoid overheating and to provide clearance for cable connections, make sure there is at least 6 in. (15 cm) of clearance at the rear of the pump.

Selecting the Site 31

2

Figure 2-1 Pump Dimensions

24 in.

12 in.

17 in. (43 cm)

7 in.

5 in.

Clearance for Optional Column

Heater

Clearance for Bottle Holder

(61 cm)

Installing the 1525 Pump 32

2

2.2 Unpacking the Pump

The pump is shipped in one carton that contains the following items:

• Waters 1525µ or 1525EF Binary HPLC Pump

• Startup Kit

• Bottle holder

• Certificate of Structural Validation

• Certificate of Functional Performance

• Waters 1525µ and 1525EF HPLC Pump Installation and Maintenance Guide

To unpack the pump:

1. Open the carton and remove the Startup Kit and other items from the top of the carton.

2. Remove the top packing tray and foam insert, then slide the box up and off the unit.

3. Lift the pump off the shipping pallet.

4. Carefully set the pump down and remove the foam packing material from both ends of the pump.

5. Check the contents of the Startup Kit against the Startup Kit parts list to confirm that all items are included.

6. Verify that the serial number on the inside left frame of the pump matches the serial number on the Certificate of Functional Performance. Keep the Certificate of Functional Performance and the Certificate of Structural Validation with this guide for future reference.

7. Inspect all items for damage. Immediately report any shipping damage to both the shipping company and your Waters representative. Contact Waters Technical Service at 800 252-4752, U.S. and Canadian customers only. Other customers, call your local Waters subsidiary or Waters corporate headquarters in Milford, Massachusetts 01757 (U.S.A.). Refer to the Waters Licenses, Warranties, and Support document for complete information on reporting shipping damages and submitting claims.

Unpacking the Pump 33

2

2.3 Making Electrical Connections

For proper operation, the pump requires:

• A grounded AC power supply with no abrupt voltage fluctuations

• A unique IEEE-488 address

• An IEEE-488 connection to a MassLynx workstation and other IEEE-488 devices

• An Inject Start signal output connection (if a manual injector is installed)

Required Materials

• 2-meter IEEE-488 cable with a GPIB connector (in Startup Kit)

• Power cord (in Startup Kit)

• Small, flat-blade screwdriver (required when connecting the Inject Start signal)

2.3.1 Connecting the Power Supply

The 1525µ and 1525EF Pumps automatically adjust for AC input voltage.

To make the power supply connection:

1. Insert the 120-V or 240-V power cord into the power cord connection on the rear of the pump.

2. Plug the male end of the power cord into a grounded power outlet.

2.3.2 Setting the IEEE-488 Address

Set a unique address for the 1525µ or 1525EF Pump (and for each device connected to the MassLynx workstation on the IEEE-488 bus). Valid IEEE-488 instrument addresses are 2 through 29, and are set using the DIP switches on the rear panel.

To set the IEEE-488 address:

1. Ensure that the power switch is off.

2. Set the DIP switches on the rear panel of the pump to a unique IEEE-488 address. Refer to Table 2-2 for DIP switch settings.


2

Table 2-2 IEEE-488 DIP Switch Settings

IEEE-488 AddressDIP Switch Settingsa

a. ON = 1 and OFF = 0.

16 8 4 2 1

2 OFF OFF OFF ON OFF

3 OFF OFF OFF ON ON

4 OFF OFF ON OFF OFF

5 OFF OFF ON OFF ON

6 OFF OFF ON ON OFF

7 OFF OFF ON ON ON

8 OFF ON OFF OFF OFF

9 OFF ON OFF OFF ON

10 OFF ON OFF ON OFF

11 OFF ON OFF ON ON

12 OFF ON ON OFF OFF

13 OFF ON ON OFF ON

14 OFF ON ON ON OFF

15 OFF ON ON ON ON

16 ON OFF OFF OFF OFF

17 ON OFF OFF OFF ON

18 ON OFF OFF ON OFF

19 ON OFF OFF ON ON

20 ON OFF ON OFF OFF

21 ON OFF ON OFF ON

22 ON OFF ON ON OFF

23 ON OFF ON ON ON

24 ON ON OFF OFF OFF

25 ON ON OFF OFF ON

26 ON ON OFF ON OFF

27 ON ON OFF ON ON

28 ON ON ON OFF OFF

29 ON ON ON OFF ON

Making Electrical Connections 35

2

2.3.3 Making IEEE-488 Connections

The 1525µ and 1525EF Pumps connect to the MassLynx workstation using IEEE-488 cables. Most chromatography devices are shipped with a 1-meter cable with dual-receptacle connectors at each end. The MassLynx workstation ships with a 2-meter cable with a dual-receptacle connector at one end and a single-receptacle connector (GPIB) at the other end.

Note: Before connecting the pump, refer to the IEEE-488 guidelines below.

IEEE-488 Guidelines

Follow these guidelines to install and use the system:

• Keep all devices powered on while your system is in use.

• While a system is active on the IEEE-488 bus, do not power on or off any other device on the bus.

• The maximum number of devices that can be connected together to form one interface system is 15 (14 instruments plus the National Instruments GPIB card).

• The maximum cable length to connect the devices and the GPIB connector in one interface system is 2 meters (6.5 feet) times the number of devices, or 20 meters (65 feet), whichever is smaller.

• The maximum cable length between two devices is 4 meters (13 feet).

• The minimum cable length between two devices is 1 meter (3.25 feet).

Connecting IEEE-488 Devices

Note: This procedure assumes that there are no other IEEE-488 devices (such as a detector) electronically connected to the MassLynx workstation. If there are other devices already connected to the workstation, connect the pump to the existing chain of devices (see Figure 2-2). The order in which you connect IEEE-488 devices is not important.

To make the IEEE-488 connections for the MassLynx system:

1. Connect the single-receptacle end of the 2-meter IEEE-488 cable to the National Instruments GPIB card in the MassLynx workstation (Figure 2-2).

STOPAttention: Cable lengths greater than the maximum values or less than the minimum values can cause IEEE-488 communication failures. Use the minimal cable lengths to ensure proper signal transmission.


2
Figure 2-2 IEEE-488 Connection Example
2. Connect the dual-receptacle end of the 2-meter IEEE-488 cable to the IEEE-488 port on one of the IEEE-488 devices (pump, column heater, autosampler, or detector).

3. Connect one end of a 1-meter IEEE-488 cable (with dual-receptacle connectors at each end) to the cable receptacle on the first device. Connect the other end of the cable to the IEEE-488 port on the next device.

4. Repeat step 3 for each additional IEEE-488 device.

5. Make sure that all IEEE-488 cable connector screws are finger tight.

2.4 Making Fluidic Connections

This section describes how to make fluidic connections to a Waters 1525 Pump by:

• Making eluent supply connections

• Installing different mixer configurations

• Installing an optional manual injector

• Installing an optional column heater

• Making outlet connections

• Connecting fluid waste lines

2.4.1 Making Eluent Supply Connections

Follow the instructions in this section to connect a 1525µ or 1525EF Pump inlet to an eluent reservoir.

GPIB Card in Workstation

Pump Detector

2-Meter IEEE-488

Cable

1-Meter IEEE-488

Cables

IEEE-488 Connectors

Column Heater

Autosampler

Making Fluidic Connections 37

2

Note: When using a Waters inline degasser, refer to the degasser operator’s guide for details on connecting the degasser to the reservoir and pump.

Required Materials

• Tefzel® ferrule and compression screw – one set per pump (in Startup Kit)

• 1/8-inch OD ETFE tubing (part number WAT270714 in Startup Kit)

• ETFE tubing (part number WAT024036 in Startup Kit)

• Reservoir containing filtered, degassed eluent – one per pump

• Bottle holder

• Bottle caps (1 L) (part number WAT062479 in Startup Kit)

• Stainless steel solvent filter – one per pump (part number WAT025531 in Startup Kit)

• Plastic tubing cutter (not included) (part number WAT031795) or razor blade

• Inlet tubing labels (part numbers WAT087186[A] and WAT087187[B])

Installing the Bottle Holder

1. Carefully position the rack along the left side of the pump so the cutouts at the bottom inside of the rack rest on their corresponding positioning screws on the pump.

2. Holding the rack flush against the surface of the pump, finger-tighten the captive screw to secure the rack.

Connecting Eluent Tubing to the Pump Inlet

1. Measure the length of 1/8-inch ETFE tubing required to connect an eluent reservoir (mounted in the bottle holder) to an inlet manifold on the pump. When using an inline degasser, refer to the degasser operator’s guide for details on connecting to the reservoir and pump.

Note: It may be necessary to substantially increase reservoir height if you use high-viscosity eluents (18 to 24 inches above the inlet manifold is not uncommon).

2. Insert the ETFE tubing into the 1/8-inch diameter hole of the tubing cutter, making sure that the tubing that extends from the metal side of the cutter is the correct length.

3. Insert the razor blade into the cutter and press down to cut the tubing. Make sure the cut end is straight and free from burrs.

4. Slide a compression screw over one end of the tubing, followed by a ferrule with its tapered end facing away from the tubing end, and its wide end flush with the tubing end, as shown in Figure 2-3.


2

Figure 2-3 Reverse Ferrule and Compression Screw Assembly

5. Firmly seat the tubing end into the inlet manifold on the pump, then finger-tighten the compression screw.

6. Repeat steps 1 to 5 for the second pump.

Connecting Eluent Tubing to an Eluent Reservoir

1. Slide an inlet tubing label onto the appropriate tube (see Figure 1-3 or Figure 1-4).

2. Insert the free end of the 1/8-inch ETFE inlet tubing into the cap of an eluent reservoir.

3. Slide one piece of ETFE tubing over the end of the 1/8-inch ETFE tubing for about 0.75 in. (1.9 cm).

4. Insert the stainless steel tubing fitting on the solvent filter into the open end of this tubing.

5. Install the cap onto the eluent reservoir and push the tubing through the cap until the filter reaches the bottom of the reservoir.

6. Repeat steps 1 through 5 for the second pump.

2.4.2 Installing Different Mixer Configurations

The Waters 1525µ and 1525EF HPLC Pumps can be used with various configurations of gradient mixers. This section describes these configurations and how to install each one.

STOPAttention: To avoid damaging the ferrule, do not overtighten the compression screw.

STOPAttention: To avoid having eluent leak from a pump outlet, position each eluent reservoir below its corresponding pump inlet until the outlet is connected to the system.

TP01170

Tubing

Compression Ferrule

Tubing End (Straight and Smooth)

(Flush with Tubing End)Screw


2

1525µ Binary HPLC Pump

The 1525µ Pump is shipped with a 50-µL mixer (part number 700002306, approximate 50-µL volume), mounted on the upper-left side of the chassis face.

For applications with low flow rates and/or steep gradients, you may decide to bypass the gradient mixer (see the procedure “Bypassing the Mixer in the 1525µ Pump” on page 42).

Note: Removing the mixer will result in an increase of the gradient ripple and baseline noise.

1525EF HPLC Pump

The 1525EF Pump is shipped with one large-volume gradient mixer (part number WAT051519, approximate 1100-µL volume), mounted on the upper-left side of the chassis face (see configuration 2 at the top of Figure 2-4).

If you want to replace the standard large-volume gradient mixer with dual GM150 Gradient Mixers (approximate 300-µL volume), see configuration 1 in Figure 2-4.


2

Figure 2-4 1525EF Pump Mixer Configurations

Installing Mixers

There are four pairs of mounting holes on the upper-left side of the chassis face. For the following configurations, these hole pairs are numbered from 1 to 4 top to bottom.

Dual GM150 Gradient Mixers

To use this configuration with the 1525EF Pump:

1. Have available two GM150 Gradient Mixers (part number WAT055847). Refer to Appendix B.

Out

In

1

2Gradient Mixers


2

2. Loosen the compression screws on the inlet and outlet tubing of the existing large-volume gradient mixer.

3. Move the tubing enough to allow repositioning of the mounting clamps.

4. Loosen the compression screw on the outlet tubing.

5. Remove the mounting screws, the large-volume gradient mixer, and mounting clamps.

To install the dual GM150 Gradient Mixers in the 1525EF Pump:

1. Put the mixers inside the mounting clamps (as shown in configuration 1 of Figure 2-4), using hole pairs 2 and 4.

2. Firmly tighten the mounting screws.

3. Reattach the pressure transducer outlet tubing to the inlet of the bottom mixer.

4. Connect the outlet of the bottom mixer to the inlet of the top mixer with the U-tube (part number 430000405 in the Startup Kit).

5. Reconnect the outlet of the top mixer to the reference valve inlet tubing.

Bypassing the Mixer in the 1525µ Pump

1. Disconnect the tubing running from the bottom of the pressure transducer to the gradient mixer.

2. Disconnect the tubing running from the mixer to the vent valve.

3. Connect the pressure transducer to the vent valve port using the tubing assembly supplied in the Startup Kit (part number 430000879).

2.4.3 Installing an Optional 1500 Series Manual Injector

Refer to the instructions and Startup Kit list included with the 1500 Series Manual Injector.

Note: For most configurations on the 1525µ and 1525EF Pumps, an autosampler will be used.

2.4.4 Installing an Optional 1500 Series Column Heater

The column heater is mounted on the right side panel with three screws. Connectors on the rear panel provide power and control to the column heater.

To install the column heater:

1. Remove the three screws from the right side panel of the pump.

2. Insert the screws through the standoffs (in the Startup Kit) so that the flat side of the standoffs are flush with the right side panel.

3. Mount the column heater on the three screws on the right side panel.


2

4. Insert the female end of the power cable to the power connector on the rear of the column heater and the male end into the power source outlet.

5. Connect the IEEE-488 signal cable to the IEEE-488 port on the rear of the column heater. The other end of this cable must be connected to a MassLynx workstation for software control.

6. See Section 2.3.2, Setting the IEEE-488 Address, to set the column heater address.

2.4.5 Connecting the Pump Outlet

Follow the instructions in this section to connect the pump outlet to the next instrument in the flow path. Making pump outlet connections involves:

• Cutting the tubing

• Attaching a compression fitting to each end of the tubing

• Connecting each end of the tubing

Required Materials

• Two stainless steel ferrules and standard compression screws (part number WAT025604 in Startup Kit)

• 1/16-inch OD stainless steel tubing (in Startup Kit)

• Circular tubing cutter (part number WAT022384) or knife-edge file

• Needle-nose pliers (two pairs if cutting tubing with a knife-edge file)

• 5/16-inch open-end wrench (part number WAT022527 in Startup Kit)

Cutting the Tubing to Length

1. Measure the length of stainless steel tubing required to connect the pump outlet to the injector or other instrument.

2. Use a knife-edge file to scribe the circumference of the tubing at the desired break. Whenever possible, use a circular tubing cutter instead of the knife-edge file to obtain a clean, square cut.

3. Grasp the tubing on both sides of the scribe mark with cloth-covered pliers (to prevent marring of the tube surface) and gently work the tube back and forth until it snaps. Ensure that the break is square and free of burrs.

Caution: To avoid having eluent leak from a pump outlet, position each eluent reservoir below its corresponding pump inlet until the outlet is connected to the system.


2

Attaching the Compression Fittings to the Tubing

1. Slide a compression screw onto one end of the tubing, then slide a ferrule onto the tubing with the large end of the taper toward the screw, as shown in Figure 2-5.

Figure 2-5 Standard Ferrule and Compression Screw Assembly

2. While firmly pressing the tubing into the pump outlet or other system component, finger-tighten the compression screw.

3. Use the 5/16-inch wrench to tighten the screw another quarter turn. This seats the ferrule against the tubing.

4. Unscrew the assembled fitting and verify that the length of tubing extending beyond the ferrule is 0.12 inch (3 mm). (See Figure 2-5.)

5. Repeat steps 1 to 4 for the other end of the stainless steel tubing.

Making the Connections

1. While pressing one end of the tubing assembly into the pump outlet fitting, finger-tighten the compression screw, then use the 5/16-inch wrench to tighten the screw another 1/8-turn.

Note: Leave the instrument end of the outlet tubing disconnected until you have primed the pump as described in Section 3.2.1, Priming the Pump.

2. After you primed the pump, press the free end of the tubing assembly into the injector or next device in your HPLC system and finger-tighten the compression screw. Then use the 5/16-inch wrench to tighten the screw another 1/8-turn.

2.4.6 Connecting Fluid Waste Lines

Follow the instructions in this section to connect the fluid waste lines. The instructions describe how to do the following:

• Connect the reference valve and optional manual injector waste lines

• Connect the drip tray waste line

Compression Screw

Ferrule

Smooth, Square EndTubing

0.12 Inch


2

Required Materials

• Tubing (part number WAT076775 in Startup Kit)

• Teflon tubing (part number WAT050693 in Startup Kit)

• Convoluted ETFE tubing (part number WAT241095 in Startup Kit)

• Plastic tubing cutter (not included) (part number WAT031795) or razor blade

Connecting Pump Waste Lines

1. Obtain a piece of convoluted tubing and stretch it apart.

2. Feed a piece of smooth Teflon tubing through the convoluted tubing until ends of both pieces are flush with one another. This dual tubing will prevent the waste line from developing kinks.

3. Mount this piece of dual tubing parallel with the drip tray (see Figure 1-3 or Figure 1-4) and secure it in the provided spring clips.

4. Insert the reference valve waste line and the optional manual injector waste lines together in the top end of the dual tubing.

5. Insert the other end of the dual tubing in a properly situated waste container.

Connecting the Drip Tray to the Waste Line

1. Attach tubing to the drip tray waste exit (see Figure 1-3 or Figure 1-4).

2. Insert the other end of the tubing in a waste container.

Connecting an Optional Manual Injector to the Column or Optional Column Heater

1. Make sure you installed the manual injector as described in the instructions included with the optional manual injector.

2. Connect the free end of the manual injector outlet tubing to the column or column heater.


3

Chapter 3Preparing for Operation

This chapter describes how to do the following:

1. Power on the 1525µ or 1525EF Pump and select it in MassLynx.

2. Prepare the 1525µ or 1525EF Pump.

3. Power off the 1525µ or 1525EF Pump.

3.1 Powering On and Selecting the Pump

1. Make sure the pump is installed according to the instructions in Chapter 2.

2. To power on the pump, find the power switch on the left side panel, and set it to the 1 (ON) position.

Note: Power on the pump and other chromatographic system devices before powering on the MassLynx workstation.

3. Power on the MassLynx workstation and log on to Windows®.

4. Select Start > Programs > MassLynx > MassLynx V4.0. The MassLynx Main window appears.

5. Click the Inlet Method button (left side of the window) to open the Inlet Editor.

6. Select Tools > Instrument Configuration. The Inlet Configuration page opens.

7. Click Configure.

8. Follow the Inlet Configuration wizard to select the 1525µ or 1525EF Pump.

9. Select the proper head size for the pump (50, 100, or 225 µL).

3.2 Preparing the Pump

Overview

The procedures for preparing the pump for operation depend on the status of the chromatography system, as indicated in Table 3-1. Use this table to determine when to perform certain preparation procedures.

Powering On and Selecting the Pump 46

3

Priming and equilibrating procedures are described in Section 3.2.1 and Section 3.2.2.

Eluent Guidelines

When preparing a 1525µ or 1525EF Pump for operation, follow these guidelines:

• Use methanol to prime a new pump, or one that has been stored dry for an extended period.

• Use an intermediate solvent when changing between eluents that are not miscible.

• Use only HPLC-grade eluents to obtain accurate, reproducible chromatography.

• To prevent air from being drawn into the pump, keep the eluent level in the reservoir at least 4 inches (10 cm) higher than the inlet manifold. Do not place eluent reservoirs on top of the pump.

• Use only filtered, degassed eluents.

• After use with a buffer, purge the pump with HPLC-grade water.

• When possible, use eluents of similar viscosity when you run a gradient.

• Know the properties of your eluents; for example, cyclohexane solidifies at 3.5 K psi.

• Consider column chemistry. If the column can be damaged by an intermediate or new eluent, remove the column from the system and substitute a union. Refer to the care and use guide for your column.

• Dedicate an eluent supply tube and filter assembly for each eluent. If this is not possible, purge the tube and filter of any remaining eluent before you use a new eluent.

• To prevent cross contamination of eluents, dedicate a set of glassware for preparing, storing and supplying each eluent. After a container is used for an eluent, do not use it for any other eluent unless it has been thoroughly cleaned.

Table 3-1 Pump Preparation Recommendations

Status Prime Equilibrate

Initiating operation

Changing eluent

Adding fresh eluent

System has been idle for some time

Caution: To avoid chemical hazards, observe safe laboratory practices when handling eluents. Refer to the Material Safety Data Sheets for the eluents in use. For more information on eluent properties, see Appendix C, Solvent Considerations.

Preparing for Operation 47

3

3.2.1 Priming the Pump

Priming the pump is necessary to ensure proper pump operation when:

• Starting the pump for the first time

• Changing eluent

• The system has been idle for some time

Required Material

Priming syringe (part number WAT010337 in Startup Kit)

Procedure

1. Power on the pump.

2. Power on the MassLynx workstation and log on to Windows.


4. Click Status to get the LC status.

5. Click the Inlet Method button (left side of the window) to open the Inlet Editor window.

6. Click the Inlet icon to go to the pump method.

7. Set the flow rate:

• For the 1525µ Pump, set the rate to 1.0 mL/min.

• For the 1525EF Pump, set the rate to 3.0 mL/min.

If the pump is dry, start with a lower flow rate.

8. Click Save in the Inlet Method window to save the method.

9. Fully depress the priming syringe plunger to remove all air from the syringe, then insert the syringe into the Luer fitting at the center of the draw-off valve handle (Figure 3-1).

STOPAttention: Do not start the pump at this point in the procedure.

Preparing the Pump 48

3

Figure 3-1 Priming the Pump

10. Turn the draw-off valve handle counterclockwise 1/2-turn to open the valve.

11. Withdraw several milliliters of eluent with the syringe, then close the valve handle. Expel the contents of the syringe into a suitable waste container.

12. Click Load Method in the Inlet Method window to download the method to the system. This will initiate the pump and flow will begin.

13. Allow the pump to run for approximately one minute, then verify normal pump pressure for the selected eluent and flow rate. If the pump pressure is not correct, repeat steps 2 through 6 to reset the flow rate as necessary.

14. Click the Start/Stop Pumping icon in the Inlet Method window to turn off the pump flow.

3.2.2 Equilibrating the Pump

Equilibrate the system when one or more of the following conditions occur:

• Starting the system for the first time

• After changing eluents or adding fresh eluent

• After leaving the system idle for more than one day

Before Equilibrating

1. Make sure the eluent is filtered and degassed.

STOPAttention: To prevent eluent flow through the pump heads due to gravity pressure, never leave the pump unattended with the draw-off valve in the open position.


3

2. Allow the eluent and the system to reach proper temperatures before pump operation.

3. Prime the pump as described in Section 3.2.1, Priming the Pump.

Equilibrating

1. Power on all MassLynx system instruments and any peripheral devices, such as a printer.

2. Power on the MassLynx workstation and log on to Windows.


4. Click the Status button to get the LC status.

5. Click the Inlet Method button (left side of the window) to open the Inlet Editor window.

6. Click the Inlet icon to go to the pump method.

7. Set the flow rate:

• For the 1525µ Pump, set the rate to 1.0 mL/min.

• For the 1525EF Pump, set the rate to 3.0 mL/min.

If the pump is dry, start with a lower flow rate.

Note: These flow rates may not be appropriate for all columns.

8. Click Save in the Inlet Method window to save the method.

9. Click Load Method in the Inlet Method window to download the method to the system. This will initiate the pump and flow will begin.

10. Allow the pump to equilibrate for several minutes (the amount of time depends on total system volume, eluent miscibility, flow rate, etc.).

11. Click the Start/Stop Pumping icon in the Inlet Method window to turn off the pump flow.

For more information about selecting an equilibration method, refer to the MassLynx Help.

3.3 Powering Off the Pump

1. If you used buffers, purge them from the pump and other HPLC system components with HPLC-grade water. If the pump will be idle for more than one day, purge it with a methanol/water solution to prevent the growth of microorganisms.

Powering Off the Pump 50

3

2. If the pump is running, powering off the pump will stop flow immediately.

3. Set the power switch to the 0 (OFF) position.

STOPAttention: Since abrupt flow changes can damage columns, it is advisable to ramp flow to 0 using an appropriate user-defined MassLynx shutdown method.


4

Chapter 4Maintaining the 1525 Pump

This chapter provides important safety and handling considerations for the Waters 1525 Pump, describes how to run two diagnostic tests and set pump calibration parameters, and explains how to replace pump components.

4.1 Maintenance Considerations

Safety and Handling

When performing maintenance procedures on your pump, keep the following safety considerations in mind.

Proper Operating Procedures

To keep the pump running smoothly, follow the operating procedures and guidelines in Section 3.2, Preparing the Pump.

Caution: To prevent injury, always observe good laboratory practices when handling eluents, changing tubing, or operating the 1525 HPLC Pump. Know the physical and chemical properties of the eluents. Refer to the Material Safety Data Sheets for the eluents in use.

Caution: To avoid possible eye injury or cuts, handle the plunger with care. Wear safety glasses and use the plunger insertion tool. Pieces of a broken plunger are very sharp.

Caution: To avoid possible electric shock, do not remove the cover. The interior of the pump does not contain user-serviceable parts.

STOPAttention: Read this entire chapter thoroughly before you begin any pump maintenance.

Maintenance Considerations 52

4

Spare Parts

Refer to Appendix B, Spare Parts and Accessories, for spare parts information. Parts not included in Appendix B are not recommended for replacement by the customer.

Contacting Waters Technical Service

If you encounter any problems replacing parts in the pump, contact Waters Technical Service at 800 252-4752, U.S. and Canadian customers only. Other customers, call your local Waters subsidiary (or Technical Service Representative) or Waters corporate headquarters in Milford, Massachusetts 01757 (U.S.A.).

4.2 Diagnostic Tests

The following diagnostic tests can help you track system performance and prevent or identify potential problems before they interfere with operation:

• Retention time stability test

• Ramp-and-decay test

4.2.1 Retention Time Stability Test

Observing retention time stability during system performance monitoring tests is useful for determining the performance of your HPLC system and its components, including the pump. Erratic or changing retention times could be a result of dirty or malfunctioning check valves, worn plungers or plunger seals, air bubbles in the lines, incorrectly set flow rate, leaks, or other pump-related problems. Be aware that other factors, such as system and column equilibration, column age, operating temperature, and so on, can also affect retention time stability. See your system suitability documentation for more information.

4.2.2 Ramp-and-Decay Test

Use the ramp-and-decay test to monitor check valve performance.

Required Materials

• Three compression plugs (part number WAT025566 in Startup Kit)

• Tissues

• 5/16-inch open-end wrench (in Startup Kit)

• HPLC-grade methanol

Maintaining the 1525 Pump 53

4

Procedure

1. Disconnect the outlet tube of the system at the reference (or vent) valve and install a compression plug in its place.

2. Purge the pumps with 100% methanol. If your eluent is not miscible with methanol, use an intermediate eluent.

3. Disconnect pump B from the tee by removing the outlet tube of the pulse dampener and install a compression plug in its place.

4. Disconnect the left pump head outlet tube of Pump A from the draw-off valve and install a compression plug in its place.

5. Place tissues under the disconnected pump head outlet tube and the disconnected pulse dampener tube to catch drips.

6. Ensure that the draw-off valves are closed and the reference valve is closed to waste.

7. Set the following pump parameter values:

• Flow parameter:

1525µ Pump: 0.2 mL/min

1525EF Pump: 0.5 mL/min

• High-pressure limit:

1525µ Pump: 5000 psi (41,370 kPa, 401 bar)

1525EF Pump: 5000 psi (34,475 kPa, 334 bar)

8. With the 100% methanol reservoir still connected to the pump inlet, click Load Method in the Inlet Method window to start the pump. Observe pump pressure in the MassLynx Main window or in the Status window of the Inlet Method window:

• If the check valves are operating properly, pressure continually rises with each plunger stroke on the connected head, holding as the plunger recedes.

• If there is a bad inlet check valve, pressure may stop at a certain point, or may not rise at all.

• If there is a bad outlet check valve, pressure may increase, then immediately decrease as the plunger recedes.

• If the pressure does not rise to the high-pressure limit, try repriming the pump or increasing the Flow parameter setting.

9. Allow the pump to reach the high-pressure limit. The pump will stop flow automatically.

STOPAttention: Before starting this procedure, move the eluent reservoir below the pump head levels. This eliminates gravity flow of the eluent.

Diagnostic Tests 54

4

10. After 2 minutes, record the pressure (P1). One minute later, record the pressure again (P2). Calculate head pressure decay with the formula (P1 – P2)/P1. Verify that head pressure decay is 0.15 or less. If the pressure decay for the pump head is greater than 0.15, you may have bad check valves. Remove and clean or replace the inlet and outlet check valves (see Section 4.4, Replacing 1525µ Check Valves).

11. Relieve head pressure by slowly opening the reference (or vent) valve.

12. Disconnect the right pump head outlet tubing from the draw-off valve. Transfer the compression plug from the left port of the draw-off valve to the right port of the draw-off valve. Reconnect the left pump head outlet tubing to the left port of the draw-off valve.

13. Repeat steps 5 through 12 to test the left pump head.

14. Remove the compression plugs from the draw-off valve and the tee. Reconnect the right pump head outlet tubing. Reconnect the tubing from the pulse dampener.

15. Repeat steps 3 through 14 on pump B. In step 3, disconnect pump A from the tee.

16. When finished, purge the pump with eluent and close the reference (or vent) valve.

17. Remove the compression plug from the reference (or vent) valve and reconnect the outlet tube to the system.

4.3 Replacing 1525µ Plunger Seals and Plungers

This section describes how to:

• Remove and replace plunger seals and plunger bearings

• Remove and replace plungers

The plungers in a Waters 1525 Pump are ultrasmooth, chemically inert sapphire rods. Salt crystals that precipitate out from the eluent can form on the plunger and cause wear on the plunger seals and on the plunger itself. The result is a slow leak and a slight cyclic pressure fluctuation, with a possible increase in retention time.

For continued high performance of the pumps:

• Replace the plunger seals and bearings twice yearly, or as needed.

• Replace the plunger twice yearly, or as needed.

• Clean and inspect the plunger twice yearly (or sooner if using abrasive eluents).

4.3.1 Preparing for Plunger Seal Replacement

Required Materials


• 5/32-inch Allen wrench (in Startup Kit)


4

• 1/2-inch adjustable wrench (in Startup Kit)

• Priming syringe (in Startup Kit)

• Fitting plug (in Startup Kit)

• Replacement seal kit, plunger seal insertion tool, and seal extraction tool (in Startup Kit) (see Appendix B for part numbers)


To access a plunger seal, first remove the pump head.

Removing the Pump Head

1. Purge the pump with methanol. If methanol is not miscible with your current eluent, use an intermediate eluent.

2. Insert the priming syringe into the Luer fitting at the center of the draw-off valve handle, then turn the handle counterclockwise about 1/2-turn to open the valve.

3. Remove the solvent inlet lines from the reservoirs.

4. Use the syringe to withdraw all methanol.

5. Using the software, navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to run the pump.

6. After the indicator rod fully retracts into the pump head, click the Stop Flow button to turn off the pump. This ensures that the weight of the pump head does not rest on the plunger while you remove the head.

7. Use the 1/2-inch adjustable wrench to hold the check valve housings in place. Use the 5/16-inch open-end wrench to remove the inlet and outlet tubings from the check valve housings on the pump.

8. Holding the pump head against the pump, use the 5/32-inch Allen wrench to remove the two pump head assembly mounting screws. Loosen the screws 1/2-turn at a time for the first two turns (Figure 4-1).

STOPAttention: Before starting this procedure, move the eluent reservoir below the pump head level. This eliminates gravity flow of the eluent.

STOPAttention: Hold the pump head against the pump while removing the screws to prevent breaking the plunger.

Replacing 1525µ Plunger Seals and Plungers 56

4

Figure 4-1 Unscrewing Pump Head Mounting Screws (1525µ Pump)

9. Carefully slide the pump head assembly off the pump.

4.3.2 Removing and Replacing the 1525µ Plunger Seal1. Remove the plunger bearing from the pump head.

2. Remove the seal from the pump head using the seal extraction tool. (Twist the tapered screw end of the tool clockwise into the seal, then pull the seal to remove it from the pump head. Do not reuse the seal.)

3. Place the new plunger seal onto the tip of the seal insertion tool. Make sure the spring in the seal is facing away from the end of the tool (you should be able to see the seal spring at the tip of the tool).

4. Moisten the seal with methanol.

5. Place the small end of the block part of the seal insertion tool into the pump head. Slide the tip of the tool (with the seal on it) through the block and into the pump head. Press the tool firmly into the pump head, then remove it.

6. Using the small tip part of the insertion tool, push the seal into the pump head as far as it will go.

7. Replace the plunger bearing on top of the seal. Make sure the grooves in the bearing are facing outward and are visible as you look at the pump head.

8. Remoisten the seal and plunger with methanol.

STOPAttention: Pull the pump head straight off to avoid breaking the plunger.

Mounting Screws


4

9. Carefully slide the pump head assembly into position over the plunger. Make sure the indicator rod is properly aligned to prevent damaging the plunger.

10. While holding the pump head assembly against the pump, reinstall the two pump head mounting screws. Alternately tighten each screw, using half turns to evenly align the assembly.

11. Using the software, navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to start the pump. Verify that the indicator rod moves freely.

12. Click the Stop Flow button to stop the pump.

13. Use the 1/2-inch adjustable wrench to hold the check valve housing in place. Reconnect the inlet and outlet tubings to the pump head.

14. Prime the pump as described in Section 3.2.1 on page 48. If you notice leaks, verify pump head and plunger seal installation.

If necessary, repeat these procedures (removing the pump head and replacing plunger seals) for the other pump heads.

4.3.3 Cleaning and Replacing 1525µ Plungers

The plungers are sapphire rods that require careful handling. Although cleaning the plungers is not difficult, it is important to follow these instructions carefully to avoid damaging the plungers. Assemble all materials and read the procedure thoroughly before you begin.

Required Materials





• Plunger insertion tool and seal extraction tool (in Startup Kit) (see Appendix B for part numbers)

• Sonicator


• Replacement plunger

• Replacement plunger seals

STOPAttention: Hold the pump head against the pump while replacing the screws to avoid breaking the plunger.


4

Removing the Plunger

1. Remove the pump head as described in Section 4.3.1, Preparing for Plunger Seal Replacement.

2. Remove the four head support screws with the 9/64-inch Allen wrench (Figure 4-2), then carefully slide the head support assembly and the indicator rod off the pump. Set the head support assembly on the benchtop.

Figure 4-2 Exposed Head Support Assembly (1525µ Pump)

3. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to run the pump. When the plunger is fully extended, click the Stop Flow button to stop the pump.

4. Remove the plunger by sliding it out of the back of the support assembly. Set the support assembly aside. Continue with “Cleaning the Plunger”.

Cleaning the Plunger

Clean the plunger by sonicating it in 50:50 methanol/water for a few minutes. The parts of the plunger assembly are shown in Figure 4-3. Continue with “Inspecting the Plunger”.


Caution: To avoid possible eye injury or cuts, handle the plunger with care. Wear safety glasses. Pieces of a broken plunger are very sharp.

Head Support Screws

Head Support Assembly


4

Figure 4-3 Disassembled Plunger (1525µ Pump)

Inspecting the Plunger

After cleaning the plunger, inspect it for damage by holding it under a bright white light and looking down its length for nicks and scratches. It is easier to see scratches under a bright light than to feel them with your fingers.

• If the plunger is not scratched or otherwise damaged, reassemble it with new seals. Continue with “Replacing the Plunger”.

• If the plunger is damaged, replace both the plunger and the seals. Continue with “Replacing the Plunger”.

Replacing the Plunger

1. Reassemble the components of the plunger assembly (as shown in Figure 4-3).

2. Lubricate the ball and seat with high-pressure grease. Make sure the brass seat is located at the bottom of the piston.

3. Reinstall the assembled plunger into the support assembly.

4. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 1.0 mL/min, click Save, then click Load Method to run the pump. When the piston is fully retracted, click the Stop Flow button to stop the pump.

5. With the indicator rod hole oriented to the left, reinstall the head support assembly onto the pump. Alternately tighten the four screws. Do not overtighten.



Plunger

Retainer Seal

Piston

Retainer

Ball Seat

Spring


4

6. Moisten the plunger seal and the plunger with methanol.

7. Carefully slide the pump head over the plunger and onto the pump. Alternately tighten the two screws.

Check for even head alignment by observing the gap between the pump head and the pump head support assembly.

8. In the Inlet Method window, set the Flow parameter to 1.0 mL/min, click Save, then click Load Method to run the pump.

9. While the pump is running, observe the movement of the indicator rod:

• If the rod does not move smoothly, the head is misaligned. Click the Stop Flow button to stop the pump, loosen the pump head, then repeat steps 7 and 8.

• If the rod snaps moves smoothly, click the Stop Flow button to stop the pump. Continue with step 10.

10. Use the 1/2-inch adjustable wrench to hold the check valve housings in place. Reconnect the inlet and outlet tubing assemblies to the pump head.

11. Prime the pump as described in Section 3.2.1 on page 48. If you notice leaks, verify pump head and plunger seal installation as described in Section 4.3.1 on page 55.

4.4 Replacing 1525µ Check Valves

This section describes how to replace inlet, outlet, and restrictor tee check valves for the 1525µ Pump. Replace the inlet, outlet, and restrictor tee check valves every 6 months, or more frequently if required by your application. Waters recommends keeping a spare set of new check valves on hand.

Required Materials




• Replacement check valves


• Forceps

STOPAttention: Hold the pump head against the pump while tightening the screws to avoid breaking the plunger.


4

Replacing the Inlet Check Valve


2. Hold the check valve housing with the 1/2-inch adjustable wrench. Use the 5/16-inch open-end wrench to loosen the compression screw and remove the tubing assembly from the inlet check valve housing.

3. Use the 1/2-inch adjustable wrench to remove the inlet check valve housing from the pump head (Figure 4-4).

Figure 4-4 Disconnecting the Inlet and Outlet Tubing (1525µ Pump)

4. Remove the cartridge from the housing and replace it with the new cartridge.

Note: To help keep the cartridge seated within the housing, moisten it with methanol during installation.

5. With the housing held upright to prevent the cartridge from falling out, finger- tighten the housing into the pump head. Tighten the check valve another 1/4-turn with the 1/2-inch adjustable wrench.


STOPAttention: The arrow printed on the check valve cartridge indicates the direction in which it will allow liquid to flow. Therefore, its direction is critical for proper operation. For all check valve assemblies, the arrow should point upward when installed on the pump.

Compression Screws Outlet Check Valve

Pump Heads

Outlet Tubing

Housings

Inlet Check ValveHousings

Inlet Tubing

Replacing 1525µ Check Valves 62

4

6. Hold the check valve housing with the 1/2-inch adjustable wrench. Reinstall the inlet tubing assembly. While tightening the compression screw, hold the housing to prevent it from turning.

7. Repeat steps 2 through 6 to replace the other inlet check valves as necessary.

8. Prime the pump as described in Section 3.2.1 on page 48. Check for leaks.

Replacing the Outlet Check Valve


2. Run the pump until the indicator rod fully retracts into the pump head, then click the Stop Flow button to turn off the pump. This ensures that the weight of the pump head does not rest on the plunger when you remove the pump head.

3. Use the 1/2-inch adjustable wrench to hold the housings in place. Use the 5/16-inch open-end wrench to loosen the compression screws and remove the inlet and outlet tubing assemblies from the check valve housings.

4. Use the 1/2-inch adjustable wrench to loosen the outlet check valve housing.

5. While holding the pump head securely against the pump, use the 5/32-inch Allen wrench to remove the two pump head assembly mounting screws. Loosen the screws 1/2-turn at a time for the first two turns.

6. Carefully slide the pump head assembly off the pump. (See the instructions on page 55.)

7. Hold the pump head upside-down in one hand with the outlet check valve housing facing the floor, then remove the housing. Remove the cartridge from the housing and replace it with the new cartridge.

Note: To help keep the cartridge seated within the housing, moisten it with methanol during installation.


STOPAttention: Pull the pump head off straight to avoid breaking the plunger.

STOPAttention: The arrow printed on the check valve cartridge indicates the direction in which it will allow liquid to flow. Therefore, its direction is critical for proper operation. For all check valve assemblies, the arrow should point upward when installed on the pump.


4

8. With the outlet check valve housing held upright to prevent the cartridge from falling out, finger-tighten the housing into the pump head.

9. Turn the pump head over and carefully slide the pump head over the plunger and onto the pump.

10. Holding the pump head securely against the pump, reinstall the two mounting screws. Alternately tighten the screws to secure the pump head to the pump. Do not overtighten.

11. Tighten the check valve housing 1/4-turn with the 1/2-inch adjustable wrench.

12. Reinstall the outlet tubing assembly. Use the 5/16-inch open-end wrench to tighten the compression screw while holding the housing with the 1/2-inch adjustable wrench.

13. Reinstall the inlet tubing assembly. Use the 5/16-inch open-end wrench to tighten the compression screw while holding the housing with the 1/2-inch adjustable wrench.

14. Prime the pump as described in Section 3.2.1. Check for leaks.

Replacing the Restrictor Tee Check Valves

To replace the restrictor tee check valves, follow the procedure in “Replacing the Inlet Check Valve” on page 62.

4.5 Replacing 1525EF Plunger Seals and Plungers

This section describes how to:

• Remove and replace plunger seals

• Remove and replace plungers

The plungers in a Waters 1525 Pump are ultrasmooth, chemically inert sapphire rods. Salt crystals that precipitate out from the eluent can form on the plunger and cause wear on the plunger seals and on the plunger itself. The result is a slow leak and a very slight cyclic pressure fluctuation, with a possible increase in retention time.

For continued high performance of the pumps:

• Replace the plunger seals twice yearly, or as needed.

• Replace the plunger once each year, or as needed.

• Clean and inspect the plunger twice yearly (or sooner if using abrasive eluents).

STOPAttention: Ensure that the pump head is straight and the indicator rod is properly aligned with the pump head to avoid damaging the plunger.

Replacing 1525EF Plunger Seals and Plungers 64

4

4.5.1 Preparing for Plunger Seal Replacement

Required Materials





• Fitting plug (in Startup Kit)

• Replacement seal kit (in Startup Kit)

• Plunger insertion tool (part number WAT011042)

• Seal extraction tool (see Appendix B for part number)


To access a plunger seal, first remove the pump head.

To remove the pump head:



3. Use the syringe to withdraw all methanol.

4. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to run the pump.

5. After the indicator rod fully retracts into the pump head, click the Stop Flow button to turn off the pump. This ensures that the weight of the pump head does not rest on the plunger while you remove the head.

6. Use the 1/2-inch adjustable wrench to hold the check valve housings in place. Use the 5/16-inch open-end wrench to remove the inlet and outlet tubings from the check valve housings on the pump.

7. Holding the pump head against the pump, use the 5/32-inch Allen wrench to remove the two pump head assembly mounting screws. Loosen the screws 1/2-turn at a time for the first two turns (Figure 4-5).



4

Figure 4-5 Unscrewing Pump Head Mounting Screws (1525EF Pump)

8. Carefully slide the pump head assembly off the pump.

4.5.2 Removing and Replacing the 1525EF Plunger Seal1. Use the seal extraction tool to remove the seal from the pump head. (Twist the

plastic screw end of the tool clockwise into the seal, then pull the seal to remove it from the pump head. Do not reuse the seal.)

2. Place the new plunger seal onto the tip of the seal insertion tool. Make sure the spring in the seal is facing away from the end of the tool (you should be able to see the seal spring at the tip of the tool).

3. Moisten the seal with methanol.

4. Place the block part of the seal insertion tool into the pump head. (The smaller end of the tapered hole should be against the pump head.) Slide the tip of the tool (with the seal on it) through the block and into the pump head.

5. Press the tool firmly into the pump head, then remove it.

6. Remoisten the seal and plunger with methanol.

7. Carefully slide the pump head assembly into position over the plunger.

STOPAttention: Hold the pump head against the pump while removing the screws to avoid breaking the plunger.




4

8. Holding the pump head securely against the pump, reinstall the two pump head mounting screws. Alternately tighten each screw, using half turns to evenly align the assembly.

9. Using the software, navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to start the pump. Verify that the indicator rod moves freely.

10. Click the Stop Flow button to stop the pump.

11. Use the 1/2-inch adjustable wrench to hold the check valve housings in place. Reconnect the inlet and outlet tubings to the pump head. Tighten the tubings using the 5/16-inch open-end wrench.

12. Prime the pump as described in Section 3.2.1 on page 48. If you notice leaks, verify pump head and plunger seal installation.

4.5.3 Cleaning and Replacing 1525EF Plungers

The plungers are sapphire rods that require careful handling. Although cleaning the plungers is not difficult, it is important to follow these instructions carefully to avoid damaging the plungers. Assemble all materials and read the procedure thoroughly before you begin.

Required Materials





• Snap-ring pliers (part number WAT025263)

• Seal extraction tool and seal insertion tool (see Appendix B for part numbers)

• Plunger insertion tool (part number WAT011042)

• Sonicator


• Replacement plunger

• Replacement plunger seals

Removing the Plunger

1. Remove the pump head as described in Section 4.5.1, Preparing for Plunger Seal Replacement.



4

2. Remove the four head support screws with the 9/64-inch Allen wrench (Figure 4-6), then carefully slide the head support assembly and the indicator rod off the pump. Set the head support assembly on the benchtop.

Figure 4-6 Exposed Head Support Assembly (1525EF Pump)

3. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 0.3 mL/min, click Save, then click Load Method to run the pump. When the piston is fully extended, click the Stop Flow button to stop the pump.

4. Use the plunger insertion tool to compress the spring as far back as it will go (see Figure 4-8).

5. Keeping the spring compressed with the plunger insertion tool, use the snap-ring pliers to remove the snap ring that holds the plunger in place.

6. Carefully remove the plunger assembly. Continue with “Cleaning the Plunger”.

Cleaning the Plunger

1. Remove the plunger from the plunger assembly.

2. Clean the plunger by sonicating it in 50:50 methanol/water for a few minutes. The parts of the plunger assembly are shown in Figure 4-7. Continue with “Inspecting the Plunger”.


Head SupportScrewsHead Support

Assembly


4

Figure 4-7 Assembled Plunger (1525EF Pump)

Inspecting the Plunger

After cleaning the plunger, inspect it for damage by holding it under a bright white light and looking down its length for nicks and scratches. It is easier to see scratches under a bright light than to feel them with your fingers.

• If the plunger is not scratched or otherwise damaged, reassemble it with new seals. Continue with “Replacing the Plunger”.

• If the plunger is damaged, replace both the plunger and the seals. Continue with “Replacing the Plunger”.

Replacing the Plunger

1. Reassemble the components of the plunger assembly as shown in Figure 4-7.

2. Lubricate the ball and seat with high-pressure grease. Make sure the brass seat is located at the bottom of the piston.

3. Use the plunger insertion tool to push the plunger assembly into the pump piston as far as it will go.

4. While holding the plunger in position with the insertion tool, use the snap-ring pliers to insert the snap ring. Make sure the snap ring is fully seated in the groove before you release the pliers (Figure 4-8).



Snap-Ring

Plunger

Washer

Spring

Retainer


4

Figure 4-8 Inserting the Plunger (1525EF Pump)

5. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 1.0 mL/min, click Save, then click Load Method to run the pump. When the plunger is fully retracted, click the Stop Flow button to stop the pump.

6. With the indicator rod hole oriented to the upper right, reinstall the head support assembly. Alternately tighten the four screws. Do not overtighten.

7. Moisten the plunger seal and plunger with methanol.

8. Carefully slide the pump head over the plunger and onto the pump. Holding the pump head securely against the pump, alternately tighten the two screws.

Check for even head alignment by observing the gap between the pump head and the pump head support assembly.

9. In the Inlet Method window, set the Flow parameter to 1.0 mL/min, click Save, then click Load Method to run the pump.

10. Pull out and release the indicator rod.


Snap-Ring Pliers

Plunger Insertion Tool


4

• If the rod does not snap back easily, the head is misaligned. Stop the pump, loosen the pump head, then repeat steps 8 and 9.

• If the rod snaps back easily, click the Stop Flow button to stop the pump. Continue with step 11.

11. Use the 1/2-inch adjustable wrench to hold the check valve housings in place. Reconnect the inlet and outlet tubing assemblies to the pump head. Tighten the tubings using the 5/16-inch open-end wrench.

12. Prime the pump as described in Section 3.2.1 on page 48. If you notice leaks, verify pump head and plunger seal installation as described in Section 4.3.1 on page 55.

4.6 Replacing 1525EF Check Valves

This section describes how to replace inlet and outlet check valves for the 1525EF Pump. Replace the inlet and outlet check valves every 6 months, or more frequently if required by your application. Waters recommends keeping a spare set of new check valves on hand.

Note: Each check valve and check valve housing for the 1525EF Pump must be replaced as a single unit. Do not separate the check valve from the housing.

Required Materials




• Replacement check valves (assembled with housings)


• Forceps

Replacing the Inlet Check Valve


2. Hold the inlet check valve housing with the 1/2-inch adjustable wrench. Use the 5/16-inch open-end wrench to loosen the compression screw and remove the tubing assembly from the check valve housing.

3. Use the 1/2-inch adjustable wrench to remove the entire check valve assembly from the pump head.



4

4. Replace the entire old check valve assembly with a new one. Finger-tighten the new check valve housing onto the pump head. Tighten the check valve with the 1/2-inch adjustable wrench.

5. Hold the check valve housing with the 1/2-inch adjustable wrench. Reinstall the tubing assembly. Tighten the compression screw with the 5/16-inch open-end wrench while holding the assembly to prevent it from turning.

6. Repeat steps 2 through 5 to replace the other inlet check valves on the pump.

7. Prime the pump as described in Section 3.2.1 on page 48. Check for leaks.

Replacing the Outlet Check Valve


2. Run the pump until the indicator rod fully retracts into the pump head, then click the Stop Flow button to turn off the pump. This ensures that the weight of the pump head does not rest on the plunger when you remove the pump head.

3. Use the 1/2-inch adjustable wrench to hold the housings in place. Use the 5/16-inch open-end wrench to loosen the compression screws and remove the inlet and outlet tubing assemblies from the check valve housings (Figure 4-9).

Figure 4-9 Disconnecting the Outlet Tubing (1525EF Pump)


Pump Heads

Inlet Check

Outlet

HousingCheck Valve

Compression Screws

Valve Housing

Replacing 1525EF Check Valves 72

4

4. Use the 1/2-inch adjustable wrench to loosen the outlet check valve housing.

5. While holding the pump head in place, use the 5/32-inch Allen wrench to remove the two pump head assembly mounting screws. Loosen the screws 1/2-turn at a time for the first two turns.

6. Carefully slide the pump head assembly off the pump. (See the procedure on page 65.)

7. Hold the pump head upside-down in one hand with the outlet check valve housing facing the floor. Remove the entire outlet check valve housing and replace it with a new one.

8. Finger-tighten the new outlet check valve housing into the pump head.

9. Turn the pump head over and carefully slide the pump head over the plunger and onto the pump.

10. Holding the pump head securely against the pump, reinstall the two screws. Alternately tighten the screws to secure the pump head to the pump. Do not overtighten.

11. Tighten the outlet check valve housing with the 1/2-inch adjustable wrench.

12. Reinstall the outlet tubing assembly. Tighten the compression screw with the 5/16-inch open-end wrench while holding the housing with the 1/2-inch adjustable wrench.

13. Reinstall the inlet tubing assembly. Tighten the compression screw with the 5/16-inch open-end wrench while holding the housing with the 1/2-inch adjustable wrench.

14. Prime the pump as described in Section 3.2.1. Check for leaks.

4.7 Replacing a Draw-Off Valve

Replace a draw-off valve if you notice that the valve leaks even after you tighten the knob.

Required Materials



STOPAttention: Ensure that the pump head is straight and the indicator rod is properly aligned with the pump head to avoid damaging the plunger.


4




• Phillips screwdriver

• Replacement draw-off valve


Removing the Draw-Off Valve



3. Use the syringe to withdraw all the methanol.

4. Use the 5/16-inch open-end wrench to disconnect the three stainless steel compression screws from the draw-off valve (Figure 4-10).

Figure 4-10 Draw-Off Valve Inlet Manifold Assembly

STOPAttention: Before continuing, lower the eluent reservoir to eliminate gravity flow of eluent.

TP01416

Eluent Line

Handle Draw-Off Valve

Bracket

Cutout in Bracket

Allen Screw (7/64-Inch)

Compression Screw

Inlet Manifold

Replacing a Draw-Off Valve 74

4

5. Use the 5/16-inch open-end wrench to disconnect the two stainless steel compression screws from the inlet manifold (these screws connect the inlet manifold to the inlet check valves). See Figure 4-10.

6. Use the 7/64-inch Allen wrench to remove the two Allen screws that hold the draw-off valve assembly and bracket to the pump. Carefully remove the assembly.

7. Remove the two Phillips screws that hold the draw-off valve to the bracket.

Installing a Draw-Off Valve

1. Attach the new draw-off valve to the bracket using the two Phillips screws. Make sure the new valve is oriented on the bracket as shown in Figure 4-10.

2. Reposition the draw-off valve inlet manifold assembly onto the pump, being careful not to pinch the stainless steel tubing behind the bracket. When correctly positioned, the stainless steel tubing extends through the cutouts on the sides of the bracket.

3. Secure the bracket to the pump with the two Allen screws.

4. Reconnect the inlet tubing to the inlet manifold.

5. Reconnect the outlet tubing to the new draw-off valve.

6. Reposition the eluent reservoir, then prime the pump (see Section 3.2.1 on page 48). Check for leaks.

4.8 Replacing Fuses

The fuse holder is located on the rear panel of the pump (Figure 4-11). The 1525 Pump is shipped with two factory-installed 3.15-A fuses.


4

Figure 4-11 Rear Panel Fuse Holder

Identifying Faulty Fuses

Suspect a faulty fuse if:

• The pump fails to power up.

• The fan does not turn on.

Required Materials

• Small flat-blade screwdriver

• Replacement fuses

Procedure

1. Power off the pump, then unplug the power cord from the pump.

2. Use the small flat-blade screwdriver to remove the fuse holder located just above the power cord connector.

3. Remove and discard the old fuses.

Caution: To avoid possible electrical shock, power down and unplug the pump before checking the fuses. For continued protection against fire hazard, replace fuses only with those of the same type and rating.

Fuse Holder

Replacing Fuses 76

4

4. Insert the new fuses into the fuse holder.

5. Insert the fuse holder into the receptacle and gently push until it locks into position.

Caution: For continued protection against fire hazard, replace fuses with those of the appropriate type and rating.


5

Chapter 5Troubleshooting

This chapter describes the following:

• Troubleshooting pump problems based on error messages and malfunctions

• Identifying and correcting noises

• Identifying pump-related chromatographic problems

Contacting Waters Technical Service

If you encounter problems troubleshooting the pump that you cannot resolve, contact Waters Technical Service at 800 252-4752, U.S. and Canadian customers only. Other customers, call your local Waters subsidiary (or Technical Service Representative) or Waters corporate headquarters in Milford, Massachusetts 01757 (U.S.A.).

5.1 Troubleshooting Pump Problems

Table 5-1 is a guide to troubleshooting pump problems. It lists pump-related symptoms and their possible causes, and suggests corrective actions.

Caution: To prevent injury, observe good laboratory practices when troubleshooting the 1525 HPLC Pump.

Troubleshooting Pump Problems 78

5

Table 5-1 Correcting Pump Problems

Symptom Possible Cause Corrective Action

High pressure Pump flow rate set too high Set the correct flow rate.

High-pressure limit set too low Set the high-pressure limit to the correct value.

Blocked column Clean the column according to the column care and use manual.

Blocked mixer Backflush or disassemble the mixer.

Viscous eluents in use Check eluent viscosity (the observed pressure may be normal for the column/eluent blend). If necessary, change to a less viscous eluent.

Defective pressure transducer Contact Waters Technical Service.

Blocked outlet tubing or fluid path (in pump outlet, detector, column, or injector)

Locate the source of the blockage. Clean or replace tubing according to the appro-priate operator’s guide.

Ambient temperature has changed

Stabilize the operating temperature.

Low pressure Pump flow rate set too low Set the correct flow rate.

Low-pressure limit set too high Set the low-pressure limit to the correct value.

Eluent reservoir empty Fill the reservoir.

Air in eluent lines Prime the pump (see Section 3.2.1, Priming the Pump).

Incorrect eluent used Use the correct eluent.

Leaks Carefully tighten any loose fittings.

Ambient temperature has changed

Stabilize the operating temperature.

Troubleshooting 79

5

Pump does not run Pump not connected to power source

Ensure the power cable is properly connected to the power source and pump.

No power at outlet Check the outlet by connecting to another electrical unit known to be working. If that unit does not work, relocate the pump to a functioning elec-trical outlet.

Blown fuse Replace fuse.

Pump not delivering eluent

Draw-off valve open or leaking Close the draw-off valve. If eluent still leaks, replace the valve (see Section 4.7, Replacing a Draw-Off Valve).

Pump low-pressure limit set higher than operating pressure

Set the correct low-pressure limit.

Flow rate set to zero Set the desired pump flow rate.

Pressure transducer out of adjustment or defective

Contact Waters Technical Service.

Pump not primed Prime the pump (see Section 3.2.1, Priming the Pump).

Table 5-1 Correcting Pump Problems (Continued)



5

Pump not delivering eluent (continued)

Immiscible eluents in pump head

Purge the pump with appro-priate eluents. Verify miscibility of eluents being used and change to more miscible eluents.

Dirty or malfunctioning inlet or outlet check valve

Replace the check valves (see Section 4.4 or Section 4.6).

Damaged plunger seal (indi-cated by eluent leaking from behind the pump head or by salt crystal build-up around the back of the pump head)

Verify if both pump heads can maintain pressure as outlined in Section 4.2.2. If a head is leaking, replace the plunger seal (see Section 4.3 or Section 4.5).

Pump cavitation due to one of the following:• Eluent reservoirs positioned

at or below pump height• Loose, bent, or blocked inlet

tubing

• Improperly degassed eluent• Dirty eluent reservoir inlet

filter• Volatile eluents in pump

head• Tubing ID too small

Perform the following correc-tive action:• Raise eluent reservoirs

above the pump.• Check tubing. Tighten,

straighten, or replace tubing.

• Degas.• Replace the filter.

• Prime the pump (see Section 3.2.1).

• Use correct tubing.

Ruptured high-pressure pulse dampener


Defective pump motor Contact Waters Technical Service.

Defective circuit board Contact Waters Technical Service.



Troubleshooting 81

5

Leak from pump head Worn pump plunger seals Replace defective plunger seals (see Section 4.3.3 and Section 4.5.3).

Worn plunger Replace plunger see Section 4.3.3 and Section 4.5.3).

Loose pump head Tighten pump head screws. Ensure that both screws are tightened equally, otherwise uneven seal wear may result. Do not overtighten.

Loose inlet or outlet check valve

Tighten loose check valve(s). Do not overtighten.

Check fittings and ferrules for wear. Replace if necessary.

Leak from draw-off valve

Draw-off valve open or broken Close the draw-off valve. If leaking continues, replace the valve (see Section 4.7, Replacing a Draw-Off Valve).

Erratic flow rate/pump pulsations

Pump not primed Prime the pump. If using a volatile eluent (such as hexane), prime the pump with a miscible, less volatile eluent such as THF or methanol. Ensure that the column is disconnected to avoid disrupting equilibrium.




5

Erratic flow rate/pump pulsations (continued)

Reservoir low or out of eluent Refill the reservoir (filter and degas eluent).

Air bubble in pump head Prime the pump to remove the bubble. Ensure there are no air bubbles in the inlet lines. Degas eluents.

Dirty or malfunctioning check valve(s)

Replace check valves (see Section 4.4 or Section 4.6).

Inlet filter or inlet lines blocked Check lines for blockages. Replace the inlet filter frit.

Pump plunger seal leaking (under pump head)

Replace the pump plunger seal (see Section 4.3 or Section 4.5).

Worn pump plunger Replace the plunger (see Section 4.3 or Section 4.5).

Immiscible eluents in pump head

Refer to the Corrective Action under the “Pump not delivering eluent” symptom earlier.

Pump cavitation Refer to the Corrective Action(s) under the “Pump not delivering eluent” symptom earlier.

Pump electronics failure Contact Waters Technical Service.

High system pressure due to pump

Pump Flow parameter set too high

Set the correct flow parameter value.

Pressure transducer out of adjustment or defective




Troubleshooting 83

5

5.2 Troubleshooting Noises

Table 5-2 is a guide to troubleshooting and correcting noises in the Waters 1525 Pump.

Table 5-2 Correcting Noises


Click or loud snap Binding plunger seal If the click does not stop and cannot be isolated to one pump head, replace the plunger seals, one at a time (see Section 4.3 or Section 4.5).

Worn indicator rod spring Replace the spring.

Squeak Plunger seals dry Moisten the plunger with appropriate eluent through pump head access holes.

Binding plunger seal Replace the plunger seal assembly (see Section 4.3 or Section 4.5).

Improper plunger seal Install the correct pump plunger seal (see Section 4.3 or Section 4.5).

Binding piston indicator rod Replace the indicator rod.

Troubleshooting Noises 84

5

5.3 Troubleshooting Chromatographic Problems

Table 5-3 is a guide to troubleshooting and correcting chromatographic problems.

Table 5-3 Correcting Chromatographic Problems


Erratic retention times Air bubble in pump head Degas all eluents and prime the pump (see Section 3.2.1, Priming the Pump).

Malfunctioning check valves Replace pump check valves (see Section 4.4 or Section 4.6).

Leaking pump seals Replace pump seals (see Section 4.3 or Section 4.5).

Leaks around fittings Tighten fittings.

Separation chemistry Check the eluent and column.

Clogged eluent filters Replace filters.

Increased retention times

Incorrect flow rate Change the flow rate.

Incorrect eluent composition Change eluent composition.

Column heater not turned on Turn the column heater on.

Incorrect eluent Use the correct eluent.

Column contaminated Clean or replace the column.

Incorrect column Use the correct column.

Air bubble in pump head Degas all eluents and prime the pump (see Section 3.2.1, Priming the Pump).

Clogged eluent filter Replace the filter.

Reduced retention times


Incorrect eluent composition Change the composition.

High column temperature Reduce the column temperature.

Incorrect eluent Use the correct eluent.


Incorrect column Use the correct column.

Eluent not properly degassed Degas the eluent.

Troubleshooting 85

5

Reproducibility errors Eluent not properly degassed Degas the eluent.

Incorrect chemistry/integration Check the chemistry/integration.

Baseline drift, rapid Column not equilibrated Equilibrate the column.

Eluent contaminated Use fresh eluent.

Eluent not properly degassed (rapid or slow drift)

Degas the eluent.

Flow fluctuations (rapid or slow drift)

Fix pump problems, replace pump seals, and check valves.

Baseline drift, slow Eluent contaminated Use fresh eluent.

Ambient temperature fluctuations

Stabilize the operating envi-ronment temperature enough to allow full equilibration.

Baseline noise cycling, short-term (30 to 60 sec)

Fluctuating flow rate Degas the eluent. Try repriming the pump(s). If necessary, check flow accu-racy of the pump(s).

Inadequate eluent blending Install a mixer as shown in Section 2.4.2, Installing Different Mixer Configurations.

Power source (short- or long- term cycling)

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

Radio frequency noise (short- or long-term cycling)

Eliminate interference.

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

Ambient temperature fluctuations

Stabilize the ambient temperature.

Integrator or recorder faulty Check the integrator or recorder for excessive base-line noise.

Table 5-3 Correcting Chromatographic Problems (Continued)


Troubleshooting Chromatographic Problems 86

5

Baseline noise, random

Eluents not properly degassed Degas eluents.

Flow erratic, pump not primed Prime the pump (see Section 3.2.1, Priming the Pump).

Check for air in the pump, or for failing seals.

Eluents contaminated Use fresh eluent.

Column contaminated Clean and/or replace column.

System improperly grounded Plug into an outlet on a different electrical circuit.

Use the power conditioner.

Recorder voltage incorrect Set the recorder to the correct voltage.

Radio frequency noise Eliminate interference.

Straight baseline, no peaks

No pump flow Set the pump flow rate.

Leak in eluent path Repair the leak.

Sensitivity loss Degraded, contaminated, or improperly prepared sample

Use fresh sample.


Loss of column efficiency Clean or replace the column.

Change in eluent composition Correct eluent pH or ionic composition.


Table 5-3 Correcting Chromatographic Problems (Continued)


Troubleshooting 87

A
Appendix ASpecifications
This appendix provides the following specifications for the Waters® 1525 Pumps:

• Physical

• Environmental

• Electrical

• Performance

Table A-1 Physical Specifications for 1525 Pumps

Item Specification

HeightDepthWidthWeight

17.0 in (43 cm)24.0 in (61.0 cm)12.0 in (30.5 cm) (not including bottle holder)60.0 lb (27.2 kg)

Wetted surface material 316 stainless steel, sapphire, reinforced fluorocarbon polymer seals (1525µ Pump), UHMWPE seals (1525EF Pump), carbon-reinforced Tefzel®

Table A-2 Physical Specifications for the Column Heater

Item Specification

HeightDepthWidthWeight

17.0 in (43 cm)14.0 in (35.5 cm)6.0 in (15.2 cm)

13 lb (5.9 kg)

Specifications 88

A
Table A-3 Environmental Specifications
Item Specification

Operating temperature 4 to 40 °C (39 to 104 °F)

Humidity 20 to 80%, noncondensing

Audible noise <70 dBA at operator position

Solvent compatibility Solvents consistent with materials of construction. Salts and buffers can reduce seal life, especially at pressures in excess of 3000 psi.

Table A-4 Electrical Specifications

Item Specification

Protection classa

a. Protection Class I – The insulating scheme used in the instrument to protect you from electrical shock. Class I identifies a single level of insulation between live parts (wires) and exposed conductive parts (metal panels), in which the exposed conductive parts are connected to a grounding system. In turn, this grounding system is connected to the third pin (ground pin) on the electrical power cord plug.

Class I

Overvoltage categoryb

b. Overvoltage Category II – Pertains to instruments that receive their electrical power from a local level such as an electrical wall outlet.

Category II

Pollution degreec

c. Pollution Degree 2 – A measure of pollution on electrical circuits, which may produce a reduction of dielectric strength or surface resistivity. Degree 2 refers to normally only nonconductive pollution. Occasionally, however, a temporary conductivity caused by condensation must be expected.

Degree 2

Moisture protectiond

d. Moisture Protection – Normal (IPXO) – IPXO means that there is no Ingress Protection against any type of dripping or sprayed water. The X is a placeholder to identify protection against dust, if applicable.

Normal (IPXO)

Power requirements (pump) 200 VA

Power requirements (column heater)

150 VA

Line voltage 120/140 VAC

Frequency 50/60 Hz, single phase

Fuses 3.15 A, 250 V, two

Specifications 89

A
Table A-5 Performance Specifications for the 1525µ Pump
Item Specification

Programmable flow rate, range 50-µL heads

0.00 to 5.00 mL/min, in 0.01-mL/min increments

Maximum operating pressure 6000 psi (41,370 kPa, 401 bars) with 50-µL heads

Pressure ripple (one pump) <2.0% at 0.5 mL/min, degassed methanol, at 1000 to 2000 psi backpressure

Flow precision 2 seconds SD, six replicates based on retention time or volumetric measures at 0.2 mL/min, 1000 to 2000 psi backpressure, PQ test (each pump individually)

Flow accuracy 2% of setting at 0.1 mL/min using degassed methanol at 1000 to 2000 psi backpressure (each pump individually)

Gradient accuracy <1% (typical) between 10 to 90%, and 1000 psi back-pressure (methanol: methanol with propylparaben and one 50-µL mixer)

Gradient precision <0.5% RSD (typical) between 10 to 90%, 0.1 mL/min, and 1000 psi backpressure (methanol: water with octanophenone and one 50-µL mixer)

Gradient delay volume <60 µL (without mixer, no manual injector and no column heater) <200 µL (with one 50-µL mixer, no manual injector, and no column heater)

Table A-6 Performance Specifications for the 1525EF Pump

Item Specification

Programmable flow rate, range 225-µL heads

0.00 to 22.5 mL/min, in 0.01-mL/min increments

Maximum operating pressure 5000 psi (34,474 kPa, 345 bars) with 225-µL heads; programmable upper and lower limits

Pressure ripple (one pump) <3.0% at 2.25 mL/min, degassed methanol, at 1000 psi backpressure

Flow precision <0.1% RSD or 2 seconds SD, six replicates based on retention time or volumetric measures at 1 mL/min, 1000 to 2000 psi backpressure, PQ test (each pump individually)

Specifications 90

A
Flow accuracy ±0.5% of setting at 1 mL/min, or 30 µL/min, whichever is larger, using degassed methanol at 1000 to 2000 psi backpressure (each pump individually)
Gradient accuracy < ±1.0% between 10 to 90%, 1 mL/min, and 1000 psi backpressure (methanol: methanol with 5.6 mg/L propylparaben > 05 AUFS @257 nm and one GM150 mixer)

Gradient precision <0.5% RSD between 10 to 90%, 1 mL/min, and 1000 psi backpressure (methanol: methanol with 5.6 mg/L propylparaben > 05 AUFS @257 nm and one GM150 mixer)

Gradient ripple <1.5% normalized to full scale between 10 to 90%, 1 mL/min, and 1000 psi backpressure (methanol: meth-anol with 5.6 mg/L propylparaben > 05 AUFS @257 nm and one GM150 mixer)

Gradient delay volume <60 µL (without mixer, no manual injector and no column heater) < 500 µL (with one GM150 mixer, no manual injector, and no column heater)

Table A-6 Performance Specifications for the 1525EF Pump (Continued)

Item Specification

Specifications 91

B

Appendix BSpare Parts and Accessories

This appendix lists the spare parts, accessories, and maintenance and operation kits for the Waters 1525 HPLC Pumps.

The spare parts in Table B-1 and Table B-2 are recommended for customer installation. Any parts not listed in these tables may require installation by a trained Waters service representative.

Table B-1 Spare Parts for the 1525µ Pump

Item Quantity Part Number

Check Valve Assemblies 2 700002275

Check Valve Housings 2 WAT055845

Plunger Kit & Rod 1 WAT031788

Plunger Seals 2 WAT034515

Pump Head Support Assembly 1 WAT023061

Pump Head Assembly 1 700002250

Draw-Off Valve Assembly 1 WAS207085

Inlet Tubing Assembly (LH) 1 WAS207081

Inlet Tubing Assembly (RH) 1 WAS207082

Outlet Tubing Assembly (LH) 1 WAS207083

Outlet Tubing Assembly (RH) 1 WAS207084

Lower Drip Tray 1 700001091

Vent Valve Drip Tray 1 700001092

Tube Assembly, Vent Valve – Waste 1 700002245

Tube Assembly, Mixer – Vent Valve 1 700002244

Tube Assembly, Mixer – Mixer 1 700001164

Fuses, F 3.15 A/250 V (20 mm) 2 WAT163-16

Vent Valve 1 WAT055841

Gradient Mixer, 50-µL 1 700002306

Spare Parts and Accessories 92

B

Use the accessories in Table B-3 to optimize the Waters 1525µ Pump for your application and to simplify common procedures.

Table B-2 Spare Parts for the 1525EF Pump


Inlet Check Valve Assemblies 1 WAT060307

Inlet B & S Check Valve Kits 2 WAT088223

Inlet Check Valve Housing 1 WAT060308

Outlet Check Valve 1 WAT025216

Outlet Check Valve Rebuild Kits 2 WAT026014

Outlet Check Valve Housing 1 WAT025207

Pump Plungers 2 WAT031788

Plunger Seals 2 700002282

Indicator Rod Kit 1 WAT069583

Pump Head Support Assemblies 2 WAT060305

Pump Head Assembly 1 700002281

Draw-Off Valve Assembly 1 WAS207085

Inlet Tubing Assembly (LH) 1 WAS207081

Inlet Tubing Assembly (RH) 1 WAS207082

Outlet Tubing Assembly (LH) 1 WAS207083

Outlet Tubing Assembly (RH) 1 WAS207084

Lower Drip Tray 1 700001091

Tube Assembly, Tee - Transducer 1 700001093

Tube Assembly, Transducer – Ref. Valve

1 700001094

Tube Assembly, Ref. Valve – Waste 1 700001095

Tube Assembly, Transducer – Mixer 1 700001162

Tube Assembly, Mixer – Ref. Valve 1 700001163

Tube Assembly, Mixer – Mixer 1 700001164

Fuses, F 3.15 A/250 V (20 mm) 2 WAT163-16

Ref. Valve 1 WAT026592


B

Use the accessories in Table B-4 to optimize the Waters 1525EF Pump for your application and to simplify common procedures.

Table B-3 Accessories for the 1525µ Pump


Seal Wash Kit 1 205000203

Eluent Reservoir Filter 1 WAT025531

Syringe, 10-mL, Polypropylene 1 WAT010337

#316 Stainless Steel Tubing1/16-inch OD x 0.009-inch ID, 10 feet (3 m)1/16-inch OD x 0.020-inch ID, 10 feet (3 m)

1

1

WAT026973

WAT026804

ETFE Tubing1/8-inch OD x 0.062-inch ID, 25 feet (7.7 m)0.08-inch OD x 0.058-inch ID, 25 feet (7.7 m)

1

1

WAT026808

WAT026974

Gradient Mixer, 150-µL 1 WAT055847

Inline Degasser (2-Channel) 1 186001273

Inline Degasser (4-Channel) 1 186001272

Table B-4 Accessories for the 1525EF Pump


Plunger Wash Kit 1 WAT030852

Extended Flow Outlet Check Valve Assemblies

2 WAT025216

Eluent Reservoir Filter 1 WAT025531

Syringe, 10-mL, Polypropylene 1 WAT010337

#316 Stainless Steel Tubing1/16-inch OD x 0.009-inch ID, 10 feet (3 m)1/16-inch OD x 0.020-inch ID, 10 feet (3 m)

1

1

WAT026973

WAT026804


B
The kits in Table B-5 contain the parts necessary for common maintenance operations.
ETFE Tubing1/8-inch OD x 0.062-inch ID, 25 feet (7.7 m)0.08-inch OD x 0.058-inch ID, 25 feet (7.7 m)

1

1

WAT026808

WAT026974

Manual Inject Valve Kit, 7725i 1 186000872

Gradient Mixer, 150-µL 1 WAT055847

Large-Volume Gradient Mixer 1 WAT051519

Table B-5 Maintenance and Operation Kits and Tools


Solvent Clarification Kit110 V/60 Hz220 V/50 Hz

11

WAT085113WAT085122

Compression Screw/Ferrule Kit 5 per package WAT025604

Plunger Insertion Tool 1 WAT011042

Snap-Ring Pliers 1 WAT025263

Seal Extraction Tool (50/100-µL) 1 WAT039803

Seal Extraction Tool (50/225-µL) 1 289001375

Seal Insertion Tool (50-µL) 1 WAT088726

Seal Insertion Tool (100-µL 1 WAT024765

Seal Insertion Tool (225-µL) 1 WAT024924

Table B-4 Accessories for the 1525EF Pump (Continued)



C

Appendix CSolvent Considerations

C.1 Introduction

Clean Solvents

Clean solvents provide:

• Reproducible results

• Operation with minimal instrument maintenance

A dirty solvent can cause:

• Baseline noise and drift

• Blockage of the solvent filters with particulate matter

Solvent Quality

Use HPLC-grade solvents to ensure the best possible results. Filter solvents through 0.45-µm filters before use. Solvents distilled in glass generally maintain their purity from lot to lot; use them to ensure the best possible results.

Preparation Checklist

The following solvent preparation guidelines help to ensure stable baselines and good resolution:

• Filter solvents with a 0.45-µm filter.

• Degas and/or sparge the solvent.

• Stir the solvent.

• Keep in a place free from drafts and shock.

Caution: To avoid chemical hazards, always observe safe laboratory practices when operating your system.

Solvent Considerations 96

C

Water

Use water only from a high-quality water purification system. If the water system does not provide filtered water, filter it through a 0.45-µm membrane filter before use.

Buffers

When using buffers, dissolve salts first, adjust the pH, then filter to remove insoluble material.

Tetrahydrofuran (THF)

When using unstabilized THF, ensure that the solvent is fresh. Previously opened bottles of THF contain peroxide contaminants, which cause baseline drift.

C.2 Solvent Compatibility

The Waters 1525 HPLC Pump is constructed of high-quality (316) stainless steel components that, with some minor restrictions, can be used with all solvents. This section lists the solvents that have and have not been approved for use with the 1525 HPLC Pump.

Solvents to Avoid

You can use any solvent with the 1525 HPLC Pump. However, long-term static exposure to halide salts (for example, fluoride, bromide, chloride, and iodide) will pit and corrode stainless steel parts. When using these salts, flush the system thoroughly with water if the pump will be idle for more than 2 days.

Solvents to Use

Materials of construction used in the 1525 HPLC Pump are nonreactive with most acids, bases, salts, and organic solvents.

The solvents in Table C-1 through Table C-4 have been approved for use with the 1525 HPLC Pump. These include salts, acids, and bases in concentrations up to 1 M (unless otherwise noted), and organic solvents in concentrations of up to 100% (unless otherwise noted). Higher concentrations can be used in many instances.

Caution: THF contaminants (peroxides) are potentially explosive if concentrated or taken to dryness.


C


Information on the use of a specific solvent or concentration that is not listed in this guide can be obtained by contacting Waters.

Table C-1 Aqueous Buffers for Use with the 1525 HPLC Pump

Aqueous Buffers

Acetate K2SO4 Na2S Perfluorobutyric acid

Al2SO4 K3Fe(CN)6 Na2CO3 NH4Cl

Ca(OCl)2 K4Fe(CN)6 Na2SO4 Phosphate

CaCl2 KBr NaCl Tartrate

Citrate KCl Sodium acetate Trilithium citrate

H2O2 up to 10% KHCO3 NaH2BO3 Tris

HIBA KMnO4 NaHCO3 4-(2-pyridylazo)resorcinol monoso-dium salt monohydrate

K2CO3 KNO3 NaHSO4

K2Cr2O3 LiCl04 NaNO3

K2S Na2B4O7 NaOCl

Table C-2 Acids for Use with the 1525 HPLC Pump

Acids

Acetic acid, glacial Hydrochloric acid Perchloric acid

Benzoic acid Lactic acid Phosphoric acid

Chromic acid Methanesulphonic acid Pyridine-2,6-dicarboxylic acid

Citric acid Nitric acid, up to 37.5% (6 N) Sulfuric acid, up to 0.20 M

Formic acid Octanesulphonic acid Trifluoroacetic acid (TFA), up to 10%

Glyceric acid Oxalic acid

Table C-3 Bases for Use with the 1525 HPLC Pump

Bases

Ba(OH)2 NaOH, up to 10 M

KOH NH4OH, up to 3 M

LiOH Tetramethylammonium hydroxide pentahydrate

C
C.3 Solvent Miscibility
Before changing solvents, refer to Table C-5 to determine the miscibility of the solvents to be used. When changing solvents, be aware that:

• Changes involving two miscible solvents may be made directly. Changes involving two solvents that are not totally miscible (for example, from chloroform to water), require an intermediate solvent (such as methanol).

• Temperature affects solvent miscibility. When running a high-temperature application, consider the effect of the higher temperature on solvent solubility.

• Buffers dissolved in water may precipitate 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 you add the organic solvent.

Table C-4 Organic Solvents for Use with the 1525 HPLC Pump

Organic Solvents

4-Cyanophenol Chloroform Ethylene glycol Methylene chloride

Acetone Cyclohexane Formaldehyde n-Propanol

Acetonitrile Cyclohexanone Heptane Phenol

Amyl acetate Dibutyl phathalate Hexane Tetrahydrofuran (THF)

Benzaldehyde Dimethyl formamide Isooctane Toluene

Benzene Dimethyl sulfoxide Isopropanol Waters PIC® Reagents

Benzyl alcohol Ethanol Lysine hydrochloride Xylene

Butanol Ethyl acetate Methanol

Carbon tetrachloride

Ethylene dichloride Methyl ethyl ketone


C

Table C-5 Solvent Miscibility

Polarity Index

SolventViscosity CP, 20 °C

Boiling Point °C (@1 atm)

Miscibility Number

(M)

λ Cutoff (nm)

–0.3 N-Decane 0.92 174.1 29 ––

–0.4 Isooctane 0.50 99.2 29 210

0.0 N-Hexane 0.313 68.7 29 ––

0.0 Cyclohexane 0.98 80.7 28 210

1.7 Butyl ether 0.70 142.2 26 ––

1.8 Triethylamine 0.38 89.5 26 ––

2.2 Isopropyl ether 0.33 68.3 –– 220

2.3 Toluene 0.59 100.6 23 285

2.4 P-Xylene 0.70 138.0 24 290

3.0 Benzene 0.65 80.1 21 280

3.3 Benzyl ether 5.33 288.3 –– ––

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 Tetrahydrofuran 0.55 66.0 17 220

4.3 Ethyl acetate 0.47 77.1 19 260

4.3 1-Propanol 2.30 97.2 15 210

4.3 2-Propanol 2.35 117.7 15 ––

4.4 Methyl acetate 0.45 56.3 15, 17 260

4.5 Methyl ethyl ketone 0.43 80.0 17 330

4.5 Cyclohexanone 2.24 155.7 28 210

4.5 Nitrobenzene 2.03 210.8 14, 20 ––

4.6 Benzonitrile 1.22 191.1 15, 19 ––

4.8 Dioxane 1.54 101.3 17 220

5.2 Ethanol 1.20 78.3 14 210

5.3 Pyridine 0.94 115.3 16 305

5.3 Nitroethane 0.68 114.0 –– ––

5.4 Acetone 0.32 56.3 15, 17 330


C

How to Use Miscibility Numbers (M-Numbers)

Use miscibility numbers (M-numbers) to predict the miscibility of a liquid with a standard solvent (see Table C-5).

To predict the miscibility of two liquids, subtract the smaller M-number value from the larger M-number value.

• If the difference between the two M-numbers is 15 or less, the two liquids are miscible in all proportions at 15 °C.

• A difference of 16 indicates a critical solution temperature between 25 and 75 °C, with 50 °C as the optimal temperature.

• If the difference is 17 or greater, the liquids are immiscible or their critical solution temperature is above 75 °C.

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 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 and 32. Two liquids with dual M-numbers are usually miscible with each other.

5.5 Benzyl alcohol 5.80 205.5 13 ––

5.7 Methoxyethanol 1.72 124.6 13 ––

6.2 Acetonitrile 0.37 81.6 11, 17 210

6.2 Acetic acid 1.26 117.9 14 ––

6.4 Dimethylformamide 0.90 153.0 12 ––

6.5 Dimethylsulfoxide 2.24 189.0 9 ––

6.6 Methanol 0.60 64.7 12 210

7.3 Formamide 3.76 210.5 3 ––

9.0 Water 1.00 100.0 –– ––

Table C-5 Solvent Miscibility (Continued)

Polarity Index

SolventViscosity CP, 20 °C

Boiling Point °C (@1 atm)

Miscibility Number

(M)

λ Cutoff (nm)


C

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.

C.4 Buffered Solvents

When using a buffer, use a good quality reagent and filter it through a 0.45-µm filter.

Do not leave the buffer stored in the system after use. Flush all fluidic pathways with HPLC-quality water before shutting down the system and leave water in the system (flush with 90% HPLC-quality water:10% methanol for shutdowns scheduled to be more than one day). Flush using a minimum of 15 mL.

C.5 Head Height

Position the solvent reservoirs in the bottle holder supplied with the 1525 HPLC Pump (with adequate spill protection).

C.6 Solvent Viscosity

Generally, viscosity is not important when you are operating with a single solvent or under low pressure. However, when you are running a gradient, the viscosity changes that 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 affect the analysis is not known, monitor the pressure during the run.

C.7 Mobile Phase Solvent Degassing

Mobile phase difficulties account for the majority of all liquid chromatographic problems. Using degassed solvents is important, especially at wavelengths below 220 nm. Degassing provides:

• Stable baselines and enhanced sensitivity

• Reproducible retention times for eluting peaks


C

• Reproducible injection volumes for quantitation

• Stable pump operation

This section presents information on the solubility of gases, solvent degassing methods, and solvent degassing considerations.

C.7.1 Gas Solubility

Only a finite amount of gas can be dissolved in a given volume of liquid. This amount depends on:

• The chemical affinity of the gas for the liquid

• The temperature of the liquid

• The pressure applied to the liquid

Changes in the composition, temperature, or pressure of the mobile phase can lead to outgassing.

Effects of Intermolecular Forces

Nonpolar gases (N2, O2, CO2, and He) are more soluble in nonpolar solvents than in polar solvents. Generally, a gas is most soluble in a solvent whose intermolecular attractive forces are similar to those in the gas (“like dissolves like”).

Effects of Temperature

Temperature affects the solubility of gases. If the heat of solution is exothermic, the solubility of the gas decreases when you heat the solvent. If the heat of solution is endothermic, the solubility increases when you heat the solvent. For example, the solubility of He in H2O decreases with an increase in temperature, but the solubility of He in benzene increases with an increase in temperature.

Effects of Partial Pressure

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 you decrease the partial pressure of the gas, the amount of that gas in solution also decreases.

C.7.2 Eluent Degassing Methods

You can degas eluents using any of the following methods:

• Vacuum degassing

• Inline degassing

• Heating

• Vacuum sonication


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Vacuum Degassing

Vacuum degassing operates on the principle of Henry’s law to remove dissolved gases from the solvent. According to Henry’s law, the mole fraction of a gas dissolved in a liquid is proportional to the partial pressure of that gas in the vapor phase above the liquid. If the partial pressure of a gas on the surface of the liquid is reduced, for example, by evacuation, then a proportional amount of that gas comes out of solution.

Note: Vacuum degassing may change the composition of mixed solvents.

Inline Degassing

Inline degassing removes gases from the eluent as it passes through a gas-permeable membrane enclosed in a vacuum chamber. The vacuum in the chamber accelerates the rate at which the dissolved gas diffuses through the gas-permeable membrane. This method provides an automatic, continuous method of removing dissolved gasses, and allows for quick eluent changeover. Waters makes two inline degassers: part numbers 186001273 (2 Eluent) and 186001272 (4 Eluent). Contact Waters for details.

Using an Inline Degasser

The longer a solvent is exposed to a vacuum, the more dissolved gases are removed. Two factors affect the amount of time the solvent is exposed to the vacuum:

• Flow rate – At low flow rates, most of the dissolved gas is removed as the solvent passes through the vacuum chamber. At higher flow rates, lesser amounts of gas per unit volume of solvent are removed.

• Surface area of the degassing membrane – The length of the degassing membrane is fixed in each vacuum chamber. To increase the length of membrane, you can connect two or more vacuum chambers in series.

Heating

Heating to remove dissolved gas is usually not effective unless you boil the eluent, and this is not practical or safe for mixed, flammable, or volatile eluents. However, raising the temperature even slightly raises the partial pressure of the eluent, and thereby reduces the rate of resolubilization of a gas.

Vacuum Sonication

Sonication in combination with a vacuum degasses eluents very quickly. This technique does not change the composition of mixed eluents appreciably.

Caution: Apply vacuum only to suitable vessels. The brown gallon bottles in which eluent is shipped are not designed for vacuum degassing. There is a high risk of implosion if these bottles are used for vacuum degassing.


C

Conclusions

With any of the above techniques, the eluent reequilibrates to air saturation in 12 to 24 hours, depending on the eluent.

Degassing by vacuum or sonication or both is often performed for improved pump performance in multipump gradient applications.

C.8 Wavelength Selection

The tables in this section provide UV cutoff values for:

• Common solvents

• Common mixed mobile phases

• Chromophores

UV Cutoffs for Common Solvents

Table C-6 shows the UV cutoff (the wavelength at which the absorbance of the solvent is equal to 1 AU) for some common chromatographic solvents. Operating at a wavelength near or below the cutoff increases baseline noise due to the absorbance of the solvent.

Table C-6 UV Cutoff Wavelengths for Common Chromatographic Solvents

Solvent UV Cutoff (nm) Solvent UV Cutoff (nm)

1-Nitropropane 380 Ethylene glycol 210

2-Butoxyethanol 220 Isooctane 215

Acetone 330 Isopropanol 205

Acetonitrile 190 Isopropyl chloride 225

Amyl alcohol 210 Isopropyl ether 220

Amyl chloride 225 Methanol 205

Benzene 280 Methyl acetate 260

Carbon disulfide 380 Methyl ethyl ketone 330

Carbon tetrachloride 265 Methyl isobutyl ketone 334

Chloroform 245 Methylene chloride 233

Cyclohexane 200 n-Pentane 190

Cyclopentane 200 n-Propanol 210

Diethyl amine 275 n-Propyl chloride 225

Dioxane 215 Nitromethane 380


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Mixed Mobile Phases

Table C-7 provides approximate wavelength cutoffs for some other solvents, buffers, detergents, and mobile phases. The solvent concentrations represented are those most commonly used. If you want to use a different concentration, you can determine approximate absorbance using Beer’s law, since absorbance is proportional to concentration.

Ethanol 210 Petroleum ether 210

Ethyl acetate 256 Pyridine 330

Ethyl ether 220 Tetrahydrofuran 230

Ethyl sulfide 290 Toluene 285

Ethylene dichloride 230 Xylene 290

Table C-7 Wavelength Cutoffs for Different Mobile Phases

Mobile PhaseUV Cutoff

(nm)Mobile Phase

UV Cutoff (nm)

Acetic acid, 1% 230 Sodium chloride, 1 M 207

Ammonium acetate, 10 mM 205 Sodium citrate, 10 mM 225

Ammonium bicarbonate, 10 mM

190 Sodium dodecyl sulfate 190

BRIJ 35, 0.1% 190 Sodium formate, 10 mM 200

CHAPS, 0.1% 215 Triethyl amine, 1% 235

Diammonium phosphate, 50 mM

205 Trifluoracetic acid, 0.1% 190

EDTA, disodium, 1 mM 190 TRIS HCl, 20 mM, pH 7.0, pH 8.0

202, 212

HEPES, 10 mM, pH 7.6 225 Triton-X™ 100, 0.1% 240

Hydrochloric acid, 0.1% 190 Waters PIC Reagent A, 1 vial/liter

200

MES, 10 mM, pH 6.0 215 Waters PIC Reagent B-6, 1 vial/liter

225

Potassium phosphate, monobasic, 10 mM dibasic, 10 mM

190190

Waters PIC Reagent B-6, low UV, 1 vial/liter

190

Sodium acetate, 10 mM 205 Waters PIC Reagent D-4, 1 vial/liter

190

Table C-6 UV Cutoff Wavelengths for Common Chromatographic Solvents (Continued)

Solvent UV Cutoff (nm) Solvent UV Cutoff (nm)


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Refractive Indices of Common Solvents

Table C-8 lists the refractive indices for some common chromatographic solvents. Use this table to verify that the solvent you intend to use for your analysis has a refractive index (RI) significantly different from the RIs of the sample components.

Table C-8 Refractive Indices for Common Chromatographic Solvents

Solvent RI Solvent RI

Fluoroalkanes 1.25 Tetrahydrofuran (THF) 1.408

Hexafluoroisopropanol (HFIP) 1.2752 Amyl alcohol 1.410

Methanol 1.329 Diisobutylene 1.411

Water 1.33 n-Decane 1.412

Acetonitrile 1.344 Amyl chloride 1.413

Ethyl ether 1.353 Dioxane 1.422

n-Pentane 1.358 Ethyl bromide 1.424

Acetone 1.359 Methylene chloride 1.424

Ethanol 1.361 Cyclohexane 1.427

Methyl acetate 1.362 Ethylene glycol 1.427

Isopropyl ether 1.368 N,N-Dimethyl Formamide (DMF)

1.428

Ethyl acetate 1.370 N,N-Dimethyl Acetamide (DMAC)

1.438

1-Pentene 1.371 Ethyl sulfide 1.442

Acetic acid 1.372 Chloroform 1.443

Isopropyl chloride 1.378 Ethylene dichloride 1.445

Isopropanol 1.38 Carbon tetrachloride 1.466

n-Propanol 1.38 Dimethyl sulfoxide (DMSO) 1.477

Methylethylketone 1.381 Toluene 1.496

Diethyl amine 1.387 Xylene ~1.50

n-Propyl chloride 1.389 Benzene 1.501

Methylisobutylketone 1.394 Pyridine 1.510

Nitromethane 1.394 Chlorobenzene 1.525

1-Nitropropane 1.400 o-Chlorophenol 1.547

Isooctane 1.404 Aniline 1.586

Cyclopentane 1.406 Carbon disulfide 1.626


I NDEX

Index

Numerics1525 Pumps 24

accessories 92, 93, 94control by MassLynx software 30dimensions 31electrical specifications 89electronic components 29environmental specifications 89equilibrating 50fluid-handling components 26installing 31maintaining 52maintenance kits 95operation 46operation kits 95performance specifications 90physical specifications 88powering off 50powering on 46preparing 46priming and purging 48selecting 46site requirements 31spare parts 92, 93tools 95unpacking 33

AAccessories 92Air bubbles 83

BBench space 31Bottle holder, installing 38Buffered solvents 102Buffers 97

CCavitation in pump head 81, 83Check valves

loose 82malfunctioning 81replacing 61, 71

Chromatographic problems 85Clearance 31Column heater 27

installing 42physical specifications 88

Conventions, documentation 22

DDegassing 81, 102–105

benefits 103considerations 104inline 104

Diagnostic tests 53Documentation

conventions 22related 21

Draw-off valve 27installing 75leaking 80, 82removing 74replacing 73

Drip tray 28

EElectrical connections 34Electromagnetic fields 31Eluent

filter 81, 83supply connections 37

Index 108

INDEX

I

Eluent tubingconnecting to eluent reservoir 39connecting to pump inlet 38

Eluentsbuffered 99considerations 96guidelines 47heating 103viscosity 102volatile 81

Equilibrating system 49

FFerrule 82Filters

eluent 81, 83high-pressure noise 81

Flow pulsations 82, 83Flow rate 80

erratic 82, 83Fluid waste lines, connecting 44Fluidic connections 37Frequency 31Fuses

indentifying faulty 76replacing 75

GGas solubility 103Gradient mixer 28Guidelines, eluents 96

HHead height 81Heating eluents 104High-pressure noise filter 81

IIEEE-488

connecting devices 36connections 36guidelines 36interface 24setting address 34

Immiscible eluents 81Inlet check valve assemblies 28Inlet check valves, replacing 62, 71Inlet manifold 28Inline degassing 104Input voltage 31

LLeaking

draw-off valves 80, 82plunger seals 83

Leaks 82

MMaintenance considerations 52Manual injector 28

connecting to column 45connecting to column heater 45installing 42waste line 28

Methanol 82Miscibility

numbers 99of eluents 81of solvents 99

Mixersbypassing 42different configurations 39GM150 Gradient (dual) 41installing 41

M-numbers. See Miscibility numbers

Index 109

INDEX

Mobile phasecompatibility 97degassing 103miscibility 99viscosity considerations 102

NNoises, troubleshooting 84

OOperating procedures 52Outlet check valves, replacing 63, 72

PPlunger indicator rods 28Plunger seals 81

cleaning 55, 64leaking 83replacing 55, 57, 64, 65, 66squeaking 84worn 82

Plungerscleaning 55, 58, 59, 64, 67, 68inspecting 60, 69removing 59, 67replacing 58, 60, 67, 69worn 82

Power requirements 31Power supply, connecting 34Pressure transducer 28, 80, 83Priming the pump 80, 82Pulsations in eluent delivery 82, 83Pulse dampeners 28Pump head

assembly 28removing 56, 65

Pump outlet 28connecting 43

Pump, priming 80, 82

I

RRamp-and-decay test 53Reference valve 28Refractive indices 107Related documentation 21Relative humidity 31Reservoirs, positioning 102Restrictor tee 28Restrictor tee check valve, replacing 64Retention time stability test 53

SSalt crystals 81Seal wash holes 28Solvent

buffered 102compatibility 97general considerations 96guidelines 96miscibility 99refractive indices 107UV cutoff 105viscosity considerations 102

Solvent reservoirs. See ReservoirsSonication 104Spare parts 53, 92Specifications

1525 Pumps 88electrical 89environmental 89performance 90

Static electricity 31

Index 110

INDEX

TTee 28Temperature 31Tests

diagnostic 53ramp-and-decay 53retention time stability 53

THF (tetrahydrofuran) 82, 97Troubleshooting

chromatographic problems 85contacting Waters 78noises 84pump problems 78

Tubingattaching compression fittings 44cutting to length 43making connections 44problems with 81

VVacuum sonication 104Vent valve 28Vibration 31Viscosity, eluent 102Volatile eluents 81, 82

WWaste line

connecting drip tray 45connecting pump 45

Wavelength selection 105

I

Index 111

Waters 1525µ and 1525EF HPLC Pump - Waters … · The Waters® 1525µ and 1525EF HPLC Pumps can be...

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