Waters 1525M Installation And Maintenance Manual
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Waters 1525M Installation And Maintenance Manual

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Waters 1525µ and
1525EF HPLC Pump
Installation and Maintenance Guide
34 Maple Street
Milford, MA 01757
71500152504, Revision A

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  • Page 1 Waters 1525µ and 1525EF HPLC Pump Installation and Maintenance Guide 34 Maple Street Milford, MA 01757 71500152504, Revision A...
  • Page 2 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.
  • Page 3 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.
  • Page 4 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.
  • Page 5 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. •...
  • Page 7 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.
  • Page 8 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.
  • Page 9 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.
  • Page 10 Commonly Used Symbols Direct current Courant continu Gleichstrom Corrente continua Corriente continua Alternating current Courant alternatif Wechselstrom Corrente alternata Corriente alterna Protective conductor terminal Borne du conducteur de protection Schutzleiteranschluss Terminale di conduttore con protezione Borne del conductor de tierra...
  • Page 11 Commonly Used Symbols (Continued) Frame or chassis terminal Borne du cadre ou du châssis Rahmen- oder Chassisanschluss Terminale di struttura o telaio Borne de la estructura o del chasis Caution or refer to manual Attention ou reportez-vous au guide Vorsicht, oder lesen Sie das Handbuch Prestare attenzione o fare riferimento alla guida Actúe con precaución o consulte la guía Caution, hot surface or high temperature...
  • Page 12 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 puncture Attention, risques de perforation de la taille d’une aiguille Vorsicht, Gefahr einer Spritzenpunktierung Precauzione, rischio di puntura con ago...
  • Page 13 Commonly Used Symbols (Continued) Fuse Fusible Sicherung Fusibile Fusible Electrical power on Sous tension Netzschalter ein Alimentazione elettrica attivata Alimentación eléctrica conectada Electrical power off Hors tension Netzschalter aus Alimentazione elettrica disattivata Alimentación eléctrica desconectada...
  • Page 14 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,”...

  • Page 15: Table Of Contents

    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.............
  • Page 16 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 .............
  • Page 17 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 ............
  • Page 18 Waters 1525EF Pump with an Optional Column Heater....25 Fluid-Handling Components in the Waters 1525µ Pump....26 Fluid-Handling Components in the Waters 1525EF Pump .... 27 Rear Panel Electronic Components in the Waters 1525 Pump ..29 Pump Dimensions................32 IEEE-488 Connection Example ............. 37 Reverse Ferrule and Compression Screw Assembly.....
  • Page 19 List of Tables Fluid-Handling Components in the Waters 1525 Pump ....27 Electronic Components in the Waters 1525 Pump ....30 Installation Site Requirements ............31 IEEE-488 DIP Switch Settings..........35 Pump Preparation Recommendations .......... 47 Correcting Pump Problems ............79 Correcting Noises ..............
  • Page 20 UV Cutoff Wavelengths for Common Chromatographic Solvents ..................105 Wavelength Cutoffs for Different Mobile Phases ....106 Refractive Indices for Common Chromatographic Solvents ..107 List of Tables...

  • Page 21: Preface

    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.
  • Page 22 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.
  • Page 23 Convention Usage 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. Notes Notes call out information that is helpful to the operator.

  • Page 24: Chapter 1 Overview

    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.

  • Page 25: Waters 1525Ef Pump With An Optional Column Heater

    Bottle Holder Column Heater TP01722 Figure 1-2 Waters 1525EF Pump with an Optional Column Heater Overview...

  • Page 26: Fluid-Handling Components

    1.1 Fluid-Handling Components Before installing a Waters 1525 Pump, become familiar with its fluid-handling components, as illustrated in Figure 1-3 Figure 1-4. Draw-Off Pump Head Assembly Valve Outlet Check Valve Assembly Indicator Rod Gradient Inlet Check Mixer Valve Assembly Vent Valve...

  • Page 27: Fluid-Handling Components In The Waters 1525Ef Pump

    Pump Module A Drip Tray Drip Tray Waste Exit 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...
  • Page 28 Table 1-1 Fluid-Handling Components in the Waters 1525 Pump (Continued) Component Description 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...

  • Page 29: Electronic Components

    1.2 Electronic Components Before installing the Waters 1525 Pump, become familiar with its electronic components as illustrated in Figure 1-5. I/O Terminal Block IEEE-488 Address Switches Power Switch (Side of Unit) IEEE-488 Port Cooling Fan Vent Fuse Holder Ethernet Port...

  • Page 30: 1525Μ And1525Ef Pump Control

    (for service use only). 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).

  • Page 31: Installing The 1525 Pump

    Chapter 2 Installing 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.

  • Page 32: Pump Dimensions

    17 in. (43 cm) Clearance for Optional Column Heater 7 in. 24 in. (61 cm) 12 in. Clearance for 5 in. Bottle Holder Figure 2-1 Pump Dimensions Installing the 1525 Pump...

  • Page 33: Unpacking The Pump

    • 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.

  • Page 34: Making Electrical Connections

    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 •...

  • Page 35: Ieee-488 Dip Switch Settings

    Table 2-2 IEEE-488 DIP Switch Settings DIP Switch Settings IEEE-488 Address a. ON = 1 and OFF = 0. Making Electrical Connections...

  • Page 36: Making Ieee-488 Connections

    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.

  • Page 37: Making Fluidic Connections

    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 •...
  • Page 38 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) •...

  • Page 39: Installing Different Mixer Configurations

    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.
  • Page 40 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...

  • Page 41: 1525Ef Pump Mixer Configurations

    Gradient Mixers 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.

  • Page 42: Installing An Optional 1500 Series Manual Injector

    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.

  • Page 43: Connecting The Pump Outlet

    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.

  • Page 44: Connecting Fluid Waste Lines

    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. Ferrule 0.12 Inch Compression Screw...
  • Page 45 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.

  • Page 46: Preparing For Operation

    Chapter 3 Preparing 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.

  • Page 47: Pump Preparation Recommendations

    Table 3-1 Pump Preparation Recommendations Status Prime Equilibrate Initiating operation Changing eluent Adding fresh eluent System has been idle for some time Priming and equilibrating procedures are described in Section 3.2.1 Section 3.2.2. Eluent Guidelines When preparing a 1525µ or 1525EF Pump for operation, follow these guidelines: •...

  • Page 48: Priming The Pump

    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.

  • Page 49: Equilibrating The Pump

    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.

  • Page 50: Powering Off The Pump

    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.
  • Page 51 2. If the pump is running, powering off the pump will stop flow immediately. Attention: Since abrupt flow changes can damage columns, it is advisable STOP to ramp flow to 0 using an appropriate user-defined MassLynx shutdown method. 3. Set the power switch to the (OFF) position.

  • Page 52: Maintaining The 1525 Pump

    Chapter 4 Maintaining 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...

  • Page 53: Diagnostic Tests

    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.).
  • Page 54 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. Attention: Before starting this procedure, move the eluent reservoir below STOP...

  • Page 55: Replacing 1525Μ Plunger Seals And Plungers

    • 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.
  • Page 56 • 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) •...

  • Page 57: Removing And Replacing The 1525Μ Plunger Seal

    Mounting Screws Figure 4-1 Unscrewing Pump Head Mounting Screws (1525µ Pump) 9. Carefully slide the pump head assembly off the pump. Attention: Pull the pump head straight off to avoid breaking the plunger. STOP 4.3.2 Removing and Replacing the 1525µ Plunger Seal 1.

  • Page 58: Cleaning And Replacing 1525Μ Plungers

    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. Attention: Hold the pump head against the pump while replacing the screws STOP to avoid breaking the plunger. 10.

  • Page 59: Exposed Head Support Assembly (1525Μ Pump)

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

  • Page 60: Disassembled Plunger (1525Μ Pump)

    Retainer Seal Spring Plunger Piston Ball Seat Retainer Figure 4-3 Disassembled Plunger (1525µ Pump) 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. 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.

  • Page 61: 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.

  • Page 62: Disconnecting The Inlet And Outlet Tubing (1525Μ Pump)

    Replacing the Inlet Check Valve Attention: Before starting this procedure, move the eluent reservoir below the pump STOP head level. This eliminates gravity flow of the eluent. 1. Purge the pump with methanol. If methanol is not miscible with your current eluent, use an intermediate eluent.
  • Page 63 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.

  • Page 64: Replacing 1525Ef Plunger Seals And Plungers

    • 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.

  • Page 65: Preparing For Plunger Seal Replacement

    4.5.1 Preparing for Plunger Seal Replacement Required Materials • 5/16-inch open-end wrench (in Startup Kit) • 5/32-inch Allen wrench (in Startup Kit) • 1/2-inch adjustable wrench (in Startup Kit) • Priming syringe (in Startup Kit) • Fitting plug (in Startup Kit) •...

  • Page 66: Removing And Replacing The 1525Ef Plunger Seal

    Figure 4-5 Unscrewing Pump Head Mounting Screws (1525EF Pump) Attention: Hold the pump head against the pump while removing the screws STOP to avoid breaking the plunger. 8. Carefully slide the pump head assembly off the pump. Attention: Pull the pump head off straight to avoid breaking the plunger. STOP 4.5.2 Removing and Replacing the 1525EF Plunger Seal 1.

  • Page 67: Cleaning And Replacing 1525Ef Plungers

    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 mL/min, click Save, then click Load Method to start the pump.

  • Page 68: Exposed Head Support Assembly (1525Ef Pump)

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

  • Page 69: Assembled Plunger (1525Ef Pump)

    Washer Snap-Ring Plunger Spring Retainer Figure 4-7 Assembled Plunger (1525EF Pump) 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. 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.

  • Page 70: Inserting The Plunger (1525Ef Pump)

    Snap-Ring Pliers Plunger Insertion Tool Figure 4-8 Inserting the Plunger (1525EF Pump) 5. Use the software to navigate to the Inlet Method window. Set the Flow parameter to 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.

  • Page 71: 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.

  • Page 72: Disconnecting The Outlet Tubing (1525Ef Pump)

    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.

  • Page 73: Replacing A Draw-Off Valve

    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.
  • Page 74 • 7/64-inch Allen wrench (in Startup Kit) • 1/2-inch adjustable wrench (in Startup Kit) • Priming syringe (in Startup Kit) • Phillips screwdriver • Replacement draw-off valve • HPLC-grade methanol Removing the Draw-Off Valve 1. Purge the pump with methanol. If methanol is not miscible with your current eluent, use an intermediate eluent.

  • Page 75: Replacing Fuses

    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.
  • Page 76 Fuse Holder Figure 4-11 Rear Panel Fuse Holder 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. Identifying Faulty Fuses Suspect a faulty fuse if: •...
  • Page 77 Caution: For continued protection against fire hazard, replace fuses with those of the appropriate type and rating. 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. Maintaining the 1525 Pump...

  • Page 78: Chapter 5 Troubleshooting

    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.).

  • Page 79: Correcting Pump Problems

    If necessary, change to a less viscous eluent. Defective pressure transducer Contact Waters Technical Service. Blocked outlet tubing or fluid Locate the source of the path (in pump outlet, detector, blockage. Clean or replace...
  • Page 80 Set the correct low-pressure higher than operating pressure limit. Flow rate set to zero Set the desired pump flow rate. Pressure transducer out of Contact Waters Technical adjustment or defective Service. Pump not primed Prime the pump (see Section 3.2.1, Priming the Pump).
  • Page 81 • Volatile eluents in pump • Prime the pump (see head Section 3.2.1). • Tubing ID too small • Use correct tubing. Ruptured high-pressure pulse Contact Waters Technical dampener Service. Defective pump motor Contact Waters Technical Service. Defective circuit board Contact Waters Technical Service.
  • Page 82 Table 5-1 Correcting Pump Problems (Continued) Symptom Possible Cause Corrective Action Leak from pump head Worn pump plunger seals Replace defective plunger seals (see Section 4.3.3 Section 4.5.3). Worn plunger Replace plunger see Section 4.3.3 Section 4.5.3). Loose pump head Tighten pump head screws.

  • Page 83: Pump Plungers

    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 Pump Flow parameter set too Set the correct flow parameter due to pump high value.

  • Page 84: Troubleshooting Noises

    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 Symptom Possible Cause Corrective Action Click or loud snap Binding plunger seal If the click does not stop and...

  • Page 85: Troubleshooting Chromatographic Problems

    5.3 Troubleshooting Chromatographic Problems Table 5-3 is a guide to troubleshooting and correcting chromatographic problems. Table 5-3 Correcting Chromatographic Problems Symptom Possible Cause Corrective Action Erratic retention times Air bubble in pump head Degas all eluents and prime the pump (see Section 3.2.1, Priming the Pump).
  • Page 86 Table 5-3 Correcting Chromatographic Problems (Continued) Symptom Possible Cause Corrective Action 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 Degas the eluent.
  • Page 87 Table 5-3 Correcting Chromatographic Problems (Continued) Symptom Possible Cause Corrective Action Baseline noise, Eluents not properly degassed Degas eluents. random 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.

  • Page 88: Appendix A Specifications

    Appendix A Specifications ® 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 Height 17.0 in (43 cm) Depth 24.0 in (61.0 cm) Width 12.0 in (30.5 cm) (not including bottle holder)

  • Page 89: Environmental Specifications

    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.

  • Page 90: Performance Specifications For The 1525Μ Pump

    Table A-5 Performance Specifications for the 1525µ Pump Item Specification Programmable flow rate, range 0.00 to 5.00 mL/min, in 0.01-mL/min increments 50-µL heads 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...
  • Page 91 Table A-6 Performance Specifications for the 1525EF Pump (Continued) Item Specification 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 >...

  • Page 92: Spare Parts And Accessories

    Waters 1525 HPLC Pumps. The spare parts in Table B-1 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...

  • Page 93: Spare Parts For The 1525Ef Pump

    Tube Assembly, Mixer – Mixer 700001164 Fuses, F 3.15 A/250 V (20 mm) WAT163-16 Ref. Valve WAT026592 Use the accessories in Table B-3 to optimize the Waters 1525µ Pump for your application and to simplify common procedures. Spare Parts and Accessories...

  • Page 94: Accessories For The 1525Μ Pump

    Inline Degasser (2-Channel) 186001273 Inline Degasser (4-Channel) 186001272 Use the accessories in Table B-4 to optimize the Waters 1525EF Pump for your application and to simplify common procedures. Table B-4 Accessories for the 1525EF Pump Item Quantity Part Number Plunger Wash Kit...

  • Page 95: Maintenance And Operation Kits And Tools

    Table B-4 Accessories for the 1525EF Pump (Continued) Item Quantity Part Number ETFE Tubing 1/8-inch OD x 0.062-inch ID, WAT026808 25 feet (7.7 m) 0.08-inch OD x 0.058-inch ID, WAT026974 25 feet (7.7 m) Manual Inject Valve Kit, 7725i 186000872 Gradient Mixer, 150-µL WAT055847 Large-Volume Gradient Mixer...

  • Page 96: Solvent Considerations

    Appendix C Solvent Considerations Caution: To avoid chemical hazards, always observe safe laboratory practices when operating your system. C.1 Introduction Clean Solvents Clean solvents provide: • Reproducible results • Operation with minimal instrument maintenance A dirty solvent can cause: • Baseline noise and drift •...

  • Page 97: Solvent Compatibility

    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.

  • Page 98: Aqueous Buffers For Use With The 1525 Hplc Pump

    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 Perfluorobutyric acid Fe(CN)

  • Page 99: Solvent Miscibility

    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 Ethylene dichloride Methyl ethyl ketone tetrachloride C.3 Solvent Miscibility...

  • Page 100: Solvent Miscibility

    Table C-5 Solvent Miscibility Boiling Miscibility λ Cutoff Polarity Viscosity Solvent Point °C Number Index CP, 20 °C (nm) (@1 atm) –0.3 N-Decane 0.92 174.1 –– –0.4 Isooctane 0.50 99.2 N-Hexane 0.313 68.7 –– Cyclohexane 0.98 80.7 Butyl ether 0.70 142.2 ––...
  • Page 101 Table C-5 Solvent Miscibility (Continued) Boiling Miscibility λ Cutoff Polarity Viscosity Solvent Point °C Number Index CP, 20 °C (nm) (@1 atm) Benzyl alcohol 5.80 205.5 –– Methoxyethanol 1.72 124.6 –– Acetonitrile 0.37 81.6 11, 17 Acetic acid 1.26 117.9 ––...

  • Page 102: Buffered Solvents

    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.

  • Page 103: Gas Solubility

    • 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: •...
  • Page 104 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.

  • Page 105: Wavelength Selection

    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: •...
  • Page 106 TRIS HCl, 20 mM, pH 7.0, 202, 212 pH 8.0 HEPES, 10 mM, pH 7.6 Triton-X™ 100, 0.1% Hydrochloric acid, 0.1% Waters PIC Reagent A, 1 vial/liter MES, 10 mM, pH 6.0 Waters PIC Reagent B-6, 1 vial/liter Potassium phosphate,...
  • Page 107 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 Solvent...

  • Page 108: Index

    Index Numerics 1525 Pumps Cavitation in pump head accessories Check valves control by MassLynx software loose dimensions malfunctioning electrical specifications replacing electronic components Chromatographic problems environmental specifications Clearance equilibrating Column heater fluid-handling components installing installing physical specifications maintaining Conventions, documentation maintenance kits operation operation kits...
  • Page 109 Eluent tubing connecting to eluent reservoir connecting to pump inlet IEEE-488 Eluents connecting devices buffered connections considerations guidelines guidelines interface heating setting address viscosity Immiscible eluents volatile Inlet check valve assemblies Equilibrating system Inlet check valves, replacing Inlet manifold Inline degassing Input voltage Ferrule Filters...
  • Page 110 Mobile phase compatibility degassing Ramp-and-decay test miscibility Reference valve viscosity considerations Refractive indices Related documentation Relative humidity Reservoirs, positioning Restrictor tee Noises, troubleshooting Restrictor tee check valve, replacing Retention time stability test Operating procedures Outlet check valves, replacing Salt crystals Seal wash holes Solvent buffered...
  • Page 111 Temperature Tests diagnostic ramp-and-decay retention time stability THF (tetrahydrofuran) Troubleshooting chromatographic problems contacting Waters noises pump problems Tubing attaching compression fittings cutting to length making connections problems with Vacuum sonication Vent valve Vibration Viscosity, eluent Volatile eluents Waste line connecting drip tray...

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