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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

and

Figure 1-4

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

Gradient
Mixer

Inlet
Manifold

Restrictor
Tee
(Patent Pending)

Pressure
Transducer

Pulse Dampeners

(C overed by S hroud)

Pump Head

Assembly

Draw-Off

Valve

Drip Tray Waste Exit

Drip Tray

Waste Line

Vent Valve

Pump Outlet

Vent Valve

Inlet Check

Valve Assembly

Outlet Check

Valve Assembly

Indicator Rod

Summary of Contents for 1525EF

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 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 Microsof...

Page 3: ...id 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 Funktionstüchtigk...

Page 4: ... 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 un...

Page 5: ...lsolfossido 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 s...

Page 6: ......

Page 7: ...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 s...

Page 8: ...n 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 ...

Page 9: ...t 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 ...

Page 10: ...e 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: ... 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 Attention surface chaude ou température élevée Vorsicht heiße Oberfläche oder hohe Temperatur Precauzione superficie calda o elevata temperatura Precaución superficie caliente o temperatura elevada Commonly Used Symbols Con...

Page 12: ...rica 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 Precaución riesgo de punción con aguja Caution ultraviolet light Attention rayonnement ultrviolet Vorsicht Ultraviolettes Licht Precauzione luce ultravioletta Precaución emisiones de luz ultravioleta Co...

Page 13: ... on Sous tension Netzschalter ein Alimentazione elettrica attivata Alimentación eléctrica conectada Electrical power off Hors tension Netzschalter aus Alimentazione elettrica disattivata Alimentación eléctrica desconectada Commonly Used Symbols Continued 1 0 ...

Page 14: ...s 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 Contro...

Page 15: ... 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 O...

Page 16: ... 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 P...

Page 17: ...ecifications 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 ...

Page 18: ...igurations 41 2 5 Standard Ferrule and Compression Screw Assembly 44 3 1 Priming the Pump 49 4 1 Unscrewing Pump Head Mounting Screws 1525µ Pump 57 4 2 Exposed Head Support Assembly 1525µ Pump 59 4 3 Disassembled Plunger 1525µ Pump 60 4 4 Disconnecting the Inlet and Outlet Tubing 1525µ Pump 62 4 5 Unscrewing Pump Head Mounting Screws 1525EF Pump 66 4 6 Exposed Head Support Assembly 1525EF Pump 68 ...

Page 19: ... Heater 88 A 3 Environmental Specifications 89 A 4 Electrical Specifications 89 A 5 Performance Specifications for the 1525µ Pump 90 A 6 Performance Specifications for the 1525EF Pump 90 B 1 Spare Parts for the 1525µ Pump 92 B 2 Spare Parts for the 1525EF Pump 93 B 3 Accessories for the 1525µ Pump 94 B 4 Accessories for the 1525EF Pump 94 B 5 Maintenance and Operation Kits and Tools 95 C 1 Aqueous...

Page 20: ...List of Tables 20 C 6 UV Cutoff Wavelengths for Common Chromatographic Solvents 105 C 7 Wavelength Cutoffs for Different Mobile Phases 106 C 8 Refractive Indices for Common Chromatographic Solvents 107 ...

Page 21: ... 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 reco...

Page 22: ...ment 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...

Page 23: ...u 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 STOP Attention 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 unp...

Page 24: ...ed 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 n...

Page 25: ...Overview 25 1 Figure 1 2 Waters 1525EF Pump with an Optional Column Heater TP01722 Bottle Holder Column Heater ...

Page 26: ... 1 3 and Figure 1 4 Figure 1 3 Fluid Handling Components in the Waters 1525µ Pump Gradient Mixer Inlet Manifold Restrictor Tee Patent Pending Pressure Transducer Pulse Dampeners Covered by Shroud Pump Head Assembly Draw Off Valve Drip Tray Waste Exit Drip Tray Waste Line Vent Valve Pump Outlet Vent Valve Inlet Check Valve Assembly Outlet Check Valve Assembly Indicator Rod ...

Page 27: ...re 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 Assembly Draw Off Gradient Manual Injector Pressure Tee Pump Outlet Inlet Check Valve Assembly Pump Module B Pump Module A Reference Valve Drip Tray Drip Tray Waste Exit Optional V...

Page 28: ...r 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 th...

Page 29: ...nents as illustrated in Figure 1 5 Figure 1 5 Rear Panel Electronic Components in the Waters 1525 Pump Ethernet Port Power 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 ...

Page 30: ... Section 3 2 1 on page 48 Table 1 2 Electronic Components in the Waters 1525 Pump Component Description 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 IEE...

Page 31: ...idity 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 electr...

Page 32: ...Installing the 1525 Pump 32 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 ...

Page 33: ... 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 it...

Page 34: ... 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 t...

Page 35: ... 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 ...

Page 36: ...able 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...

Page 37: ...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 lin...

Page 38: ...crews 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...

Page 39: ...g 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 desc...

Page 40: ...passing 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 standar...

Page 41: ...ng 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 2 Gradient Mixers ...

Page 42: ...xer 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 ...

Page 43: ... 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...

Page 44: ...less 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 t...

Page 45: ... 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 e...

Page 46: ...vices 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 wizar...

Page 47: ...uents 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 u...

Page 48: ...ars 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 Fu...

Page 49: ...essure 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 ...

Page 50: ...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 depen...

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

Page 52: ...ating 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 We...

Page 53: ...ime 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 lin...

Page 54: ...sure 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 eac...

Page 55: ...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 a...

Page 56: ...te 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 p...

Page 57: ...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 in...

Page 58: ...ed 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 inst...

Page 59: ...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 ...

Page 60: ...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 h...

Page 61: ...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...

Page 62: ...ge 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 STOP Attention Before starting this procedure move the eluent reservoir below the pump head level...

Page 63: ...ings 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 do...

Page 64: ...e 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 a...

Page 65: ...1 2 turn to open the valve 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 wh...

Page 66: ...e 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...

Page 67: ...g 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 5 16 inch open end wrench in Startup Kit 5 32 inch Allen wrench in Startup Kit 9 64 inch Allen...

Page 68: ...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 plu...

Page 69: ...ssembly 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...

Page 70: ... 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 ...

Page 71: ... 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 5 16 inch open end wrench in Startup Kit 1 2 inch adjustable wrench in Startup Kit Priming syringe in Startup Kit Replacement check valves assemb...

Page 72: ...pump with methanol If methanol is not miscible with your current eluent use an intermediate eluent 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...

Page 73: ...he 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 comp...

Page 74: ...nto 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 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 STOP Attention Before continuing low...

Page 75: ...lips 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 ...

Page 76: ...ocedure 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...

Page 77: ...t 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 ...

Page 78: ...p 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...

Page 79: ...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 ...

Page 80: ...ace 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 Se...

Page 81: ... 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 straighte...

Page 82: ... 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 le...

Page 83: ...p 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 Techn...

Page 84: ...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 se...

Page 85: ... 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 Sectio...

Page 86: ...n 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...

Page 87: ...r 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 Column contaminated Clean or replace the column Loss of column efficiency Clean or replace the column...

Page 88: ...cation Height Depth Width Weight 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 Height Depth Width Weight 17 0 in 43 cm 14 0 in 35...

Page 89: ... connected to the third pin ground pin on the electrical power cord plug Class I Overvoltage category b 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 degree c c Pollution Degree 2 A measure of pollution on electrical circuits which may produce a reduction of dielectric strength or sur...

Page 90: ...ck 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 f...

Page 91: ... 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 5...

Page 92: ...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 ...

Page 93: ...s 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 Tub...

Page 94: ... feet 3 m 1 1 WAT026973 WAT026804 ETFE Tubing 1 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 Item Quantity Part Number Plunger Wash Kit 1 WAT030852 Extended Flow Outlet Check Valve Assem...

Page 95: ...519 Table B 5 Maintenance and Operation Kits and Tools Item Quantity Part Number Solvent Clarification Kit 110 V 60 Hz 220 V 50 Hz 1 1 WAT085113 WAT085122 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 Sea...

Page 96: ... 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...

Page 97: ... 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 cons...

Page 98: ...BA 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 t...

Page 99: ...fers 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 Propano...

Page 100: ... 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 Cyclohe...

Page 101: ...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 nu...

Page 102: ...tle 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 ...

Page 103: ...hose 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 wit...

Page 104: ...gasser 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 Th...

Page 105: ...me 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 A...

Page 106: ...obile Phase UVCutoff 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...

Page 107: ...yl 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 dichlor...

Page 108: ...1 spare parts 92 93 tools 95 unpacking 33 A Accessories 92 Air bubbles 83 B Bench space 31 Bottle holder installing 38 Buffered solvents 102 Buffers 97 C Cavitation in pump head 81 83 Check valves loose 82 malfunctioning 81 replacing 61 71 Chromatographic problems 85 Clearance 31 Column heater 27 installing 42 physical specifications 88 Conventions documentation 22 D Degassing 81 102 105 benefits ...

Page 109: ...s 96 H Head height 81 Heating eluents 104 High pressure noise filter 81 I IEEE 488 connecting devices 36 connections 36 guidelines 36 interface 24 setting address 34 Immiscible eluents 81 Inlet check valve assemblies 28 Inlet check valves replacing 62 71 Inlet manifold 28 Inline degassing 104 Input voltage 31 L Leaking draw off valves 80 82 plunger seals 83 Leaks 82 M Maintenance considerations 52...

Page 110: ...ons in eluent delivery 82 83 Pulse dampeners 28 Pump head assembly 28 removing 56 65 Pump outlet 28 connecting 43 Pump priming 80 82 R Ramp and decay test 53 Reference valve 28 Refractive indices 107 Related documentation 21 Relative humidity 31 Reservoirs positioning 102 Restrictor tee 28 Restrictor tee check valve replacing 64 Retention time stability test 53 S Salt crystals 81 Seal wash holes 2...

Page 111: ...g chromatographic problems 85 contacting Waters 78 noises 84 pump problems 78 Tubing attaching compression fittings 44 cutting to length 43 making connections 44 problems with 81 V Vacuum sonication 104 Vent valve 28 Vibration 31 Viscosity eluent 102 Volatile eluents 81 82 W Waste line connecting drip tray 45 connecting pump 45 Wavelength selection 105 ...

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