Download
Share
Print this page
  1. Manuals
  2. Brands
  3. Waters Manuals
  4. Laboratory Equipment
  5. 2535
  6. Operator's manual

Waters 2535 Operator's Manual

Quaternary gradient module
Hide thumbs

Advertisement

Quick Links

Download this manual
Waters 2535/2545/2555
Quaternary Gradient
Module
Operator's Guide
Revision B
−
Copyright © Waters Corporation 2008
2009
All rights reserved

Advertisement

loading
Need help?

Need help?

Do you have a question about the 2535 and is the answer not in the manual?

Questions and answers

Related Manuals for Waters 2535

Summary of Contents for Waters 2535

  • Page 1 Waters 2535/2545/2555 Quaternary Gradient Module Operator’s Guide Revision B − Copyright © Waters Corporation 2008 2009 All rights reserved...
  • Page 2 Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, its use.
  • Page 3 Customer comments Waters’ Technical Communications department invites you to tell us of any errors you encounter in this document or to suggest ideas for otherwise improving it. Please help us better understand what you expect from our documentation so that we can continuously improve its accuracy and usability.
  • Page 4 Contacting Waters ® Contact Waters with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail. Waters contact information Contacting medium Information Internet The Waters Web site includes contact information for Waters locations worldwide.
  • Page 5 Considerations specific to the quaternary gradient module High voltage hazard To avoid electric shock, do not remove the mass spectrometer’s Warning: protective panels. The components they cover are not user-serviceable. Safety advisories Consult Appendix A for a comprehensive list of warning and caution advisories.
  • Page 6 Intended use of the quaternary gradient module Waters designed the quaternary gradient module to be used as a research tool in purification systems. It is not for use in diagnostic procedures. ISM classification ISM Classification: ISM Group 1 Class B This classification has been assigned in accordance with CISPR 11 Industrial Scientific and Medical (ISM) instruments requirements.
  • Page 7 ISM Classification: ISM Group 1 Class B ............vi EC Authorized Representative ................. vi 1 Setting up the Quaternary Gradient Module ........1-1 Overview ......................1-2 2535 QGM major components ................. 1-3 2535 QGM flow ....................1-4 2545 QGM major components ................. 1-5 2545 QGM flow ....................1-6 2555 QGM major components .................
  • Page 8 Making plumbing connections ..............1-15 Installing solvent inlet tubes ................ 1-15 Installing seal-wash solvent inlet tubes ............1-18 Installing the vent-valve waste tube ............1-20 Installing the system outlet tube ..............1-21 Installing the drip tray waste line ..............1-21 Making the sparge system connections .............
  • Page 9 Reinstalling the pump head ................3-33 Replacing an inlet check valve cartridge ..........3-36 Replacing an outlet check valve cartridge ..........3-39 Replacing the 2535 QGM’s selector-valve rotor-seal ......3-42 Replacing the 2545 and 2555 QGM’s vent-valve rotor-seal ....3-44 Replacing solvent filters ................3-46...
  • Page 10 Warning symbols ....................A-2 Task-specific hazard warnings................ A-2 Specific warnings ..................... A-3 Caution symbol ....................A-4 Warnings that apply to all Waters instruments ......... A-4 Electrical and handling symbols ..............A-11 Electrical symbols ..................A-11 Handling symbols ..................A-12 B Specifications ..................B-1 C Solvent Considerations ...............
  • Page 11 Buffered solvents ..................... C-6 Solvent stabilizers ................... C-6 Solvent viscosity ....................C-6 Mobile phase solvent degassing ..............C-7 Solvent degassing methods ................C-8 Wavelength selection ..................C-9 UV cutoffs for common solvents..............C-10 Mixed mobile phases..................C-11 Refractive indices of common solvents ............C-12 Index .....................
  • Page 12 Table of Contents...
  • Page 13 Setting up the Quaternary Gradient Module Contents Topic Page Overview Before you begin 1-11 Installing the QGM 1-12 Making plumbing connections 1-15 Making the sparge system connections 1-23 Making signal connections 1-28 Connecting to the electricity source 1-35...
  • Page 14 The Waters Quaternary Gradient Module (QGM) is a pump that delivers solvents from their reservoirs to the HPLC system. Three QGM models (2535, 2545, and 2555) can operate over the flow ranges shown in the table below. QGM operating ranges...
  • Page 15 2535 QGM major components The following diagram shows the 2535 QGM’s major components. Stop flow switch and LCD LCD screen Power switch contrast adjustment STOP FLOW Transducer connectors PRIME Prime button Outlet check valves Manifold outlet check valve Mixer, 1.1-mL...
  • Page 16 2535 QGM flow The following screen shows the 2535 QGM’s flow. Setting up the Quaternary Gradient Module...
  • Page 17 2545 QGM major components The following diagram shows the 2545 QGM’s major components. Stop flow switch and LCD LCD screen Power switch contrast adjustment STOP Transducer FLOW connectors Prime button PRIME Outlet check valves Manifold outlet check valve Mixer, 1.1-mL Pressure Pressure transducer...
  • Page 18 2545 QGM flow The following screen shows the 2545 QGM’s flow. Setting up the Quaternary Gradient Module...
  • Page 19 2555 QGM major components The following diagram shows the 2555 QGM’s major components. Stop flow switch and LCD LCD screen Power switch contrast adjustment STOP Transducer FLOW connectors Prime button PRIME Outlet check valves Manifold outlet check valve Pressure Pressure transducer transducer (left (right actuator)
  • Page 20 2555 QGM flow The following screen shows the 2555 QGM’s flow. QGM major components Component Description 2535 2545 2555 Drip tray waste Connects the waste tube to the tube connector leak management system. Filter Filters the solvent before it goes to the pump outlet.
  • Page 21 Mixer, 7.4-mL A component that mixes the solvents downstream of pumps, improving compositional ripple. For the 2535 QGM, the Note: 7.4 mL mixer is in line only when the QGM is in the large flow scale state.
  • Page 22 QGM major components (Continued) Component Description 2535 2545 2555 Power switch A switch that powers-on and powers-off the QGM. (See page 2-2.) Pressure A transducer that monitors the transducer (left head pressure of the right-side actuator) pump. Pressure A transducer that monitors the...
  • Page 23 If you discover any damage or discrepancy when you inspect the contents of the cartons, immediately contact the shipping agent and your local Waters representative. Customers in the USA and Canada can report damage and discrepancies to Waters Technical Service (800 252-4752).
  • Page 24 QGM. Collapse hazard. Do not configure systems in which any one Warning: Waters “base” module, such as the QGM, physically supports more than two vertically oriented “accessory” Waters modules, such as 515 pumps, photodiode array detectors, evaporative light-scattering detectors, or dual-wavelength absorbance detectors.
  • Page 25 Managing solvent and waste reservoirs Explosion hazard. Ensure all containers, columns, and Warning: instruments are properly grounded. Electric shock hazard. To avoid electrostatic discharge from Warning: the waste container, be sure to connect the stainless-steel tubing included in the startup kit to the vent-valve waste, and route it to the waste container.
  • Page 26 To install the detector drip tray Insert the Tygon tubing onto the fitting on the bottom of the detector drip tray. Adhesive tape Fitting on bottom of drip tray Peel the adhesive tape, removing it from the bottom of the detector drip tray.
  • Page 27 Place the drip tray on top of the QGM, along its front. Detector Sparge tube routed through hole (optional) Detector drip tray Making plumbing connections Inspect all fittings and tighten them if necessary. Fittings can Caution: loosen during shipment, particularly if the module has undergone temperature extremes.
  • Page 28 Gloves: clean, powder-free, chemical-resistant • Solvent inlet tubing assembly • Wrench, 7/16-inch (for 2535 QGM) • Wrench, 9/16-inch (for 2545 and 2555 QGM) To make the initial plumbing connections Install the solvent filters on the ends of the solvent inlet tubing as shown below.
  • Page 29 Slide the tubing through the compression nut, and tighten the nut using the 9/16-inch wrench (2545 and 2555 QGM) or the 7/16-inch wrench (2535 QGM). Tighten the fitting 3/4-turn beyond finger-tight. Tighten fittings in the order B, C, A, and D.
  • Page 30 Solvent manifold fittings Repeat step 4 through step 7 for the remaining solvent tubes. You can shorten the solvent inlet tubes to accommodate the solvent container locations. Installing seal-wash solvent inlet tubes The seal-wash solvent inlet tube uses a PEEK™ compression screw and 2-piece ferrule.
  • Page 31 Compression screw and 2-piece ferrule Chamfered side of stainless- steel ferrule ring Compression screw Ferrule Required material Seal-wash inlet tube assembly To install the seal-wash solvent supply tubes Insert the seal-wash solvent supply tube into the bottom port of the seal-wash pump.
  • Page 32 Route the seal-wash supply tube through the hole at the upper, right-hand side of the QGM. Hole on upper, right side of QGM Installing the vent-valve waste tube Required materials • Open-end wrench, 1/4-inch • Waste tube assembly (stainless-steel tube, gold compression screw) To install the vent-valve waste tube Install the waste tube onto the front panel’s vent-valve port, “Waste”.
  • Page 33 Installing the system outlet tube Required materials • System outlet tube assembly • Wrench, 1/4-inch To install the system outlet tube Insert the system outlet tube assembly into the filter outlet, and tighten the tube’s compression nut using the 1/4-inch wrench. QGM outlet tube (left side orientation shown) 1/4-inch wrench Filter outlet...
  • Page 34 To install the drip tray waste line Caution: • To avoid damaging the drip tray, support the barbed drain fitting while you slide the waste line over it. • To avoid damaging the barbed drain fitting, ensure that the front of the QGM aligns with the front of the workbench. Support the barbed drain fitting with your hand, slide the waste line over the fitting, and then route it to a suitable waste container.
  • Page 35 Route the drip tray waste line to a suitable waste container. Correct positioning of drip tray waste line and vent-valve waste tube Drip tray Drip tray waste line waste line Vent-valve waste tube Vent-valve waste tube Correct Incorrect—will likely TP02709 cause fluid backup The vent-valve waste tube can be immersed in waste solvent.
  • Page 36 System sparging overview Helium sparging reduces the total dissolved gas in the eluent reservoirs and maintains that condition during operation. Minimizing dissolved gases decreases the gas that can be released when different mobile phases are mixed in the gradient proportioning valve. Use an ultra-pure, carrier (UPC) grade of helium to prevent eluent contamination.
  • Page 37 Installing the solvent sparge outlet lines To install the solvent sparge outlet lines Insert the solvent sparge outlet lines in their corresponding sparge ports on the front of the QGM. Sparge ports Insert the corresponding vent tubes, solvent lines, and sparge lines into the solvent reservoir.
  • Page 38 Tips: • A stainless-steel sparge diffuser is attached to the end of each sparge line. • A solvent reservoir filter is attached to the end of each solvent line. Sparge tube To fume hood 3-inch minimum from top of bottle to arc in sparge tube Vent tube (optional kit)
  • Page 39 Before performing a run, sparge the solvent(s), at 100%, Recommendation: for 10 minutes per liter of solvent. Then sparge for an additional 10 minutes per liter of solvent, at 20%. Continue to sparge at 20% during an actual run. To attach compression fittings to the helium line Slide the compression screw onto an end of the 1/16-inch OD, 0.040 ID, stainless-steel tubing.
  • Page 40 To connect the helium sparge supply to the QGM Slide a 1/16-inch, gold, compression screw and brass ferrule onto the 1/16-inch OD, 0.040 ID, stainless-steel tubing from the helium supply, push the tubing into the sparge inlet fitting until it bottoms, and tighten the compression screw using a 1/4-inch open-end wrench.
  • Page 41 To make signal connections Connect one end of the Ethernet communication cable (RJ-45) to the rear panel and the other end according to the instructions in the Ethernet Instrument Getting Started Guide. Connect the event input/output cables from an external device (for example, column-switcher or fraction collector) to the appropriate connector block on the rear panel.
  • Page 42 QGM rear panel major components Power input Fuse holder RS-232 diagnostics (for Waters RS-232 technical service use only) ETHERNET Ethernet (RJ-45) QSPI QSPI Fan vents Chassis ground A Function Switch 1 (Out) Switch 1 (Out) Switch 2 (Out) Switch 2...
  • Page 43 Making I/O signal connections To avoid electric shock, power-off and unplug the QGM before Warning: making signal connections. Refer to the signal connection location shown on a label on the rear panel of each instrument. The rear panel includes two removable connectors that hold the pins for the I/O signals (see the following figure).
  • Page 44 I/O signals for the QGM (Continued) Signal Connections Description Ground A5, A10, B5, Connected to signal ground and used as and B10 reference for outputs. Gradient A6 and A7 An input that initiates the pumps to begin Start gradient operation by either contact closure input or TTL (transistor-transistor logic) low.
  • Page 45 Through software, you can select one of the following signals as the chart out signal: • Flow rate • System pressure • Composition (%A, %B) 2535 QGM chart-out signal conditions Parameter Setting at 0 Parameter Setting at Signal Volts (Minimum) 2.000 Volts (Maximum)
  • Page 46 2555 QGM chart-out signal conditions Parameter Setting at 0 Parameter Setting at Signal Volts (Minimum) 2.000 Volts (Maximum) Flow Rate 0.000 mL/min 300 mL/min System Pressure 0 kPa (0 bar, 0 psi) 20,700 kPa (207 bar, 3000 psi) Composition 0.0% 100.0% To make signal connections Attach the positive and negative leads of the signal cable to the...
  • Page 47 Avoid electrical shock: Warning: • Use power cord SVT-type in the United States and HAR-type or better in Europe. For other countries, contact your local Waters distributor. • Power-off and unplug the QGM before performing any maintenance operation on the instrument.
  • Page 48 To connect to the electricity source Use a line conditioner or an uninterruptible power supply Recommendation: (UPS) for optimum long-term input voltage stability. Move the off/on ( ) switch to the “off” ( ) position. Connect the female end of the power cord to the receptacle on the rear panel of the QGM.
  • Page 49 Preparing for Operation This chapter explains how to configure the QGM as a chromatographic system pump. Contents Topic Page Powering-on Adjusting the LCD contrast Configuring in Empower software Starting the console from Empower software Configuring in MassLynx software Starting the console from MassLynx software 2-11 Enabling and disabling the leak sensor 2-11...
  • Page 50 Powering-on When you complete all electrical, plumbing, and signal connections, you can power-on the QGM. In the event of a solvent leak, to avoid contacting solvent, Warning: ensure the QGM’s front door is closed when operating under normal conditions. To avoid damaging electrical parts, do not power-on the QGM Caution: until you complete all electrical, plumbing, and signal connections.
  • Page 51 Configuring in Empower software W25X5Q is used to denote the 2535, 2545, and 2555 quaternary Note: gradient modules in Empower software. Perform these tasks to configure in Empower software: • Install the W25X5Q instrument control software (for installation instructions, see the 2535/2545/2555 Quaternary Gradient Module Release Notes).
  • Page 52 In the Empower Node Properties dialog box, click the Instruments tab. The system instruments that are successfully communicating with Tip: your system are indicated by a Yes in the “OK?” column. Instruments tab Ensure that a quaternary gradient module appears in the instrument list and is successfully communicating with your system, and then click Right-click Systems, and then select New >...
  • Page 53 In the System Type area of the New Chromatographic System Wizard dialog box, select Create New System, and then click Next. System Selection dialog box In the System Selection dialog box, drag the name of the instrument(s) you want to include in the new system from the Available Instruments pane to the New System Instruments pane, and click Next.
  • Page 54 Empower. When MassLynx software controls the system, the QGM's control panel appears on the Additional Status tab of the Inlet Editor window. QGM control panel Pump model number appears here (2535, 2545, or 2555) Status indicator Instrument state Flow rate...
  • Page 55 Displays the Prime Seal Wash dialog box. (See page 2-15.) Set sparge Displays the Set Sparge dialog box. (See page 2-18.) Set flow scale (2535 QGM only) Displays the Set Flow Scale dialog box. (See page 2-20.) Configuring in Empower software...
  • Page 56 Additional functions in the QGM control panel (Continued) Control panel function Description Reset QGM Resets the QGM to recover from an error condition. Help Displays the QGM online Help. Launch Instrument Console Launches the instrument console software. Starting the console from Empower software To start the console from Empower software In the Run samples window, right-click in the QGM control panel, and select Launch Instrument Console.
  • Page 57 Configuring in MassLynx software W25X5Q and 25X5Q are used to denote the 2535, 2545, and 2555 Note: quaternary gradient modules in MassLynx software. Perform these tasks to configure in MassLynx software: • Install the W25X5Q instrument control software and the Waters Pump Control software (for installation instructions, see the 2535/2545/2555 Quaternary Gradient Module Release Notes).
  • Page 58 12. Click Finish, and then click Finish again. 13. In the Inlet Method window, click the Inlet icon. 14. In the Modify Instrument window, click Config. 15. In the Waters Pump Control Configuration window, click Scan. A list of connected instruments appears, among them the Result: Quaternary Gradient Module.
  • Page 59 19. In the Modify Instrument Method window, click OK. 20. In the Inlet Method window, select LC > Reset Communications. Starting the console from MassLynx software To start the console from MassLynx software In the MassLynx window, click Inlet Method. In the Inlet Method window, click the Additional Status tab.
  • Page 60 Click Close. Operating from the standalone console The standalone software can control the pump and a compatible Waters detector. The software monitors various pressure readings generated by the QGM, but saves them for only 96 hours. The software does not collect data from the detector.
  • Page 61 To launch the standalone console: Locate the icon for the Standalone ACQUITY Console in the C:\Program Files\Waters Instruments\Bin folder. Create a shortcut for the icon, and copy the shortcut to the desktop. Click the icon to launch the standalone console.
  • Page 62 Preparing sample lists Use the Samples window to create a sample list. To access the Samples window In the console, select Samples from the instrument tree. The Samples window appears. For information about preparing sample lists, consult the console’s online Tip: Help.
  • Page 63 Priming the seal-wash pump Caution: • Ensure the seal wash solvent is fully miscible with the mobile phase. • To avoid reducing the life of the seal-wash pump and plunger seals, do not use a nonvolatile buffer as the seal-wash solvent. •...
  • Page 64 Connect the tubing adapter to the syringe, and then connect the syringe assembly to the outlet tubing from the seal-wash system (the tube from the left side of the left actuator). In the console, select Quaternary Gradient Module from the instrument tree.
  • Page 65 The default prime flow rate for the 2545 QGM and 2555 QGM is 75 mL/min. The default prime flow rate for the 2535 QGM is 50 mL/min.
  • Page 66 To manually prime the QGM using a syringe (for a dry system only) Open the module’s front door. Connect the short priming tube (startup kit) to the open port on the left side of the inlet manifold valve. (See page 1-3 through page 1-7.)
  • Page 67 In the Normal Rate box, specify the normal duty cycle, and click Set. The normal duty cycle begins once the system has sparged at initial Tip: duty cycle for the duration specified in the Initial Rate pane. To set the sparge parameters in the Instrument Method Editor Click the Sparge tab.
  • Page 68 Setting the 2535 QGM flow scale The flow scale option is available for the 2535 QGM only. When you select small flow scale, flow is routed through the small-scale mixer tee directly to the 2535 QGM outlet. When you select large flow scale, flow is routed first through the small-scale mixer tee, then through an additional large-scale mixer, and finally to the 2535 QGM outlet.
  • Page 69 To set the 2535 QGM flow scale in the control panel for instruments controlled by MassLynx software In the Inlet Method Editor, right-click in the control panel, and then select Set flow scale. In the Set Flow Scale dialog box, select the flow scale: Large or Small.
  • Page 70 In the Set Flow Scale dialog box, select the flow scale: Large or Small. Large Default: Click Set. To set the 2535 QGM flow scale in the Instrument Method Editor Click the General tab. Select the flow scale from the drop-down list: Large or Small. Large Default: Click Set.
  • Page 71 Using the manual sample inlet line for large-sample injections The sample inlet line (startup kit) must be attached to the front of the pump as shown below. Inlet manifold Sample inlet line valve To prime the inlet line Route the length of sample inlet line tubing from the inlet manifold valve to a reservoir containing the initial solvent.
  • Page 72 You must ensure mobile phase passes through the pump Requirement: before sample enters the system, taking into account that, for the first injection, the sample inlet line contains mobile phase. Turn the inlet manifold valve to the Run position, selecting the mobile phase.
  • Page 73 Creating gradients You create a gradient to change the composition ratio of solvents during a run. To access the gradient table In the Instrument Method Editor, click 25X5Q > General tab > Gradient table. For information about creating gradients, see the Empower or MassLynx Tip: online Help.
  • Page 74 2-26 Preparing for Operation...
  • Page 75 Replacing the plunger 3-27 Replacing an inlet check valve cartridge 3-36 Replacing an outlet check valve cartridge 3-39 Replacing the 2535 QGM’s selector-valve rotor-seal 3-42 Replacing the 2545 and 2555 QGM’s vent-valve rotor-seal 3-44 Replacing solvent filters 3-46 Performing the static leak test...
  • Page 76 Waters Technical Service (800 252-4752). Otherwise, phone the Waters corporate headquarters in Milford, Massachusetts (USA), or contact your local Waters subsidiary. The Waters’ Web site includes phone numbers and e-mail addresses for Waters locations worldwide. Visit www.waters.com. When you contact Waters, be prepared to provide this information: •...
  • Page 77 AC outlet. Wait 10 seconds thereafter before you disconnect an assembly. Spare parts Replace only parts mentioned in this document. For spare parts details, see the Waters Quality Parts Locator on the Waters Web site’s Services & Support page. Maintenance considerations...
  • Page 78 Resolving leak sensor errors All fluid-handling areas of the QGM are designed to route spilled solvent to the drip tray (fitted with a leak sensor). The solvent then flows to the waste tube below the front panel. After approximately 3.0 mL of liquid accumulates in the leak sensor reservoir, an alarm sounds, indicating that the leak sensor detected a leak.
  • Page 79 To resolve a leak sensor error View the Leak Sensors dialog box to confirm that the QGM leak sensor detected a leak. If a leak is detected, a “Leak Detected” error message appears. Tip: To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to the QGM.
  • Page 80 Use a nonabrasive, lint-free wipe to dry the leak sensor prism. Prism Lint-free wipe TP02891 Roll up a nonabrasive, lint-free wipe, and use it to absorb the liquid from the leak sensor reservoir and its surrounding area. Leak sensor reservoir TP03127 Rolled up lint-free wipe...
  • Page 81 Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place. TP02892 T-bar TP03130 Slot in leak sensor reservoir Leak sensor installed in reservoir If you detached the connector from the front of the instrument, reattach 10.
  • Page 82 Replacing the QGM’s leak sensor To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM. Consult the Material Safety Data sheets for the physical and chemical properties of solvents you use. The leak sensor can be contaminated with biohazardous Warning: and/or toxic materials.
  • Page 83 Remove the leak sensor from its reservoir by grasping it by its serrations and pulling upward on it. Serrations Unpack the new leak sensor. Align the leak sensor’s T-bar with the slot in the side of the leak sensor reservoir, and slide the leak sensor into place. TP02892 T-bar TP03130...
  • Page 84 In the console, select Quaternary Gradient Module from the instrument tree. 10. In the Quaternary Gradient Module window, click Control > Reset 25X5Q, to reset the module. Cleaning the door’s air filter Required material ® T10 TORX driver To clean the air filter Using the T10 TORX driver, remove the 4 screws that secure the air filter frame and air filter to the inside of the module’s door.
  • Page 85 Align the air filter with the air filter frame. Using the T10 TORX driver and the 4 screws, attach the air filter and frame to the inside of the door. Replacing the door’s air filter If you cannot clean the air filter, replace it. Required materials •...
  • Page 86 Remove the old air filter from the air filter frame and discard it. Align the new air filter with the air filter frame. Using the T10 TORX driver and the 4 screws, attach the air filter and frame to the inside of the door. Replacing the transducer To prevent injury, always observe Good Laboratory Practices Warning:...
  • Page 87 To replace the transducer Flush the QGM with methanol. If the solvent you are using is not miscible with methanol, Requirement: use an intermediate solvent. Disconnect the transducer signal cable from the front panel. Transducer signal cable Using the 5/16-inch open-end wrench, disconnect the two stainless-steel solvent lines on the top and bottom of the transducer.
  • Page 88 Using the T10 TORX driver, tighten the two TORX screws to hold the transducer in place. Reconnect the transducer signal cable to its connector on the front panel, ensuring the connector clicks securely into place. Using the 5/16-inch open-end wrench, reconnect the two stainless-steel solvent lines to the top and bottom of the transducer.
  • Page 89 • Replacement seal kit • Seal insertion/extraction tool • Tweezers, plastic To avoid damaging the sealing surfaces, use the seal Caution: insertion/extraction tool. Do not use a sharp or metallic tool to remove or install seals. To remove the pump head and seal-wash assembly Flush the QGM with non-hazardous solvent.
  • Page 90 Squeeze the tabs of the pressure transducer cable, and pull gently, disconnecting the cable from the bulkhead. Tabs Pressure transducer cable connector Disconnect the solvent inlet tube from the inlet check valve at the bottom of the pump head. Pump head Inlet check valve Solvent inlet tube 10.
  • Page 91 12. Supporting the pump head’s weight, use the T27 TORX driver to remove the 4 screws from the support plate. Actuator housing T27 TORX driver Seal-wash port Support plate Screws (4) Pressure transducer To avoid damaging the plunger, support the head from Caution: below as you remove it.
  • Page 92 To replace the plunger seal Use the T27 TORX driver to remove the 2 screws securing the support plate to the pump head assembly, and then remove the support plate. 1. Remove the screws that secure the support plate to the pump head assembly.
  • Page 93 Remove the seal-wash housing. Plunger wash seal Seal-wash housing High pressure seal Pump head assembly While holding the pump head, insert the seal insertion/extraction tool into the head, and with a rocking motion, lift the seal from the head. Seal insertion/extraction tool Replacing the pump head seals and O-ring 3-19...
  • Page 94 To avoid scratching surfaces, use plastic tweezers when Caution: removing the O-ring. Using the plastic tweezers, remove the O-ring. High pressure seal Plastic tweezers O-ring Pump head Wet the seal cavity and new plunger seal with methanol. Seal cavity 3-20 Maintaining the Quaternary Gradient Module...
  • Page 95 Using the seal insertion/extraction tool, insert the new plunger seal into the pump head. Ensure the flange side of the plunger seal faces outward. Requirement: Seal insertion/extraction tool Insert the new O-ring into the O-ring groove in the pump head. O-ring Replacing the pump head seals and O-ring 3-21...
  • Page 96 Use plastic tweezers to remove the plunger wash seal. Caution: Using the plastic tweezers, remove the existing plunger wash seal. Plastic tweezers Plunger wash seal Wet the new plunger wash seal with methanol, and insert it into the seal-wash housing. Seal-wash housing Plunger wash seal 10.
  • Page 97 11. Reassemble the pump head, seal-wash assembly, and support plates, and secure them with screws tightened evenly using the T27 TORX driver. Ensure that the drain fitting and check valve housing Requirement: align. T27 TORX driver Support plate 12. If you are replacing the plunger assembly, proceed to “Replacing the plunger”...
  • Page 98 Wetting the seal cavity Seal cavity Wetting the plunger Plunger To avoid damaging the plunger, ensure it is retracted Caution: before you push the pump head onto it. See step 2 through step 5 under “To remove the pump head and seal-wash assembly” on page 3-15.
  • Page 99 To avoid damaging the plunger, support the head from Caution: below as you install it. 15. Supporting the pump head’s weight, attach the 4 T27 TORX screws to the support plate, and tighten them 1/8-turn beyond finger-tight. Tighten the screws evenly. Support plate T27 TORX driver Pump head...
  • Page 100 18. Reconnect the transducer outlet tube to the transducer outlet, using a 1/4-inch open-end wrench. Transducer outlet 1/4-inch wrench 19. Connect the pressure transducer cable to the bulkhead. The left-hand pressure cable connects to the leftmost port. Requirement: The right-hand pressure cable connects to the rightmost port. The two center ports are empty.
  • Page 101 Replacing the plunger To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM. Consult the Material Safety Data sheets for the physical and chemical properties of solvents you use. To prevent contamination, wear clean, powder-free, Caution: chemical-resistant gloves when replacing the plungers.
  • Page 102 To avoid damaging electrical parts, never disconnect an Caution: electrical assembly while power is applied to the QGM. To completely interrupt power, set the module’s power switch to “off”, and then unplug the power cord from the AC outlet. Wait 10 seconds thereafter before you disconnect an assembly.
  • Page 103 Disconnect the solvent inlet tube from the inlet check valve at the bottom of the pump head. Pump head Inlet check valve Solvent inlet tube 10. Using the 1/4-inch open-end wrench, disconnect the tube connection at the outlet of the transducer. Transducer outlet 1/4-inch wrench 11.
  • Page 104 12. Supporting the pump head’s weight, use the T27 TORX driver to remove the 4 screws from the support plate. T27 TORX driver Support plate Screws (4) To avoid damaging the plunger, support the head from Caution: below as you remove it. 13.
  • Page 105 To replace the plunger Stand the head assembly upright on a clean surface, and set it aside. Replace the head seals when you replace the Recommendation: plunger. With the plunger in its forward position, carefully slide the 13/16-inch open-end wrench over the plunger, loosen its retaining nut, and remove the plunger.
  • Page 106 For the 2535 and 2545 QGM, install the flat washer and then the 3 wave washers, in order stated. Ensure the wave washers are bowed toward the plunger Requirement: tip. For the 2555 QGM, install the flat washer and 1 wave washer.
  • Page 107 Insert the new plunger into the actuator piston, and tighten the plunger-retaining nut with a 13/16-inch open-end wrench. Align the new plunger so that it points straight out from the piston. Aligning the plunger facilitates installing the head without Tip: damaging the plunger.
  • Page 108 Carefully align the pump head with the plunger, and gently push the pump head onto the plunger until the 4 screw holes align with the holes in the support plate. To avoid damaging the plunger, support the head from Caution: below as you install it.
  • Page 109 10. Connect the solvent inlet tube to the inlet check valve at the bottom of the pump head. Pump head Inlet check valve Solvent inlet tube 11. Reconnect the transducer outlet tube to the transducer outlet, using a 1/4-inch open-end wrench. Transducer outlet 1/4-inch wrench Replacing the plunger...
  • Page 110 12. Reconnect the transducer cable to the appropriate connector on the front panel, making sure it clicks into place. Tabs Pressure transducer cable connector 13. Prime the QGM (see page 2-16). Replacing an inlet check valve cartridge The QGM has one inlet check valve per pump head. The check valves, made of synthetic ruby and sapphire, are contained in easy-to-replace cartridges.
  • Page 111 Required materials • Open-end wrench, 1/2-inch • Gloves: clean, powder-free, chemical-resistant • Methanol (or other suitably miscible solvent) in a wash bottle • Replacement check valve cartridge • Tweezers, plastic To replace an inlet check valve cartridge Remove the tube fitting at the inlet check valve cartridge holder. Use the 1/2-inch open-end wrench to remove the check valve cartridge holder from the head.
  • Page 112 Wet the new check valve cartridge with an appropriately miscible solvent. Check valve cartridge Insert the replacement cartridge into the check valve cartridge holder with the arrow pointed away from the hex nut. Replacement cartridge Check valve TP03114 cartridge holder Insert the check valve cartridge holder in the head, and finger tighten the fitting.
  • Page 113 Reattach the fitting from the solvent inlet tube to the check valve cartridge holder. Pump head Inlet check valve Solvent inlet tube 10. Prime the QGM (see page 2-16). Replacing an outlet check valve cartridge Within the module’s flow path are two outlet check valves per pump manifold. The check valves, made of synthetic ruby and sapphire, are contained in easy-to-replace cartridges.
  • Page 114 Required materials • Open-end wrenches, 1/2-inch, 1/4-inch • Gloves: clean, powder-free, chemical-resistant • Methanol (or other suitably miscible solvent) in a wash bottle • Replacement check valve cartridge • Tweezers, plastic To replace an outlet check valve cartridge Using the 1/4-inch open-end wrench, remove the fitting on the holder for the outlet check valve cartridge.
  • Page 115 Use the 1/2-inch, open-end wrench to remove the holder for the outlet check valve cartridge from the manifold. Holder for outlet check valve cartridge 1/2-inch wrench To avoid damaging the manifold, always use plastic Caution: tweezers to remove the outlet check valve cartridge. Use the plastic tweezers to remove the existing outlet check valve cartridge from inside the manifold.
  • Page 116 2-18). 11. Prime the QGM (see page 2-16). Replacing the 2535 QGM’s selector-valve rotor-seal To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM. Consult the Material Safety Data sheets for the physical and chemical properties of solvents you use.
  • Page 117 To replace the selector-valve rotor-seal on the 2535 QGM Using the 9/64-inch Allen wrench, remove the two screws on the front of the selector-valve, and then remove the stator. Screws Using the plastic tweezers, remove the rotor-seal from inside the valve.
  • Page 118 Tighten the two 9/64-inch Allen screws on the front of the selector-valve until you encounter some resistance, then alternate tightening each screw 1/8-turn until it bottoms out. Prime the QGM. Replacing the 2545 and 2555 QGM’s vent-valve rotor-seal To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM.
  • Page 119 To replace the vent-valve rotor-seal on the 2545 QGM and 2555 QGM Using the 5/8-inch open-end wrench, loosen the head assembly at the front of the vent valve, and then remove it. Waste line Vent valve 5/8-inch open-end wrench Head assembly Using the plastic tweezers, remove the rotor-seal assembly from inside the valve.
  • Page 120 Replacing solvent filters To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM. Consult the Material Safety Data sheets for the physical and chemical properties of solvents you use. Caution: • Wear clean, powder-free, chemical-resistant gloves when handling the solvent filter.
  • Page 121 To replace a solvent filter Remove the filtered end of the solvent tubing from the solvent bottle. Remove the old solvent filter from the solvent tubing. Insert the new solvent filter into the solvent tubing. Solvent filter Solvent tubing Insert the filtered end of the solvent tubing into the solvent bottle. Shake the filter to remove any air from it.
  • Page 122 In the Test Parameters area, specify your typical operation pressure and select the solvent. Range: • 3,450 to 34,500 kPa (34.5 to 345 bar, 500 to 5,000 psi), for the 2535 and 2545 QGM • 3,450 to 17,240 kPa (34.5 to 172.4 bar, 500 to 2500 psi), for the 2555 Default: •...
  • Page 123 If the right and left pump heads fail, evidencing the same decay rate, a leak occurring in the areas common to both pump heads, is likely: • Tubing between the outlet check valve and vent valve • Fittings between the outlet check valve and vent valve •...
  • Page 124 Grasp the two small tabs on the fuse holder drawer and pull the drawer out. Small tabs The fuse holder drawer can only be partially withdrawn. Note: Open the fuse cover. Fuse holder drawer Fuse cover Insert the tip of a small screwdriver through the hole at the bottom of the fuse holder drawer under each fuse, and push each fuse up.
  • Page 125 Discard the fuses. For continued protection against fire, replace fuses with Warning: those of the appropriate type and rating. Insert the new fuses into the holders and the holders into the fuse holder drawer. Close the fuse holder drawer cover and fuse holder drawer. Reconnect the power cord to the power entry module.
  • Page 126 3-52 Maintaining the Quaternary Gradient Module...
  • Page 127 Troubleshooting Contents Topic Page Proper operating procedures System troubleshooting Troubleshooting mechanical, electrical, and electronic components 4-3 Chromatography troubleshooting...
  • Page 128 Proper operating procedures So that your system always runs optimally, follow the operating procedures and guidelines in Chapters 2 and 3. Contact Waters Technical Service to report malfunctions or other problems. (See page 3-2). System troubleshooting To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM.
  • Page 129 Troubleshooting mechanical, electrical, and electronic components This table contains suggestions for resolving hardware problems in the QGM. If the suggested solutions do not solve the problem, contact Waters Technical Service. Consult the column's care and use instructions before Requirement: performing any of the corrective actions listed in this table.
  • Page 130 3-46). Plunger seal leaking Replace plunger seal assembly (see page 3-14). Defective gradient Contact Waters Technical proportioning valve. Service (see page 3-2). Backpressure too Apply a small amount of low. backpressure on the flow cell using a restrictor tube...
  • Page 131 Troubleshooting the hardware (Continued) Symptom Possible Cause Corrective Action Fans do not run Open (spent) or Contact Waters Technical defective fuse Service (see page 3-2). Power supply fan 1. Power-off and power-on wiring or motor again. problem 2. If the problem persists,...
  • Page 132 Leak in QGM. 1. Power-off the QGM. internal drip tray 2. Place the solvent supply bottles on the floor to reduce pump head pressure. 3. Contact Waters Technical Service (see page 3-2)). Priming problems; will Solvent connect 1. Ensure solvent...
  • Page 133 Troubleshooting the hardware (Continued) Symptom Possible Cause Corrective Action Unit does not power-on Power cord not Check power cord. connected No power at outlet Test line voltage. Power supply fuse Replace power supply blown or missing fuses. Chromatography troubleshooting To prevent injury, always observe Good Laboratory Practices Warning: when you handle solvents, change tubing, or operate the QGM.
  • Page 134 (see “Erratic retention drift) times” symptom later in this table). Incorrect Ensure that solvent does not have wavelength for absorbance at the wavelength used. solvent Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Baseline drift, Solvent Use fresh solvent.
  • Page 135 2-20). cycling, short gradient mixing. term (30 to 60 seconds) (continued) Radio frequency Eliminate interference. noise (short- or long-term cycling) Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Baseline noise Ambient Stabilize ambient temperature. cycling, long-term temperature...
  • Page 136 Operate unit with covers in place. properly Ensure there is adequate air space behind it. Radio frequency Eliminate interference. noise Defective detector Troubleshoot detector. Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Insufficient Add gradient mixing (see page 2-20).
  • Page 137 Incorrect column. Use correct column. Sample diluent is • Dilute sample in a weaker stronger than the solution. initial mobile • Inject less. phase. Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Insufficient Add gradient mixing (see page 2-20).
  • Page 138 Incorrect detector Verify wavelength setting. wavelength Leak in solvent Inspect fittings. path Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Flat-topped peaks Detector not Zero the detector baseline. zeroed Incorrect recorder...
  • Page 139 Incorrect column. Use correct column. Fluid leak (causes • Inspect fittings for leaks. lower flow rate). • Perform static decay test (see console online Help). Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Sparging of mixed Use external vacuum degassing for solvents.
  • Page 140 Possible cause Corrective action Increase in Tubing is Systematically inspect tubing by system pressure plugged. making and breaking connections. (continued) Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Loss of column System not Equilibrate column using a...
  • Page 141 Change wash solvents and/or errors (continued) solvents or volumes. volumes. Incorrect volume Change volume range. range for injection method or type. Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. Sample energy Contaminated Use fresh mobile phase. decreased,...
  • Page 142 Correct mobile phase pH or ionic (continued) phase composition. composition Leak in detector Repair leak. If problem persists, flow cell replace detector flow cell. Incorrect flow Set correct flow rate. rate Defective Contact Waters Technical Service gradient (see page 3-2). proportioning valve. 4-16 Troubleshooting...
  • Page 143 Safety Advisories Waters instruments display hazard symbols designed to alert you to the hidden dangers of operating and maintaining the instruments. Their corresponding user guides also include the hazard symbols, with accompanying text statements describing the hazards and telling you how to avoid them.
  • Page 144 Heed all warnings when you install, repair, and operate Waters instruments. Waters assumes no liability for the failure of those who install, repair, or operate its instruments to comply with any safety precaution.
  • Page 145 Biohazard warning This warning applies to Waters instruments that can be used to process material that might contain biohazards: substances that contain biological agents capable of producing harmful effects in humans.
  • Page 146 To avoid damage, do not use abrasives or solvents to clean the Caution: instrument’s case. Warnings that apply to all Waters instruments When operating this device, follow standard quality control procedures and the equipment guidelines in this section. Safety Advisories...
  • Page 147 注意：未經有關法規認證部門允許對本設備進行的改變或修改,可能會使使用者喪失操作該設 備的權利。 注意：未经有关法规认证部门明确允许对本设备进行的改变或改装，可能会使使用者丧失操 作该设备的合法性。 주의: 규정 준수를 책임지는 당사자의 명백한 승인 없이 이 장치를 개조 또는 변경할 경우, 이 장치를 운용할 수 있는 사용자 권한의 효력을 상실할 수 있습니다. 注意：規制機関から明確な承認を受けずに本装置の変更や改造を行うと、本装置のユー ザーとしての承認が無効になる可能性があります。 Warnings that apply to all Waters instruments...
  • Page 148 Warning: Use caution when working with any polymer tubing under pressure: • Always wear eye protection when near pressurized polymer tubing. • Extinguish all nearby flames. • Do not use tubing that has been severely stressed or kinked. • Do not use nonmetallic tubing with tetrahydrofuran (THF) or concentrated nitric or sulfuric acids.
  • Page 149 • Hay que tener en cuenta que el cloruro de metileno y el sulfóxido de dimetilo dilatan los tubos no metálicos, lo que reduce la presión de ruptura de los tubos. 警告：當在有壓力的情況下使用聚合物管線時，小心注意以下幾點。 • 當接近有壓力的聚合物管線時一定要戴防護眼鏡。 • 熄滅附近所有的火焰。 • 不要使用已經被壓癟或嚴重彎曲管線。 • 不要在非金屬管線中使用四氫呋喃或濃硝酸或濃硫酸。 • 要了解使用二氯甲烷及二甲基亞楓會導致非金屬管線膨脹，大大降低管線的耐壓能力。 Warnings that apply to all Waters instruments...
  • Page 150 警告：当有压力的情况下使用管线时，小心注意以下几点： • 当接近有压力的聚合物管线时一定要戴防护眼镜。 • 熄灭附近所有的火焰。 • 不要使用已经被压瘪或严重弯曲的管线。 • 不要在非金属管线中使用四氢呋喃或浓硝酸或浓硫酸。 • 要了解使用二氯甲烷及二甲基亚枫会导致非金属管线膨胀，大大降低管线的耐压能力。 경고: 가압 폴리머 튜브로 작업할 경우에는 주의하십시오. • 가압 폴리머 튜브 근처에서는 항상 보호 안경을 착용하십시오. • 근처의 화기를 모두 끄십시오. • 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오. • 비금속(Nonmetallic) 튜브를 테트라히드로푸란(Tetrahydrofuran: THF) 또는 농축...
  • Page 151 警告：使用者必須非常清楚如果設備不是按照製造廠商指定的方式使用，那麼該設備所提供 的保護將被消弱。 警告：使用者必须非常清楚如果设备不是按照制造厂商指定的方式使用，那么该设备所提供 的保护将被削弱。 경고: 제조업체가 명시하지 않은 방식으로 장비를 사용할 경우 장비가 제공하는 보호 수단이 제대로 작동하지 않을 수 있다는 점을 사용자에게 반드시 인식시켜야 합니다. 警告： ユーザーは、製造元により指定されていない方法で機器を使用すると、機器が提供 している保証が無効になる可能性があることに注意して下さい。 Warnings that apply to all Waters instruments...
  • Page 152 To protect against fire, replace fuses with those of the type Warning: and rating printed on panels adjacent to instrument fuse covers. Attention: pour éviter tout risque d'incendie, remplacez toujours les fusibles par d'autres du type et de la puissance indiqués sur le panneau à...
  • Page 153 Electrical and handling symbols Electrical symbols These can appear in instrument user manuals and on the instrument’s front or rear panels. Electrical power on Electrical power off Standby Direct current Alternating current Protective conductor terminal Frame, or chassis, terminal Fuse Recycle symbol: Do not dispose in municipal waste.
  • Page 154 Handling symbols These handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments. Keep upright! Keep dry! Fragile! Use no hooks! A-12 Safety Advisories...
  • Page 155 Specifications Physical specifications Attribute Specification Height 43.2 cm (17 inches) Depth 66.0 cm (26 inches), without drip tray Width 28.0 cm (11 inches) Weight 39.5 kg (87 pounds), without solvent or sparge tubing Environmental specifications Attribute Specification Operating temperature 4 to 40 °C (39.2 to 104 °F) Operating humidity 20 to 80%, noncondensing Shipping and storage temperature...
  • Page 156 Electrical specifications (Continued) Attribute Specification Grounded AC Line voltages, nominal Power requirements 6.5 A Line voltage 100 to 240 VAC ±10% Frequency 50/60 Hz Fuse 10 A, 5 × 20 mm, slow-blow, IEC type Power consumption 650 W a. Protection Class I – The insulating scheme used in the instrument to protect from electri- cal shock.
  • Page 157 2535 QGM performance specifications Attribute Specification Number of eluents One to four Modes of operation Gradient, isocratic, and flow programming Operating flow range Programmable from 0 to 50 mL/min, in 0.01 increments Flow accuracy ±1.0% of required flow. Flow rate 1.0 to 50.0 mL/min using water or...
  • Page 158 2535 QGM performance specifications (Continued) Attribute Specification Composition precision Retention time variation 0.15 min SD, based on 6 repeat injections. Test conditions: Mobile phases: 70:30 water/methanol dial-a-mix (possibly use methanol and no modifiers). Flow rates (mL/min): 1, 5, 20, 50...
  • Page 159 2535 QGM performance specifications (Continued) Attribute Specification Delay volume (dwell volume) Small-scale: <6.0 mL, measured to the outlet of the pump at 1 mL/min, 50% point of the step gradient. Large-scale: <13.0 mL, measured to the outlet of the pump at 10 mL/min, 50% point of the step gradient.
  • Page 160 2545 QGM performance specifications Attribute Specification Number of eluents One to four Modes of operation Gradient, isocratic, and flow programming Operating flow range Programmable from 0 to 150 mL/min, in 0.01 increments Flow accuracy ±1.0% of required flow. Flow rate 1.0 to 150.0 mL/min using water or methanol, backpressure approximately 4140 kPa (41.4 bar,...
  • Page 161 2545 QGM performance specifications (Continued) Attribute Specification Composition precision Retention time variation 0.15 min SD, based on 6 repeat injections. Test conditions: Mobile phases: 70:30 water/methanol dial-a-mix (possibly use methanol and no modifiers). Flow rates (mL/min): 1, 5, 20, 50, 150 Sample: Uracil/caffeine Injection volumes (µL):...
  • Page 162 2545 QGM performance specifications (Continued) Attribute Specification Delay volume (dwell volume) <6.5 mL, measured to the outlet of the pump at 1 mL/min, 50% point of the step gradient. Pump seal wash Pump and seals required to wash the rear of the high pressure seal (pulsed type flow, about 20 µL per pulse).
  • Page 163 2555 QGM performance specifications (Continued) Attribute Specification Composition range 0 to 100% programmable in 0.1% increments for each of four reservoirs: A, B, C, and D. Total composition must sum to 100%. Composition accuracy ±3.0% absolute (full scale) from 4 to 260 mL/min using water/water or methanol/methanol spiked with dye or marker, backpressure approximately...
  • Page 164 2555 QGM performance specifications (Continued) Attribute Specification Maximum operating pressure 20,700 kPa (207 bar, 3000 psi) up to 200 mL/min (linear roll-off) to 17,200 kPa (172 bar, 2500 psi), at 300 mL/min. Programmable pressure limits Lower: 20,700 kPa (207 bar, 3000 psi) Higher: 20,700 kPa (207 bar, 3000 psi) Delay volume (dwell volume) <13.0 mL, measured to the outlet of...
  • Page 165 Solvent Considerations To avoid chemical hazards, always observe Good Laboratory Warning: Practices when operating your system. The information in this appendix applies to the 2535, 2545, and 2555 quaternary gradient modules. Contents Topic Page Introduction Solvent miscibility Solvent stabilizers Solvent viscosity...
  • Page 166 Preventing contamination For information on preventing contamination, refer to Controlling Contamination in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307), or visit www.waters.com. Clean solvents Clean solvents ensure reproducible results and permit you to operate with minimal instrument maintenance.
  • Page 167 Use water only from a high-quality water purification system. If the water system does not deliver filtered water, filter the water through a 0.45-µm membrane filter. Using 100% water can cause microbial growth. Waters Caution: recommends changing 100% water solutions daily. Adding a small amount of an organic solvent (approximately 10%) prevents microbial growth.
  • Page 168 When you switch from a strong buffer to an organic solvent, thoroughly flush the system with distilled water before you add the organic solvent. Solvent miscibility Viscosity Boiling Miscibility λ Cutoff Polarity Solvent cP, 20 °C point °C number index (nm) (@1 atm) (@1 atm)
  • Page 169 Solvent miscibility (Continued) Viscosity Boiling Miscibility λ Cutoff Polarity Solvent cP, 20 °C point °C number index (nm) (@1 atm) (@1 atm) Pyridine 0.94 115.3 Nitroethane 0.68 114.0 –– –– Acetone 0.32 56.3 15, 17 Benzyl alcohol 5.80 205.5 –– Methoxyethanol 1.72 124.6...
  • Page 170 For information on preventing contamination, refer to Controlling See also: Contamination in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307), or visit www.waters.com. Solvent stabilizers Do not leave solvents containing stabilizers, like THF with butylated hydroxytoluene (BHT), to dry in the system’s flow path. A dry flow path,...
  • Page 171 viscosity changes that occur as the solvents are mixed in different proportions can effect pressure changes during the run. For example, a 1:1 water/methanol mixture produces twice the pressure of either water or methanol alone. If you do not know the extent to which pressure changes affect the analysis, monitor the pressure during the run.
  • Page 172 The mass of gas dissolved in a given volume of solvent is proportional to the partial pressure of the gas in the vapor phase of the solvent. If the gas pressure decreases, the amount of that gas in solution also decreases. Dissolved oxygen affects UV-VIS detector performance in several ways Oxygen dissolved in solvents can form a UV absorbing complex, the amount of which is considerably different in different solvents.
  • Page 173 Helium sparging combines the convenience of short initial degassing time, ease of maintaining the solvent condition during operation, and complete control within the framework of the Waters preparative systems. The degassing operation must be efficient. To remove the gas as quickly as possible, you must consider these factors: •...
  • Page 174 UV cutoffs for common solvents The table below shows the UV cutoff (the wavelength at which the absorbance of the solvent equals 1 AU) for some common chromatographic solvents. Operating at a wavelength near or below the cutoff increases baseline noise because of solvent absorbance.
  • Page 175 10 mM, pH 7.6 Hydrochloric acid, 0.1% ® Waters PIC Reagent A, 1 vial/liter Morpholinoethanesulfonic acid Waters PIC Reagent B-6, (MES), 10 mM, pH 6.0 1 vial/liter Potassium phosphate, Waters PIC Reagent B-6, monobasic, 10 mM low UV, 1 vial/liter...
  • Page 176 Refractive indices of common solvents The following table 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.
  • Page 177 Refractive indices for common chromatographic solvents (Continued) Solvent Solvent 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 Wavelength selection C-13...
  • Page 178 C-14 Solvent Considerations...
  • Page 179 1-20 electricity source 1-36 helium sparge supply 1-26 signal cables 1-34 flow rate sparge system 1-24 flow scale, 2535 QGM contacting Waters Technical Service setting in console 2-20 contamination, preventing in control panel under control panel, using Empower 2-20 in control panel under...
  • Page 180 ISM classification outlet check valve cartridges, replacing 3-39 output, signals 1-31 overview contrast, adjusting display solvent considerations leak detector signals 1-32 leak problems, resolving 3-48 leak sensor performance specifications disabling 2-11 2535 QGM Index-2...
  • Page 181 2545 QGM safety considerations, maintenance 2555 QGM seals, replacing 3-14 physical specifications seal-wash pump, priming 2-15 power cord 1-35 selecting, system instruments powering on, QGM selector-valve rotor-seal assembly, pressure, system replacing 3-43 prime seal wash control signal prime solvents control cables, connecting 1-34 prime with syringe, performing...
  • Page 182 MassLynx software specifications outlet tube 1-21 electrical pressure environmental troubleshooting helium 1-24 operation terminals performance auxiliary 2 1-32 2535 QGM ground 1-32 2545 QGM start gradient 1-32 2555 QGM stop flow 1-32 physical switch 1-31 stabilizers, solvent tetrahydrofuran start gradient, terminals...
  • Page 183 3-45 viscosity considerations warning symbols waste tube, vent-valve 1-20 water, as solvent Waters Technical Service, contacting wavelength cutoffs C-11 selection C-10 Index-5...
  • Page 184 Index-6...

This manual is also suitable for:

25452555