Waters Reagent Manager Operator s Guide

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

Customer comments Waters Technical Communications department invites you to tell us of any errors you encounter in this document or to suggest ideas for otherwise improving it. Please help us better understand what you expect from our documentation so that we can continuously improve its accuracy and usability. We seriously consider every customer comment we receive. You can reach us at tech_comm@waters.com. 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. Visit www.waters.com. Telephone and fax From the USA or Canada, phone 800 252-HPLC, or fax 508 872 1990. For other locations worldwide, phone and fax numbers appear in the Waters Web site. Conventional mail Waters Corporation 34 Maple Street Milford, MA 01757 USA iii

Safety considerations Some reagents and samples used with Waters instruments and devices can pose chemical, biological, and radiological hazards. You must know the potentially hazardous effects of all substances you work with. Always follow Good Laboratory Practice, and consult your organization s safety representative for guidance. When you develop methods, follow the Protocol for the Adoption of Analytical Methods in the Clinical Chemistry Laboratory, American Journal of Medical Technology, 44, 1, pages 30 37 (1978). This protocol addresses good operating procedures and the techniques necessary to validate system and method performance. Considerations specific to the Reagent Manager Pinch-point hazard Warning: To avoid hazards associated with the reciprocating or rotating parts in the source, keep clear of the regions marked with yellow and gray labels. High temperature hazard Warning: To avoid burn injuries, avoid touching the source enclosure with your hand when operating or servicing the instrument. High voltage hazard Warning: To avoid electric shock, do not remove the instrument s protective panels. The components they cover are not user-serviceable. To avoid nonlethal electric shock when the instrument is in Operate mode, avoid touching the areas marked with the high voltage warning symbol. To touch those areas, first put the instrument in Standby mode. iv

Safety advisories Consult Appendix A for a comprehensive list of warning and caution advisories. Environmental information This product complies with the European Union RoHS Directive 2002/95/EC. Operating this device When operating this device, follow standard quality-control (QC) procedures and the guidelines presented in this section. Applicable symbols Symbol Definition Authorized representative of the European Community Confirms that a manufactured product complies with all applicable European Community directives ABN 49 065 444 751 Australia C-Tick EMC Compliant Confirms that a manufactured product complies with all applicable United States and Canadian safety requirements This product has been tested to the requirements of CAN/CSA-C22.2 No. 61010-1, second edition, including Amendment 1, or a later version of the same standard incorporating the same level of testing requirements v

Audience and purpose This guide is intended for use by individuals who need to operate, maintain, and/or troubleshoot the Waters Reagent Manager. You should be familiar with HPLC terms, practices, and basic HPLC system operations such as connecting tubing. Intended use of the Reagent Manager Waters designed the Reagent Manager as a dependable metering pump for general laboratory or industrial use. It is a single-piston, pulse-dampened pumping system for post-column reagent delivery. 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. Group 1 products apply to intentionally generated and/or used conductively coupled radio-frequency energy that is necessary for the internal functioning of the equipment. Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply network. vi

EC Authorized Representative Waters Corporation (Micromass UK Ltd.) Floats Road Wythenshawe Manchester M23 9LZ United Kingdom Telephone: +44-161-946-2400 Fax: +44-161-946-2480 Contact: Quality manager vii

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Table of Contents Copyright notice... ii Trademarks... ii Customer comments... iii Contacting Waters... iii Safety considerations... iv Considerations specific to the Reagent Manager... iv Safety advisories... v Environmental information... v Operating this device... v Applicable symbols... v Audience and purpose... vi Intended use of the Reagent Manager... vi ISM classification... vi ISM Classification: ISM Group 1 Class B... vi EC Authorized Representative... vii 1 Overview... 1-1 Introduction... 1-2 Features... 1-3 Polymeric flow path... 1-3 Automatic seal wash... 1-4 Front control panel... 1-4 Priming and purging... 1-4 Filters... 1-4 Spill protection... 1-5 Remote Start/Stop feature... 1-5 Fluid-handling components... 1-5 Flow paths... 1-7 Pump cycle... 1-8 Table of Contents ix

Pulse damper... 1-9 Electronic components... 1-10 Remote Start/Stop feature... 1-12 2 Installation... 2-1 Installation tasks... 2-2 Site requirements... 2-2 Unpacking... 2-3 Making electrical connections... 2-5 AC Voltage input... 2-5 Connecting to the power supply... 2-5 Connecting for remote start/stop operation... 2-5 Making fluidic connections... 2-13 Connecting the solvent reservoir to the pump inlet... 2-15 Connecting the pump outlet to a device... 2-17 Connecting the overflow tubing... 2-19 3 Operating the Waters Reagent Manager... 3-1 Using the front control panel... 3-2 Digital display... 3-2 Operation keys... 3-2 Status LEDs... 3-4 Preparing the Reagent Manager... 3-5 Solvent guidelines... 3-5 Priming the pump... 3-6 Purging the pump... 3-7 Priming the seal wash tubing... 3-8 Operating as a stand-alone pump... 3-8 Before operating... 3-8 Operating the pump... 3-8 After operating... 3-9 Routine tasks... 3-12 x Table of Contents

Remote start/stop operation... 3-12 Using the Waters ACQUITY binary solvent manager... 3-12 Using the Waters 2695 separations module... 3-13 Using the Waters 600E controller... 3-13 4 Maintaining the Waters Reagent Manager... 4-1 Maintenance Considerations... 4-2 Safety and handling... 4-2 Proper operating procedures... 4-2 Spare parts... 4-2 Routine maintenance procedures schedule... 4-3 Contacting Waters Technical Service... 4-4 Filters... 4-4 Replacing the inlet filter... 4-5 Replacing the outlet filter... 4-5 Replacing the high-pressure inline filter... 4-6 Check valves... 4-8 Cleaning the check valves... 4-9 Replacing the pump head check valves... 4-10 Check valve and pressure ramp test... 4-12 Pump head, piston, and components... 4-13 Removing the pump head... 4-14 Cleaning and inspecting the piston... 4-16 Replacing the seal-wash components... 4-17 Installing the piston seal... 4-17 Installing the pump head... 4-18 Prime/purge valve... 4-19 Inspecting the prime/purge valve... 4-19 Removing the prime/purge valve... 4-20 Replacing the prime/purge valve seal... 4-21 Installing the prime/purge valve... 4-21 Table of Contents xi

Restrictor coil... 4-22 Fuse... 4-24 Fan... 4-25 5 Troubleshooting... 5-1 Overview... 5-2 Contacting Waters Technical Service... 5-2 General troubleshooting... 5-2 Troubleshooting fluidics... 5-3 Troubleshooting leaks... 5-8 Troubleshooting electronics... 5-9 Identifying and correcting audible noises... 5-12 A Safety Advisories... A-1 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-5 Electrical and handling symbols... A-11 Electrical symbols... A-11 Handling symbols... A-12 B Specifications... B-1 C Spare Parts... C-1 D Solvent Considerations... D-1 Solvent preparation... D-2 Solvent outgassing and sparging... D-2 Solvent quality... D-2 Using buffers... D-3 xii Table of Contents

Solvent miscibility... D-3 Solvent viscosity... D-6 Solvent degassing... D-6 Gas solubility... D-6 Solvent degassing methods... D-7 Preventing cavitation... D-8 Filtration... D-9 Index... Index-1 Table of Contents xiii

xiv Table of Contents

1 Overview Topic Page Introduction 1-2 Features 1-3 Fluid-handling components 1-5 Electronic components 1-10 Remote Start/Stop feature 1-12 1-1

Introduction The Waters Reagent Manager is a high-performance pump designed for use as a dependable metering pump for general laboratory or industrial use. As a single-piston, pulse-dampened pumping system for post-column reagent delivery, the Waters Reagent Manager combines the most important aspects of solvent delivery: High precision and accuracy Reliability Smooth solvent flow Ease of use The reagent manager is designed for optimum performance, operational convenience, and safety. The following figure shows the front view of a Waters Reagent Manager. Waters Reagent Manager Front control panel Mode Fault Run ml/min PSI Hi Pr Lo Pr PRIME/PURGE Pump head with seal wash Prime Run/ Stop FILTER/OUTLET Waters Reagent Manager Power switch I/O TP01561 1-2 Overview

Features The reagent manager includes a full array of important features, such as pulse damping, a nonmetallic flow path, automatic washing of the pump piston seal, and an easy-to-use front control panel. Other features include a convenient priming/purging system, a high-pressure inline filter, spill protection, and a remote start/stop feature. The Waters Reagent Manager operates from 0.05 to 2.00 ml/min up to 2500 psi. The flow rate can be set in 0.01-mL increments with a precision of 0.5%. The pump automatically shuts down at user-settable high- or low-pressure limits. The pump contains a pulse damper to minimize detector baseline noise. Low flow modulation of the reciprocating, single-piston pump is achieved with an advanced electronic rapid-refill design, programmed stepper motor acceleration, and a pulse damper. The rapid refill design reduces pulsations in the pump cycle and provides constant, stable flow from the pump at high pressure. Polymeric flow path The materials that make up the fluidic pathway of the Waters Reagent Manager are nonmetallic and biocompatible. They include: polyetheretherketone (PEEK) polytetrafluoroethylene (PTFE) Tefzel sapphire ruby polyethylene fluoroethylene propylene copolymer (FEP) ultra-high molecular weight polyethylene (UHMWPE) Wetted surfaces in the pump head include ruby, sapphire, and polymer (PEEK). Features 1-3

Automatic seal wash During pump operation, the pump head is continuously washed in the region behind the primary high pressure seal. The seal washing action helps to reduce seal wear by washing away the microscopic layer of solvent that wets the area between the piston and the seal. If you use buffered solvents, the solvent layer can evaporate behind the seal and deposit crystals on the piston. Crystals on the piston can abrade the pump seal and cause premature seal failure, leakage, and damage to the pump. The seal-washing pump head washes away the film of solvent before crystals can precipitate. Front control panel A keypad and LED display on the front panel allow you to quickly enter parameters before and during a run. The solvent-resistant membrane keypad has touch keys, while the display provides status and monitoring information for the pump: The four-digit LED front panel display shows the flow rate or pressure. The status LED indicators show the operational status of the pump. Priming and purging Filters The prime/purge valve and the Prime key on the front panel provide quick and easy solvent introduction and rapid solvent changeover using the priming and purging feature. To help maintain the solvent quality through the pump flow path, the reagent manager uses the following filters: An inlet filter In the solvent reservoir (external, slip-on type) A high-pressure inline filter Between the pulse damper and the restrictor coil An outlet filter At the filter/outlet port 1-4 Overview

Spill protection The reagent manager has internal spill protection in case of leaks in the flow path. In the event of a leak, solvent drips to the drip pan and is routed to the waste fluid drain connector on the bottom right side of the pump. Recommendation: Provide a solvent bottle tray for the top of the pump if you decide to place solvents or any instrument with fluidic lines on top of the pump. For example, a solvent bottle tray can provide storage for solvent reservoirs and a seal wash reservoir, with leak protection for a spill. Remote Start/Stop feature You can easily integrate the reagent manager into your HPLC system using remote stop/start inputs and stop output capabilities. To accomplish this, the reagent manager accepts remote run/stop input (contact closure) signals from other instruments. It also generates a remote stop output signal to an instrument when a high- or low-pressure fault is triggered. This capability requires a remote start/stop cable, which is included in the startup kit. Fluid-handling components The fluid-handling components of the Waters Reagent Manager are shown in the following figure. Fluid-handling components 1-5

Pump fluid-handling components Pump outlet Pump head Seal wash outlet Seal wash inlet Pump inlet Prime/purge valve Filter/outlet valve TP01563 Caution: To prevent solvent leaks from the pump outlet, position the solvent reservoir below the pump inlet before you connect the pump outlet fitting. The following table describes the functions of the major fluidics components. Fluid-handling components Component Pump head Pump inlet and outlet Function Draws in and delivers solvent. Maintains flow direction of solvent through the pump using check valves that open in one direction only. 1-6 Overview

Fluid-handling components (Continued) Component Prime/purge valve Filter/outlet valve Seal wash inlet and outlet Function When the valve is open, allows priming and purging of the pump. Considerations: Connecting a syringe to the valve allows you to draw solvent through the pump for priming. Operating the pump allows solvent to flow through the pump and out the valve for purging. When the valve is closed, high-pressure flow is directed to the filter/outlet valve. Routes solvent out of the pump to a fitting, injector, column, or detector. Washes the piston seal with seal wash solvent using check valves to maintain flow of seal wash in one direction. Flow paths The flow path of a solvent through the pump is as follows: During priming and purging Solvent flows from the solvent reservoir through the pump head to the pulse damper and out the prime/purge valve. The flow rate ramps up to 10 ml/min during priming and purging. During normal operation Solvent flows from the inlet filter in the solvent reservoir through the pump head to the pulse damper, prime/purge valve, inline filter, restrictor coil, and out the filter/outlet. Tip: As shown in the following figure, the seal wash solvent (dotted lines) continuously flushes the pump in the area behind the piston seal. Fluid-handling components 1-7

Flow paths of solvents through pump Reagent manager Pulse damper Pump Head Seal wash reservoir Outlet check valve for seal wash Solvent outlet check valve Prime/purge valve High-pressure inline filter Fluid waste Solvent reservoir with filter Inlet check valve for seal wash Solvent inlet check valve Restrictor coil Filter/outlet Next device Pump cycle The pump cycle consists of two phases, the delivery phase and the refill phase. During the delivery phase, the pump piston moves forward at a constant linear speed, and is eccentrically driven by the motor. This results in a constant, stable flow from the pump at high pressure. During the refill phase, the piston quickly retracts, refilling the pump head with solvent. Although the pump head output flow completely stops during refill, the pulse damper transiently maintains some flow. In addition, the motor speed is adjusted by the microprocessor to facilitate an efficient refill phase. The combination of increased motor speed and the rapid refill design of the cam generates a refill time of less than 5% of the pump cycle at 1 ml/min. 1-8 Overview

Pulse damper The patented diaphragm pulse damper helps to maintain the flow rate and pressure. The pulse damper consists of a compressible fluid (isopropanol) held in an isolated cavity by a flexible diaphragm. During the delivery phase, the pressure of the solvent displaces the diaphragm, compressing the fluid in the cavity and storing energy. During the refill phase, the pressure of the pumped solvent on the diaphragm is reduced and the compressed fluid expands, releasing its stored energy and continuing to pressurize the solvent path. Because the amount of solvent in contact with the pulse damper is small (0.9 ml at 2500 psi), and the flow past the damper is completely swept, solvent changeover is rapid. The pulse damper requires a back pressure of approximately 500 psi or greater. A length of small-bore, restrictor tubing (the restrictor coil) between the pulse damper and the filter/outlet ensures that this minimum back pressure is maintained. Fluid-handling components 1-9

Electronic components The Waters Reagent Manager electronics are illustrated in the following figures. Pump electronics front view LED display Fault ml/min PSI Hi Pr Lo Pr Status LEDs Keypad Mode Prime Run Run/ Stop Waters Reagent Manager Power switch I/O TP01560 1-10 Overview

Pump electronics back view The following table describes the functions of the electronic components. Electronic components Component Location Function LED display Front Provides pump status and monitoring information. Keypad with operation keys Front Use to select pump operating parameters and values. Status LEDs Front Provides status information. Power switch Front Powers the pump on and off. Power entry module Back Provides the power cord connector and a receptacle for a fuse. Cooling fan Back Exhausts air to cool the internal electronics. Terminal strip Back Provides connection through a remote start/stop cable to an external control device for remote start/stop signal control. Also provides a remote stop output on pressure fault. See Connecting for remote start/stop operation on page 2-5 for more information. RS-232 connector Back Reserved for future use. Electronic components 1-11

Remote Start/Stop feature When the reagent manager is configured for remote start/stop operation, the external control device triggers the pump on or off by contact closure signals sent over a cable. If a fault occurs in the pump during operation, the reagent manager stops running and sends a signal to the external device. The reagent manager startup kit contains the remote start/stop cable for remote start/stop operation. See Connecting for remote start/stop operation on page 2-5, for instructions on how to install the remote start/stop cable. 1-12 Overview

2 Installation This chapter describes how to unpack and install your Waters Reagent Manager. Topic Page Installation tasks 2-2 Site requirements 2-2 Unpacking 2-3 Making electrical connections 2-5 Making fluidic connections 2-13 2-1

Installation tasks The following figure outlines the installation tasks. Flowchart for installing the reagent manager Start installation Make signal and power connections Select a site Make fluidic connections Unpack Prime the pump Inspect Installation complete Site requirements Install the reagent manager on a stable, flat, clean surface at a site that meets the specifications indicated in the following table. Warning: To avoid overheating and to provide clearance for cable connections, ensure that the rear panel of the reagent manager has at least 3 inches (7.6 cm) of clearance. Installation site requirements Factor Temperature Relative humidity Requirement 5 to 35 C (41 to 95 F) 20% to 90%, noncondensing 2-2 Installation

Installation site requirements (Continued) Factor Size Clearance Input voltage ranges Input voltage frequency Electromagnetic fields Requirement Width: 9.1 in. (23.1 cm) Depth: 17.4 in. (44.2 cm) Height: 6.5 in. (16.5 cm) At least 3 in. (7.6 cm) at rear for ventilation and cable connections 100 to 240 VAC Universal Input - No fuse change required 50 to 60 Hz No nearby source of electromagnetic noise, such as arcing relays or electric motors Note: The unit is only for indoor use, at a maximum altitude of 2000m. The unit is intended for use in a POLLUTION DEGREE 2 ENVIRONMENT. Unpacking The Waters Reagent Manager is shipped in one carton that contains the following items: Waters Reagent Manager Reagent Manager Startup Kit (see Appendix C for a list of contents) Prior to opening the shipping container, inspect it for damage or evidence of mishandling. If it has been damaged or mishandled, notify the carrier before opening the container. See the Waters Licenses, Warranties, and Support Services document (on the documentation CD) for more information. Tip: Save the shipping container for future use. To unpack the reagent manager 1. Open the carton and remove the startup kit and other items from the slot at one end of the carton. 2. Using both hands, lift the pump and its foam packing material out of the carton. Unpacking 2-3

3. Carefully set the pump down, then remove the foam packing material from the ends of the pump. 4. Check the contents of the startup kit against the startup kit parts list to confirm all items are included. 5. Verify that the serial number on the back of the pump matches the serial number on the Certificate of Functional Performance. Keep the Certificate of Functional Performance for future reference. 6. For more information about the warranty, see the Waters Licenses, Warranties, and Support Services document (on the documentation CD). 7. Inspect all items for damage. Immediately report any shipping damage to both the shipping company and your Waters representative. Contact Waters Technical Service at 800-252-4752, U.S. and Canadian customers only. Other customers, call your local Waters subsidiary or Technical Service Representative, or consult the Waters web site (www.waters.com) for worldwide phone and fax numbers. See the Waters Licenses, Warranties, and Support Services document for complete information on reporting shipping damages and submitting claims. 2-4 Installation

Making electrical connections For proper operation, the reagent manager requires a grounded AC power supply with no abrupt voltage fluctuations. AC Voltage input The reagent manager allows voltage input from 100 to 240 VAC at 50 to 60 Hz. No fuse change is required. Connecting to the power supply The reagent manager automatically adjusts for AC input voltage and frequency. To connect the power supply 1. Ensure that the power switch on the front panel is in the Off (O) position. 2. Insert the power cord into the power connector on the back of the pump. 3. Insert the other end of the power cord into a grounded power outlet. Connecting for remote start/stop operation For optional remote start and stop operation, the remote start/stop cable is required. This cable is included in the startup kit. This cable connects the Waters Reagent Manager to a Waters ACQUITY Binary Solvent Manager, a Waters 2695 Separations Module, a Waters 600E Controller, or other external device equipped with TTL-logic or contact closure outputs. Making electrical connections 2-5

The following table describes the connections on the reagent manager terminal strip. Reagent Manager terminal strip connections Term Number Description STOP OUT - NO 8 Stop output signal, normally open; contact closure signal from reagent manager to external device a STOP OUT - NC (None) Stop output signal, normally closed; contact closure signal from reagent manager to external device b OUTPUT COM 6 Ground for output connection RUN INPUT 3 Contact closure signal from external device to start pumping INPUT COM 1 Ground for input connection a. Waters devices are normally open b. Other devices may require normally closed for TTL closure To connect the reagent manager to a Waters ACQUITY binary solvent manager 1. Remove the terminal strip from the back of the reagent manager. 2. Attach the leads on the end of the start/stop cable to the reagent manager terminal strip as shown in Figure Connecting to a Waters ACQUITY binary solvent manager on page 2-8. Use the Table titled Reagent Manager terminal strip connections on page 2-6 as a guide. 3. Reinstall the terminal strip on the back of the reagent manager. 4. Remove the terminal strip from the Waters ACQUITY binary solvent manager. 2-6 Installation

5. Connect the leads on the end of the start/stop cable to the terminal strip of the Waters ACQUITY binary solvent manager as shown in Figure Connecting to a Waters ACQUITY binary solvent manager on page 2-8. 6. Reinstall the terminal strip on the Waters ACQUITY binary solvent manager. Making electrical connections 2-7

Connecting to a Waters ACQUITY binary solvent manager Remote Start/Stop Cable Waters Reagent Manager Terminal Strip Green Blue White Red 10 9 8 STOP OUT - NO 7 STOP OUT - NC 6 OUTPUT COM 5 4 3 RUN INPUT 2 STOP INPUT 1 INPUT COM Waters ACQUITY Binary Solvent Manager 1 2 3 4 + Stop Flow - Stop Flow 1 2 3 4 Green Blue Run Stopped Out Run Stopped Out 5 6 7 8 White Red 5 6 7 8 9 9 10 11 10 11 12 12 I II 2-8 Installation

To connect the reagent manager to a Waters 2695 separations module 1. Remove the terminal strip from the back of the reagent manager. 2. Attach the leads on the end of the start/stop cable to the reagent manager terminal strip as shown in Figure Connecting to a Waters 2695 separations module on page 2-10. Use the Table titled Reagent Manager terminal strip connections on page 2-6 as a guide. 3. Reinstall the terminal strip on the back of the reagent manager. 4. Remove the terminal strip from the Waters 2695 separations module. 5. Connect the leads on the end of the start/stop cable to the terminal strip of the Waters 2695 separations module as shown in Figure Connecting to a Waters 2695 separations module on page 2-10. 6. Reinstall the terminal strip on the Waters 2695 separations module. Making electrical connections 2-9

Connecting to a Waters 2695 separations module Remote Start/Stop Cable Waters Reagent Manager Terminal Strip Green Blue White Red 10 9 8 STOP OUT - NO 7 STOP OUT - NC 6 OUTPUT COM 5 4 3 RUN INPUT 2 STOP INPUT 1 INPUT COM Waters 2695 Separations Module Green Blue 1 2 3 4 5 6 White Red + Stop Flow - Stop Flow 1 2 3 4 5 6 Switch 1 Switch 1 7 8 7 8 9 9 10 11 10 11 12 12 Connector B Connector A 2-10 Installation

To connect the Reagent Manager to a Waters 600E Controller 1. Remove the terminal strip from the back of the reagent manager. 2. Attach the leads on the end of the start/stop cable to the reagent manager terminal strip as shown in Figure Connecting to a Waters 600E controller on page 2-12. Use the Table titled Reagent Manager terminal strip connections on page 2-6 as a guide. 3. Reinstall the terminal strip on the back of the reagent manager. 4. Remove the terminal strip from the Waters 600E Controller. 5. Connect the leads on the end of the start/stop cable to the terminal strip of the Waters 600E Controller as shown in Figure Connecting to a Waters 600E controller on page 2-12. 6. Reinstall the terminal strip on the back of the Waters 600E Controller. Making electrical connections 2-11

Connecting to a Waters 600E controller Reagent Manager Remote Start/Stop Cable Terminal Strip Green Blue White Red 10 9 8 STOP OUT - NO 7 STOP OUT - NC 6 OUTPUT COM 5 4 3 RUN INPUT 2 STOP INPUT 1 INPUT COM White: Switch 1 Green: Stop 1 2 3 4 5 6 7 8 9 Terminal Strips Waters 600E Controller Blue: Ground Red: Ground 2-12 Installation

Making fluidic connections As shown in the Figure Connecting the tubing on page 2-14, making fluidic connections to the Waters Reagent Manager involves: Connecting the solvent reservoir to the pump inlet Connecting the pump outlet to a device Installing the seal wash tubing that washes the piston seal on the pump head Connecting tubing from the waste fluid drain under the pump to a waste container Making fluidic connections 2-13

Connecting the tubing Note: Ensure that the reservoirs are at least 4 in. (10 cm) above the pump head during operation. Reagent Reservoir Reagent Tubing Seal Wash Reservoir Waters Reagent Manager Seal Wash Tubing Priming Syringe Outlet Tubing to Next Device Seal Wash Tubing to Waste Container TP01565e Recommendation: Degas, filter, and cover all solvents. See Appendix D for solvent considerations. Required Materials Startup Kit Inlet filter, inlet ferrule, and inlet nut (one each) Outlet ferrule and outlet nut (three each) Inlet tubing, 48 in. (122 cm), 0.085-inch i.d. 1/8-inch o.d. 2-14 Installation

Seal wash tubing, PTFE, 72 in. (183 cm), 1/8-inch i.d. 1/4-inch o.d. Seal wash fittings (two each) Priming syringe (30 ml) Outlet tubing, PEEK, 60 in. (152 cm), 0.020-inch i.d. 1/16-inch o.d. Overflow tubing, PTFE, 72 in. (183 cm, cut in two 36-in [91.44 cm] lengths), 1/4-inch i.d. 3/8-inch o.d. Polymeric tubing cutter Wrench, 5/16-in. open-end Reservoir containing filtered, degassed solvent Reservoir containing filtered seal wash solvent (20% (v/v) methanol in water, or 20% (v/v) acetonitrile in water) Waste container Connecting the solvent reservoir to the pump inlet Caution: To avoid a solvent leak from the pump outlet during installation, position the reservoir below the pump inlet until the pump outlet is connected to the system. During pump operation, position the bottom of the reservoir about 4 inches (10 cm) higher than the pump head. If you use high-viscosity solvents, you may need to elevate the reservoirs as much as 18 to 24 inches (46 to 61 cm) above the inlet to prevent bubble formation as the pressure drops during the refill phase. Making fluidic connections 2-15

To connect the solvent reservoir to the pump solvent inlet 1. Measure the length of inlet tubing required to connect the solvent reservoir to the pump inlet. The inlet tubing should be as short as practical to minimize pressure losses on the inlet side of the pump during the refill phase (to prevent bubble formation). 2. Use the tubing cutter to cut the PEEK tubing to the measured length. Make sure the cut end is straight and free from burrs. 3. Slide a nut and then a ferrule over one end of the tubing, as shown in the following figure. Attaching a ferrule and nut to tubing Nut Tubing Ferrule Tubing End (straight and smooth) Caution: To avoid damaging the ferrule, do not overtighten the nut. 4. Firmly seat the tubing end into the inlet on the pump, then finger-tighten the nut. 5. Insert the free end of the inlet tubing through the cap of the solvent reservoir. 6. Cut a 1.5-inch (3.8-cm) section of outlet tubing and slide half of its length over the end of the inlet tubing. 7. Insert the tubing fitting of the inlet filter into the outlet tubing sleeve at the free end of the inlet tubing. 8. Install the cap onto the solvent reservoir (containing filtered, degassed solvent) and push the outlet tubing through the cap until the filter reaches the bottom of the reservoir. 2-16 Installation

Connecting the pump outlet to a device To connect the pump outlet to a device 1. Measure the length of outlet tubing required to connect the pump outlet to a device. Use a polymeric tubing cutter to cut the tubing to the measured length. Ensure that the tubing is cut squarely with no burrs or crimps and that the center hole is open. 2. Remove the fitting plug from the pump filter/outlet fitting. 3. Slide an outlet nut and then a ferrule over one end of the tubing (see the above figure). 4. Firmly seat the tubing into the filter/outlet fitting, then finger-tighten the nut. 5. Unscrew the assembled fitting and verify that the length of tubing extending beyond the ferrule is 0.12 inch (3 mm). 6. Finger-tighten the nut again. 7. Repeat steps 3 through 6 for the other end of the tubing only after you have primed the pump and flushed the system, as described in Priming the pump on page 3-6. Installing and priming the seal wash tubing The pump head seal wash requires 250 ml to 500 ml of seal wash solvent. The seal wash solvent drains to a fluid waste container below the pump. Use an aqueous seal wash solvent with enough organic content to inhibit bacterial growth. To install the seal wash tubing 1. Cut the seal wash tubing at its midpoint. 2. Install seal wash fittings on one end of each length of seal wash tubing. 3. Screw the fitting on one length of tubing to the seal wash inlet as shown in the following figure. Ensure that the fitting and tubing fit tightly to prevent leaks. Making fluidic connections 2-17

Connecting the seal wash tubing Tubing from seal wash outlet to waste container Tubing from seal wash reservoir to seal wash inlet TP01563 4. Install the other end of the tubing in the seal wash reservoir. Secure the tubing in the reservoir to ensure that the tubing stays immersed during pump operation. Position the reservoir at least 4 in. (10 cm) higher than the pump head. 5. Screw the fitting on the other length of tubing to the seal wash outlet. Ensure that the fitting and tubing fit tightly. 6. To prime the seal wash tubing, connect the seal wash fitting to the priming syringe, then connect the syringe to the free end of the seal wash outlet tubing. 7. Draw 10 ml to 20 ml of seal wash solvent through the seal wash outlet. Ensure that the tubing is filled with solvent and that no bubbles remain in the lines. 8. Disconnect the syringe, then install the end of the seal wash outlet tubing in a waste container located at a level below the pump so that fluid drains by gravity. Secure the tubing to ensure that the tubing stays in the container during pump operation. 2-18 Installation

Connecting the overflow tubing To install the overflow tubing 1. Connect one end of the overflow tubing to the drip tray connector underneath the reagent manager. 2. Connect the other end of the tubing to a waste container located at a level below the pump. Making fluidic connections 2-19

2-20 Installation

3 Operating the Waters Reagent Manager This chapter describes how to prepare and operate the Waters Reagent Manager either as a stand-alone pump using the front control panel or under remote start/stop operation. Topic Page Using the front control panel 3-2 Preparing the Reagent Manager 3-5 Operating as a stand-alone pump 3-8 Remote start/stop operation 3-12 3-1

Using the front control panel The front control panel of the Reagent Manager consists of a four-digit LED display, the operation keys, and status LEDs, as shown in the following figure. Reagent Manager front control panel Display Fault ml/min PSI Hi Pr Lo Pr Status LEDs Operation Keys Mode Prime Run Run/ Stop Waters Reagent Manager Power Switch To power-on the pump, locate the power switch on the lower left front panel and set it to the On (I) position. Digital display The four-digit LED display can show the pump flow rate (ml/min), system pressure (psi), or the high- or low-pressure settings (psi). You use the Mode key to choose which parameter to display. Operation keys I/O TP01560 The operation keys provide access to all pump operating parameters. Use the Mode, Up, Down, and Run/Stop operation keys to navigate through the parameters. 3-2 Operating the Waters Reagent Manager

The following table describes the functions of the operation keys on the Reagent Manager control panel. Functions of operation keys Key Mode Function Cycles through the four (parameter) modes: flow rate in ml/min, pressure in psi, high-pressure limit, and low-pressure limit. The status LEDs to the right of the digital display indicate the active mode. Up - increases the value for the selected mode (parameter). Down - decreases the value for the selected mode (parameter). Prime Initiates priming at 10 ml/min. To stop priming, press any key. Run/ Stop When the unit is not pumping, starts the pump at all current parameter settings. The maximum flow rate during operation is 2.0 ml/min. When the unit is pumping, immediately stops the pump. Tip: Also clears any high- or low-pressure faults that may be occurring within the Reagent Manager. Functions of and details on all Status LED parameters are provided in Table titled Status LEDs on page 3-4. Tip: You can display the software version of the Reagent Manager using the Run/Stop and Up arrow keys. To display the software version, prior to turning on the instrument, press and hold both the Run/Stop and Up arrow keys, power-on the Reagent Manager, and then release the two keys when User appears in the LED display. The Reagent Manager software version appears in the LED display (in decimal point format). To exit this test mode, press the Run/Stop key. Using the front control panel 3-3

Status LEDs The status LEDs display the real-time status of the pump. This table describes all Status LEDs. Status LEDs Parameter Status LED Description Parameter value Flow Rate ml/min Lights when the flow rate is displayed. Pressure PSI Lights when the current system pressure is displayed. High- Pressure Limit Low- Pressure Limit Hi Pr Lo Pr Lights when the user-selected high- pressure limit (in psi) is displayed. Lights when the user-selected low- pressure limit (in psi) is displayed. Fault Fault Lights when a fault occurs and stops the pump. Run Run Lights when the pump is running. The flow rate in ml/min at which the pump is currently operating. The flow rate range is 0.05 ml/min to 2.00 ml/min. The pressure in psi at which the pump is currently set to operate. The pressure range is 0 psi to 2500 psi. The pressure is a read-only value. Select this mode to track the pressure during priming, operating, and troubleshooting tasks. The pressure at or above which the pump is to shut down. The Fault LED also lights. You can set the Hi Pr in 10-psi increments. The maximum value is 2500 psi. The minimum value is the low-pressure limit plus 100 psi. The pressure at or below which the pump is to shut down. The Fault LED also lights. You can set the Lo Pr in 10-psi increments. The maximum value is the upper limit minus 100 psi. The minimum value is 0 psi. On indicates a problem that must be cleared before operation. Off indicates that the pump is ready to operate. On indicates that the pump is running or priming. Off indicates that the pump is not running. 3-4 Operating the Waters Reagent Manager

Preparing the Reagent Manager The procedure to prepare the Reagent Manager for operation depends on the status of the pump. If the pump is dry, you must prime the pump (see Priming the pump on page 3-6). If you are changing solvent, restarting the pump after a long idle time, or adding fresh solvent, and the pump contains fluid, you must purge the pump (see Purging the pump on page 3-7). Solvent guidelines Required Materials Priming syringe Solvents for the system, filtered and degassed Waste tubing Waste container Seal wash solvent Selection and preparation of solvents to use in the Waters Reagent Manager are critical factors for successful operation. See Appendix D for detailed solvent consideration information. Caution: Do not use any of the following solvents in the Waters Reagent Manager because they damage PEEK components. Methylene chloride Tetrahydrofuran (THF) at greater than 10% (v/v) Freon Concentrated sulfuric acid (H 2 SO 4 ) Concentrated nitric acid (HNO 3 ) Preparing the Reagent Manager 3-5

Also keep in mind the following solvent guidelines: Use methanol to prime a new pump or a pump that has been stored for an extended period. Use an intermediate solvent when you change between solvents that are not miscible. Always keep the solvent level in the reservoir at least 4 inches (10 cm) higher than the pump inlet. Do not place solvent reservoirs on top of the Reagent Manager. Use only filtered, degassed solvents. After using a buffer, purge the pump with HPLC-grade, distilled, or deionized water to prevent crystallization and damage to the piston or seal. Know the properties of the solvents under different pressures. Dedicate a solvent supply tube and filter assembly for each solvent. If this is not possible, purge the tube and filter of any remaining solvent before you use a new solvent. To prevent cross-contamination of solvents, dedicate a set of glassware for preparing, storing, and supplying each solvent. Once a container has been used for a solvent, do not use it for any other solvent unless it has been thoroughly cleaned. Warning: To avoid chemical hazards and prevent injury, always observe Good Laboratory Practices when you handle solvents. Know the physical and chemical properties of the solvents you use. See the Material Safety Data Sheets for the solvents in use. For more information on solvent properties, see Appendix D. Priming the pump To prime the pump 1. Power on the pump. 2. Raise the reservoir so that its base is above the level of the pump inlet. 3. Open the prime/purge valve by turning it fully counterclockwise. 4. Connect the fully discharged priming syringe to the prime/purge valve (see the following figure). 3-6 Operating the Waters Reagent Manager

Priming the Reagent Manager Prime/Purge Valve Priming Syringe 5. Press Prime to run the pump. The pump automatically ramps up to 10 ml/min. 6. Draw at least 20 ml of solvent (and any air bubbles) through the pump and into the syringe. 7. Press Run/Stop or any key to stop priming. 8. Close the prime/purge valve by turning it fully clockwise, and remove the syringe. Dispose of solvent properly. 9. Connect the free end of the outlet tubing to the inlet fitting of the next device in your HPLC system and finger-tighten the nut. Purging the pump Purging ensures that all solvent in the fluidic path is replaced with fresh solvent. Caution: When you change between two solvents that are not totally miscible, follow the purging procedure described below, first using an intermediate solvent, then the final solvent. See Appendix D for more information. Before you press the Prime key, be sure the prime/purge valve is open. To prevent damage to the pump, always draw fluid from the prime/purge valve. Do not inject fluid into the prime/purge valve with a syringe. To prevent solvent flow through the pump head under gravity pressure, never leave the pump unattended with the prime/purge valve in the open position. Preparing the Reagent Manager 3-7

To purge the pump 1. Perform steps 1 through 9 in Priming the pump on page 3-6. 2. Press Run/Stop to start the pump. 3. Let the pump run for about one minute to verify that the pump pressure is normal for the selected solvent and flow rate. If the pump pressure is not correct, repeat steps 1 and 2, as necessary. 4. Press Run/Stop to stop the pump. Priming the seal wash tubing If you see air bubbles in the seal wash tubing, prime the seal wash tubing as described in Installing and priming the seal wash tubing on page 2-17. Operating as a stand-alone pump You operate the Reagent Manager as a stand-alone pump using only the operation keys on the front control panel. Before operating Before you operate the Reagent Manager, ensure that: You have already completed any required priming and purging, as described in Preparing the Reagent Manager on page 3-5. Solvent reservoirs are filled with filtered, degassed solvent. Caution: To avoid chemical hazards and prevent injury, always observe Good Laboratory Practices when you handle solvents. See the Material Safety Data Sheets for the solvents you use. For more information on solvent properties, see Appendix D. Operating the pump The operation keys provide access to all pump operating parameters. Use the Mode, Up, Down, and Run/Stop operation keys to navigate through the parameters. You can change pump parameters while the pump is running. 3-8 Operating the Waters Reagent Manager

To operate the Reagent Manager 1. Press Mode to display the flow rate (as indicated by the illuminated ml/min status LED). Press the Up or Down arrow key to increase or decrease the value. 2. Press Mode to display the high-pressure limit (as indicated by the illuminated Hi Pr status LED). Press the Up or Down arrow key to increase or decrease the value. 3. Press Mode to display the low-pressure limit (as indicated by the illuminated Lo Pr status LED). Press the Up or Down arrow key to increase or decrease the value. 4. Once all parameters values are correctly set, press Mode to display the operating pressure. 5. Press Run/Stop to start the pump. Operation continues until you press Run/Stop to stop the pump. Tip: To immediately stop the pump, press Run/Stop. After operating After operating the Reagent Manager, you can power-off the pump for storage. The procedures to follow before you power-off the pump depend on the status of your chromatography system and the expected length of time until the next use. Use Table titled Pump storage recommendations on page 3-10 as a guide for determining when and in what sequence to perform certain storage procedures. Purging the pump on page 3-7 describes the purging procedure. Caution: To avoid damaging the pump, ensure that the pump always has solvent. Do not allow the pump to run dry. Operating as a stand-alone pump 3-9

Pump storage recommendations Status Purge with Water Purge with 20% Methanol Purge with Isopropanol For overnight storage For short-term storage For long-term storage For shipping Short-term storage Required Materials HPLC-grade, distilled, or deionized water 20% (v/v) methanol/water To prepare for short-term storage (up to a week) 1. Use HPLC-grade, distilled, or deionized water to purge the pump (see Purging the pump on page 3-7). 2. If the pump is to be idle for more than one day, use 20% (v/v) methanol/water as the final solvent to purge the pump. If 20% methanol/water is not fully miscible with the current solvent, purge with an intermediate solvent before switching to 20% methanol/water. 3. Press Run/Stop, then turn the power switch to the Off (O) position. Tip: All Reagent Manager parameter values are saved at their current settings when power is turned off. 3-10 Operating the Waters Reagent Manager

Long-term storage Required Materials HPLC-grade, distilled, or deionized water 20% (v/v) methanol/water Isopropanol Priming syringe Fitting plug or plastic film Small plastic bag and elastic band To store the pump for more than a week 1. If you use buffers, purge the pump with HPLC-grade, distilled, or deionized water (see Purging the pump on page 3-7). 2. Purge the pump with 20% (v/v) methanol/water. If 20% methanol/water is not fully miscible with the current solvent, purge with an intermediate solvent before switching to 20% methanol/water. 3. Disconnect the outlet tubing from the pump. 4. Place the inlet filter in isopropanol. 5. Open the prime/purge valve, then use the priming syringe to draw a minimum of 50 ml of isopropanol through the pump. 6. Pump a minimum of 5 ml of solvent through the filter/outlet. 7. Press Run/Stop, then turn the power switch to the Off (O) position. 8. With the inlet tubing connected to the pump, place the inlet filter in a small plastic bag and seal the bag against the tubing with an elastic band. 9. Plug the outlet port with the fitting plug, leave a length of outlet tubing on the pump, or cover the outlet port with plastic film. Operating as a stand-alone pump 3-11

Routine tasks Perform the following tasks on a routine basis to keep your Reagent Manager running smoothly: Replenish the seal wash solvent weekly or as needed. If the exterior requires cleaning, wipe the surface with a lint-free cloth moistened with water. Wipe up leaks and spills immediately. Empty and rinse the waste fluid containers when they are full or at least weekly. Remote start/stop operation For optional remote start and stop operation, the remote start/stop cable is required. This cable is included in the startup kit. See Connecting for remote start/stop operation on page 2-5 for instructions on how to install the remote start/stop cable. You can remotely start and stop the Reagent Manager using the Waters ACQUITY Binary Solvent Manager, the Waters 2695 Separations Module, or the Waters 600E Controller. Using the Waters ACQUITY binary solvent manager To start the Reagent Manager 1. Power-on the Waters Reagent Manager. 2. On the Waters ACQUITY binary solvent manager front panel, press Menu/Status. 3. Press Switch 1. 4. Press 1, and then press Enter. To stop the Reagent Manager 1. On the Waters ACQUITY binary solvent manager front panel, press Menu/Status. 2. Press Switch 1. 3-12 Operating the Waters Reagent Manager

3. Press 0, and then press Enter. 4. Power-off the Waters Reagent Manager. For more information, see the Waters ACQUITY UPLC System Operator s Guide. Using the Waters 2695 separations module To start the Reagent Manager 1. Power-on the Waters Reagent Manager. 2. On the Waters 2695 Separations Module front panel, press Menu/Status. 3. Press Switch 1. 4. Press 1, and then press Enter. To stop the Reagent Manager 1. On the Waters 2695 Separations Module front panel, press Menu/Status. 2. Press Switch 1. 3. Press 0, and then press Enter. 4. Power-off the Waters Reagent Manager. For more information, see the Waters 2695 Separation Module Operator s Guide. Using the Waters 600E controller To start the Reagent Manager 1. Power-on the Waters Reagent Manager. 2. On the Waters 600E controller front panel, press Direct Screen. 3. Press Switch 1. 4. Press 1, and then press Enter. Remote start/stop operation 3-13