Waters 432 Conductivity Detector Operator s Guide

Transcription

1 Waters 432 Conductivity Detector Operator s Guide /Revision B Copyright Waters Corporation 2010 All rights reserved

2 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 Alliance, Millennium, and Waters are registered trademarks of Waters Corporation, and Empower, LAC/E, PowerLine, SAT/IN, Sep-Pak, UltraWISP, WISP and THE SCIENCE OF WHAT S POSSIBLE. are trademarks of Waters Corporation. Other registered trademarks or trademarks are the sole property of their owners. ii

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. We seriously consider every customer comment we receive. You can reach us at 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 Telephone and fax From the USA or Canada, phone HPLC, or fax For other locations worldwide, phone and fax numbers appear in the Waters Web site. Conventional mail Waters Corporation 34 Maple Street Milford, MA USA 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 iii

4 Good Laboratory Practice, and consult your organization s safety representative for guidance. Safety advisories Consult Appendix A for a comprehensive list of warning and caution advisories. Operating this instrument When operating this instrument, follow standard quality-control (QC) procedures and the guidelines presented in this section. Applicable symbols Symbol Definition Manufacturer location Authorized representative of the European Community Confirms that a manufactured product complies with all applicable European Community directives Australia C-Tick EMC Compliant Confirms that a manufactured product complies with all applicable United States and Canadian safety requirements Consult instructions for use Audience and purpose This guide is intended for personnel who install, operate, and maintain the Waters 432 Conductivity Detector. iv

5 Intended use of the Waters 432 Conductivity Detector Use the Waters 432 Conductivity Detector, as a standalone module or configured as part of an HPLC system, to determine changes in the thermal conductivity of column eluent as compared with that from a reference flow. The Waters 432 Conductivity Detector is for research use only. Calibrating To calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards should include the entire range of QC samples, typical specimens, and atypical specimens. When calibrating mass spectrometers, consult the calibration section of the operator s guide for the instrument you are calibrating. In cases where an overview and maintenance guide, not operator s guide, accompanies the instrument, consult the instrument s online Help system for calibration instructions. Quality-control Routinely run three QC samples that represent subnormal, normal, and above-normal levels of a compound. Ensure that QC sample results fall within an acceptable range, and evaluate precision from day to day and run to run. Data collected when QC samples are out of range might not be valid. Do not report these data until you are certain that the instrument performs satisfactorily. 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. v

6 EC authorized representative Waters Corporation (Micromass UK Ltd.) Floats Road Wythenshawe Manchester M23 9LZ United Kingdom Telephone: Fax: Contact: Quality manager vi

7 Table of Contents Copyright notice... ii Trademarks... ii Customer comments... iii Contacting Waters... iii Safety considerations... iii Safety advisories... iv Operating this instrument... iv Applicable symbols... iv Audience and purpose... iv Intended use of the Waters 432 Conductivity Detector... v Calibrating... v Quality-control... v ISM classification... v ISM Classification: ISM Group 1 Class B... v EC authorized representative... vi 1 Introduction Features Method of operation Measurement technique Flow cell design Ion detection theory Installing the Detector Selecting the installation site Operating environment Required space Power requirements Table of Contents vii

8 Unpacking and inspection Unpacking Inspection AC power connection Power cord Required material Procedure I/O signal connections I/O signal descriptions PowerLine controller connections Empower and Millennium 32 connections Data module connections Chart recorder connections Chart marker input connections Auto Zero input connections Alliance Separations Module connections Making fluidic connections Cutting stainless steel tubing Cutting polymeric tubing Assembling compression fittings Connecting to the 432 Detector Installing the pulse dampener Passivating the system Verifying the detector Calibration procedure Operating the Detector Controls and indicators Power switch Display Keypad Beep function viii Table of Contents

9 Startup and shutdown Startup procedure Standby setup Long-term storage Operating recommendations Temperature equilibration Base range Integrator output Recorder output Polarity Eluent handling Performing Ion Analysis Fundamental considerations Water Containers High-pH eluents Sample preparation Configuring the system Pulse dampener Eluents for ion analysis Preparing anion eluent Preparing cation eluent Standards for ion analysis Preparing anion standards Injecting anion standards Preparing cation standards Injecting cation standards Required materials Injecting the standard Maintenance Routine maintenance Replacing the fuse Table of Contents ix

10 Maintaining the flow cell Cleaning the detector exterior Troubleshooting When you call Waters service Detector does not turn on Startup diagnostics Power supply Error messages Troubleshooting procedure Removing bubbles A Safety Advisories... A-1 Warning symbols... A-2 Task-specific hazard warnings... A-2 Specific warnings... A-3 Caution symbol... A-5 Warnings that apply to all Waters instruments... A-6 Electrical and handling symbols... A-12 Electrical symbols... A-12 Handling symbols... A-13 B Specifications... B-1 C Spare Parts... C-1 D Ion Chromatography Methods... D-1 General anion analysis using conductivity and UV detection... D-2 Preparing eluent... D-3 Preparing standards... D-3 Preparing a sample... D-4 Empower data processing method... D-5 Method validation... D-5 Method linearity... D-6 Quantitation precision... D-8 x Table of Contents

11 Method detection limits... D-9 Quantitation accuracy... D-10 Analyte recovery... D-10 Example of use... D-11 Using direct UV detection... D-11 Preparing lithium borate/gluconate 50X stock concentrate... D-13 Preparing lithium borate/gluconate eluent... D-14 Alkali and alkaline earth cations, ammonium, and amines... D-15 Preparing eluent... D-16 Preparing standards... D-16 Preparing a sample... D-17 Empower data processing method... D-18 Method detection limits... D-19 Examples of use... D-20 Preparing stock reagent... D-21 E Validation Support... E-1 Validation regulation overview... E-2 Waters regulatory compliance support... E-2 Basic operation... E-2 Instrument maintenance... E-3 Additional Waters support... E-3 Index... Index-1 Table of Contents xi

12 xii Table of Contents

13 1 Introduction Contents Topic Page Features 1-2 Method of operation 1-3 Ion detection theory

14 Features The Waters 432 Conductivity Detector is specifically designed to be integrated into chromatographic systems. The following features contribute to its performance in measuring the conductivity of column eluents: Unique 5-electrode flow cell design Heat exchanger and a built-in automatic temperature control system for stable operation Auto baseline/auto zero External recorder/integrator and chart mark connections Three time constant selections Leak-detected alarm signal Waters 432 conductivity detector Waters 432 Conductivity Detector IN OUT 1-2 Introduction

15 Method of operation This section discusses the method of operation of the 432 Detector. Additional descriptive information appears in these sections: I/O signal descriptions on page 2-8 Controls and indicators on page 3-2 Appendix B Measurement technique The 432 Detector responds to all ions present in the flow cell, since all ions in solution conduct electricity. This allows the 432 Detector to detect a wide variety of sample ions. The 432 Detector eliminates the eluent s contribution to conductivity with an electronic technique called baseline suppression. The detector measures the eluent conductivity and assigns it a value of zero. Thus, any sample ions appear as positive or negative measurements, relative to the baseline. The temperature of an ionic solution affects the conductivity of the ions. Generally, a solution s conductivity rises about 2% for every degree Celsius of temperature increase. The special flow cell heater in the 432 Detector minimizes the effect of ambient temperature fluctuations on measurement accuracy. Flow cell design The flow cell in the 432 Detector contains five electrodes connected in a measuring circuit: two reference electrodes, two detection electrodes, and a guard electrode that provides a local electrical ground. Column eluent flows through the heater to attain the set temperature, and then flows through the cell, directly contacting the electrodes. The 5-electrode design permits measurement of conductivity to be made with a very low current at the detection electrodes. The low current employed eliminates impedance and other problems associated with simpler designs, and results in a stable baseline and an extended range of linearity. Method of operation 1-3

16 Flow cell schematic =Reference electrodes 2=Detection 1= Reference electrodes Electrodes 3=Guard 2= Detection electrode Electrodes 3= Guard Electrode Flow cell Cell block Block (heated) (heated) Fluid outlet Outlet Ion detection theory The conductance of a solution of known concentration can be calculated using the following equation: G = λc 10 3 G = measured conductance of the solution, in Siemens (1 S = ohm 1 ) C = concentration in equivalents per 1000 cm 3 K = length/area of cell (the cell constant) λ = equivalent conductance in S cm 2 equiv Introduction

17 The table below lists the equivalent conductances of some common ions. 1 Concentrations above 10-5 to 10-3 N, generally exhibit decreased equivalent conductance due to interionic effects. Limiting equivalent conductance of ions in water at 25 C Cations l + Anions λ H OH Li F 55.4 Na Cl 76.4 K Br 78.1 Rb I 76.8 Ag NO NH ClO (CH 3 ) 2 NH ClO Hg IO Mg Formate 54.6 Ca Acetate 40.9 Ba Benzoate 32.4 Cu SO Zn CO La Fe(CN) Ce Henry H. Bauer et al., eds. Instrumental Analysis, Allyn and Bacon, Boston (1978), p Reprinted with permission from the publisher. Ion detection theory 1-5

18 1-6 Introduction

19 2 Installing the Detector This chapter guides you through the following steps in preparing the 432 Detector for operation in a chromatographic system: Selecting an installation site that satisfies the detector s power and environmental requirements Unpacking and inspecting the 432 Detector and accompanying items Connecting the detector to your AC power supply Connecting the detector electrically to the other components of your chromatographic system Connecting the detector inlet to the column and the detector outlet to a waste receptacle (and, if required, installing the pulse dampener) Passivating the detector and other post-column fluid path components After you have successfully completed this chapter, familiarize yourself with the information in Controls and indicators on page 3-2. When you are ready to operate the detector, perform the startup procedure described in Startup and shutdown on page 3-8. Contents Topic Page Selecting the installation site 2-2 Unpacking and inspection 2-3 AC power connection 2-4 I/O signal connections 2-7 Making fluidic connections 2-23 Passivating the system 2-29 Verifying the detector

20 Selecting the installation site Operating environment The 432 Detector operates in any standard laboratory environment that provides suitable electrical power and remains within the following ranges: Temperature: 5 to 35 C (40 to 95 F) Humidity: 20 to 80%, noncondensing Install the instrument in a clean area that is free from exposure to: Temperature or humidity extremes, which can be found near direct sunlight, heat registers, and air conditioning vents Strong electromagnetic radiation, such as from large motors or arcing contacts Appreciable shock or vibration Required space The 432 Detector requires bench space that measures approximately: 10 inches (25 cm) high 14 inches (34 cm) wide 24 inches (60 cm) deep Caution: Make sure that air can circulate freely through the ventilation slots on both side panels. Power requirements The 432 Detector requires: One properly grounded AC voltage outlet. Correct voltage and fuse selections as shown in the table titled Nominal operating voltage on page Installing the Detector

21 Unpacking and inspection Unpacking The 432 Detector is shipped in one carton that contains the following items: Waters 432 Conductivity Detector Startup Kit Validation certificate Waters 432 Conductivity Detector Operator s Guide Packing list Declaration of conformity Tip: If you purchased the 432 Detector as part of an ion/liquid chromatograph system, a Waters representative will perform the installation and startup. Inspection To unpack the 432 Detector: 1. Locate the packing list. 2. Unpack the contents of the shipping carton and check the contents against the packing list to make sure that you received all items. 3. Check the contents of the Startup Kit against the Waters 432 Conductivity Detector Startup Kit List. 4. Save the shipping carton for future transport or shipment. Inspect all items. If you find any damage or discrepancy, immediately contact the shipping agent and Waters. For more information about the instrument warranty, refer to Waters Licenses, Warranties, and Support. If the shipment is complete and undamaged, record the installation date and serial number of the 432 Detector in the spaces provided in Appendix C. Unpacking and inspection 2-3

22 AC power connection Warning: To avoid a potential fire hazard and damage to the 432 Detector, make sure that the voltage selector in the power connector is set correctly to match the available AC power source, and that the correct fuses are installed before you apply AC power. Power cord The power connector is located on the lower-right corner of the rear panel. If a power plug other than the one supplied is needed for your location, consult the table titled Power cord wire identification on page 2-5 and observe the existing applicable regulations. 2-4 Installing the Detector

23 Rear panel IEEE DIP switch cover Power cord wire identification Wire (USA) Wire (International) Connection Black Brown Hot White Blue Neutral Green Green/Yellow Ground (Earth) The 432 Detector can be adapted to operate within two voltage ranges at 50 or 60 Hz. The table below describes these voltage ranges and the fuse value that is appropriate to each. AC power connection 2-5

24 Nominal operating voltage Nominal Voltage (VAC) Fuse 100/120 T 2A 220/240 T 1A Required material Procedure You need a flat-blade screwdriver to perform this procedure. Warning: To avoid the possibility of electrical shock, turn off the front panel power switch and unplug the power cord. To change the operating voltage setting: 1. Remove the power cord from its connector on the rear panel of the controller and pry open the power connector cover with a flat-blade screwdriver. 2. Remove the voltage selection barrel and locate the correct voltage setting. 3. Reinstall the voltage selection barrel so the desired voltage setting appears through the window when you close the power connector cover. 2-6 Installing the Detector

25 Changing the voltage setting Voltage setting TP Determine if you need to change the fuses (see the table titled Nominal operating voltage on page 2-6). All units are supplied with two 2-A fuses installed for 100/120 volt operation. If you operate the unit on 220/240 volt power, change the fuse as outlined in Replacing the fuse on page Reinstall the power connector cover and the power cord. I/O signal connections The 432 Detector is usually installed as an integral part of a data collection system. You can control the 432 Detector either locally from the keypad on the front panel or remotely from a PowerLine controller, such as the Waters 600S. This section describes the detector s I/O signals and how they connect to the following devices: PowerLine controller Empower or Millennium 32 software Data module SAT/IN module Chart recorder I/O signal connections 2-7

26 Device signalling the Chart Marker input Device signalling the Auto Zero input Caution: To meet the regulatory requirements of immunity from external electrical disturbances that may affect the performance of this instrument, do not use cables longer than 9.8 feet (3 meters) when connecting to the screw-type barrier terminal strips. In addition, ensure you always connect the shield of the cable to chassis ground at one instrument only. I/O signal descriptions The 432 Detector rear panel has an IEEE-488 connector for communication with a PowerLine controller, and a terminal strip for the input/output signals. These signals are described in the table titled I/O signal descriptions on page Installing the Detector

27 I/O terminal strip + + Int INT Rec REC LEAK Leak + MARKER Marker in IN MARKER Marker out OUT + AUTO Auto zero ZERO I/O signal descriptions Terminal pairs Rec (+ and ) Int (+ and ) Marker Out Function Recorder output A 10-mV full-scale analog output signal appears on these terminals. The measurement range is determined by the product of the Base Range and Sensitivity settings: for example, 500 μs (base range) x (sensitivity) = 2.5 μs full scale. Integrator output A 1-V full-scale analog output signal appears on these terminals. The measurement range is selectable: 10, 50, or 100 μs full scale. Marker output A 1-second contact closure signal appears on these terminals when either of the following events occurs: The Chart Mark key on the keypad is pressed A contact closure signal occurs between the Marker In terminals I/O signal connections 2-9

28 I/O signal descriptions (Continued) Terminal pairs Leak Auto Zero (+ and ) Marker In (+ and ) Function Leak Alert output A contact closure signal appears on these terminals if a leak is detected inside the detector. Auto Zero input The voltage at the Recorder and Integrator output terminals is set to the user-selected balance offset level when a contact closure occurs between these terminals. Marker input A chart mark (~0.5 mv for 3 seconds) is added to the Recorder output signal when a contact closure signal appears between these terminals. Required material To connect cables to the I/O terminals, use a small flat-blade screwdriver. Other rear panel connections and DIP switch In addition to the I/O terminal strip, the rear panel also contains the following items: IEEE-488 connector Communication bus for use with a Waters PowerLine system controller, such as the Waters 600S. DIP switch Sets the IEEE-488 address seen by the system controller. Ground lugs Used to connect the 432 Detector to an earth ground connection and also used as a chassis ground connection to other system instruments. PowerLine controller connections The 432 Detector can be programmed remotely by a PowerLine controller (such as the Waters 600S) via the IEEE-488 data communications bus. Required material You need a 2.5-mm Allen wrench to connect to the 432 Detector Installing the Detector

29 Procedure To connect the 432 Detector to a PowerLine controller: 1. Turn off the PowerLine controller and the 432 Detector. 2. Plug one end of the IEEE-488 cable (included in the Startup Kit) into the bus connector on the rear panel of the 432 Detector (see the figure Rear panel on page 2-5) and the other end of the cable into the bus connector on the PowerLine controller. 3. Remove the DIP switch cover (see the figure Rear panel on page 2-5) using a 2.5-mm Allen wrench. 4. Refer to the table titled IEEE-488 DIP switch setting on page 2-12 to set the DIP switches on the rear panel of the 432 Detector (see the figure IEEE-488 address switch on page 2-11) to a unique IEEE-488 address between 2 and After you set the DIP switches, reinstall the DIP switch cover. Tip: To operate the 432 Detector in local mode, press the front panel Remote key. The illuminated light above the key will go out. IEEE-488 address switch Switch 1 OFF (Address 8 Shown) Switch 5 I/O signal connections 2-11

30 The IEEE-488 address DIP switch employs positive logic to determine the address of the 432 Detector from the switch settings. The table below shows the settings for valid addresses. IEEE-488 DIP switch setting IEEE-488 DIP switch settings address OFF ON OFF OFF OFF 3 ON ON OFF OFF OFF 4 OFF OFF ON OFF OFF 5 ON OFF ON OFF OFF 6 OFF ON ON OFF OFF 7 ON ON ON OFF OFF 8 OFF OFF OFF ON OFF 9 ON OFF OFF ON OFF 10 OFF ON OFF ON OFF 11 ON ON OFF ON OFF 12 OFF OFF ON ON OFF 13 ON OFF ON ON OFF 14 OFF ON ON ON OFF 15 ON ON ON ON OFF 16 OFF OFF OFF OFF ON 17 ON OFF OFF OFF ON 18 OFF ON OFF OFF ON 19 ON ON OFF OFF ON 20 OFF OFF ON OFF ON 21 ON OFF ON OFF ON 22 OFF ON ON OFF ON 23 ON ON ON OFF ON 24 OFF OFF OFF ON ON 25 ON OFF OFF ON ON 26 OFF ON OFF ON ON 2-12 Installing the Detector

31 IEEE-488 DIP switch setting (Continued) IEEE-488 address 27 ON ON OFF ON ON 28 OFF OFF ON ON ON 29 ON OFF ON ON ON PowerLine operation DIP switch settings Under PowerLine control, the 432 Detector is recognized as a 431 Detector and it retains the functionality of the 431 Detector with the following differences: The Balance field on the detector setup page of the PowerLine controller affects the Integrator Balance and the Integrator Output only. When you press the Setup key on the controller, the selected Balance value is sent to the 432 Detector from the PowerLine controller. However, the 432 Detector output does not change to the selected balance until the detector is autozeroed by a contact closure at the Auto Zero input terminals on the rear panel (remote or local mode) or when you press the Auto Zero key on the front panel (local mode only). Under PowerLine control, the 432 Detector retains the full functionality of local mode operation, except for the following differences: The Recorder Sensitivity ranges of and are not accessible. The Integrator Sensitivity ranges are not accessible. The 432 Detector does not automatically perform an Auto Zero after an Auto Base routine has occurred. Empower and Millennium 32 connections Empower and Millennium 32 software perform data acquisition, processing, and management of chromatographic information. This software requires the detector s analog signal to be converted to a digital form. I/O signal connections 2-13

32 Empower and Millennium 32 are menu-driven applications specifically designed by Waters for chromatographers. Use the software to: Acquire data Process data Generate and print reports Store information (or data) in a central area and share this information with users who have proper security access To connect the 432 Detector to an Empower or Millennium 32 computer, be sure to: Connect the Bus Satellite Interface (SAT/IN) module to the Bus Laboratory Acquisition and Control/Environment (LAC/E ) card in the Empower computer, Millennium 32 computer, acquisition client, or LAC/E 32. Connect the 432 Detector to the Bus SAT/IN module (Channel 1 or 2). Remove the IEEE-488 cable from the rear panel of the 432 Detector, if it is connected. The 432 Detector is in local mode when it is connected to an Empower and Millennium 32 computer. Bus SAT/IN module The Waters Bus SAT/IN module translates analog signals into digital form. It then transmits these digital signals to the Bus LAC/E card inside the workstation, acquisition client, or LAC/E Installing the Detector

33 Bus SAT/IN module (front panel) Waters SAT/IN Module EVENTS CH1 CH2 CHANNEL 1 CHANNEL 2 IN + OUT IN + OUT CH 1 CH 2 OK Tip: To prevent damage to the unit, always disconnect the power cord at either the wall outlet or the power supply before you attach or remove the power connection to the Bus SAT/IN module. The Bus SAT/IN module does not have a power switch. Connecting the Bus SAT/IN module to the Bus LAC/E card The Bus SAT/IN module connects to the Bus LAC/E through an I/O distribution box. To connect the Bus SAT/IN module to the Bus LAC/E card: 1. Use the I/O distribution cable to connect the I/O distribution box to the 9-pin I/O distribution port on the Bus LAC/E card at the back of the Millennium 32 computer. 2. Use a serial cable to connect the data terminal on the back of the Bus SAT/IN to a port of the I/O distribution box. 3. Configure the serial port for the Bus SAT/IN module as described in the Empower or Millennium 32 installation and configuration guides. I/O signal connections 2-15

34 Bus SAT/IN to Bus LAC/E connections I/O Distribution Por t (9-pin) of Bus LAC/E Card I/O Distribution Cable Modified Modular Jack Connections I/O Distribution Box Serial Cable SAT/IN Module Rear Panel Connect SAT/IN to Port 1 on the I/O Distribution Box BCD DATA PWR AC to DC Converter 2-16 Installing the Detector

35 Connecting the Bus SAT/IN module to the 432 Detector The Bus SAT/IN module connects to the 432 Detector as shown below. Refer to the procedure following the figure and the table titled Bus SAT/IN cable connections on page 2-18 for complete details. Caution: To prevent damage to the unit, do not plug in the power cord of the Bus SAT/IN module until you perform all of the procedures described in the Waters Bus SAT/IN Module Installation Guide. Bus SAT/IN to 432 Detector connections Waters 432 Detector Red Black + + INT REC LEAK Waters SAT/IN Module CHANNEL 1 CHANNEL 2 EVENTS CH1 CH2 IN OUT IN OUT CH CH 1 2 OK + + MARKER IN MARKER OUT AUTO ZERO TP01264 To connect the 432 Detector to the Bus SAT/IN module: 1. Connect the white wire of the analog cable (included with the Bus SAT/IN module) to the Int + terminal on the rear panel of the 432 Detector. Connect the black wire to the Int terminal. I/O signal connections 2-17

36 2. Connect the other end of the cable to either the Channel 1 or Channel 2 connector on the front panel of the Bus SAT/IN module. 3. Connect the Event In terminals of the channel you chose in the previous step to the Inject Start output signal of the Waters Alliance solvent delivery system or the Waters 717plus (or equivalent) Autosampler. 4. Remove the IEEE-488 cable from the rear panel of the 432 Detector, if it is connected. The connections from the 432 Detector to the Bus SAT/IN are summarized below. Bus SAT/IN cable connections 432 Detector I/O connector terminal Bus SAT/IN cable Bus SAT/IN connector Int (+) White wire Int ( ) Black wire Channel 1 or 2 Data module connections This section describes how to connect the analog output signal from the 432 Detector to the Waters 746 Data Module. Caution: Remember to meet the regulatory requirements of immunity from external electrical disturbances that may affect the performance of this instrument, do not use cables longer than 9.8 feet (3 meters) when connecting to the screw-type barrier terminal strips. In addition, ensure you always connect the shield of the cable to chassis ground Installing the Detector

37 Analog signal To send the analog output signal from the 432 Detector to a Waters data module, connect the signal cable in the 432 Detector Startup Kit as described below. Data module signal cable connections Wire 432 Detector I/O connector terminal 746 terminal Red Int (+) (+) Black Int ( ) ( ) Shield Ground lug None Marker out signal The Marker Out terminals of the 432 Detector provide a contact closure output signal when either of the following events occurs: Chart Mark key is pressed Marker In terminals are shorted together Use the signal to start a Waters 746 Data Module by connecting a signal cable to the module s data cable. Data module chart mark cable connection Wire 432 Detector I/O connector terminal 746 cable Either wire Marker Out Join to both Remote Start wires (white and red) Other wire Marker Out Green wire Chart recorder connections To connect the 432 Detector to a chart recorder: 1. Attach the Recorder cable (see Appendix C) to the 432 Detector REC output terminals, as indicated in the table titled Chart recorder cable connections on page I/O signal connections 2-19

38 2. Connect the cable shield to the ground lug on the 432 Detector rear panel. 3. Connect the other end of the cable to the 10-mV input terminals on the chart recorder. Chart recorder cable connections Wire 432 Detector I/O connector terminal Chart recorder terminal Red Rec (+) Pen (+) Black Rec ( ) Pen ( ) Chart marker input connections The 432 Detector accepts a chart mark (start inject) signal from the following devices: Waters 717plus Autosampler Any other device that provides a compatible switch closure Waters 717plus Autosampler To connect the 432 Detector to a Waters 717/717plus Autosampler, connect a signal cable as indicated in the table below. Autosampler chart mark cable connections 432 Detector I/O connector terminal Marker In (+) Marker In ( ) Autosampler terminal Either Inject Start terminal of a pair Other Inject Start terminal of the same pair 2-20 Installing the Detector

39 Auto Zero input connections The voltage at the Recorder and Integrator outputs is set to the user-selected balance offset level when a contact closure occurs between the Auto Zero terminals. This section describes how to connect the 432 Detector to the following devices (so that an auto zero occurs at the injection point): Waters 717plus Autosampler Any other device that provides a compatible switch closure Waters 717plus Autosampler To connect the 432 Detector to a Waters 717plus Autosampler, connect a signal cable as indicated in the table below. Autosampler Auto Zero cable connections 432 Detector I/O connector terminal Auto Zero (+) Auto Zero ( ) Autosampler terminal Either Inject Start terminal of a pair Other Inject Start terminal of the same pair Alliance Separations Module connections Connect the detector to Waters Alliance Separations Modules, when it is not under the control of the Millennium 32 software, to perform the following tasks: Auto-Zero on inject Chart mark on inject Method start Generating Auto-Zero on inject To generate the Auto-Zero function on the 432 Detector at the start of an injection, make the connections summarized in the table titled Connections for generating Auto-Zero on inject on page 2-22 and illustrated in the figure I/O signal connections 2-21

40 Alliance Separations Module connections to the 432 Detector Auto-Zero on inject on page Connections for generating Auto-Zero on inject Alliance Separations Modules (B inputs and outputs) 432 Detector (A inputs) Pin 1 Inject Start Auto-Zero (+) Pin 2 Inject Start Auto-Zero ( ) Before you can generate an Auto-Zero from an Alliance Separations Module, you must configure the Auto-Zero signal at the 432 Detector front panel. The default Auto-Zero signal is Low. Alliance Separations Module connections to the 432 Detector Auto-Zero on inject Waters Alliance B (inputs and outputs) Inject start + Inject start Ground Stop flow + Stop flow Hold inject 1+ Hold inject 1 Hold inject 2 + Hold inject 2 Ground Chart out + Chart out Waters 432 Detector A (inputs) + Int 1 Inject Start + 2 Inject Start Ground Rec 4 Lamp On/Off + 5 Lamp On/Off - Leak 6 Chart Mark + 7 Chart Mark Ground Marker in 9 Auto-Zero + 10 Auto-Zero - Marker out + Auto Zero Generating chart mark on inject To generate the chart mark function at the start of an injection, make the connections summarized in the table below and illustrated in the figure 2-22 Installing the Detector

41 Alliance Separations Module connections to the 432 Detector chart mark on inject on page Connections for generating chart mark on inject Alliance Separations Modules (B inputs and outputs) 432 Detector (A Inputs) Pin 1 Inject Start Marker In (+) Pin 2 Inject Start Marker In ( ) Before you can generate a chart mark from an Alliance Separations Module, you must configure the chart mark signal at the front panel. The default chart mark signal is Low. Alliance Separations Module connections to the 432 Detector chart mark on inject Waters Alliance B (inputs and outputs) Inject start + Inject start Ground Stop flow + Stop flow Hold inject 1+ Hold inject 1 Hold inject 2 + Hold inject 2 Ground Chart out + Chart out Waters 432 Detector A (inputs) + Int INT 1 Inject Start + 2 Inject Start Ground REC Rec 4 Lamp On/Off + 5 Lamp On/Off - LEAK Leak 6 Chart Mark + 7 Chart Mark + - MARKER 8 Ground IN Marker in 9 Auto-Zero + 10 Auto-Zero - MARKER Marker out OUT + AUTO Auto Zero ZERO Making fluidic connections Fluid lines to a column and waste container connect to the front of the 432 Detector, as shown in the figure Fluid connections on page To make these connections: Making fluidic connections 2-23

42 Cut the tubing. Assemble compression fittings and ferrules. Connect the tubing to the detector. This section will guide you through each of these procedures. Caution: Conductivity detection is sensitive to flow rate fluctuations. If you use a non-waters pump or a Waters pump without the SILK microflow compensation algorithm, you must install the pulse dampener kit supplied in the Startup Kit for optimum performance. Refer to the installation procedure in this section. Fluid connections Waters 432 Conductivity Detector In In from From column Column IN OUT Out To to Waste waste (18 inches. (18 inches, inch inch I.D.) ID) Cutting stainless steel tubing You need the following tools to cut stainless steel tubing: A file with cutting edge Two cloth- or plastic-covered pliers 2-24 Installing the Detector

43 To cut the tubing: 1. Measure the length of 1/16-inch OD, inch ID, stainless steel tubing you need to make the following connections: Column to the detector inlet Detector outlet to a suitable waste container 2. Use a file with a cutting edge to scribe the circumference of the tubing at the desired length. 3. Grasp the tubing on both sides of the scribe mark with cloth-covered pliers. Gently work the tubing back and forth until it separates. 4. File the ends smooth. Cutting polymeric tubing Waters chromatography systems are supplied with a tubing cutter (similar to the one in the figure Cutting polymeric tubing on page 2-26) to facilitate cutting polymeric tubing. This section presents the recommended procedure for using the tubing cutter. Tip: To avoid bandspreading caused by angled cuts, always use a tubing cutter. Angled cuts leave unswept dead volumes at the connection junction due to the poor fit of the tubing against the connector or port. To cut a length of polymeric tubing: 1. Estimate the length of tubing required to connect the components. Allow slack so that the tubing is not pulled tightly around sharp corners. 2. Insert the tubing into the cutter so that the tubing extending from the metal side is the length required. Use the proper hole to have a snug enough fit so that the tubing is not flexed by the blade when you cut it. Making fluidic connections 2-25

44 Cutting polymeric tubing 3. Press down on the razor blade to cut the tubing. Discard the excess tubing that extends from the clear side of the cutter. 4. Inspect the cut for burrs or scratches and for the perpendicularity of the cut. Assembling compression fittings To assemble each compression fitting: 1. Slide the compression screw over the tubing end, followed by the ferrule. 2. Mount the ferrule with its taper end facing the end of the tubing. Ferrule and compression screw assembly Compression Compression Screw screw Ferrule inch ID tubing inch (0.23 mm) I.D. Tubing 0.23 mm) 2-26 Installing the Detector

45 Connecting to the 432 Detector To make connections at the column outlet and detector inlet, and at the detector outlet: 1. Install a compression screw and then a ferrule on the length of inch tubing from the column outlet. Use stainless steel fittings on stainless steel tubing and PEEK fittings on PEEK tubing. Tip: If you are using a column with 1/4 28 end fittings and there is a length of tubing with 1/4 28 fittings on each end, use the 1/4 28 to Z-detail adapter (included in the Startup Kit) to connect this tubing to the tubing that leads to the detector inlet. The Waters IC-Pak C column comes supplied with a length of tubing that has a 1/4 28 fitting on one end (column outlet) and a Waters compression screw and ferrule on the other end (detector inlet). 2. Push the free end of the tubing as far as it will go into the IN fitting on the 432 Detector. While you hold it there, use a 5/16-inch open-end wrench to tighten the compression screw 3/4-turn past finger-tight. Tip: The 432 Detector and IC-Pak series of columns have very deep ferrules. 3. Remove the compression screw and tubing from the connection and verify that fluid can flow freely. 4. Reconnect the tubing to the IN fitting, making sure to push the tubing all the way into the fitting. 5. Install a ferrule on an 18-inch length of inch tubing and connect it to the OUT connection on the 432 Detector. Use stainless steel fittings on stainless steel tubing and PEEK fittings on PEEK tubing. 6. Place the other end of the tube in a waste container. If you are using any Teflon tubing, attach it after the stainless steel or PEEK tubing. Installing the pulse dampener To achieve the best performance from the 432 Detector in a chromatographic system with a non-waters pump, Breeze software, or Waters HPLC 515 Pump, you must install the pulse dampener kit supplied in the Startup Kit. The pulse dampener is not required if you are using a Waters 2695 Separations Module. Making fluidic connections 2-27

46 To install the pulse dampener between the pump and the injector: 1. Assemble the pulse dampener (see the figure Pulse dampener on page 2-28) using the instructions in the pulse dampener kit. 2. Connect the large-id (0.020-inch) tubing to the pump outlet using a stainless steel compression screw and ferrule. 3. Connect the small-id (0.009-inch) tubing to the injector inlet using a stainless steel compression screw and ferrule. 4. Disconnect the tubing from the injector inlet. 5. Pump ASTM Type I reagent water at 2 ml/min through the pulse dampener assembly until you see a constant stream exiting from the restrictor assembly outlet line. 6. Reconnect the tubing to the injector inlet. Pulse dampener inch I.D inch Tubing ID tubing inch I.D inch Tubing ID tubing From pump Pum p Union To To injector Injector Low Low pressure Pressure FilterA filter assembly ssembly Restrictor Restrictor assembly Assembly 2-28 Installing the Detector

47 Passivating the system Passivating the system removes potential contamination from the wetted surfaces of all system components. Perform passivation on a new system, and subsequently, whenever you suspect that contamination may have occurred. See Troubleshooting on page 5-6, for help diagnosing performance problems. Use this procedure for Waters hardware only. For other equipment, check with the manufacturer before you continue with this procedure. Caution: If you are installing the 432 Detector into an existing Waters system, replace the pump seals before you passivate. Use the new pump seals supplied in the Startup Kit and refer to the replacement procedure in the pump manual. To passivate the system: 1. Replace the column with a union fitting. 2. If the system is not new, flush it thoroughly with ASTM Type I reagent water to remove any residual solvents or salts. 3. Connect the power cord to the 432 Detector and plug the other end into an AC power outlet. Push the 432 Detector power switch to turn on the instrument. Warning: To avoid chemical hazards, always wear safety glasses and gloves when you are using solvents. 4. Prime the pump with 6 N nitric acid (HNO 3 ) and run it at a flow rate of 1.2 ml/min for 20 minutes to passivate all the wetted parts of the detector. Press the Clear key to stop the overrange alarm. 5. Stop the pump. 6. Remove the inlet line from the nitric acid and place it in ASTM Type I reagent water. 7. Flush the system using one of the following methods: Prime and start the pump, then flush it with ASTM Type I reagent water at 1.2 ml/min until you observe a consistent reading of less than 20 μs (base range set to 50 μs). Passivating the system 2-29

48 Flush the system overnight with 100% methanol at a reduced flow rate. By the next morning the system will be passivated and ready for use. Tip: If you are using a pump with seal-wash capability, skip step Use a syringe to flush the back of the pump seals and pistons by slowly running about 5 ml of water into the top hole in the baseplate of the pump heads. Place a tissue under the baseplates to absorb the water. 9. Set the pump flow rate to 0.0 ml/min. It is not necessary to turn off the 432 Detector unless it will be idle for an extended period (14 days). For best results, always leave the power on to maintain cell temperature; it takes a minimum of 2 to 3 hours once the detector is turned on to equilibrate the flow cell at the selected operating temperature. Verifying the detector This procedure is a guideline for verifying that the detector works correctly within its expected operational range. The detector is calibrated before shipping, and recalibration is not normally required. Verify the detector when any of these conditions apply: When you replace the flow cell To verify accuracy When you make adjustments Calibration procedure Tip: You need solution of 1 mm potassium chloride (KCl) to calibrate the detector. Tip: Waters suggests one of its Technical Service Representatives perform this procedure. To perform the calibration procedure: 1. Turn on the 432 Detector and set the temperature control to 35 C. Allow 2 to 3 hours for the temperature in the flow cell to equilibrate. 2. Set the base range to 200 μs. 3. Set the Filter Time Response to Fast Installing the Detector

49 4. Pump 1 mm KCl solution through the detector (without a column in place). 5. Verify that the front panel output is 147 μs ± 5 μs. Verifying the detector 2-31

50 2-32 Installing the Detector

51 3 Operating the Detector This chapter contains: A description of front panel controls and displays Procedures for starting up, shutting down, and long-term storage Recommended operating practices Contents Topic Page Controls and indicators 3-2 Startup and shutdown 3-8 Operating recommendations

52 Controls and indicators The figure below illustrates the controls and indicators on the front panel of the 432 Detector. Front panel Waters 432 Conductivity Detector CONDUCT ( S/cm) BASE ( S/cm/FS) SENS Remote Temp. Pol. Chart Mark Auto Base Base Range Auto Zero Sens. Range Resp. Bal. 0. Clear Shift Enter IN OUT ON OFF Power switch Display The power switch (located in the lower-right corner of the front panel) controls power to the 432 Detector. Upon startup, an initialization routine verifies the data in ROM memory, tests RAM memory function, and checks for any internal leakage or an eluent conductivity over-range condition. The display shows instrument status and parameter values in two 20-character lines of text. Upon startup, Waters 432 Self Check appears 3-2 Operating the Detector

53 Keypad briefly. If any error conditions are detected during startup or normal operation, the appropriate error message is displayed. The main screen shows the measured conductivity, as well as the base range and sensitivity settings. When you set an operating parameter, the display shows the selected or entered value. Error messages A corresponding error message is displayed if one of the following conditions occurs: ROM/RAM error (checked during startup only) Error: ROM/RAM Leakage detected Error: Leak Temperature control failure Error: Temp Over-range (above base range setting) Error: Over Range Overflow (above 10,000 μs) Error: Over Flow Press the Clear key to clear an error alarm and message. For a continuing error condition, the error message remains after the audio alarm is cleared. Use the keypad to control the operation of the 432 Detector. The table titled Key descriptions on page 3-4 describes the function of each key. Controls and indicators 3-3

54 Tip: Three keys (Balance, Sensitivity Range, and the numeral 1) perform an alternate function when they are preceded by the Shift key. Key descriptions Key Remote Pol. Base Range Sens. Range Function Remote key: Toggles between local and remote operating modes. In remote mode, the light above the key is on and all other front panel controls are disabled. Polarity key: Toggles the polarity of the signal to the external chart recorder and integrator. When positive polarity is selected, the light above the key is illuminated. Base Range key: Sets the base sensitivity range of the 432 Detector to the appropriate value for the eluent being used. The base sensitivity is set to one of ten steps, from 10 μs (maximum gain) to 10,000 μs, using the Up and Down keys or the numeric keypad. Sensitivity Range key: Sets the sensitivity range multiplier of the 432 Detector. The sensitivity range has twelve steps, from (maximum sensitivity) to 1.0 (available only with 100 μs multiplier setting), and is set using the Up and Down keys or the numeric keypad. The 10-mV full-scale recorder response is calculated by multiplying the Base Range by the Sensitivity Range to obtain a value of x μs / 10 mv FS. The recorder range is 1 to for the 100 μs setting and 0.1 to for the two lower settings. Shift key then Sensitivity Range key: Sets the sensitivity range multiplier of the integrator to 100, 50, or 10 μs using the Up and Down keys or the numeric keypad; the integrator output is 100, 50, or 10 μs/1 V, respectively. 3-4 Operating the Detector

55 Key descriptions (Continued) Key Bal. Temp. Resp. Auto Zero Function Balance key: Manually sets the offset (%) of the signal to the external chart recorder. (Use the numeric keypad or Up and Down keys.) Shift key then Balance key: Manually sets the offset (%) of the signal to the integrator. (Use the numeric keypad or Up and Down keys.) Shift key after Balance key: Changes the polarity of the offset. Allowable values are 100 to +100%. Temperature key: Sets the temperature of the detection cell. Use the Up and Down keys or the numeric keypad to turn temperature control off (Setting 0) or select one of the following eight settings: 30, 35, 40, 45, 50, 55, 60, or 65 C. The light above the key is illuminated when the temperature control is on. Response key: Sets the response time constant of the 432 Detector to optimize signal-to-noise ratio. Use the Up and Down keys or the numeric keypad to choose Setting 1 (Fast, 0.25 sec) for very narrow peaks, Setting 2 (Standard, 0.5 sec), or Setting 3 (Slow, 1.0 sec) to detect wider peaks. Setting 2 is used for most applications. Auto Zero key: Automatically zeros the Recorder and Integrator signals to the specified Recorder Balance and Integrator Balance offsets, respectively. Auto Base Auto Base key: Automatically sets the base range of the 432 Detector to the appropriate value for the eluent being used. This is the next highest setting above the actual background conductivity of the eluent. Controls and indicators 3-5

56 Key descriptions (Continued) Key Shift Function Shift key: Press the Shift key before, not along with, other keys to access additional functions and also to change polarity when you set balance offset values. When the Shift key is pressed, an asterisk (*) appears at the right side of the display; press Shift again to return to normal mode. Shift key then Balance key: Displays the integrator balance offset value. When setting the balance offset, press Shift to change polarity. Shift key then Sensitivity Range key: Displays integrator range value. Chart Mark Enter Clear Shift key then 1 key: Displays the current, actual value of the chart recorder balance offset. Press Enter to return to the main screen. Chart Mark key: When this key is pressed, a 1-second, 1-mV signal is sent to the Recorder terminals and a 1-second contact closure is sent to the Marker Out terminals. Enter key: When you manually set offsets, sensitivity range, or base range, pressing Enter records the displayed value and returns the display to the main screen. The Enter key is also used to access the beep setting function. Clear key: Erases a value input from the keypad. The Clear key is also used to clear an error alarm and message. For a continuing error condition, the error message remains after the audio alarm is cleared. Up key: Increments the current parameter setting. 3-6 Operating the Detector

57 Key descriptions (Continued) Key Function Down key: Decrements the current parameter setting. Beep function You can set the 432 Detector to beep when a key is pressed and/or an error condition is detected. Use the Clear key to stop an error alarm. For a continuing error condition, the error message remains after the beep is cleared. To set the beep function, follow the procedure in the table below. Setting the beep function Keystroke Key Function 1 Enter Accesses the beep function parameters. 2. (decimal point) 3 Enter 4 1 Turns on the beep-on-error function. 0 Turns off the beep-on-error function. 5 1 Turns on the beep-on-keystroke function. 0 Turns off the beep-on-keystroke function. 6 Enter Saves settings and returns to the main screen. Tip: Since the void volume in many separations contains highly conductive counter-ions, the error alarm sounds upon each injection. See the table titled Setting the beep function on page 3-7 to silence the beep-on-error alarm. Controls and indicators 3-7

58 Startup and shutdown Startup procedure Perform the following procedure to start the 432 Detector. Typically, this procedure is done at the beginning of each workday. Tip: This procedure assumes that the flow cell has stabilized at the selected temperature (minimum 2 to 3 hours). Standard practice is to leave the detector powered and with the temperature control on unless the instrument will be unused for several days. Set the temperature at least 5 C above the highest ambient temperature expected for the duration of the application. 1. Prime the pump with properly degassed eluent and set the flow rate to 1.2 ml/min or to the flow rate recommended for your particular column or application. Do not sparge eluents, since sparge gasses may contain ionic contaminants. 2. Set the response (time constant) to the desired setting by pressing the Response key. A standard setting (0.5 seconds) is preferred for most applications. 3. Set the base value by pressing the Auto Base key or by manually entering the base range that is the next highest setting above the eluent s background conductivity. 4. Turn on the recorder/integrator and run the system until the baseline stabilizes. 5. Depending on whether you are using a recorder or an integrator, do one of the following actions: If you are using a 10-mV recorder connected to the Recorder terminals on the rear panel, select the desired sensitivity by pressing the Sensitivity Range key, then the appropriate Up or Down arrow key. If you are using an integrator connected to the Integrator terminals on the rear panel, select the desired sensitivity by pressing the Shift and Sensitivity Range keys, then the appropriate Up or Down arrow key. 3-8 Operating the Detector

59 6. Zero the recorder/integrator by pressing the Auto Zero key or have the Auto Zero terminals of the rear panel I/O terminal strip connected to your manual injector or autosampler. The 432 Detector is now ready for operation. Standby setup To eliminate the need to allow time for the flow cell temperature to equilibrate, leave the 432 Detector turned on at the end of the workday or workweek. Set the temperature control to the operating temperature and the pump flow rate to 0.01 to 0.1 ml/min (depending on the pump). Long-term storage If the 432 Detector is to be removed from a system for storage or if the system itself is to be stored for a long time, flush the detector/system with 100% water, then 100% HPLC-grade methanol. Leave the methanol in the system after shutdown. If you are removing the 432 Detector from the system, seal the inlet and outlet bulkheads with dead-end fittings or a loop of tubing. Operating recommendations Observe the following recommendations for best detector performance. Temperature equilibration The 432 Detector should be powered up and set at the desired operating temperature for two to three hours before use. Select a temperature at least 5 C above the highest ambient temperature expected during the duration of the application. The detector is usually set at 35 C, but it should be set higher if the ambient temperature will exceed 30 C. You may choose to leave the 432 Detector powered up overnight at a flow rate of ml/min (depending on the pump) to minimize the daily reequilibration time. A drifting baseline is one indication that the temperature of the flow cell is not uniform across the flow cell or stable over time. Operating recommendations 3-9

60 Base range The Base Range is normally set at the next setting above the background conductivity of the eluent. For example, if the conductivity of borate/gluconate eluent is 270 μs, set the Base Range to 500 μs. Integrator output The Integrator output is not attenuated; signals should be below 1 V. Set the Integrator output to 10 μs/v for small signals or to 50 μs/v when you expect a signal greater than 10 μs. If you are using the 432 Detector with chemical suppression, set the integrator output to 100 μs/v. Recorder output Polarity The Recorder output is attenuated and the Sensitivity Range should be adjusted to provide the appropriate output level. Signal polarity depends on eluent conductivity. If necessary, press the Polarity key to obtain peaks rather than dips. Eluent handling Replace your eluent reservoir filter regularly. When you analyze cations, use an all-plastic eluent reservoir filter. Filter and degas eluents to prolong column life, reduce pressure fluctuations, and decrease baseline noise. When you change eluents, flush the flow cell and associated tubing thoroughly with the new eluent Operating the Detector

61 4 Performing Ion Analysis This chapter provides essential information about ion analysis techniques. Two representative columns serve as typical examples: the Waters IC-Pak A for anions and the IC Pak C M/D for cations. The following topics are discussed: Fundamental considerations Configuring the system Selecting and preparing eluents Preparing and injecting standards Also refer to the manufacturer s manual for the particular column you are using. A recommended source for more information about ion analysis in general is Ion Chromatography: Principles and Applications by Paul R. Haddad and Peter E. Jackson, Elsevier Science Publishing, New York, Contents Topic Page Fundamental considerations 4-2 Configuring the system 4-5 Eluents for ion analysis 4-6 Standards for ion analysis

62 Fundamental considerations Water To obtain full performance from the 432 Detector in chromatographic applications, observe the guidelines presented in this section regarding: Water Containers High-pH eluents Sample preparation Water purity (the absence of conductivity) is the most important consideration in ion analysis. Tip: Using water other than ASTM Type I reagent water will compromise the accuracy of your results. Freshly drawn ASTM Type I reagent water, conforming to ASTM specification D1193, with total organic carbon <100 ppb is recommended. Sophisticated laboratory water systems that use a combination of reverse osmosis, mixed bed ion exchange, and carbon adsorption cartridges produce ASTM Type I reagent water, and are recommended for ion chromatography applications. Do not use HPLC-grade water or distilled water. Caution: To avoid damage to the detector flow cell, do not allow the flow cell to dry out. Containers Use plastic to contain all anion and cation samples, cation standards, and cation eluents. When you analyze trace level ions in water, polystyrene containers such as tissue culture flasks are recommended; polypropylene or polymethylpentene containers suit most other applications. Use polystyrene tissue culture flasks for long-term storage. If your system operates on Breeze software or contains a 2695 Separations Module, use 4-mL polycarbonate vials to hold your samples and standards. Caution: Avoid glass containers (which tend to leach sodium cations) when you are analyzing for cations. 4-2 Performing Ion Analysis

63 Preparing containers for low-level analysis To prepare plastic containers for low-level analysis: 1. Soak all containers for 5 hours with a 1:1 solution of nitric acid (HNO 3 ) and ASTM Type I reagent water. 2. Rinse with plenty of ASTM Type I reagent water. The containers are ready for analysis in the ppm range. 3. For analysis in the ppb range, fill each container completely with ASTM Type I reagent water and let soak overnight. Certain applications that involve ppb level analysis may require container considerations beyond the scope of this manual. For further instructions on trace metal cleaning of plasticware, see Selection and Cleaning of Plastic Containers for Storage of Trace Element Samples, JR Moody and RM Lindstrom, Analytical Chemistry, v. 49, Dec 1977, pp , or contact the Waters Technical Services Department. Cleaning syringes To avoid contamination, always rinse a syringe two to three times with ASTM Type I reagent water before you draw standards or samples for injection. High-pH eluents High-pH eluents (such as hydroxide eluent) absorb atmospheric CO 2, which slowly acidifies the eluent causing baseline drift and retention time changes. To minimize carbonate absorption, connect a soda lime (Ascarite ) tube to the eluent bottle as follows: 1. Insert a 3/4-inch (2-cm) piece of glass wool in one end of a polyethylene tube with end fittings. Attach the end fitting. Warning: To avoid chemical burns, wear gloves, lab coat, and eye glasses when you are handling soda lime. 2. Fill the tube with soda lime (Ascarite) until it reaches 3/4 inches (2 cm) from the top. 3. Add another piece of glass wool to the other end of the tube and attach the end fitting. Fundamental considerations 4-3

64 4. Drill a hole in the cap of the reagent bottle. The hole should be large enough to accommodate the end fitting. Drill a second hole for the pump inlet line. 5. Pass the pump inlet line through the hole. Seal the hole with paraffin film. 6. Change the soda lime in the tube when it is exhausted. Soda lime tube End fittings Fittings Glass Wool wool Soda Lime lime Polypropylene Tube tube Pump inlet Inlet line Line Reagent bottle Bottle 4-4 Performing Ion Analysis

65 Sample preparation Sample preparation is very important in ion analysis. Contact the Waters Technical Services Department, if you need assistance. As a general rule, to analyze a sample of completely unknown ionic concentration, initially prepare at least a 1:100 dilution and inject 100 μl. For best results, injections should contain a total anion concentration of no more than 300 ppm for the IC-Pak A column or a total cation concentration of no more than 10 ppm per ion for the IC-Pak C M/D column. The sample volume (usually 100 μl) generally equilibrates to the ph of the eluent upon injection. However, for samples with ph values that differ greatly from that of the eluent (for example, strong acids and bases), bring the sample ph close to that of the eluent before you inject the sample, if possible. Do not inject concentrated samples directly into the mobile phase. Direct injection may cause precipitation of the salts in the sample. Dissolve (or dilute) samples in an appropriate volume of the mobile phase first. If you must use other solvents, watch for precipitation upon injection into the eluent. Always filter samples before you use them. Cationic samples that contain organic amines may exhibit hydrophobic interaction between the mobile phase and packing. You may use a water-miscible organic mobile phase, such as acetonitrile, as a modifier to reduce this. Pretreat the sample with a Sep-Pak C 18 cartridge to remove hydrophobic compounds. Configuring the system The figure System configuration for ion analysis on page 4-6 shows a typical system configuration. Refer to Making fluidic connections on page 2-23, for the procedures to cut tubing and assemble fittings. Configuring the system 4-5

66 System configuration for ion analysis Waters in-line In-Line degasser* Degasser* Eluent reservoir Eluent Reservoir Pump *Optional * Optional Guard column Column Holder holder* * Column Injector Pulse Dampener dampener t Waters 432 Detector To waste To Waste Required t for Waters pumps without SILK or non-waters pumps Required for non-waters pumps or Waters pumps with Breeze software, such as the HPLC 515 TP01269 Pulse dampener If your system uses a non-waters pump or a Waters pump with Breeze software, such as the HPLC 515, use a pulse dampener (supplied in the Startup Kit) to achieve the best performance from the 432 Detector. Install the pulse dampener between the pump and the injector, as described in Installing the pulse dampener on page Eluents for ion analysis This section describes how to select, prepare, and use eluents for ion analysis. Caution: Never recirculate eluents. Ions from sample and standard injections progressively contaminate a recirculating eluent. General guidelines Observe the following guidelines when you prepare eluents for ion analysis: Use only ASTM Type I reagent water with total organic carbon <100 ppb. Use the highest purity salts and reagents available. 4-6 Performing Ion Analysis

67 A ph meter is recommended for checking the ph of eluents; care should be taken to avoid cross contamination. Adjust the ph with potassium hydroxide (KOH) or lithium hydroxide (LiOH). For eluents such as octane sulfonate, test an aliquot of the eluent with ph paper. Never immerse ph paper directly into a batch of eluent. Use the following formula to prepare eluents: Formula Wt of Salt x Molarity = g/l Salt Eluent filtering and degassing The Waters Solvent Clarification Kit is recommended for eluent filtration and preliminary degassing. Durapore 0.22-mm filters can be used for all ion chromatography eluents. Millipore 0.45-mm HATF filters may be used for aqueous eluents containing no organic modifier. For eluents containing organic modifier, use the Durapore filters. Caution: To avoid contamination when you analyze for cations, minimize the time that the eluent is in contact with the glass filtration apparatus and transfer the eluent to a suitable pre-cleaned plastic container as soon as possible. After you install a new filter, pass 20 to 30 ml of eluent through the filter under vacuum. Turn off the vacuum, swirl the eluent throughout the flask and discard. Reattach the flask to the filter apparatus and filter the remaining eluent. As soon as filtration is complete, transfer the eluent to a precleaned plastic container, introducing the least possible amount of bubbles in the process. The Waters In-line Degasser is recommended for continuous online degassing. Preparing anion eluent This section presents the procedure for the preparation of sodium borate/gluconate concentrate and eluent. Consult the manufacturer s manual for your column (IC-Pak Column and Guard Column Care and Use Manual included with Waters columns) for additional instructions on the selection and preparation of eluents. A recommended source for more information about ion analysis in general is Ion Chromatography: Principles and Applications, by Paul R. Haddad and Peter E. Jackson, Elsevier Science Publishing, New York, Eluents for ion analysis 4-7

68 Preparing lithium borate/gluconate concentrate To prepare sodium borate/gluconate concentrate, refer to Preparing lithium borate/gluconate 50X stock concentrate on page D-13 and Preparing lithium borate/gluconate eluent on page D-14. Preparing cation eluent To prepare 1 L of cation eluent, refer to Preparing eluent on page D-3. Standards for ion analysis This section describes how to prepare and inject ion standards. Tip: It is recommended to purchase certified 1000-ppm anion standards instead of preparing them manually. Certify all manual standards against National Institute of Science and Technology traceable standards. Standard concentrations in this manual are defined in terms of mass. For example, 1 mg of sample per liter of water equals a 1 ppm concentration, since 1 L of water has a nominal mass of 1 kg (0.997 kg at 25 C). 1 part per thousand = 1 mg/ml = 1 g/l = 1000 ppm 1 part per million (ppm) = 1 μg/ml = 1 mg/l = 1000 ppb 1 part per billion (ppb) = 1 ng/ml = 1 μg/l = 1000 ppt 1 part per trillion (ppt) = 1 pg/ml = 1 ng/l Storing standards For accurate quantitative results, do not store standards beyond the approximate periods listed in the table below. Be aware that shelf-life depends on many factors and may be significantly shorter than shown here. Shelf-life of standards Standard Carbonate, ppm Chloride, ppm All, ppb Nitrite and carbonate concentrates Shelf-life 1 day 3 weeks 1 day 1 week 4-8 Performing Ion Analysis

69 Shelf-life of standards (Continued) Standard All other anion concentrates Cation standards Cation concentrates Shelf-life 1 to 2 months 1 month 6 months Cation standards must be stored in properly prepared plasticware. See Containers on page 4-2. Preparing anion standards This section presents the procedure for preparing a 7-anion standard. If a simpler standard suffices, follow the procedure, but select only three or four salts, such as sodium chloride, sodium nitrate, and sodium sulfate. Always use salts of at least reagent-grade purity. If you require quantitative results or you use hygroscopic salts, dry the salts overnight at 80 C before you make solutions. Store the dried salts in a desiccator. Preparing a 7-anion standard To prepare a 7-anion standard: 1. Weigh out the amounts of dry salts shown in the table below or use the following formula to calculate the amount for a salt not listed: (Mol. Wt. Salt / Mol. Wt. Cation) x 0.1 = g Salt Salts for anion standard concentrates Salt (anion) Weight (mg) Sodium fluoride (F ) Sodium chloride (Cl ) Sodium nitrite (NO 2 ) Potassium bromide (Br ) Sodium nitrate (NO 3 ) Potassium phosphate, monobasic (HPO 4 ) 2 Sodium sulfate (SO 4 ) Standards for ion analysis 4-9

70 2. Place each salt in a separate plastic100-ml volumetric flask and dilute to the mark with ASTM Type I reagent water. Each concentrate contains 1000 ppm of the anion. 3. Combine the amounts listed in the table below in a 100-mL volumetric flask with ASTM Type I reagent water. Anion concentrate dilutions Anion Amount (μl) Final Concentration (ppm) Fluoride 100 μl 1 ppm Chloride 200 μl 2 ppm Nitrite 400 μl 4 ppm Bromide 400 μl 4 ppm Nitrate 400 μl 4 ppm Phosphate 600 μl 6 ppm Sulfate 400 μl 4 ppm Injecting anion standards Required materials To inject a standard, obtain the following materials: Borate/gluconate eluent Refer to Preparing lithium borate/gluconate 50X stock concentrate on page D-13 and Preparing lithium borate/gluconate eluent on page D cc disposable plastic syringe Pharmaseal Stylex disposable syringe with a Luer Slip tip, or equivalent. Autoinjector or manual injector with 100-μL loop Ion chromatography commonly uses a 100-μL injection volume. When you use a fixed loop, overfill a minimum of three times Performing Ion Analysis

71 Injecting the standard To inject the standard: 1. Set up the 432 Detector as follows: Base Sensitivity = 500 μs Integrator Sensitivity = 10 μs/v Recorder Sensitivity = 0.01 (strip chart) Response = STD (0.5 seconds) Temperature = 35 C Polarity = + 2. Equilibrate the 432 Detector as described in Startup procedure on page Rinse a 1-cc disposable plastic syringe two or three times with ASTM Type I reagent water, then load the standard. 4. Place the syringe tip into the sample loading port and overfill the 100 μl loop at least three times (that is, 300 μl). 5. Inject the sample. The figure Chromatogram of a 7-anion standard on page 4-12 shows a representative chromatogram of the 7-anion standard run on an IC-Pak A (4.6 mm x 5.0 cm) column with borate/gluconate eluent at 1.2 ml/min flow rate. The separation of the standard typically takes 12 to 15 minutes with this setup. Standards for ion analysis 4-11

72 Chromatogram of a 7-anion standard Preparing cation standards This section presents the procedure for preparing an 8-cation standard. If a simpler standard suffices, follow this procedure selecting only those salts that you want in the standard. For accurate quantitative results, use only properly prepared plasticware and do not store standards beyond the recommended shelf-lives listed in the table titled Shelf-life of standards on page 4-8. Preparing cation standard concentrates Tip: It is recommended that you use certified 1000-ppm cation standards not prepared in acid with this method Performing Ion Analysis

73 To prepare concentrated stock solutions for an 8-cation standard (prepare fewer types of cations, if a simpler standard suffices): 1. Weigh out the amounts of dry salts shown in the table titled Salts for cation standard concentrates on page 4-13 or use the following formula to calculate the amount for a salt not listed. (Mol. Wt. Salt / Mol. Wt. Cation) = g Salt If you choose to use other salts, be sure to avoid any combinations that will form a precipitate. Salts for cation standard concentrates Salt (cation) Weight (g) Lithium hydroxide monohydrate (Li + ) Sodium chloride (Na + ) Ammonium chloride (NH 4 + ) Potassium chloride (K + ) Magnesium nitrate hexahydrate (Mg 2+ ) Calcium nitrate tetrahydrate (Ca2+) Strontium nitrate tetrahydrate (Sr2+) Barium chloride dihydrate (Ba 2+ ) Place each salt in a separate plastic1-l volumetric flask and dilute to the mark with reagent-grade water. Each concentrate contains 1000 ppm of the cation. Standards for ion analysis 4-13

74 Preparing an 8-cation standard To prepare 1 liter of 8-cation standard: 1. Add the volume of stock (concentrate) standard listed in the table below to a plastic 1-L volumetric flask. Cation concentrate dilutions Cation Amount (ml) Final concentration (ppm) Lithium Sodium Ammonium Potassium Magnesium Calcium Strontium Barium Fill the flask to the mark with ASTM Type I reagent water. Injecting cation standards Required materials To inject the standard, obtain the following materials: 0.1 mm EDTA/ 3 mm HNO 3 cation eluent Refer to Preparing cation eluent on page cc disposable plastic syringe Pharmaseal Stylex disposable syringe with a Luer Slip tip, or equivalent. Injector or autosampler with a 100-μL loop Ion chromatography commonly uses a 100-μL injection volume. When you use a fixed loop, overfill a minimum of three times Performing Ion Analysis

75 Injecting the standard Tip: You can substitute the method described in Alkali and alkaline earth cations, ammonium, and amines on page D-15, for the following procedure. To inject the standard. 1. Set up the 432 Detector as follows: Base Sensitivity = 2000 μs Integrator Sensitivity = 50 μs/v Recorder Sensitivity = 0.01 (strip chart) Response = STD (0.5 seconds) Temperature = 35 C Polarity = (negative) 2. Equilibrate the 432 Detector as described in Startup procedure on page Rinse a 1-cc disposable plastic syringe two or three times with ASTM Type I reagent water, then load the standard. 4. Place the syringe tip into the sample loading port and overfill the 100-μL loop at least three times (that is, 300 μl). 5. Inject the sample. The figure Chromatogram of an 8-cation standard on page 4-16 shows a representative chromatogram of an 8-cation standard run on an IC-Pak C M/D column with 0.1 mm EDTA/3 mm HNO 3 eluent at 1.0 ml/min flow rate. The separation of the standard typically takes 20 to 25 minutes with this setup. Standards for ion analysis 4-15

76 Chromatogram of an 8-cation standard 4-16 Performing Ion Analysis

77 5 Maintenance This chapter contains information about maintaining the 432 Detector and troubleshooting charts to help you isolate and correct problems.. Contents Warning: To avoid the possibility of electric shock, power off the detector and disconnect the power cord before you service the instrument. Topic Page Routine maintenance 5-2 Cleaning the detector exterior 5-6 Troubleshooting

78 Routine maintenance This section contains information designed to help you maintain the 432 Detector. Routine maintenance for the 432 Detector includes: Replacing the fuse Calibrating the detector Maintaining the flow cell Waters service specialists provide maintenance for the 432 Detector on a corrective, as required, basis. Contact the Waters Technical Services Department if you have questions regarding the repair or performance of your instrument. Follow these suggestions to help you maintain the 432 Detector: Stock the recommended spare parts listed in Appendix B to reduce downtime. Contact the Waters Service Department for assistance. Record the initial installation date and serial number of your instrument in Appendix C for easy reference. Keep a file of typical chromatograms for comparison when you suspect problems. Replacing the fuse To change the operating voltage fuse: 1. Turn off the front panel power switch and remove the power cord from its connector on the rear panel of the detector. Warning: To avoid the possibility of electric shock, turn off the front panel power switch, and unplug the power cord from the rear panel. 2. Pry open the power connector cover with a screwdriver. 3. To change the AC power fuses, pull out each fuse holder as though opening a drawer. Spare fuses are included in the System Startup Kit. For ordering information, see Appendix C. 4. The table titled Nominal operating voltage on page 2-6 lists the operating voltage fuses (for use in either North America or Europe). 5-2 Maintenance

79 5. Install the correct fuse in the holder and slide it back into place. The arrow on each fuse holder points up when in the correct position. Installing operating voltage fuses 6. Close the power connector cover. Then plug the power cord into its connector on the rear panel of the detector. Maintaining the flow cell Maintenance for the 432 Detector consists of ensuring the flow cell is free of foreign material. Foreign material in the flow cell may cause baseline drift, cycling, or noise. Caution: To avoid damaging the column, remove it before you flush the system. Do not reconnect the column until the eluent has returned to approximately ph 7. To clean the cell: 1. Flush the system with ASTM Type I reagent water. 2. Flush the system with 20 ml of 6 N nitric acid (HNO 3 ). 3. Flush the system again with ASTM Type I reagent water. Do not reconnect the column until the eluent has returned to about ph 7. Routine maintenance 5-3

80 Refer to Appendix C, to order a replacement flow cell. The following tools are required to replace the flow cell: Phillips-head screwdriver 5/16-inch open-end wrench Knife or flat-blade screwdriver Warning: To avoid electrical hazards, always unplug the power cord before you perform any of the following replacement procedures. To replace the flow cell: 1. Unplug the 432 Detector from the power source, and completely disconnect all electrical cables and fluid connections. 2. Remove the 432 Detector cover (four Phillips-head screws, two on each side). 3. Remove the two pins and pin holders that hold the cell block cover in place (see figure Flow cell assembly on page 5-5). Use a knife or flat-blade screwdriver to gently pry the pins and holders out. 4. Pull off the cover of the flow cell unit and remove the top layer of insulation. 5. Remove the four Phillips-head screws from the upper plate of the cell block, and remove the plate. Note the orientation of the plate: a notch is cut into the underside to clear one of the components within the cell block. 6. Carefully disconnect the inlet and outlet fittings from the flow cell. 7. Remove the two Phillips-head screws from the flow cell mounting bracket. 8. Unplug the flow cell cable connector from its socket in the cell block. 9. Remove the flow cell assembly. 10. Install the new flow cell by following steps 2 through 9 in reverse order. Be sure to orient the upper plate of the cell block properly before you install the four screws. 5-4 Maintenance

81 Flow cell assembly Cell Cell block block cover cover Pin Pin holder Pin holder Pin Insulation Upper Upper plate plate screws Upper plate of cell cell block Cell Cell mounting bracket Flow cell Cell Cell mounting mounting bracket screws Connector Cell block Routine maintenance 5-5

82 Cleaning the detector exterior To clean the outside of the 432 Detector, use only a soft lint-free paper or cloth dampened with mild soap and water. Troubleshooting This section contains troubleshooting charts to help you isolate and correct problems with the 432 Detector. Keep in mind that the source of apparent detector problems may lie within the chromatography or hardware of your system. The Waters Guide to Successful Operation of Your LC System contains detailed chromatographic troubleshooting tables. (Contact your nearest Waters office for information on how to get a copy.) If you cannot correct a problem, contact the Waters Technical Services Department for assistance. When you call Waters service To expedite your request for service, have the following information available when you call Waters regarding a 432 Detector problem: Symptom Type of column Eluent(s) Flow rate Operating pressure Base Range setting Integrator Sensitivity setting Recorder Sensitivity setting Type of injector (automatic or manual) Type of data integrator Detector does not turn on If your detector is completely inoperative (for example, the lights do not illuminate and the display remains completely blank when the unit is turned 5-6 Maintenance

83 on), the fuse may require replacement. Refer to Replacing the fuse on page 5-2. Startup diagnostics The 432 Detector performs startup diagnostics that check the internal memory (both RAM and ROM), and the associated processing circuitry. Power supply The following factors can adversely affect the operation of the 432 Detector: Power surges Line spikes Transient energy sources Be sure that the electrical supply used for the 432 Detector is properly grounded and free from any of these conditions. Error messages The error messages displayed by the 432 Detector are listed below along with the recommended action for each: Error: ROM/RAM ROM/RAM error (checked during startup only) Call Waters service. Error: Leak Leakage detected Check flow cell and associated plumbing connections. Error: Temp Temperature control failure Call Waters service. Error: Over Range Base over-range condition Set Base Range to the next setting above the background conductivity of the eluent. Error: Over Flow Conductivity overflow (above 10,000 μs) Dilute eluent to remain within measurable range. Troubleshooting 5-7

84 Press the Clear key to clear an error alarm and message. For a continuing error condition, the error message remains after the audio alarm is cleared. Troubleshooting procedure To troubleshoot a problem: 1. Visually examine the integrity of the electrical and fluid connections as you verify proper system configuration and installation. 2. If the results of previous runs are available, compare the current system operation with the system operation before you identified the problem. For example, if your system usually runs at a certain pressure with a certain method: Is the system pressure in the same range, or is it drastically higher (possibly caused by a blocked line) or lower (possibly caused by a leak)? Are pressure fluctuations in the same range as during normal operation? 3. Isolate the parameter that varies from normal operation. The parameters to observe include: Baseline noise Peak retention time Peak resolution Qualitative/quantitative chromatographic results System pressure Evaluate the parameters in the order presented above to rule out simple causes of the problem. 4. Refer to the table titled Troubleshooting guide on page 5-9 to determine corrective actions for the problems that you identify. Removing bubbles Bubbles in the flow cell are evident when the noise is equal to or greater than 2 μs. Use this method to remove bubbles. 5-8 Maintenance

85 To remove bubbles: 1. Disconnect the tubing from the inlet and outlet of the 432 Detector. 2. Attach a 1-mL tuberculin syringe to a priming syringe cannula which is screwed into the inlet of the detector. 3. Flush four times with 1-mL portions of ASTM Type I reagent water. 4. Flush four times with 1-mL portions of HPLC-grade methanol. 5. Flush four times with 1-mL portions of ASTM Type I reagent water. 6. Reattach the tubing from the 432 Detector outlet to a waste receptacle (18-inch length of inch ID stainless steel). 7. Start eluent flowing through the system at a flow rate of at least 1 ml/min. 8. With the eluent flowing, reattach the detector inlet tubing to the column. 9. Allow a few minutes for temperature reequilibration, then check the noise level. If it is not reduced, repeat steps 1 through 4, then proceed to steps 10 through Attach a dead-end fitting to the 432 Detector outlet. 11. Remove the priming syringe cannula and attach a dead-end fitting to the 432 Detector inlet. 12. Allow the detector to stand overnight (>12 hours) with temperature on and with methanol in the flow cell. 13. Repeat steps 5 through 9. Troubleshooting guide Symptom Possible cause Solution Noisy baseline Pulse dampener not installed See Installing the pulse dampener on page Pulsing pump Check the pump; see the pump manual. Bubbles in flow cell Remove bubbles and degas the solvent. Troubleshooting 5-9

86 Troubleshooting guide (Continued) Symptom Possible cause Solution Noisy baseline (Continued) Voltage fluctuation Use the voltage regulator. Spikes on baseline Dirty flow cell Clean the cell. Flow cell leak Check flow cell fittings and tighten. If leak continues, replace the flow cell. Bubbles in flow cell Remove bubbles and degas the solvent. Irregular noise on baseline Excessive baseline drift Temperature changes in room Cell temperature set lower than ambient Defective column Unstable temperature control Defective cell heater Temperature changes in room Cell temperature set lower than ambient Bubbles in flow cell Control ambient temperature, locate drafts, and insulate tubing and column, if necessary. Set the cell temperature to a minimum of 5 C above ambient. Replace the column. Make sure the temperature control is turned on. Call Waters service. Control ambient temperature, locate drafts, and insulate tubing and column, if necessary. Set the cell temperature to a minimum of 5 C above ambient. Remove bubbles and degas the solvent Maintenance

87 Troubleshooting guide (Continued) Symptom Possible cause Solution Excessive baseline drift (Continued) Detector cannot be zeroed Continuous noise at high sensitivity (<1μS) Solvent changeover Flow cell leak Solvent changeover Bubbles in flow cell Pump crossover noise Wait until baseline stabilizes (purge autosampler a few times). Check flow cell fittings and tighten. If leak continues, replace the flow cell. Wait until the baseline stabilizes. Remove bubbles and degas the solvent. Install a high-sensitivity noise filter on the pump. Troubleshooting 5-11

88 5-12 Maintenance

89 A 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. This appendix presents all the safety symbols and statements that apply to the entire line of Waters products. Contents Topic Page Warning symbols A-2 Caution symbol A-5 Warnings that apply to all Waters instruments A-6 Electrical and handling symbols A-12 A-1

90 Warning symbols Warning symbols alert you to the risk of death, injury, or seriously adverse physiological reactions associated with an instrument s use or misuse. 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. Task-specific hazard warnings The following warning symbols alert you to risks that can arise when you operate or maintain an instrument or instrument component. Such risks include burn injuries, electric shocks, ultraviolet radiation exposures, and others. When the following symbols appear in a manual s narratives or procedures, their accompanying text identifies the specific risk and explains how to avoid it. Warning: (General risk of danger. When this symbol appears on an instrument, consult the instrument s user documentation for important safety-related information before you use the instrument.) Warning: (Risk of burn injury from contacting hot surfaces.) Warning: (Risk of electric shock.) Warning: (Risk of fire.) Warning: (Risk of sharp-point puncture injury.) Warning: (Risk of hand crush injury.) Warning: (Risk of exposure to ultraviolet radiation.) Warning: (Risk of contacting corrosive substances.) Warning: (Risk of exposure to a toxic substance.) Warning: (Risk of personal exposure to laser radiation.) A-2 Safety Advisories

91 Warning: (Risk of exposure to biological agents that can pose a serious health threat.) Warning: (Risk of tipping.) Warning: (Risk of explosion.) Warning: (Risk of eye injury.) Specific warnings The following warnings can appear in the user manuals of particular instruments and on labels affixed to them or their component parts. Burst warning This warning applies to Waters instruments fitted with nonmetallic tubing. Warning: Pressurized nonmetallic, or polymer, tubing can burst. Observe these precautions when working around such tubing: Wear eye protection. Extinguish all nearby flames. Do not use tubing that is, or has been, stressed or kinked. Do not expose nonmetallic tubing to incompatible compounds like tetrahydrofuran (THF) and nitric or sulfuric acids. Be aware that some compounds, like methylene chloride and dimethyl sulfoxide, can cause nonmetallic tubing to swell, which significantly reduces the pressure at which the tubing can rupture. Warning symbols A-3

92 Mass spectrometer flammable solvents warning This warning applies to instruments operated with flammable solvents. Warning: Where significant quantities of flammable solvents are involved, a continuous flow of nitrogen into the ion source is required to prevent possible ignition in that enclosed space. Ensure that the nitrogen supply pressure never falls below 690 kpa (6.9 bar, 100 psi) during an analysis in which flammable solvents are used. Also ensure a gas-fail connection is connected to the LC system so that the LC solvent flow stops if the nitrogen supply fails. Mass spectrometer shock hazard This warning applies to all Waters mass spectrometers. Warning: To avoid electric shock, do not remove the mass spectrometer s protective panels. The components they cover are not user-serviceable. This warning applies to certain instruments when they are in Operate mode. Warning: High voltages can be present at certain external surfaces of the mass spectrometer when the instrument is in Operate mode. To avoid non-lethal electric shock, make sure the instrument is in Standby mode before touching areas marked with this high voltage warning symbol. A-4 Safety Advisories

93 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. Warning: Waters instruments and software can be used to analyze or process potentially infectious human-sourced products, inactivated microorganisms, and other biological materials. To avoid infection with these agents, assume that all biological fluids are infectious, observe Good Laboratory Practices, and consult your organization s biohazard safety representative regarding their proper use and handling. Specific precautions appear in the latest edition of the US National Institutes of Health (NIH) publication, Biosafety in Microbiological and Biomedical Laboratories (BMBL). Chemical hazard warning This warning applies to Waters instruments that can process corrosive, toxic, flammable, or other types of hazardous material. Warning: Waters instruments can be used to analyze or process potentially hazardous substances. To avoid injury with any of these materials, familiarize yourself with the materials and their hazards, observe Good Laboratory Practices (GLP), and consult your organization s safety representative regarding proper use and handling. Guidelines are provided in the latest edition of the National Research Council's publication, Prudent Practices in the Laboratory: Handling and Disposal of Chemicals. Caution symbol The caution symbol signifies that an instrument s use or misuse can damage the instrument or compromise a sample s integrity. The following symbol and its associated statement are typical of the kind that alert you to the risk of damaging the instrument or sample. Caution: To avoid damage, do not use abrasives or solvents to clean the instrument s case. Caution symbol A-5

94 Warnings that apply to all Waters instruments When operating this device, follow standard quality control procedures and the equipment guidelines in this section. Attention: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user s authority to operate the equipment. Important: Toute modification sur cette unité n ayant pas été expressément approuvée par l autorité responsable de la conformité à la réglementation peut annuler le droit de l utilisateur à exploiter l équipement. Achtung: Jedwede Änderungen oder Modifikationen an dem Gerät ohne die ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen. Avvertenza: qualsiasi modifica o alterazione apportata a questa unità e non espressamente autorizzata dai responsabili per la conformità fa decadere il diritto all'utilizzo dell'apparecchiatura da parte dell'utente. Atencion: cualquier cambio o modificación efectuado en esta unidad que no haya sido expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equipo. 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 : 규정준수를책임지는당사자의명백한승인없이이장치를개조또는변경할경우, 이장치를운용할수있는사용자권한의효력을상실할수있습니다. 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 A-6 Safety Advisories

95 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. Be aware that methylene chloride and dimethyl sulfoxide cause nonmetallic tubing to swell, which greatly reduces the rupture pressure of the tubing. Attention: Manipulez les tubes en polymère sous pression avec precaution: Portez systématiquement des lunettes de protection lorsque vous vous trouvez à proximité de tubes en polymère pressurisés. Eteignez toute flamme se trouvant à proximité de l instrument. Evitez d'utiliser des tubes sévèrement déformés ou endommagés. Evitez d'utiliser des tubes non métalliques avec du tétrahydrofurane (THF) ou de l'acide sulfurique ou nitrique concentré. Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le gonflement des tuyaux non métalliques, ce qui réduit considérablement leur pression de rupture. Vorsicht: Bei der Arbeit mit Polymerschläuchen unter Druck ist besondere Vorsicht angebracht: In der Nähe von unter Druck stehenden Polymerschläuchen stets Schutzbrille tragen. Alle offenen Flammen in der Nähe löschen. Keine Schläuche verwenden, die stark geknickt oder überbeansprucht sind. Nichtmetallische Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte Salpeter- oder Schwefelsäure verwenden. Durch Methylenchlorid und Dimethylsulfoxid können nichtmetallische Schläuche quellen; dadurch wird der Berstdruck des Schlauches erheblich reduziert. Warnings that apply to all Waters instruments A-7

96 Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione: Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati. Spegnere tutte le fiamme vive nell'ambiente circostante. Non utilizzare tubi eccessivamente logorati o piegati. Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido solforico o nitrico concentrati. Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano rigonfiamenti nei tubi non metallici, riducendo notevolmente la pressione di rottura dei tubi stessi. Advertencia: se recomienda precaución cuando se trabaje con tubos de polímero sometidos a presión: El usuario deberá protegerse siempre los ojos cuando trabaje cerca de tubos de polímero sometidos a presión. Si hubiera alguna llama las proximidades. No se debe trabajar con tubos que se hayan doblado o sometido a altas presiones. Es necesario utilizar tubos de metal cuando se trabaje con tetrahidrofurano (THF) o ácidos nítrico o sulfúrico concentrados. 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 A-8 Safety Advisories

97 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 : 가압폴리머튜브로작업할경우에는주의하십시오. 가압폴리머튜브근처에서는항상보호안경을착용하십시오. 근처의화기를모두끄십시오. 심하게변형되거나꼬인튜브는사용하지마십시오. 비금속 (Nonmetallic) 튜브를테트라히드로푸란 (Tetrahydrofuran: THF) 또는농축질산또는황산과함께사용하지마십시오. 염화메틸렌 (Methylene chloride) 및디메틸술폭시드 (Dimethyl sulfoxide) 는비금속튜브를부풀려튜브의파열압력을크게감소시킬수있으므로유의하십시오. 警告 : 圧力のかかったポリマーチューブを扱うときは 注意してください 加圧されたポリマーチューブの付近では 必ず保護メガネを着用してください 近くにある火を消してください 著しく変形した または折れ曲がったチューブは使用しないでください 非金属チューブには テトラヒドロフラン (THF) や高濃度の硝酸または硫酸などを流さないでください 塩化メチレンやジメチルスルホキシドは 非金属チューブの膨張を引き起こす場合があり その場合 チューブは極めて低い圧力で破裂します Warnings that apply to all Waters instruments A-9

98 Warning: The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. Attention: L utilisateur doit être informé que si le matériel est utilisé d une façon non spécifiée par le fabricant, la protection assurée par le matériel risque d être défectueuses. Vorsicht: Der Benutzer wird darauf aufmerksam gemacht, dass bei unsachgemäßer Verwenddung des Gerätes die eingebauten Sicherheitseinrichtungen unter Umständen nicht ordnungsgemäß funktionieren. Attenzione: si rende noto all'utente che l'eventuale utilizzo dell'apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall'apparecchiatura. Advertencia: el usuario deberá saber que si el equipo se utiliza de forma distinta a la especificada por el fabricante, las medidas de protección del equipo podrían ser insuficientes. 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 : 제조업체가명시하지않은방식으로장비를사용할경우장비가제공하는보호수단이제대로작동하지않을수있다는점을사용자에게반드시인식시켜야합니다. 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 A-10 Safety Advisories

99 Warning: To protect against fire, replace fuses with those of the type 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 proximité du couvercle de la boite à fusible de l'instrument. Vorsicht: Zum Schutz gegen Feuer die Sicherungen nur mit Sicherungen ersetzen, deren Typ und Nennwert auf den Tafeln neben den Sicherungsabdeckungen des Geräts gedruckt sind. Attenzione: per garantire protezione contro gli incendi, sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento. Advertencia: Para evitar incendios, sustituir los fusibles por aquellos del tipo y características impresos en los paneles adyacentes a las cubiertas de los fusibles del instrumento. 警告 : 為了避免火災, 更換保險絲時, 請使用與儀器保險絲蓋旁面板上所印刷之相同類型與規格的保險絲 警告 : 为了避免火灾, 应更换与仪器保险丝盖旁边面板上印刷的类型和规格相同的保险丝 경고 : 화재의위험을막으려면기기퓨즈커버에가까운패널에인쇄된것과동일한타입및정격의제품으로퓨즈를교체하십시오. 警告 : 火災予防のために ヒューズ交換では機器ヒューズカバー脇のパネルに記載されているタイプおよび定格のヒューズをご使用ください Warnings that apply to all Waters instruments A-11

100 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. A-12 Safety Advisories