Waters 2487 Dual A Manual

Waters 2487 Dual λ
Absorbance Detector
Operator’s Guide
34 Maple Street
Milford, MA 01757
WAT048740, Revision 2

NOTICE
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 guide is believed to be
complete and accurate at the time of publication. In no event shall Waters Corporation be
liable for incidental or consequential damages in connection with or arising from the use of
this guide.
1997–2000 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF
AMERICA. ALL RIGHTS RESERVED. THIS BOOK OR PARTS THEREOF MAY NOT
BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE
PUBLISHER.
Alliance, Millennium³², and Waters are registered trademarks, and LAC/E, PowerLine,
SAT/IN, and TaperSlit are trademarks of Waters Corporation.
Microsoft, MS, MS-DOS, Windows, and Windows NT are registered trademarks of
Microsoft Corporation.
PEEK is a trademark of Victrex Corporation.
Teflon is a registered trademark of E.I. du Pont de Nemours and Company.
All other trademarks are the sole property of their respective owners.

Note: The Installation Category (Overvoltage Category) for this instrument is Level II. The
Level II category pertains to equipment that receives its electrical power from a local level,
such as an electrical wall outlet.
STOP
Attention: The Waters 2487 Dual λ Absorbance Detector can be used for in vitro
diagnostic applications. This is a highly sensitive instrument. Read the accompanying
operator’s guide before using.
When using the instrument, follow generally accepted procedures for quality control and
methods development.
If you observe a change in the retention of a particular compound, in the resolution
between two compounds, or in peak shape, immediately take steps to determine the
reason for the changes. Until you determine the cause of a change, do not rely upon the
results of the separations.
STOP
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.
STOP
Attention: 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 you make connections to the terminal strips on the rear
panel of the unit. In addition, ensure you always connect the shield of the cable to chassis
ground at one instrument only.
Caution: To protect against fire hazard, replace fuses with those of the same type and
rating.
Caution: To avoid the possibility of electrical shock, power off the 2487 Detector and
disconnect the power cord before you service the instrument.

Symbols Used on Rear Panel of the 2487 Detector
Direct current
Alternating current
Protective conductor
terminal
Frame or chassis
terminal
Caution, risk of electrical
shock (high voltage)
Caution or refer to guide
Caution, hot surface or
high temperature

2487 Detector Information
Intended Use
You can use the Waters 2487 Dual λ Absorbance Detector for in-vitro diagnostic testing to
analyze many compounds, including diagnostic indicators and therapeutically monitored
compounds. When developing methods, follow the “Protocol for the Adoption of Analytical
Methods in the Clinical Chemistry Laboratory,” American Journal of Medical Technology,
44, 1, pages 30–37 (1978). This protocol covers good operating procedures and
techniques necessary to validate system and method performance.
Biological Hazard
When analyzing physiological fluids, take all necessary precautions and treat all
specimens as potentially infectious. Precautions are outlined in the “CDC Guidelines on
Specimen Handling,”
CDC–NIH Manual, 1984.
Calibration
Follow acceptable methods of calibration with pure standards to calibrate methods. Use a
minimum of five standards to generate the standard curve. Concentration range should
cover the entire range of quality control samples, typical specimens, and atypical
specimens.
Quality Control
Routinely run three quality-control samples and monitor them daily. Quality-control
samples should represent subnormal, normal, and above-normal levels of a compound.
Ensure that quality-control sample results are within an acceptable range, and evaluate
precision from day to day and run to run. Data collected when quality-control samples are
out of specification may not be valid. Do not report the data until you ensure that
chromatographic system performance is acceptable. Conduct a failure investigation to
determine the cause(s) for the out-of-specification results.

Table of Contents 6
How to Use This Guide..................................................................... 18
Chapter 1
2487 Detector Theory of Operation ................................................. 25
1.1 2487 Detector Description .................................................... 25
1.2 Principles of Operation.......................................................... 27
1.2.1 Waters 2487 Detector Optics..................................... 27
1.2.2 Wavelength Verification and Test............................... 29
1.2.3 Waters TaperSlit Flow Cell......................................... 30
1.2.4 Waters 2487 Dual λ Absorbance Detector
Electronics................................................................. 33
1.3 Operational Modes................................................................ 34
1.3.1 Single Wavelength Mode........................................... 34
1.3.2 Dual Wavelength Mode ............................................. 35
1.3.3 Spectrum Scanning ................................................... 37
1.3.4 Cuvette Operations.................................................... 37
1.3.5 RatioPlot.................................................................... 38
1.3.6 MaxPlot...................................................................... 38
Chapter 2
Installing the 2487 Detector ............................................................. 39
2.1 Introduction........................................................................... 39
2.2 Site Selection and Power Requirements .............................. 41
2.2.1 Site Selection............................................................. 41
2.2.2 Power Requirements................................................. 42
Table of Contents

Table of Contents 7
2.3 Unpacking and Inspecting..................................................... 42
2.3.1 Unpacking.................................................................. 43
2.3.2 Inspecting .................................................................. 43
2.4 Making Fluidic Connections.................................................. 44
2.5 Making Electrical Power Connections................................... 47
2.6 Making Signal Connections .................................................. 49
2.6.1 Making I/O Signal Connections ................................. 50
2.6.2 Connecting the 2487 Detector to a 2690 Separations
Module in a Stand-Alone Configuration..................... 52
2.6.3 Making IEEE-488 Connections in Breeze and
Millennium³²............................................................... 58
2.7 Connecting to Other Devices................................................ 64
2.7.1 Connecting the 2487 Detector to the Millennium³²
System Using the busSAT/IN Module ....................... 65
2.7.2 Connecting the 2487 Detector to a 745/745B/746 Data
Module....................................................................... 68
2.7.3 Connecting the 2487 Detector to a Chart Recorder .. 71
2.7.4 Connecting the 2487 Detector to the Waters 600 Series
Pump......................................................................... 74
2.7.5 Connecting the 2487 Detector to the Waters 717plus
Autosampler .............................................................. 79
2.7.6 Connecting the 2487 Detector to a Fraction
Collector .................................................................... 81

8 Table of Contents
Chapter 3
Using the 2487 Detector .................................................................. 83
3.1 Starting Up the 2487 Detector .............................................. 83
3.1.1 Initializing the 2487 Detector ..................................... 83
3.1.2 Diagnostics Failure.................................................... 85
3.2 Using the Operator Interface................................................. 85
3.2.1 Using the Display....................................................... 85
3.2.2 Using the Keypad ...................................................... 88
3.2.3 Navigating the User Interface.................................... 93
3.2.4 Primary and Secondary Functions............................. 95
3.2.5 Operating the Trace and Scale Functions ............... 102
3.2.6 Operating Other Detector Functions........................ 104
3.3 Operating the 2487 Detector............................................... 109
3.3.1 Overview of Detector Operation .............................. 109
3.3.2 Verifying the Detector ...............................................110
3.3.3 Wavelength Calibration.............................................113
3.3.4 Operating the Detector in Single Wavelength
Mode ........................................................................114
3.3.5 Operating the Detector in Dual Wavelength
Mode ........................................................................115
3.3.6 Programming Timed Events, Threshold Events, and
Methods....................................................................118
3.4 Scanning Spectra................................................................ 126
3.4.1 Scanning New Spectra............................................ 130
3.4.2 Storing a Spectrum.................................................. 138
3.4.3 Getting Information About a Stored Spectrum......... 138

Table of Contents 9
3.4.4 Reviewing a Stored Spectrum ................................. 139
3.4.5 Subtracting a Spectrum ........................................... 139
3.4.6 Replaying a Spectrum ............................................. 140
3.4.7 Scanning Using the Cuvette.................................... 140
3.4.8 Scanning Using a Static Flow Cell........................... 144
3.4.9 Conserving Lamp Life.............................................. 144
3.4.10 Shutting Down the 2487 Detector.......................... 146
Chapter 4
Maintenance Procedures ............................................................... 148
4.1 Introduction......................................................................... 148
4.2 Routine Maintenance.......................................................... 150
4.3 Inspecting, Cleaning, and Replacing Parts of the Flow
Cell .................................................................................... 151
4.3.1 Flushing the Flow Cell ............................................. 151
4.3.2 Removing and Cleaning the Flow Cell..................... 152
4.3.3 Disassembling and Reassembling the Flow Cell..... 152
4.4 Replacing the Lamp............................................................ 160
4.4.1 Removing the Lamp................................................. 163
4.4.2 Installing the New Lamp .......................................... 165
4.4.3 Recording the New Lamp Serial Number................ 167
4.5 Replacing Fuses ................................................................. 169
Chapter 5
Error Messages, Diagnostics, and Troubleshooting ...................... 172
5.1 Error Messages................................................................... 172
5.1.1 Startup Error Messages........................................... 172
5.1.2 Error Messages Preventing Operation .................... 175

10 Table of Contents
5.2 User-Selected Diagnostics.................................................. 179
5.2.1 Overview.................................................................. 179
5.2.2 Using the Diagnostics.............................................. 181
5.2.3 Service Diagnostics................................................. 189
5.3 Troubleshooting .................................................................. 189
5.3.1 Introduction.............................................................. 189
5.3.2 Hardware Troubleshooting ...................................... 190
Appendix A
2487 Detector Specifications.......................................................... 193
Appendix B
Spare Parts..................................................................................... 197
Appendix C
Warranty Information...................................................................... 199
C.1 Limited Express Warranty .................................................. 199
C.2 Shipments, Damages, Claims, and Returns ...................... 203
Appendix D
Solvent Considerations................................................................... 204
D.1 Introduction......................................................................... 204
D.2 Solvent Miscibility............................................................... 205
D.3 Buffered Solvents............................................................... 208
D.4 Head Height ....................................................................... 209
D.5 Solvent Viscosity ................................................................ 209
D.6 Mobile Phase Solvent Degassing....................................... 209
D.6.1 Gas Solubility.......................................................... 209
D.6.2 Solvent Degassing Methods ................................... 210

Table of Contents 11
D.6.3 Solvent Degassing Considerations..........................211
D.7 Wavelength Selection......................................................... 212
Index .......................................................................................... 217

Table of Contents 12
1-1 Waters 2487 Dual λAbsorbance Detector.................................... 25
1-2 Waters 2487 Dual λ Absorbance Detector Optics Assembly ........28
1-3 Comparison of Flow Cell Characteristics.......................................31
1-4 Time Constant (Filter Setting) Comparison ...................................33
2-1 Major Steps in Installing the 2487 Detector...................................39
2-2 2487 Detector Dimensions ............................................................ 40
2-3 2487 Detector Fluidic Connections................................................45
2-4 Ferrule and Compression Screw Assembly...................................46
2-5 2487 Detector Rear Panel Electrical Connections.........................47
2-6 Overview of Making Signal Connections.......................................50
2-7 I/O Signal Inputs and Outputs........................................................51
2-8 2690 Separations Module Connections to the 2487 Detector for
Auto Zero on Inject ........................................................................53
2-9 2690 Separations Module Connections to the 2487 Detector for
Chart Mark on Inject ......................................................................55
2-10 2690 Separations Module Connections to the 2487 Detector for
Starting a Method ..........................................................................56
2-11 2690 Separations Module Connections to the 2487 Detector for
Turning the Lamp On or Off........................................................... 57
2-12 IEEE-488 Connections to the 2487 Detector in a Waters
Millennium32 System .....................................................................60
2-13 IEEE-488 Connections to the 2487 Detector in an Alliance
System...........................................................................................61
2-14 PowerLine Controller IEEE-488 Connections................................ 62
2-15 busSAT/IN Module (Front Panel)...................................................65
2-16 busSAT/IN Module Channel 1 Connection to 2487 Detector
Channel A......................................................................................67
2-17 Waters 745/745B/746 Data Module Connections to 2487 Detector
Channels A and B..........................................................................70
List of Figures

Table of Contents 13
2-18 Chart Recorder 2 VOutput Connections on 2487 Detector Channels
A and B..........................................................................................72
2-19 Chart Recorder 10 mV Output Connections on 2487 Detector
Channels A and B..........................................................................74
2-20 Waters 600 Series Pump Lamp On/Off Connections ....................75
2-21 Waters 600 Series Pump Auto Zero Connections.........................76
2-22 Waters 600 Series Pump Chart Mark Connections....................... 77
2-23 Waters 600 Series Pump Inject Start Connections........................78
2-24 717plus Auto Zero Connections to the 2487 Detector................... 80
2-25 Waters 2487 Detector Inject Start Connections............................. 81
3-1 Waters 2487 Detector Startup Diagnostics Display.......................84
3-2 2487 Detector Absorbance Screen................................................85
3-3 2487 Detector Keypad................................................................... 89
3-4 Secondary Functions of the Absorbance Screen ..........................96
3-5 Scaled Trace of Continuous Injections with T1 Changed to
–60.............................................................................................. 102
3-6 Scaled Trace for 5 Minutes of Continuous Injections with T1
Changed to –5............................................................................. 103
3-7 Scaled Trace for 60 Minutes of Continuous Injections with AU1
Changed to 1............................................................................... 103
3-8 Scaled Trace Changing T1 to –45............................................... 103
3-9 Configuration Screens................................................................. 104
3-10 Setting the Pulse Period or Signal Width Using SW1 or SW2..... 107
3-11 Display Contrast Screen.............................................................. 107
3-12 System Info Screen Examples..................................................... 108
3-13 Help Screen................................................................................. 108
3-14 Sample and Reference Energy Diagnostic Display......................112
3-15 Cuvette Wavelength Calibration Message....................................113
3-16 Calibration Successful Message...................................................113
3-17 Wavelength Mode Change Message (for Single Wavelength).....114
3-18 Wavelength Mode Change Message (for Dual Wavelength)........116

14 Table of Contents
3-19 Dual Wavelength 370 nm Threshold Warning Message...............117
3-20 Method Choice List......................................................................120
3-21 Timed Events Screen .................................................................. 121
3-22 Threshold Events Screen ............................................................ 123
3-23 Storing a Method, Method Number Field..................................... 124
3-24 Reset Method Message............................................................... 125
3-25 Clear Events Message................................................................. 126
3-26 Scan of Anthracene at 100 nm/min and 1000 nm/min................. 128
3-27 Scan of Erbium Standard in a Cuvette from 190 nm to 600 nm
at a Pace of 200 nm/min with Tickmarks Every 20 nm................129
3-28 Scan of Erbium Standard in a Cuvette from 190 nm to 600 nm
at a Pace of 200 nm/min Without Tickmarks...............................129
3-29 Scan Choice List..........................................................................130
3-30 Zero and Sample Scan Screens.................................................. 133
3-31 Scanning Progress Bar................................................................ 134
3-32 Sample Erbium Scan Graphical Display......................................135
3-33 Sample Erbium Scan Four Highest Peaks .................................. 135
3-34 Sample Erbium Scan with λ1 Changed to 225 nm and λ2
Changed to 600 nm..................................................................... 136
3-35 Four Highest Peaks from the Scaled Sample Erbium Scan........ 136
3-36 Series of Scans of Anthracene in Acetonitrile.............................. 137
3-37 Storage Slot Number Box............................................................138
3-38 2487 Detector Cuvette Holder with the Cuvette Inserted ............ 141
3-39 Removing the Cuvette Holder...................................................... 142
3-40 Replacing the Cuvette Holder...................................................... 143
3-41 Lamp Control Screen...................................................................145
3-42 Lamp Off/On Sequence...............................................................145
4-1 View of the Waters 2487 Dual λ Absorbance Detector with
the Left Front Panel Cover Removed..........................................150
4-2 Unscrewing the Flow Cell Assembly Captive Screws.................. 153
4-3 Removing the Flow Cell Assembly..............................................154

Table of Contents 15
4-4 2487 Detector Flow Cell Assembly with the Cuvette Subassembly
Removed ..................................................................................... 155
4-5 Waters TaperSlit Flow Cell........................................................... 156
4-6 Waters TaperSlit Flow Cell, Exploded View................................. 157
4-7 Deuterium Lamp Sample Beam Intensity Profile.........................161
4-8 Lamp Assembly and Power Connector ....................................... 163
4-9 Loosening the Captive Screws at the Lamp Housing Base......... 164
4-10 Removing the Lamp..................................................................... 165
4-11 Aligning the Lamp........................................................................ 167
4-12 Change Lamp Screen..................................................................168
4-13 Lamp Serial Number OK to Store Message ................................ 168
4-14 Lamp Serial Number Confirmation Message............................... 169
4-15 2487 Detector Rear Panel Fuse Holder....................................... 169
4-16 Removing and Replacing the Rear Panel Fuses and Fuse
Holder.......................................................................................... 171
5-1 Fatal Error Display on the Absorbance Screen ........................... 175
5-2 Diagnostics Choice List ............................................................... 179
5-3 Absorbance Screen with Sticky Diagnostic Active....................... 180
5-4 Sample and Reference Energy Diagnostic.................................. 182
5-5 Input and Output Diagnostic Choice List ..................................... 183
5-6 Auto Zero Offsets Diagnostic....................................................... 183
5-7 Fix Absorbance Diagnostic.......................................................... 184
5-8 Fix Voltage Diagnostic................................................................. 184
5-9 Contact Closures & Events Diagnostic........................................ 185
5-10 Change Lamp Diagnostic............................................................ 185
5-11 Keypad Diagnostic.......................................................................186
5-12 Other Diagnostics Choice List ..................................................... 187
5-13 Generate Test Peaks Diagnostic ................................................. 187
5-14 Optical Filter Override Diagnostic................................................ 188

Table of Contents 16
2-1 Installation Site Requirements ......................................................41
2-2 I/O Signals for the 2487 Detector ............................................ 51
2-3 2487 Detector Connections to a Waters 2690 Separation Module
to Generate Auto Zero on Inject ...................................................53
2-4 2487 Detector Connections to a Waters 2690 Separation Module
to Generate Chart Mark on Inject .................................................54
2-5 2487 Detector Connections to a Waters 2690 Separation
Module to Start a Method........................................................ 55
2-6 2487 Detector Connections to a Waters 2690 Separation
Module to Turn the Detector Lamp On or Off........................... 57
2-7 2487 Detector Connections to a busSAT/IN Module................ 68
2-8 2487 Detector Connections to a Waters 745/745B/746 Data
Module.................................................................................... 68
2-9 2487 Detector Connections to a Chart Recorder..................... 71
2-10 Waters 600 Series Pump and 2487 Detector Lamp On/Off
Connections............................................................................ 74
2-11 Waters 600 Series Pump and 2487 Detector Auto Zero
Connections............................................................................ 75
2-12 Waters 600 Series Pump and 2487 Detector Chart Mark
Connections............................................................................ 76
2-13 Waters 600 Series Pump and 2487 Detector Inject Start
Connections............................................................................ 78
2-14 Waters 717plus Autosampler and 2487 Detector Auto Zero
Connections............................................................................ 79
2-15 Waters 717plus Autosampler and 2487 Detector Inject Start
Connections............................................................................ 80
2-16 Fraction Collector to 2487 Detector Connections .................... 82
3-1 Absorbance and Message Screen Icons ......................................86
3-2 2487 Detector Keypad Description.......................................... 90
List of Tables

Table of Contents 17
3-3 Primary and Secondary Function (Method) Parameters ........ 100
3-4 Timed Event Parameters....................................................... 119
3-5 Threshold Events “Set” Parameters ...................................... 122
3-6 Threshold Events “To” Parameters........................................ 122
3-7 Pace and Sampling Resolution Examples............................. 127
3-8 Sample and Zero Scan Parameters....................................... 131
5-1 Startup, Calibration, and Operating Error Messages .................. 173
5-2 Possibly Fatal Error Messages.............................................. 176
5-3 2487 Detector Diagnostics.................................................... 180
5-4 General System Troubleshooting .......................................... 190
A-1 Operational Specifications .......................................................... 193
A-2 Optical Specifications ................................................................. 196
A-3 Optional Waters TaperSlit Flow Cell Specifications .................... 196
B-1 Spare Parts ................................................................................. 197
C-1 2487 Detector Warranty Periods ........................................... 202
D-1 Solvent Miscibility .......................................................................206
D-2 UV Cutoff Wavelengths for Common Chromatographic
Solvents................................................................................ 212
D-3 Wavelength Cutoffs for Different Mobile Phases.................... 213
D-4 Electronic Absorption Bands of Representative
Chromophores ...................................................................... 215

18 How to Use This Guide
How to Use This Guide
Purpose of This Guide
The Waters 2487 Dual λ Absorbance Detector Operator’s Guide describes the
procedures for unpacking, installing, operating, verifying, maintaining, and troubleshooting
the Waters®2487 Dual λ Absorbance Detector. It also includes appendixes describing
instrument specifications, spare parts, warranty and service information, and solvent
considerations.
Audience
This guide is for use by anyone who installs, maintains, operates, or troubleshoots the
Waters 2487 Dual λ Absorbance Detector. All personnel who use this guide should be
familiar with HPLC terms and practices, and be capable of basic HPLC system operations
such as making fluidic connections.
Structure of This Guide
The Waters 2487 Dual λ Absorbance Detector Operator’s Guide is divided into
chapters and appendixes. Each page is marked with a tab and a footer to help you access
information.
The following table defines each chapter and appendix in this guide.
Chapter/Appendix Description
Chapter 1, 2487 Detector
Theory of Operation Summarizes the features of the Waters 2487 Detector
and describes the theory and principles of operation.
Chapter 2, Installing the 2487
Detector Describes how to install the Waters 2487 Detector,
make fluidic and power connections, and connect the
detector to other devices.
Chapter 3, Using the 2487
Detector Describes how to operate the Waters 2487 Detector
from power-on to power-off, and use the operator
interface.
Chapter 4, Maintenance
Procedures Describes how to clean and replace various parts of
the Waters 2487 Detector.

How To Use This Guide 19
Related Documents
The following table lists other documents related to the operation of the Waters 2487 Dual
λ Absorbance Detector.
Chapter 5, Error Messages,
Diagnostics, and
Troubleshooting
Describes the error messages, diagnostics, and
recommended actions used in troubleshooting the
Waters 2487 Detector.
Appendix A, 2487 Detector
Specifications Contains operational, optical, optional flow cell, and
voltage specifications for the Waters 2487 Detector.
Appendix B, Spare Parts Lists recommended and optional spare parts for the
Waters 2487 Detector.
Appendix C, Warranty
Information Includes warranty, serial number, and service
information for the Waters 2487 Detector.
Appendix D, Solvent
Considerations Provides information about the solvents used with
Waters detectors.
Title Description
Waters 2690 Separations
Module Operator’s Guide Describes the procedures for unpacking, installing,
using, maintaining, and troubleshooting the Waters
2690 Separations Module.
Waters 600E Multisolvent
Delivery System User’s Guide Describes the procedures for installing, using,
maintaining, and troubleshooting the Waters 600E
Multisolvent Delivery System.
Waters Bus SAT/IN Module
Installation Guide Provides the procedures for installing the Waters
busSAT/IN Module.
Waters Alliance Dissolution
System Operator’s Guide Describes the procedures for unpacking, installing,
using, maintaining, and troubleshooting the Waters
Alliance Dissolution System.
Waters 2790 Separations
Module Operator’s Guide Describes the procedures for installing, using,
maintaining, and troubleshooting the Waters 2790
Separations Module.
Chapter/Appendix Description

20 How to Use This Guide
Related Documentation
Online Documentation
The following table describes the online documentation that supports the Millennium32
software.
Title Description
Millennium32 Online Help Describes all Millennium32 windows, menus, menu
selections, and dialog boxes for the base software and all
the software options. Also includes reference information
and procedures for performing all tasks required to use
the Millennium32 software. Included as part of the
Millennium32 software.
Millennium32 Read Me File Describes product features and enhancements, helpful
tips, installation and/or configuration considerations, and
changes since the previous Millennium software version.
Millennium32 LIMS Online
Help Describes how to use the Millennium32 LIMS Interface to
export results and import worklists.
Millennium32 Toolkit
Professional Online Help Describes how to use the common object model,
message-based protocol to communicate with the
Millennium32 software from a third-party application.
Millennium32 WebServer
Online Help Describes how to use the Millennium32 WebServer
application.