λ Waters 2487 Dual 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. Attention: The Waters 2487 Dual λ Absorbance Detector can be used for in vitro STOP 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. Attention: Changes or modifications to this unit not expressly approved by the party STOP responsible for compliance could void the user’s authority to operate the equipment. Attention: To meet the regulatory requirements of immunity from external electrical STOP 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. 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. 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 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 6 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 Table of Contents 7 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 8 Table of Contents 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 Table of Contents 9 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 10 Table of Contents