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Waters 2489 Operator's Manual

Uv/visible detector
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2489 UV/Visible
Detector
Operator's Guide
715003800 / Revision A
This operator's guide is intended for use with the 2489 UV/Visible detector,
part number 186248900, as pictured above. For the documentation to support
the 2489 UV/Visible detector, part number 186002489, see the operator's
guide, part number 71500142102.
To locate the part number for your device, see the serial number label on
Note:
the back of the instrument.
Copyright © Waters Corporation 2012
All rights reserved

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Summary of Contents for Waters 2489

  • Page 1 Detector Operator’s Guide 715003800 / Revision A This operator's guide is intended for use with the 2489 UV/Visible detector, part number 186248900, as pictured above. For the documentation to support the 2489 UV/Visible detector, part number 186002489, see the operator's guide, part number 71500142102.
  • Page 2 November 9, 2012, 715003800 Rev. A...

  • Page 3: General Information

    Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, its use.

  • Page 4: Safety Considerations

    Contact 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...

  • Page 5: Safety Advisories

    Good Laboratory Practice, and consult your organization’s safety representative for guidance. When you develop methods, follow the “Protocol for the Adoption of Analytical Methods in the Clinical Chemistry Laboratory,” American Journal of Medical Technology, 44, 1, pages 30–37 (1978). This protocol addresses good operating procedures and the techniques necessary to validate system and method performance.

  • Page 6: Equipment Misuse Notice

    Equipment misuse notice If the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. Operate this instrument When operating this instrument, follow standard quality-control (QC) procedures and the guidelines presented in this section. November 9, 2012, 715003800 Rev.

  • Page 7: Applicable Symbols

    2012/19/EU, contact Waters Corporation for the correct disposal and recycling instructions. Audience and purpose This guide is intended for personnel who install, operate, and maintain 2489 UV/Visible detectors. Intended use of the 2489 UV/Visible detector Waters designed the 2489 UV/Visible detector to analyze and monitor many compounds.

  • Page 8: Calibrate

    Calibrate 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.

  • Page 9: Ec Authorized Representative

    EC Authorized Representative Waters Corporation (Micromass UK Ltd.) Floats Road Wythenshawe Manchester M23 9LZ United Kingdom Telephone: +44-161-946-2400 Fax: +44-161-946-2480 Contact: Quality manager November 9, 2012, 715003800 Rev. A...
  • Page 10 November 9, 2012, 715003800 Rev. A...

  • Page 11: Table Of Contents

    Equipment misuse notice ................... vi Operate this instrument ..................vi Applicable symbols ................... vii Audience and purpose..................vii Intended use of the 2489 UV/Visible detector ..........vii Calibrate......................viii Quality-control ....................viii ISM classification ....................viii ISM Classification: ISM Group 1 Class B ............viii EC Authorized Representative .................
  • Page 12 Operational modes ..................... 25 Single-wavelength mode..................25 Dual-wavelength mode ..................27 Spectrum scanning .................... 28 Cuvette operations..................... 29 RatioPlot......................29 MaxPlot ......................30 Thermal-wander management................30 2 Install the Detector ................. 31 Prepare to install the detector ............... 32 Site selection and power requirements ............33 Site selection ......................
  • Page 13 Scan using a flow cell and a syringe ............... 112 Conserve lamp life ................... 113 Shutting down the detector ................115 4 Maintaining the Detector ..............117 Contact Waters technical service ..............117 Maintenance considerations ................. 118 Safety and handling..................118 Spare parts ....................... 119 Proper operating procedures ................
  • Page 14 Warning symbols ....................162 Task-specific hazard warnings................ 162 Specific warnings ..................... 163 Caution advisory ....................166 Warnings that apply to all Waters instruments ........167 Warnings that address the replacing of fuses ........... 172 Electrical symbols .................... 173 Handling symbols .................... 174...
  • Page 15 B Specifications ..................175 Operational specifications ................176 Optional Waters TaperSlit flow cell specifications ......... 178 C Solvent Considerations ............... 181 Introduction ...................... 181 Preventing contamination ................181 Clean solvents ....................181 Solvent quality ....................182 Preparation checklist..................182 Water ........................ 182 Using buffers ....................
  • Page 16 November 9, 2012, 715003800 Rev. A...

  • Page 17: Theory And Principles Of Operation

    Appendix C See also: information on high-performance liquid chromatography (HPLC) solvent considerations. Detector description The Waters 2489 UV/Visible Detector is a two-channel ultraviolet/visible (UV/Vis) detector designed for high-performance liquid chromatography (HPLC) applications. November 9, 2012, 715003800 Rev. A...
  • Page 18 UV/Vis functionality. • Cuvette qualification – Facilitates qualification of the detector by insertion of a standard in a cuvette without breaking any plumbing connections. Waters qualification kits, available in cuvette form, support November 9, 2012, 715003800 Rev. A...

  • Page 19: Principles Of Operation

    This section describes the following parts and functions of the detector: • Optics • Wavelength verification and test • Flow cell • Electronics Detector optics The Waters 2489 UV/Visible detector optics are based on a Fastie-Ebert monochromator and include these components: November 9, 2012, 715003800 Rev. A...
  • Page 20 Entrance slit • Blazed, plane-holographic, diffraction grating • Beamsplitter • Sample and reference photodiodes • Waters TaperSlit™ Flow Cell (its entrance is the exit slit of the monochromator) • Cuvette holder Waters 2489 UV/Visible detector optics assembly: lamp Spherical mirror...
  • Page 21 As shown in the figure below, in a conventional cell, light bends and hits the wall of the flow cell. Four beams go in, but only two come out. In the Waters TaperSlit analytical cell, the combination of the lens and TaperSlit bore geometry prevents light from hitting the cell walls.
  • Page 22 1 Theory and Principles of Operation Comparison of flow cell characteristics: Conventional cell Window UV light Window TaperSlit analytical cell Window UV light Window The standard analytical, inert, and LC/MS cells have a path length of 10 mm. The semi-prep and microbore cell path length is 3 mm. The autopurification cell path length is 1.0 mm.
  • Page 23 Principles of operation value is greater than 30, you should specify a lower sampling rate in the instrument method. Set the sampling rate to the lowest value required to achieve 15 or more points across the narrowest peak. Excessively high sampling rates can slow the system with more data than you need for your analysis.

  • Page 24: Wavelength Verification And Test

    1 Theory and Principles of Operation Filter time constant comparison: 0 sec 1 sec 2 sec Time (minutes) Although the peak shape shows some distortion and the signal output is Tip: delayed with different time constants, the peak area remains the same. Wavelength verification and test The detector’s deuterium arc lamp and integral erbium filter exhibit peaks in the transmission spectrum at known wavelengths.

  • Page 25: Operational Modes

    521.5 nm The verification tests for the detector require 5 minutes of lamp warmup time. If you run the detector continuously, Waters recommends that you perform wavelength verification weekly by turning off the detector, then turning it on again. See “Wavelength calibration”...
  • Page 26 1 Theory and Principles of Operation mode, so you can use channel B to obtain additional information about the wavelength selected on channel A. In single-wavelength mode, the detector automatically engages the second-order filter for wavelengths 370 nm and above and removes it for wavelengths under of 370 nm.

  • Page 27: Dual-Wavelength Mode

    Operational modes • Absorbance offset (in mV) • Auto zero on inject • Auto zero on λ changes “Primary and secondary functions” on page 70 and the table on page 61 explain the functions, ranges, and defaults of these parameters. Dual-wavelength mode In dual-wavelength mode, the detector can monitor two wavelengths, one on channel A and one on channel B.

  • Page 28: Spectrum Scanning

    1 Theory and Principles of Operation channels that can be controlled independently, even in single-wavelength mode. • MaxPlot – This mode results in the output of the larger of the two absorbance values scaled to the selected AU setting. Use this mode when observing, with one data channel, multiple compounds that exhibit absorbancies at two separate wavelengths.

  • Page 29: Cuvette Operations

    Operational modes To obtain a spectrum of a sample using the detector, run a zero scan first, followed by a sample scan. Typically, you run the zero scan using pure solvent. The sample scan is a scan of the analyte dissolved in that solvent. Spectra can be simultaneously charted on the channel A output, or acquired and stored in memory for later playback.

  • Page 30: Maxplot

    1 Theory and Principles of Operation MaxPlot The MaxPlot function monitors absorbance at two selected wavelengths and plots the maximum absorbance value for each sample component. To obtain a MaxPlot, you must operate the detector in dual-wavelength mode. The MaxPlot outputs the greater of the two absorbance values on the selected channel.

  • Page 31: Install The Detector

    Install the Detector The detector requires connections to electrical power and to sample and waste lines to operate in any standard laboratory environment. This chapter describes how to install the detector, and connect it to the electrical supplies and to other equipment in an HPLC system. Contents: Topic Page...

  • Page 32: Prepare To Install The Detector

    2 Install the Detector Prepare to install the detector Major steps in installing the detector: Start installation procedure Make power connections Select appropriate site Make signal connections Unpack and inspect Make connections to other devices Install detector Installation complete Make plumbing connections After installing the detector, verify its function, and keep the verified chart output (if applicable) on file.

  • Page 33: Site Selection And Power Requirements

    Site selection and power requirements Detector dimensions: 20.8 cm (8.2 inches) 61.0 cm (24.0 inches) 34.3 cm (13.5 inches) Do not access the instrument through the top cover but Caution: through the front door, where the lamp housing, flow cell assembly, and cuvette holder are located.
  • Page 34 • HAR type (or better), in Europe For information regarding the type of cord to use in other countries, contact your local Waters distributor. You must mount the detector on a level surface to allow proper Requirement: function of the drip management system (drain tube), which you can connect to a waste reservoir to divert solvent leaks from the flow cell.

  • Page 35: Power Requirements

    Unpack and inspect The detector is packed and shipped in one carton, which contains the following items: • Waters 2489 UV/Visible detector startup kit • Power cord Unpack To unpack the detector: Unpack the contents of the shipping carton.

  • Page 36: Make Plumbing Connections

    2 Install the Detector Inspect If you discover any damage or discrepancy when you inspect the contents of the cartons, immediately contact the shipping agent and your local Waters representative. Customers in the USA and Canada can report damage and discrepancies to Waters Technical Service (800 252-4752).
  • Page 37 Make plumbing connections • Install a waste reservoir connected to the drain tube and located next to the rubber foot on the front, lower, left-hand section of the detector. ® • Use Tygon tubing to connect the drain tube to the waste reservoir. The detector, as shipped, has a standard analytical flow Caution: cell, pressure-rated at 6895 kPa (69 bar, 1000 psi).

  • Page 38: Make Electrical Power Connections

    (see the figure below). Plug the other end of the power cord into a properly grounded AC power source. The detector connects to other Waters components through rear panel electrical connections. November 9, 2012, 715003800 Rev. A...
  • Page 39 Make electrical power connections Detector rear panel electrical connections: Inputs and outputs Power input Fuse holder Rear panel connections enable the following signals: • Analog outputs – There are two pairs of attenuated analog channel outputs. Each pair supports 2-V output to external devices or data systems.

  • Page 40: Make Signal Connections From Rear Panel And Ethernet Connectors To The Detector

    • Ethernet interface – The Ethernet connection on the rear panel of the detector provides remote control operation and direct data acquisition from Waters Empower and MassLynx workstations. Make signal connections from rear panel and Ethernet connectors to the detector...
  • Page 41 Make electrical power connections Overview of making signal connections: Start signal connection procedure Install Ethernet and Connect to inject start cables Ethernet bus? Connect to other Install event and devices? I/O cable(s) if Signal connections complete Make I/O signal connections The rear panel includes two removable connectors that hold the pins for the I/O signals, as shown in the figure below.
  • Page 42 2 Install the Detector I/O signal inputs and outputs: Connector I Connector II Analog 1 + Inject Start + Analog 1 − Inject Start − Ground Ground Analog 2 + Lamp On + Analog 2 − Lamp On − Switch 1 Chart-mark + Chart-mark −...

  • Page 43: Make Ethernet Connections

    When controlling the detector from a Waters data system or controller (Empower or MassLynx workstation), you can use the Ethernet interface to send and receive information from the data system. When connecting via the Ethernet to these Waters data systems, you should be aware of the following: •...
  • Page 44 Empower allows the detector to operate, in both single and dual-wavelength, modes with a wavelength range of 190 to 700 nm. To connect Ethernet cables from the detector to a Waters data system: Connect the single receptacle end of the Ethernet cable to your data system by attaching the cable to your network card or Ethernet switch.
  • Page 45 Make electrical power connections Separations module connections to detector for starting a method: e2695 separations module (B inputs and 2489 detector (II) outputs) Pin 1 Inject Start Pin 1 Inject Start + Pin 2 Inject Start Pin 2 Inject Start –...
  • Page 46 (lamp on or off): e2695 separations 2489 detector (II) module (A outputs) Pin 1 Switch 1 Pin 4 Lamp On/Off + Pin 2 Switch 1 Pin 5 Lamp On/Off –...
  • Page 47 (B inputs and 2489 detector (II) outputs) Pin 1 Inject Start Pin 9 Auto Zero + Pin 2 Inject Start Pin 10 Auto Zero –...
  • Page 48 2 Install the Detector e2695 separations module connections to the detector for auto-zero on inject: e2695 connector B 2489 connector II Inject Start + Inject Start − Inject Start + Ground Inject Start − Lamp On/Off + Ground Lamp On/Off −...

  • Page 49: Connect To Other Devices

    Connect to other devices e2695 separations module connections to the detector for making a chart-mark on injection: e2695 connector B 2489 connector II Inject Start + Inject Start − Inject Start + Ground Inject Start − Lamp On/Off + Ground Lamp On/Off −...

  • Page 50: Connect The Detector To Empower Using An E-Sat/In Module

    Ethernet satellite interface (e-SAT/IN) module e-SAT/IN module The Waters e-SAT/IN module shown in the figure below translates analog signals from devices such as the detector into digital form. It then transmits these digital signals using an ethernet card installed in the Empower workstation.
  • Page 51 • To ensure proper startup of the e-SAT/IN module, do not power-on the module until you perform all procedures in the Waters e-SAT/IN Module Installation Guide. Improper startup can damage the unit and void the warranty. Connect the detector to the e-SAT/IN module...
  • Page 52 1 on I (Analog 1 +). • connect the black wire to pin 3 on I (Ground). e-SAT/IN module channel 1 connection to detector: 2489 connector I Analog 1 + Analog 1 − Ground...

  • Page 53: Connect The Detector To A 745/745B/746 Data Module

    Pin 8 Ground (black) Connect the detector to a 745/745B/746 data module You can connect the detector to a Waters 745/745B/746 data module using the analog output connector on the rear panel of the detector. The analog connector provides 2-V output that is scaled to the AU sensitivity setting and the voltage offset setting.
  • Page 54 To send the analog output signal from the detector to the data module, use the cable provided in the 2489 Detector Startup Kit to make the connections summarized in the table below and illustrated in the figure below: Detector connections to a data module:...

  • Page 55: Connect The Detector To A Fraction Collector

    Connect to other devices Data module connections to detector channels A and B: 2489 connector I Analog 1 + Analog 1 − Ground Analog 2 + Black Analog 2 − Switch 1 Switch 1 Ground 745/745B/746 terminals Switch 2 Switch 2...
  • Page 56 The table below indicates the detector-to-fraction collector and autoinjector-to-fraction collector connections. Detector connections to the fraction collector: 2489 connection Fraction collector I Pin 3 Ground Pin 1 Detector In –...

  • Page 57: Prepare The Detector

    Initialize the detector Prepare the Detector Contents: Topic Page Initialize the detector............... 57 Use the operator interface ............... 59 Scan spectra ..................97 After you install the detector, you can set it up and operate it as a stand-alone instrument or as part of a an Empower or MassLynx data system. •...
  • Page 58 3 Prepare the Detector Startup Diagnostics screen: STARTUP DIAGNOSTICS GPIB QSPI After it displays the Startup Diagnostics screen, the detector displays the following series of messages in sequence—a process that takes about five minutes: • Initializing grating • Initializing system •...

  • Page 59: Idle Mode

    Idle mode When the detector is successfully started, it defaults to idle mode (see the figure “2489 detector idle mode screen:” on page 59). When it is not performing any function requiring the shutter to be open (local methods, scans, noise test, and so on), the shutter is closed, and the detector remains in idle mode with the lamp lit.
  • Page 60 3 Prepare the Detector Detector absorbance screen: Absorbance Lamp on/off Shift on/off Single/dual-wavelength Channel selector Keypad lock/unlock Local (method #)/Remote control Wavelength Run time (minutes) Sensitivity Sticky diagnostic test on/off Next You can recall the absorbance screen at any time by pressing the HOME key. When you first use the detector, the absorbance screen shows the factory-set defaults.
  • Page 61 Use the operator interface Absorbance and message screen icons: Icon or field Icon/field name Function Field Sensitivity or AU Selects the chart sensitivity in requiring absorbance units (AU) for the selected entry channel (Ethernet signal is not affected). Field Wavelength Selects the wavelength monitored on the requiring selected channel.
  • Page 62 3 Prepare the Detector Absorbance and message screen icons: (Continued) Icon or field Icon/field name Function Keypad unlock Indicates unrestricted keypad entry. Keypad lock Indicates parameter changes are not allowed. The keypad locks when the instrument is under control of an external data system (remote mode only).

  • Page 63: Using The Keypad

    Use the operator interface Absorbance and message screen icons: (Continued) Icon or field Icon/field name Function Message screen Indicates that you must standby. icon. Using the keypad The detector keypad (shown below) consists of 24 keys providing, • full numeric entry (10 digits plus a decimal point); •...
  • Page 64 3 Prepare the Detector Keypad: λ/λλ SCAN Reset HOME Chart-mark Auto Zero Run/Stop METHOD Lamp Lock Calibrate System Info Previous CONFIGURE Contrast Next DIAG Scale TRACE Enter Clear Field Cancel +/− Shift Primary function keys take effect immediately, with no further entry required.
  • Page 65 Use the operator interface Detector keypad description: Unshifted Shifted HOME – Displays the ? – Displays context-sensitive absorbance screen containing help when available. HOME the icons, and the Wavelength and AU fields. Chart-mark – Causes a SCAN – Displays the list of SCAN momentary pulse to the options for generating and...
  • Page 66 3 Prepare the Detector Detector keypad description: (Continued) Unshifted Shifted Next – Displays a screen with Previous – When the Next key Previous additional options related to a is available, use Previous to Next current screen. Repeatedly navigate through the screens pressing this key brings back in the reverse order.
  • Page 67 Use the operator interface Detector keypad description: (Continued) Unshifted Shifted 1 – See 0-9, above. Lamp – Displays a lamp use Lamp statistics for the currently installed lamp and provides the means for you to turn the lamp on or off. The current state of the lamp is indicated by an icon on the absorbance screen.

  • Page 68: Navigate The User Interface

    3 Prepare the Detector Detector keypad description: (Continued) Unshifted Shifted • – Enters a decimal point. +/– – Some edit fields accept +/– Also positions the cursor at negative-number entries. Use • the last entry in a list. this function to invert the sign of the number in an active field.
  • Page 69 Use the operator interface • Press the corresponding number key to select an item immediately. • Use the up and down arrow keys to scroll through the list, and then press Enter. You can press the numeral that corresponds to a desired choice Tip: without first pressing Enter.

  • Page 70: Primary And Secondary Functions

    3 Prepare the Detector • Time constant • Analog output sensitivity Depending on the other functions that you can perform during a run, several other parameters can require programming. See page 70 and the table on page 75 for the function descriptions, fields, screen number, type of function, display units, allowable ranges, and the default settings for the absorbance screen and the secondary function screens.
  • Page 71 Use the operator interface Detector functions: (Continued) Function Description Analog out In addition to the selections for single λ above, you (dual λ) can chart the same parameters on the other channel, at a different wavelength, and you can chart the following parameters: •...
  • Page 72 3 Prepare the Detector Detector functions: (Continued) Function Description – Maximum ratio: An actual ratio equal to the maximum ratio results in a full-scale output of 2 V. Absorbance offset is ignored with this selection. For a RatioPlot, the actual voltage charted is Volts out = 0 V if Absorbance A and B <...
  • Page 73 Use the operator interface Detector functions: (Continued) Function Description Auto-zero on inject Selected by default, this parameter results in an auto-zero each time the detector receives an inject-start signal via contact closure, Ethernet, or through the front panel. You can disable this parameter by pressing any numerical key.
  • Page 74 3 Prepare the Detector Secondary functions of the absorbance screen: Voltage offset and chart Absorbance screen (HOME) polarity (available for Press Next. both λ and λλ) Press Next. Minimum AU, minimum ration, Analog rate and filter and maximum ration (available time constant (available only for λλ) for both λ...
  • Page 75 Use the operator interface Primary and secondary function (method) parameters: Function Screen Type Units Range Default λ Numeric Integer 190 nm to 254 nm (Wavelength) (Absorbance 700 nm screen) AUFS Numeric AUFS 0.0001 to 4.0000 2.0000 Analog rate 2 (of 4) or Choice (λ): 10, 20, 40, 80 2 (of 5)
  • Page 76 3 Prepare the Detector Primary and secondary function (method) parameters: (Continued) Function Screen Type Units Range Default Voltage offset 4 (of 4) or Numeric Integer –2000 to 4 (of 5) 2000 Chart polarity 4 (of 4) or Choice None 4 (of 5) –...
  • Page 77 Use the operator interface • For AU1 and AU2, press CE to reset to auto. • T1 represents the left-hand side of the trace, or ending time to be displayed. The default is –30. • T2 represents the right-hand side of the trace, or starting time. The default is 0.

  • Page 78: Operate Other Detector Functions

    3 Prepare the Detector Scaled trace for 60 minutes of continuous injections, with AU1 changed to 1: The figure below shows a 60-minute trace on channel B scaled to the last 45 minutes. T1 is changed to –45. Scaled trace changing T1 to –45: As you modify the output using the Scale function, the Trace function continues to display the detector output in real time on either or both channels.
  • Page 79 Use the operator interface Configuration screens: Configuration screen 1 of 3 Configuration screen 2 of 3 Configuration screen 3 of 3 To prevent acquisition of incorrect data when operating the Requirement: detector in dual-wavelength mode under the control of the Empower software or a MassLynx system, you must select a data sampling rate of 1 point per second.
  • Page 80 3 Prepare the Detector – Ignore – Do not respond to chart-mark input. • Auto-zero – You can configure the auto-zero input to auto-zero absorbance readings on channel A and/or channel B. You determine the response of the channel using the enable auto-zero function explained in the table on page page 75 and shown in the figure on...
  • Page 81 Use the operator interface Set the pulse period or signal width using SW1 or SW2: n Seconds Single pulse n Seconds Rectangular wave Set display contrast The Contrast function allows you to adjust the contrast of the detector display screen. When you press the Contrast key (Shift 6), the Display Contrast screen appears.

  • Page 82: Operate The Detector

    3 Prepare the Detector System info screen example: Start of message End of message Use Help The detector has limited context-sensitive help. When you press ? (Shift HOME) from a point in the program that has a Help screen associated with it, the appropriate screen appears.

  • Page 83: Verify That The Detector Operates Properly

    After you have installed the detector, verify that it is operating properly by performing the procedures in this section. Requirements: • For complete validation procedures, you must obtain the Waters accuracy and linearity cuvette kits and system qualification tool for the detector. November 9, 2012, 715003800 Rev. A...
  • Page 84 3 Prepare the Detector • For spare parts details, see the Waters Quality Parts Locator on the Services & Support page of the Waters’ Web site. Tip: Before you pump solvent or mobile phase through the system: Flush the lines with filtered, degassed, and sparged HPLC-grade methanol.
  • Page 85 Use the operator interface • the flow cell is contaminated; • the lamp needs to be replaced; • there is an air bubble in flow cell. To record the sample and reference beam energies: On the absorbance screen, use the arrow keys to highlight the λ field. Enter 230 into the λ...

  • Page 86: Wavelength Calibration

    Press Calibrate (Shift 3) from the detector keypad. A message appears asking whether you removed the cuvette and Result: flushed the flow cell with a transparent solvent (Waters recommends methanol or water). Press Enter to continue the calibration cycle or Cancel to invoke the absorbance screen without calibrating the detector.

  • Page 87: Operate The Detector In Dual-Wavelength Mode

    Use the operator interface The detector displays a message indicating it is switching to Result: single-wavelength operation. Enter the wavelength and sensitivity on the absorbance screen, as well as any secondary parameters and timed or threshold events. Changing the sensitivity (AUFS) setting affects the 2-V Caution: output.
  • Page 88 3 Prepare the Detector Enter the wavelength to be monitored in the λ field, and then press Enter. Enter the other operating parameters and timed or threshold events, if desired. Press the A/B key to switch channels. The absorbance screen for the other channel appears.
  • Page 89 Use the operator interface The RatioPlot provides a plot of the ratio of the absorbances of two wavelengths from 0 V to 2 V. The minimum and maximum ratio parameters are measured ratio units, not absorbances. (See page 70.) To obtain a RatioPlot: Be sure the detector is operating in dual-wavelength mode (see the previous discussion).

  • Page 90: Program Timed Events, Threshold Events, And Methods

    3 Prepare the Detector Program timed events, threshold events, and methods You can retrieve as many as five methods, which the detector references as numbers 1 to 5. An asterisk in the method number icon (see the table on page 61) tells you the current conditions are not stored.
  • Page 91 Use the operator interface Changing the sensitivity (AUFS) setting affects the 2-V output. Caution: For example, 1 AU gives 0.5 AU/V, and 2 AU gives 1 AU/V. Timed event parameters: Specify Number Event Units Range or default channel Wavelength 190 to 700 Filter time Seconds 0: Disable filter...
  • Page 92 3 Prepare the Detector Method choice list: Press 1, Timed events. Enter the time for the event. Timed events screen: Press Enter to enter the time. To advance to the Set field (Events choice list), press the t key. Tip: Press Enter again to display the choice list, or, if you know the event number, press the number for the event you are programming.
  • Page 93 Use the operator interface 10. Press HOME to return to the absorbance screen, and then press Run/Stop. 11. Press Reset. Tips: • If the detector is under the control of an external device, the Inject Start programmed from that device runs the method. •...
  • Page 94 3 Prepare the Detector To define the pulse period, or the frequency of a wave, see “Configure the detector” on page To program a threshold event: Press the METHOD (Shift A/B) key on the detector keypad. Press 2, Threshold events. Threshold events screen: Press Enter to advance to the next (Set) field, or press the s and t keys to move among the three fields on the threshold events screen.
  • Page 95 Use the operator interface Store a method, method number field: No warning message appears when the method number Caution: you select is already assigned to a previously stored method. Pressing Enter stores the current method conditions, overwriting any previous method stored in the same slot. Enter a number from 1 to 5 and press Enter.
  • Page 96 3 Prepare the Detector Once you specify a method number to retrieve, the method Result: choice list appears, and the method number appears within the method number icon. Press 1 to view the timed events or 2 to view threshold events within the displayed method.

  • Page 97: Scan Spectra

    Scan spectra When you press HOME, the absorbance screen’s method number icon displays an asterisk. Clear events You can clear only timed or threshold events without resetting any other operating parameters. To clear all active timed or threshold events: Return to the Method choice list by pressing METHOD (Shift A/B). Press 6, Clear events.

  • Page 98: Before You Begin

    3 Prepare the Detector Before you begin Before you run a spectrum scan, specify values for the following parameters: • λ1 – Starting wavelength. Scanning begins at this wavelength. • λ2 – Ending wavelength. Scanning ends at this wavelength. • Pace –...
  • Page 99 Scan spectra The higher the number you enter in the Pace field, the lower the Tip: resolution of the scan. • Tick marks – This value allows tick marks (check marks) to be generated at the specified wavelength increment help interpret charted data.

  • Page 100: Scan New Spectra

    3 Prepare the Detector When you select a zero scan, the detector displays three additional screens, labeled 2 of 4, 3 of 4, and 4 of 4. You can change all parameters on these screens, including starting and ending wavelength, and the pace parameter. When you select a sample scan, the detector displays two additional screens, labeled 2 of 3 and 3 of 3 (see the figure on page...
  • Page 101 Scan spectra Press 1, New scan, or use the s and t keys to move through the Scan choice list. The detector displays the first of three parameter screens for a Result: sample scan or four parameter screens for a zero scan (figure “Zero and sample scan screen:”...
  • Page 102 3 Prepare the Detector To program a zero scan: Press SCAN, then 1, New scan, and press 2, Zero scan. Press Next to advance to the second Zero scan parameter screen. Enter the starting wavelength for the zero scan, and then press Enter. Enter the ending wavelength for the zero scan, and then press Enter.
  • Page 103 Scan spectra Zero and sample scan screen: Sample scan (screen 1 of 3) Zero scan (screen 1 of 4) Zero scan (screen 2 of 4) Sample scan (screen 2 of 3) Sample scan (screen 3 of 3) Zero scan (screen 3 of 4) Zero scan (screen 4 of 4) November 9, 2012, 715003800 Rev.
  • Page 104 3 Prepare the Detector Run the sample scan: Run the zero scan before running the sample scan. To ensure identical flow cell and solvent conditions, run the sample scan for the corresponding zero scan within 15 minutes of running the zero scan. To run a sample scan: Set up the zero (or reference) scan by following the steps in the zero scan procedure on...
  • Page 105 Scan spectra To return to the Scan choice list, at the end of the scan, press SCAN Tip: (Shift Chart-mark). Press Next. Doing so displays as many as four of the highest peaks scanned Result: within the specified range. The figure below shows the four highest peaks for the erbium scan shown in the graphical display (above).
  • Page 106 3 Prepare the Detector Sample erbium scan with l1 changed to 225 nm and l2 changed to 600 nm: Once you change one or more scaling parameters, press Enter to reformat the graphical display. 10. Once the scan reappears, press Next to display the four highest peaks of the scaled scan.

  • Page 107: Store A Spectrum

    Scan spectra Series of scans of anthracene in acetonitrile: Sample scan 200 nm to 400 nm –0.001 AU to 0.5 AU anthracene Zoom of sample scan 200 nm to 300 nm –0.001 AU to 0.5 AU anthracene, 230 nm to 270 nm λ2 changed to 300 nm Zoom of sample scan 230 nm to 270 nm...
  • Page 108 3 Prepare the Detector To store a spectrum: From the graphic display of a sample scan, return to the first Scan screen by pressing SCAN (Shift Chart-mark). Press 2, Store last scan. When you select Store last scan, you are storing the zero scan and Tip: the sample scan as a pair.

  • Page 109: Review A Stored Spectrum

    Scan spectra Review a stored spectrum Once you store a spectrum, you can retrieve it for review from one of the three available storage slots by selecting the Review option from the Scan choice list. To review a spectrum: Press SCAN (Shift Chart-mark) to view the Scan choice list. Press 4, Review.

  • Page 110: Replay A Spectrum

    3 Prepare the Detector Replay a spectrum You can play back the current spectrum or a stored spectrum in real time using the Real-time replay function on the Scan choice list. The detector plays back the selected spectrum in real time both on the detector display and out the analog connector to the chart or A/D device in a data collection system.
  • Page 111 Scan spectra Detector cuvette holder with the cuvette inserted: Aperture Frosted sides of cuvette facing up and down Because the scan is actually a composite of both the contents of Restriction: the cuvette and the flow cell, you need to perform cuvette scans under identical flow cell conditions.

  • Page 112: Scan Using A Flow Cell And A Syringe

    3 Prepare the Detector To begin a cuvette scan: Open the detector’s front door. Remove the cuvette holder, sliding it toward you. With the spring guide facing you, gently insert the cuvette (containing eluent) up under the guide, with the cap facing upward (into the holder) and a frosted side of the cuvette facing up.

  • Page 113: Conserve Lamp Life

    Tip: controller to turn the lamp off and on without using the front panel. Waters recommends that you program the lamp to shut off Recommendation: or turn the lamp off manually only if the value of the “Lamp off” parameter is more than 4 hours.
  • Page 114 3 Prepare the Detector To turn off the lamp manually from the detector front panel: Press Lamp (Shift 1) from the keypad. The lamp control screen appears. Lamp control screen: Press Lamp (Shift 1) again to turn the lamp off. Lamp off/on sequence: Lamp off indicator Lamp on indicator...

  • Page 115: Shutting Down The Detector

    Scan spectra • “Program timed events, threshold events, and methods” on page and the table on page 91 for more information on programming the lamp to turn on or off using a timed event. • “Configure event inputs (contact closures)” on page 79 for more information on programming the lamp through the external contact closure.
  • Page 116 3 Prepare the Detector November 9, 2012, 715003800 Rev. A...

  • Page 117: Maintaining The Detector

    Waters Technical Service (800 252-4752). Otherwise, phone the Waters corporate headquarters in Milford, Massachusetts (USA), or contact your local Waters subsidiary. Our Web site includes phone numbers and e-mail addresses for Waters locations worldwide. Go to www.waters.com. When you contact Waters, be prepared to provide this information: •...

  • Page 118: Maintenance Considerations

    • Software version and serial number For complete information on reporting shipping damages and submitting claims, see Waters Licenses, Warranties, and Support Services. Maintenance considerations Perform the procedures in this chapter when you discover a problem with a 2489 detector component or during preventive maintenance.

  • Page 119: Spare Parts

    AC outlet. Wait 10 seconds before you disconnect an assembly. Spare parts Replace only parts mentioned in this document. For spare parts details, see the Waters Quality Parts Locator on the Waters Web site’s Services & Support page. Proper operating procedures Caution: •...

  • Page 120: Maintain The Flow Cell

    4 Maintaining the Detector • filter and degas solvents to prolong column life, reduce pressure fluctuations, and decrease baseline noise; • flush buffered mobile phases out of the detector with HPLC-grade water followed by a 5 to 10% methanol solution each time the detector is powered-off.
  • Page 121 Maintain the flow cell • noise is higher than expected; • noise test results are not meeting specifications; • the detector fails to normalize. To avoid damaging the flow cell during reverse flushing, do not Caution: overpressure the cell. If you use buffered mobile phase, flush it from the detector before powering-off.

  • Page 122: Remove And Clean The Flow Cell

    4 Maintaining the Detector If the mobile phase is not miscible in water, first use an Caution: intermediary solvent. Resume pumping mobile phase. Reattach the column. If the flow cell remains dirty or blocked, reverse flush it. Remove and clean the flow cell If flushing the flow cell is not effective, follow the procedures to remove the flow cell and inspect for dirty or broken windows or dirty gaskets.
  • Page 123 Maintain the flow cell Always replace the flow cell gaskets when cleaning, Caution: rebuilding, or replacing other flow cell components. Required materials You need the following items to remove, clean, and replace the flow cell: • 1/4-inch flat-blade screwdriver • Ethyl alcohol or methanol •...
  • Page 124 To prevent contamination, use powder-free finger cots or Caution: gloves when disassembling, inspecting, cleaning, or replacing parts within the Waters TaperSlit™ Flow Cell or when removing or replacing the flow cell within its assembly. The TaperSlit flow cell consists of these components: •...
  • Page 125 Maintain the flow cell Waters TaperSlit flow cell: Cell exit Cell entrance For replacement parts for the TaperSlit flow cell, use the Flow Cell Rebuild Kit. Use nitrogen to clean the flow cell. Use ethanol or methanol to clean the Tip: lenses and window.
  • Page 126 Caution: gloves when disassembling, inspecting, cleaning, or replacing parts within the Waters TaperSlit Flow Cell or when removing or replacing the flow cell within its assembly. Work on a clean flat surface, such as a nonparticulating cloth or similar surface.
  • Page 127 Maintain the flow cell Using ethanol or methanol, clean the contaminated part. Blow dry with nitrogen. Using the Flow Cell Rebuild Kit, replace any flow cell parts that are scratched, burred, damaged, or do not come clean using nitrogen. Rebuild the flow cell by following the procedure in the next section. Rebuild the flow cell After cleaning or replacing other flow cell parts, rebuild the flow cell.
  • Page 128 To prevent contamination, use powder-free finger cots or Caution: gloves when disassembling, inspecting, cleaning, or replacing parts within the Waters patented TaperSlit Flow Cell or when removing or replacing the flow cell within its assembly. The detector is shipped with a standard analytical flow cell installed. Replace the flow cell when, •...

  • Page 129: Replace The Lamp

    Reconnect the inlet/outlet tubing to your LC system. Reconnect the power cord, and then power-on the detector. Replace the lamp Waters recommends that you change the detector lamp Recommendation: when it repeatedly fails to ignite, or when the detector fails to calibrate.
  • Page 130 4 Maintaining the Detector Deuterium lamp sample beam intensity profile: Wavelength (nm) Lamp energy and performance As the lamps used in traditional detectors age, the signal-to-noise performance of the instrument degrades. Determining the longevity of useful lamp life can be difficult because of varying user requirements and individual lamp behavior.

  • Page 131: Removing The Lamp

    Rule: energies” on page 84 when you change the detector lamp. Waters warrants the lamp to ignite and pass startup diagnostic tests for 2000 hours or one year from the date of purchase, whichever comes first. Removing the lamp The lamp housing gets extremely hot during operation. To...
  • Page 132 4 Maintaining the Detector power-off the lamp using a timed event, see the instructions in the Empower or MassLynx Help. Power-off the detector, and disconnect the power cord. After extinguishing the lamp, allow it to cool for at least 30 minutes. Open the front door.
  • Page 133 Replace the lamp Loosening the captive screws at the lamp housing base: Withdraw the lamp assembly from the lamp housing. Lamp gas is under slight negative pressure. To prevent Caution: shattering the glass, use care when disposing of the lamp. Cushion the old lamp by placing it in the packaging material of the new lamp before disposal.

  • Page 134: Install The New Lamp

    4 Maintaining the Detector Removing the lamp: Install the new lamp To avoid exposing your eyes to harmful ultraviolet radiation, Warning: never ignite a lamp while it is outside the instrument or if it is not properly secured in place. Do not touch the glass bulb of the new lamp.
  • Page 135 Replace the lamp Make sure the detector is powered-off and the power cord is Requirement: disconnected. To install the new lamp: When you change the lamp, always power-off the Caution: detector. After installing a new lamp, power-on the detector and always wait at least 5 minutes for the new lamp to warm up.

  • Page 136: Record The New Lamp Serial Number

    4 Maintaining the Detector Aligning the lamp: Cutout in 1:00 position Record the new lamp serial number Caution: • Always run the Change Lamp diagnostic test after you install a new lamp (see “Use the lamp, display, and keypad diagnostic tests” on page 154).
  • Page 137 Replace the lamp Be sure to enter the 9-digit lamp serial number and not the lamp Tip: part number when performing this procedure. Enter the 9-digit serial number of the new lamp in the active field. Change lamp screen: Press Enter to store the serial number and to move to the “Install date” field.

  • Page 138: Set The Lamp Threshold

    4 Maintaining the Detector Set the lamp threshold You can set an alarm threshold for the lamp. When the number of hours meets or surpasses the threshold, an alarm message appears. The default alarm threshold is 2000 hours. The alarm message also appears when you first power-on the instrument. The lamp threshold screen displays the total hours of lamp use since the lamp’s installation.
  • Page 139 Replace fuses Detector rear panel fuse holder: Fuse holder To avoid electric shock, power-off and unplug the instrument Warning: before inspecting the fuses. The detector requires two 100 to 240 VAC, 50 to 60 Hz, F 3.15 A, 250 V fast blo, 5 ×...
  • Page 140 4 Maintaining the Detector Removing and replacing the rear panel fuses and fuse holder: Fuse holder Remove and discard both fuses. Make sure that the new fuses are properly rated, and then insert them into the holder and the holder into the power entry module, gently pushing until the assembly locks into position.

  • Page 141: Error Messages, Diagnostic Tests, And Troubleshooting

    Messages requiring you to perform corrective action including messages encountered at startup and during calibration or operation. • Messages requiring you to cycle power, and then contact Waters Technical Service personnel if an error persists (see “Contact Waters technical service” on page 117).
  • Page 142 1. Cycle power to the performs a complete detector. verification, which 2. Perform manual includes calibration. remeasurement of all 3. Contact Waters calibration points. Technical Service. New calibration points are compared to stored information from the most recent manual calibration.
  • Page 143 Error messages Startup, calibration, and operating error messages: (Continued) Error Message Description Corrective Action Lamp failure Lamp indicates Off 1. Check lamp icon. when it should be On. 2. Cycle power to the detector. 3. Replace lamp. Lamp lighting failure The lamp failed to 1.

  • Page 144: Error Messages Preventing Operation

    5 Error Messages, Diagnostic Tests, and Troubleshooting Startup, calibration, and operating error messages: (Continued) Error Message Description Corrective Action Wavelengths span In dual-wavelength Select wavelengths that are 370 nm: Order filter mode: each above or below 370 not in use •...
  • Page 145 • the flow cell is clean; • the front door is shut securely. Cycle power to the detector. If the terminating error persists, contact Waters Technical Service. Instrument error messages: Error Message Description Corrective Action...
  • Page 146 Description Corrective Action Dark current too low: 0 The dark energy level 1. Cycle power to the equals 0. detector. 2. Contact Waters Technical Service. Electronic A/D failure Lamp optimization is Cycle power. adjusted at the minimum level. Data acquisition via A/D 1.
  • Page 147 Corrective Action Filter initialization Unit sensors cannot find 1. Cycle power to the failure: Shutter position the shutter position. detector. 2. Contact Waters Technical Service. Grating initialization Backlash is the 1. Cycle power to the failure: Backlash too difference between the detector.

  • Page 148: User-Selected Diagnostic Tests

    1. Cycle power to the from nonvolatile detector. memory is not valid. 2. Perform manual calibration. 3. Contact Waters Technical Service. User-selected diagnostic tests Overview You can run several diagnostic tests for troubleshooting the detector and verifying that its electronics and optics perform correctly.
  • Page 149 User-selected diagnostic tests You can disable all active sticky diagnostic tests by pressing DIAG and then selecting “1, Reset instrument.” If no sticky diagnostic tests are active, the wrench icon does not appear on the absorbance screen. When you power-off the detector, sticky diagnostic tests become inactive.

  • Page 150: Use The Diagnostic Tests

    Use the diagnostic tests The detector employs both user-selectable and service diagnostic tests. You access user diagnostic tests by pressing the DIAG key. Only qualified Waters service personnel can access service diagnostic tests. Use the sample and reference energy diagnostic tests...
  • Page 151 User-selected diagnostic tests AU time trace. The current sample and reference energy readings appear in nanoamps (nA). Sample and reference energy diagnostic test: To use the sample and reference energy diagnostic test: Press DIAG, and then press 2. Change the wavelength, if necessary. Press Enter.
  • Page 152 5 Error Messages, Diagnostic Tests, and Troubleshooting Input & output diagnostic test choice list: Display auto-zero offsets From the Input & Output choice list, press 1 (Auto-zero offsets). This diagnostic test also allows you to display and reset the offset to zero on both channels by pressing Cancel (Shift 0).
  • Page 153 User-selected diagnostic tests This test sets the voltages on the analog output channels based on the current AU setting. This is a sticky diagnostic test. Set fixed voltage output From the Input & Output choice list, press 3 (Fix voltage) to select a voltage for the analog output.
  • Page 154 5 Error Messages, Diagnostic Tests, and Troubleshooting dotted-line border). Press any numerical key to change the status of the switch (ON to OFF or vice versa). Press Enter to select the second switch. Use the lamp, display, and keypad diagnostic tests To access the lamp, display, and keypad diagnostic tests, press DIAG and then press 4.
  • Page 155 The display fills from top to bottom and from right to left, then Result: returns to the Lamp, display & keypad choice list. If the display does not completely fill, either horizontally or vertically, contact your Waters service representative. From the Lamp, display & keypad choice list, press 4.
  • Page 156 5 Error Messages, Diagnostic Tests, and Troubleshooting • Generate test peaks – Specifies the generation of test peaks to calibrate your data system. • Manually override the optical filter – Selects a filter different from that specified as part of the detector’s normal operating mode. To run either test, press DIAG and then press 5 (Other diagnostics).

  • Page 157: Service Diagnostic Tests

    Most detector problems are relatively easy to correct. If you are unable to correct a problem or a failed condition, contact Waters Technical Service. November 9, 2012, 715003800 Rev. A...

  • Page 158: When You Contact Waters

    System configuration (other components) The detector can be configured as part of a system with a 2695 Tip: Separations Module, Empower or MassLynx software, or a non-Waters product. Diagnostic tests The detector performs some user-selected diagnostic tests to help you troubleshoot basic system problems.
  • Page 159 Faulty lamp Replace the lamp. does not light Lamp not plugged Plug in the lamp connector. Bad lamp power Contact Waters Technical supply board Service. Lamp switch in the Inspect the rear panel “off” position connections or timed event within a method.
  • Page 160 1 point per second. Keypad not Keypad defective 1. Cycle power to the detector functioning and run the keypad diagnostic test. 2. Contact Waters Technical Service. No sample and Lamp burned out Attempt to reignite using the reference energy lamp key.

  • Page 161: A Safety Advisories

    Safety Advisories Waters instruments display hazard symbols designed to alert you to the hidden dangers of operating and maintaining the instruments. Their corresponding user guides also include the hazard symbols, with accompanying text statements describing the hazards and telling you how to avoid them.

  • Page 162: Warning Symbols

    Heed all warnings when you install, repair, and operate Waters instruments. Waters assumes no liability for the failure of those who install, repair, or operate its instruments to comply with any safety precaution.

  • Page 163: 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. November 9, 2012, 715003800 Rev. A...
  • Page 164 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. To avoid electric shock, do not remove the mass spectrometer’s Warning: protective panels.
  • Page 165 Biohazard warning This warning applies to Waters instruments that can be used to process material that can contain biohazards: substances that contain biological agents capable of producing harmful effects in humans.

  • Page 166: Caution Advisory

    A Safety Advisories Waters instruments can be used to analyze or Warning: process potentially hazardous substances. To avoid injury with any of these materials, familiarize yourself with the materials and their hazards, observe Good Laboratory Practice (GLP), and consult your organization’s safety representative regarding proper use and handling.

  • Page 167: Warnings That Apply To All Waters Instruments

    Warnings that apply to all Waters instruments Warnings that apply to all Waters instruments When operating this device, follow standard quality-control procedures and the equipment guidelines in this section. Changes or modifications to this unit not expressly approved by the Attention: party responsible for compliance could void the user’s authority to operate the...
  • Page 168 A Safety Advisories Use caution when working with any polymer tubing under pressure: Warning: • Always wear eye protection when near pressurized polymer tubing. • Extinguish all nearby flames. • Do not use tubing that has been severely stressed or kinked. •...
  • Page 169 Warnings that apply to all Waters instruments fare attenzione quando si utilizzano tubi in materiale polimerico Attenzione: sotto pressione: • Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati. • Spegnere tutte le fiamme vive nell'ambiente circostante.
  • Page 170 A Safety Advisories 警告：当有压力的情况下使用管线时，小心注意以下几点： • 当接近有压力的聚合物管线时一定要戴防护眼镜。 • 熄灭附近所有的火焰。 • 不要使用已经被压瘪或严重弯曲的管线。 • 不要在非金属管线中使用四氢呋喃或浓硝酸或浓硫酸。 • 要了解使用二氯甲烷及二甲基亚枫会导致非金属管线膨胀，大大降低管线的耐压能力。 경고: 가압 폴리머 튜브로 작업할 경우에는 주의하십시오. • 가압 폴리머 튜브 근처에서는 항상 보호 안경을 착용하십시오. • 근처의 화기를 모두 끄십시오. • 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오. •...
  • Page 171 Warnings that apply to all Waters instruments The user shall be made aware that if the equipment is used in a Warning: manner not specified by the manufacturer, the protection provided by the equipment may be impaired. L’utilisateur doit être informé que si le matériel est utilisé d’une Attention: façon non spécifiée par le fabricant, la protection assurée par le matériel risque...

  • Page 172: Warnings That Address The Replacing Of Fuses

    A Safety Advisories Warnings that address the replacing of fuses The following warnings pertain to instruments equipped with user-replaceable fuses. If the fuse types and ratings appear on the instrument: To protect against fire, replace fuses with those of the type Warning: and rating printed on panels adjacent to instrument fuse covers.

  • Page 173: Electrical Symbols

    Warnings that address the replacing of fuses 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 November 9, 2012, 715003800 Rev.

  • Page 174: Handling Symbols

    A Safety Advisories Handling symbols These handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments Keep upright! Keep dry! Fragile! Use no hooks! November 9, 2012, 715003800 Rev. A...

  • Page 175: B Specifications

    Specifications This appendix lists individual operating specifications for the Waters 2489 UV/Visible Detector. All performance specifications are measured following a warm-up Note: period of one hour (ΔΤ < +2 °C/hr). Contents: Topic Page Operational specifications ............... 176 Optional Waters TaperSlit flow cell specifications ......178...

  • Page 176: Operational Specifications

    B Specifications Operational specifications Operational specifications: Condition Specification Wavelength range 190 to 700 nm Bandwidth <5 nm Wavelength accuracy +1.0 nm (via patented Erbium filter) Wavelength +0.1 nm repeatability <5 x 10 -6 AU Baseline noise, single wavelength (dry) (measured at 230 nm, 10 points/s, 30 second segments, dry analytical cell) Baseline wet noise, <8 x 10 -6 AU...
  • Page 177 Operational specifications Operational specifications: (Continued) Condition Specification Optical Component Specifications Light source Deuterium arc lamp, 2000-hour warranty or 1 year (whichever comes first), front accessible Photodiodes 2 silicon photodiodes (matched pair) Second-order filter Automatic for wavelengths > 370nm Wavelength Erbium filter, used at startup or on demand calibration filter Flow cell design Patented TaperSlit™...

  • Page 178: Optional Waters Taperslit Flow Cell Specifications

    61.0 cm (24.0 inches) Weight 13.6 kg (30.0 pounds) a. US Patent Numbers: 6,423,249 and 6,783,705 b. ASTM E1657-983 c. US Patent Numbers: US 5,883,721 Optional Waters TaperSlit flow cell specifications Waters Patented TaperSlit flow cell specifications: Sample tube Path Volume internal...
  • Page 179 Optional Waters TaperSlit flow cell specifications Optional Waters TaperSlit flow cell specifications Waters Patented TaperSlit flow cell specifications: (Continued) Sample tube Path Volume internal Pressure length (μl) diameter rating (mm) (Inches) (psi/bar) Microbore cell 0.005 0.005 1000/70 Inert (titanium) cell 16.3...
  • Page 180 B Specifications November 9, 2012, 715003800 Rev. A...

  • Page 181: C Solvent Considerations

    Preventing contamination For information on preventing contamination, refer to Controlling Contamination in Ultra Performance LC/MS and HPLC/MS Systems (part number 715001307), or visit www.waters.com. Clean solvents Clean solvents provide reproducible results and permit you to operate with minimal instrument maintenance.

  • Page 182: Solvent Quality

    C Solvent Considerations Solvent quality Use HPLC-grade solvents for the best possible results. Filter solvents through 0.45-µm filters before their use. Solvents distilled in glass generally maintain their purity from lot to lot; use them to ensure the best possible results. Preparation checklist The following solvent preparation guidelines help to ensure stable baselines and good resolution:...

  • Page 183: Solvent Miscibility

    Solvent miscibility Tetrahydrofuran contaminants (peroxides) are potentially Warning: explosive if concentrated or taken to dryness. Solvent miscibility Before you change solvents, refer to the table below to determine the miscibility of the solvents to be used. When you change solvents, be aware that, •...
  • Page 184 C Solvent Considerations Solvent miscibility: (Continued) Boiling Miscibility λ Cutoff Polarity Viscosity Solvent point °C number index CP, 20 °C (nm) (1 atm) P-xylene 0.70 138.0 Benzene 0.65 80.1 Benzyl ether 5.33 288.3 –– –– Methylene chloride 0.44 39.8 Ethylene chloride 0.79 83.5 ––...

  • Page 185: How To Use Miscibility Numbers

    Solvent miscibility Solvent miscibility: (Continued) Boiling Miscibility λ Cutoff Polarity Viscosity Solvent point °C number index CP, 20 °C (nm) (1 atm) Formamide 3.76 210.5 –– Water 1.00 100.0 –– –– How to use miscibility numbers Use miscibility numbers (M-numbers) to predict the miscibility of a liquid with a standard solvent (see “Solvent miscibility”...

  • Page 186: Buffered Solvents

    C Solvent Considerations Buffered solvents When using a buffer, use a good quality reagent and filter it through a 0.45-µm filter. Do not leave the buffer stored in the system after use. Flush all fluid paths with HPLC-quality water before shutting the system down and leave distilled water in the system (flush with 90% HPLC-quality water: 10% methanol for shutdowns scheduled to be more than one day).

  • Page 187: Gas Solubility

    Solvent degassing methods • stable pump operation. Gas solubility Only a finite amount of gas can be dissolved in a given volume of liquid. This amount depends on, • the chemical affinity of the gas for the liquid; • the temperature of the liquid; •...

  • Page 188: Sparging

    C Solvent Considerations You can use either of the following methods to degas solvents: • Sparging with helium • Vacuum degassing Sparging Sparging removes gases from solution by displacing dissolved gases in the solvent with a less soluble gas, usually helium. Well-sparged solvent improves pump performance.

  • Page 189: Wavelength Selection

    To increase the length of membrane, you can connect two or more vacuum chambers in series. The in-line degasser is available as an option or factory-installed in the Waters 2695 Separations Module, XE model. Wavelength selection This section includes UV cutoff ranges for, •...

  • Page 190: Mixed Mobile Phases

    C Solvent Considerations UV cutoff wavelengths for common chromatographic solvents: (Continued) UV cutoff UV cutoff Solvent Solvent (nm) (nm) Amyl alcohol Isopropyl ether Amyl chloride Methanol Benzene Methyl acetate Carbon disulfide Methyl ethyl ketone Carbon tetrachloride 265 Methyl isobutyl ketone Chloroform Methylene chloride Cyclohexane...

  • Page 191: Wavelength Selection For Chromophore Detection

    Triton-X™ 100, 0.1% Hydrochloric acid, 0.1% ® Waters PIC Reagent A, 1 vial/liter MES, 10 mM, pH 6.0 Waters PIC Reagent B-6, 1 vial/liter Potassium phosphate, Waters PIC Reagent B-6, monobasic, 10 mM low UV, 1 vial/liter dibasic, 10 mM...
  • Page 192 C Solvent Considerations Electronic absorption bands of representative chromophores*: λ ∈ λ ∈ Chemical Chromophore configuration (nm) (L/m/cm) (nm) (L/m/cm) Ether —O— 1000 Thioether —S— 4600 1600 Amine —NH 2 2800 Thiol —SH 1400 Disulfide —S—S— 5500 Bromide —Br Iodide —I Nitrile —C≡N...
  • Page 193 Wavelength selection Electronic absorption bands of representative chromophores*: (Continued) λ ∈ λ ∈ Chemical Chromophore configuration (nm) (L/m/cm) (nm) (L/m/cm) Allene —(C=C) 2 — 210-230 21,000 (acyclic) Allene —(C=C) 3 — 35,000 Allene —(C=C) 4 — 52,000 Allene —(C=C) 5 — 118,000 Allene —(C=C) 2 —...
  • Page 194 C Solvent Considerations November 9, 2012, 715003800 Rev. A...

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