Waters 2489 Overview And Maintenance Manual
  1. Manuals
  2. Brands
  3. Waters Manuals
  4. Security Sensors
  5. 2489
  6. Overview and maintenance manual

Waters 2489 Overview And Maintenance Manual

Uv/visible detector
Hide thumbs Also See for 2489:
Table of Contents

Advertisement

Quick Links

2489 UV/Visible
Detector
Overview and Maintenance Guide
715004752/Revision A
Copyright © Waters Corporation 2015
All rights reserved

Advertisement

Table of Contents
loading
Need help?

Need help?

Do you have a question about the 2489 and is the answer not in the manual?

Questions and answers

Related Manuals for Waters 2489

Summary of Contents for Waters 2489

  • Page 1 2489 UV/Visible Detector Overview and Maintenance Guide 715004752/Revision A Copyright © Waters Corporation 2015 All rights reserved...
  • Page 2 June 11, 2015, 715004752 Rev. A Page ii...

  • Page 3: Trademarks

    General Information Copyright notice © 2015 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA AND IN IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

  • Page 4: Contacting Waters

    Contacting Waters Contact Waters with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail. Waters contact information Contacting medium Information Internet The Waters Web site includes contact information for Waters locations worldwide.

  • Page 5: Fcc Radiation Emissions Notice

    FCC radiation emissions notice Changes or modifications not expressly approved by the party responsible for compliance, could void the users authority to operate the equipment. This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions: (1) this device may not cause harmful interference, and (2) this device must accept any interference received, including interference that may cause undesired operation.

  • Page 6: Intended Use Of The 2489 Uv/Visible Detector

    Serial number Part number catalog number Audience and purpose This guide is intended for personnel who install, operate, and maintain 2489 UV/Visible (UV/Vis) detectors. Intended use of the 2489 UV/Visible detector Waters designed the 2489 UV/Visible detector to analyze and monitor many compounds.

  • Page 7: Emc Considerations

    Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage, power-supply network. EC authorized representative Waters Corporation Stamford Avenue Altrincham Road Wilmslow SK9 4AX UK Telephone:...
  • Page 8 June 11, 2015, 715004752 Rev. A Page viii...

  • Page 9: Table Of Contents

    Customer comments ................... iii Contacting Waters ..................iv Safety considerations .................. iv Safety hazard symbol notice.............. iv Considerations specific to the 2489 UV/Visible detector ......iv FCC radiation emissions notice............v Electrical power safety notice............. v Equipment misuse notice ..............v Safety advisories ................
  • Page 10 1.3.6 MaxPlot..................26 1.3.7 Thermal-wander management............26 2 Installing the Detector ............. 27 Before you begin ..................27 Unpacking and inspecting ................27 2.2.1 Unpacking the detector ..............27 2.2.2 Inspecting the detector ..............28 Selecting a laboratory site ................. 28 Stacking system modules ................
  • Page 11 3.3.9 Conserving lamp life ................ 90 3.3.10 Shutting down the detector ............. 92 4 Maintaining the Detector ............93 Contacting Waters technical service ............. 93 Maintenance considerations ................ 93 4.2.1 Safety and handling ................. 94 4.2.2 Spare parts ..................94 Proper operating procedures ..............
  • Page 12 Specific warnings ................126 Notices ....................127 Bottles Prohibited symbol ................ 128 Required protection ................128 Warnings that apply to all Waters instruments and devices ......128 Warnings that address the replacing of fuses ..........131 Electrical and handling symbols ..............133 A.7.1 Electrical symbols................
  • Page 13 B Specifications ................ 137 Physical specifications ................137 Environmental specifications ..............137 Electrical specifications ................138 Performance specifications ............... 139 Optical component specifications .............. 139 Flow cell specifications ................140 C Solvent Considerations ............141 Introduction ................... 141 C.1.1 Preventing contamination ............... 141 C.1.2 Clean solvents ................
  • Page 14 June 11, 2015, 715004752 Rev. A Page xiv...

  • Page 15: Theory And Principles Of Operation

    Theory and Principles of Operation This chapter summarizes the 2489 UV/Visible Detector features and describes the theory and principles of operation. See also: Appendix B for system specifications and Appendix C for information on high-performance liquid chromatography (HPLC) solvent considerations.
  • Page 16 ® ® The detector can be configured with either Empower or MassLynx chromatography data software. Table 1–1: Detector capabilities Capability Description Stand-alone programmability Stores as many as five user-defined programs (or methods) consisting of as many as 50 programmable, timed events and two threshold events each.

  • Page 17: Principles Of Operation

    Wavelength verification and test • Flow cell • Electronics 1.2.1 Detector optics The 2489 UV/Vis detector optics are based on a Fastie-Ebert monochromator and include these components: • High brightness deuterium (D ) lamp • Two mirrors: one off-axis, ellipsoidal mirror and one spherical mirror •...
  • Page 18 1.2.1.2 Waters TaperSlit flow cell The Waters TaperSlit flow sell used in this detector renders the detector baseline less sensitive to changes in mobile phase refractive index (RI). RI changes occur during gradient separations or result from temperature or pump-induced pressure fluctuations.
  • Page 19 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 20 If the value for the narrowest peak of interest is less than 15, you must specify a higher sampling rate in the instrument method. If the value is greater than 30, 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.

  • Page 21: Wavelength Verification And Test

    Figure 1–4: Filter time constant comparison 0 sec 1 sec 2 sec Time (minutes) Tip: Although the peak shape shows some distortion and the signal output is delayed with different time constants, the peak area remains the same. 1.2.2 Wavelength verification and test The detector’s deuterium arc lamp and integral erbium filter exhibit peaks in the transmission spectrum at known wavelengths.

  • Page 22: 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 cycling power to the detector. See “Wavelength calibration” on page 1.3 Operational modes...
  • Page 23 1.3.1.1 Primary parameters Table 1–5: Major parameters applicable to single-wavelength mode Parameter Description Wavelength, in nm Specifies a wavelength for channel A, from 190 nm to 700 nm, settable in 1-nm increments. Sensitivity, in AU Specifies the scaling factor for the analog-output channels and corresponds to the AU value where the analog outputs become saturated at full-scale values.

  • Page 24: Dual-Wavelength Mode

    1.3.2 Dual-wavelength mode In dual-wavelength mode, the detector can monitor two wavelengths, one on channel A and one on channel B. The sampling frequency is reduced to 1 or 2 Hz, limiting the use of this mode to more standard chromatography where peak widths span at least 20 seconds to enable full characterization of a peak.

  • Page 25: Spectrum Scanning

    Table 1–6: Additional options for analog output in dual-wavelength mode Mode option Description of mode RatioPlot (A/B) Produces the ratio of absorbance from two wavelengths. Theoretically, the ratio is constant for a pure chromatographic peak and variable for an impure peak, which results in a nonsquared response.

  • Page 26: Cuvette Operations

    1.3.4 Cuvette operations Note: This section pertains only to flow cells that require a cuvette. Detector cuvette option is used to measure the absorbance spectrum of a sample in a cuvette. To generate and store a spectrum: Acquire a zero scan, which measures the absorbance of the contents of the cuvette and flow cell over the desired wavelength range.

  • Page 27: Installing The Detector

    HPLC system. 2.1 Before you begin Requirement: To install the 2489 UV/Vis detector, you should know how, in general, to set up and operate laboratory instruments and computer-controlled devices and also how to handle solvents. Before installing the detector, ensure that •...

  • Page 28: Inspecting The Detector

    Waters representative. Customers in the USA and Canada can report damage and discrepancies to Waters Technical Service (800 252-4752). Others can phone their local Waters subsidiary or Waters corporate headquarters in Milford, Massachusetts (USA), or visit www.waters.com.

  • Page 29: Stacking System Modules

    • HAR type (or better), in Europe For information regarding the type of cord to use in other countries, contact your local Waters distributor. Requirement: You must mount the detector on a level surface to allow for proper functioning of the drip management system (drain tube), which you connect to a waste reservoir to divert solvent leaks away from the flow cell.

  • Page 30: Connecting To The Electricity Source

    Warning: Avoid electrical shock: • Use power cord SVT-type in the United States and HAR-type or better in Europe. For other countries, contact your local Waters distributor. • Power-off and unplug the detector before performing any maintenance on the instrument.

  • Page 31: Connecting Signal Cables

    LED blinks green. The lamp LED shows steady green when the lamp is ignited. 2.6 Connecting signal cables See also: Waters Ethernet Instrument Getting Started Guide. The following figure shows the rear panel location of the connectors used to operate the detector with external devices.

  • Page 32: Connecting I/O Cables

    The signal connections necessary for your detector depend on the signal connections available on the other instruments in the HPLC system. 2.6.1 Connecting I/O cables The rear panel includes two removable connectors that hold the pins for the I/O signals, as shown in the figure below. These connectors are keyed so that you can insert them one way only.

  • Page 33: Connecting Signal Cables From The Rear Panel And Ethernet Connectors To The Detector

    Table 2–2: I/O signals for the detector (continued) Signal Description 2 V full-scale analog output signal of channel B Analog 2 (scaled to the current AU setting). Switch 1 (2) Used to connect to a fraction collector. Can be controlled by threshold and timed events. Switch 2 (2) 1.
  • Page 34 20 meters. The maximum recommended cable length between two Ethernet devices is 4 meters. Figure 2–5: Ethernet connections to the detector in a chromatography data workstation Ethernet Ethernet cable switch Ethernet connector chromatography data workstation 2489 UV/Vis detector June 11, 2015, 715004752 Rev. A Page 34...

  • Page 35: Starting A Method

    Table 2–3: Separations module connections to detector for starting a method 2695 separations module (B 2489 UV/Vis detector (II) inputs and outputs) Pin 1 Inject Start Pin 1 Inject Start + Pin 2 Inject Start Pin 2 Inject Start –...

  • Page 36: Turning The Detector Lamp On Or Off

    Table 2–3: 2695 separations module connections to the detector (lamp on or off) 2695 separations module (A 2489 UV/Vis detector (II) outputs) Pin 1 Switch 1 Pin 4 Lamp On/Off + Pin 2 Switch 1 Pin 5 Lamp On/Off –...
  • Page 37 Table 2–4: 2695 separations module connections to the detector, to generate autozero 2695 separations module (B 2489 UV/Vis detector (II) inputs and outputs) Pin 1 Inject Start Pin 9 Auto Zero + Pin 2 Inject Start Pin 10 Auto Zero –...

  • Page 38: Connecting The Detector To A Chromatography Data Workstation Using An E-Sat/In Module

    2.6.7.1 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 chromatography data workstation.
  • Page 39 Notice: To avoid damaging the e-SAT/IN module (and thus void its warranty) and to ensure its proper startup, do not power-on the module until you perform all procedures in the Waters e-SAT/IN Module Installation Guide. 2.6.7.2 Connecting the detector to the e-SAT/IN module The e-SAT/IN module connects to the detector through the B (inputs and outputs) terminal on the rear panel of the detector, as shown in the figure below.
  • Page 40 The following table summarizes the detector connections to the e-SAT/IN module: Table 2–6: Detector connections to a e-SAT/IN module e-SAT/IN 2489 UV/Vis detector (I) connector Pin 1 Analog 1 + (white) Channel 1 or 2 Pin 3 Ground (black)

  • Page 41: Connecting The Detector To A 745/745B/746 Data Module

    To prevent oversaturation of the signal from the detector to the integrator, do not exceed the input voltage rating of the integrator. The analog-output signal is transmitted via the cable provided in the 2489 UV/Vis detector startup kit. Make the connections summarized in the table below and illustrated in the figure below: Table 2–7:...

  • Page 42: Connecting The Detector To A Fraction Collector

    Figure 2–13: 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 2489 connector I Analog 1 + Analog 1 −...

  • Page 43: Plumbing The Detector

    The table below indicates the detector-to-fraction collector and autoinjector-to-fraction collector connections. Table 2–8: Detector connections to the fraction collector 2489 UV/Vis detector Fraction collector connection I Pin 3 Ground Pin 1 Detector In – II Pin 6 Chart-mark + Pin 10 Event Marker + II Pin 7 Chart-mark –...

  • Page 44: Connecting Columns In Hplc Systems

    Notice: To avoid damaging the flow cell or tubing, do not connect any tubing or device that can cause the backpressure to exceed the pressure rating of those components. The pressure rating of a standard analytical flow cell is 6895 kPa (69 bar, 1000 psi). 2.7.1 Connecting columns in HPLC systems Warning: To prevent injury, always observe Good Laboratory Practices when...

  • Page 45: Assembling The Fittings (Hplc Only)

    2.7.3 Assembling the fittings (HPLC only) To assemble each fitting: Slide the compression screw over the tubing end, followed by the ferrule. Mount the ferrule with its tapered end facing the end of the tubing. Figure 2–14: Ferrule and compression screw assembly Compression screw Tubing end (cut...
  • Page 46 June 11, 2015, 715004752 Rev. A Page 46...

  • Page 47: Prepare The Detector

    Prepare the Detector After you install the detector, you can set it up and operate it as a stand-alone instrument or with chromatography data software. Table 3–1: Detector configuration Configuration Programming required for operation The detector is a stand-alone instrument Program the detector using the front within a system, or used with any panel (see...

  • Page 48: Diagnostic Test Failure

    When the detector is successfully started, it defaults to idle mode (see the figure “2489 UV/Vis detector idle mode screen” on page 48). 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 49: Using The Operator Interface

    3.2 Using the operator interface 3.2.1 Using the display × The detector employs a 128 64-bitmap graphic display and a 24-key membrane keypad for the operator interface. After the startup diagnostic tests are run successfully, the detector displays the absorbance, or HOME, screen. Figure 3–3: Detector absorbance (HOME) screen Absorbance...
  • Page 50 Table 3–2: Absorbance and message screen icons (continued) Icon or Icon/field name Function field Field Wavelength Selects the wavelength monitored on the requiring selected channel. In single-wavelength entry mode, you cannot independently control wavelength on channel B. Channel selector Changes the channel when you press A/B.

  • Page 51: Using The Keypad

    Table 3–2: Absorbance and message screen icons (continued) Icon or Icon/field name Function field Local method Indicates the detector is not controlled number by a data system. The icon’s cursive “m” and the current method number or an asterisk (*) indicate current conditions are not stored as a method.
  • Page 52 Figure 3–4: Keypad: λ/λλ SCAN Reset HOME Chart-mark Auto Zero Run/Stop METHOD Lamp Lock Calibrate System Info Previous CONFIGURE Contrast DIAG Next Scale TRACE Enter +/− Clear Field Cancel Shift Primary function keys take effect immediately, with no further entry required. Secondary function keys require you to enter information into parameter fields and to press Enter for the functions to take effect.
  • Page 53 Table 3–3: Detector keypad description (continued) Unshifted Shifted Chart-mark – Causes a SCAN – Displays the list of momentary pulse to the options for generating and SCAN analog output (A, B, or both manipulating spectra. Chart-mark depending upon the current settings).
  • Page 54 Table 3–3: Detector keypad description (continued) Unshifted Shifted DIAG – Displays the choice list CONFIGURE – Displays the of diagnostic tests. first Configuration screen. CONFIGURE DIAG TRACE – Displays the Scale – When the wavelength absorbance monitor trace for trace or spectrum screen is Scale channel A or B.

  • Page 55: Navigating The User Interface

    Table 3–3: Detector keypad description (continued) Unshifted Shifted 6 – See 0-9, above. Contrast – Permits contrast adjustment (viewing angle) of Contrast the liquid crystal display. 0 – See 0-9, above. Cancel – In some modes, backs out of a prompt without Cancel completing the task.
  • Page 56 To display a choice list, press Enter. Then take one of these actions: • 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.

  • Page 57: Primary And Secondary Functions

    3.2.4 Primary and secondary functions You can access the following functions directly from the absorbance screen or by pressing the Next key on that screen. Table 3–1: Detector functions Function Description Wavelength Defines the operating wavelength for the channel. AUFS (absorbance Defines the relationship between the absorbance and units full-scale) the output voltage.
  • Page 58 Table 3–1: 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 59 Table 3–1: 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 60 Table 3–1: Detector functions (continued) Function Description Autozero on λ This function results in an autozero each time a changes wavelength change is requested. If you disable this function, significant changes in measured absorbance can occur after each wavelength change. Selecting “to zero”...
  • Page 61 Table 3–2: Primary and secondary function (method) parameters Function Screen Type Units Range Default Numeric Integer 190 nm to 254 nm λ (Wavelength) (Absorbanc 700 nm e screen) AUFS Numeric AUFS 0.0001 to 4.0000 2.0000 Analog rate 2 (of 4) or Choice (λ): 10, 20, 40, 2 (of 5)
  • Page 62 You can change the ending time parameter to any number from 1 to 60. You can use the Scale function to “zoom” in on a particular section of the trace. To display the Scale parameters after you press the Scale key: Press Next to display T2 (starting time).

  • Page 63: Operating Other Detector Functions

    Figure 3–8: 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. Figure 3–9: 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 64 3.2.5.2 Configuring event inputs (contact closures) You use the CONFIGURE key to edit event input settings and specify switched output settings. Using the Enter key and the numeric keypad (or the s and t keys) to select the appropriate entry, you can edit four fields on the second Configuration screen. The default for Inject, Chart-mark, and Autozero is Low;...
  • Page 65 3.2.5.3 Setting pulse periods You use the third Configuration screen (see page 63) to set pulse or signal width or to activate a pulse or rectangular wave on SW1 or SW2. • Single pulse (in seconds) – If SW1 or SW2 is programmed to generate a pulse as a timed or threshold event, then the period of the signal (single pulse width) is as specified in this field (range is 0.1 to 60 seconds).

  • Page 66: Operating The Detector

    Tip: Use the scroll bar to see all of the message. The Firmware Rev and ChkSm values shown here are examples only. They do not indicate the released version’s information. 3.2.5.6 Using 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 67: Verifying That The Detector Operates Properly

    Note: If your flow cell does not require a cuvette, you can obtain an analytical flow cell to perform the validation procedures. See the Waters Quality Parts Locator on the Services and Support page of the Waters’ web site. Requirements: •...
  • Page 68 Allow the detector to warm up for at least 30 minutes before operating. Tip: If the startup verification diagnostic tests fail, note the error message to determine the corrective action, and refer to Chapter 3.2.7.2 Recording sample and reference beam energies To determine baseline values on your detector for future reference, and to monitor lamp aging (decreased lamp output energy), you must record the baseline sample and reference beam energies to compare against future readings.

  • Page 69: Wavelength Calibration

    Press Calibrate (Shift 3) from the detector keypad. Result: A message appears asking whether you removed the cuvette and 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 70: Operating The Detector In Dual-Wavelength Mode

    Enter the wavelength and sensitivity on the absorbance screen, as well as any secondary parameters and timed or threshold events. Note: Changing the sensitivity (AUFS) setting affects the 2-V output. Select a second sensitivity setting while in single-wavelength mode, by pressing A/B and specifying the appropriate AUFS values on the channel B screen.
  • Page 71 Recommendation: In dual-wavelength mode, select a wavelength pair below or above 370-nm. If one or both of the selected wavelengths spans the 370 nm threshold, the detector beeps three times, and the warning message shown below appears. Because of possible UV light interference (second-order effects), you can observe additional peaks and inaccurate peak areas.
  • Page 72 In the Data out field, press 5, MaxPlot A,B. Press Enter to select the MaxPlot function. Return to the absorbance screen by pressing the HOME key. 3.2.10.3 Programming timed events, threshold events, and methods You can retrieve as many as five methods, which the detector references as numbers 1 to 5.
  • Page 73 Table 3–3: Timed event parameters (continued) Specify Number Event Units Range or default channel Chart-mark Does not Does not apply (10% of full apply scale) Polarity 1. + 2. – Auto-zero Does not Does not apply apply Lamp 1. Off 2.
  • Page 74 Press Enter to enter the time. Tip: To advance to the Set field (Events choice list), press the t key. Press Enter again to display the choice list, or, if you know the event number, press the number for the event you are programming. If the To field appears, enter the appropriate wavelength, in nanometers.
  • Page 75 Table 3–13: Threshold events “To” parameters (continued) Below Number Set to threshold switch state Pulse Rect wave (rectangular wave) To define the pulse period, or the frequency of a wave, see “Configuring the detector” on page To program a threshold event: Press the METHOD (Shift A/B) key on the detector keypad.
  • Page 76 Figure 3–20: Store a method, method number field Note: No warning message appears when the method number 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.
  • Page 77 To clear one or more methods: Return to the Method choice list by pressing METHOD (Shift A/B). Press 5, Reset method *. Result: A message appears, asking whether you approve of setting current conditions to factory defaults. Tip: If you press Enter, these events take place: •...

  • Page 78: Scanning Spectra

    Note: If your flow cell does not require a cuvette, you can obtain an analytical flow cell to perform the spectral scanning. See the Waters Quality Parts Locator on the Services and Support page of the Waters’ Web site. The detector must make two scans to produce an absorbance spectrum, as shown in the table below.
  • Page 79 Figure 3–21: Scan of anthracene at 100 nm/min and 1000 nm/min Pace 100 Pace 1000 Wavelength (nm) Tip: The higher the number you enter in the Pace field, the lower the 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 80: Scanning New Spectra

    Figure 3–23: Scan of erbium standard in a cuvette from 190 nm to 600 nm at 200 nm/min without tick marks Wavelength (nm) Specify these parameter values when you select a scan: zero or sample. When you select a zero scan, the detector displays three additional screens, labeled 2 of 4, 3 of 4, and 4 of 4.
  • Page 81 Figure 3–24: Scan choice list Press 1, New scan, or use the s and t keys to move through the Scan choice list. Result: The detector displays the first of three parameter screens for a sample scan or four parameter screens for a zero scan (figure “Zero and sample scan screen”...
  • Page 82 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. Enter a value in the Pace field for the rate at which the detector scans the specified wavelength range.
  • Page 83 Figure 3–25: 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) 3.3.2.1 Running the sample scan...
  • Page 84 Press Next to advance to the second sample scan screen. Tip: You can change the entries in the AUFS and Mark each fields. Press Next to advance to the third sample scan screen, and then press Run. Result: A brief message (“Initializing”) appears. The Scanning screen shows the progress of the scan, in nanometers, across a progress bar.
  • Page 85 • AU1 – minimum absorbance displayed. The default is Auto. • AU2 – maximum absorbance displayed. The default is Auto. Using this function, you zoom in on various sections (artifacts) of the spectrum. The scale of the spectrum is affected by the AUFS setting. Press Next to advance through the four scaling parameters.

  • Page 86: Storing A Spectrum

    Figure 3–31: 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 87: Reviewing A Stored Spectrum

    Figure 3–32: Storage slot number box In the slot number box, enter a number from 1 to 3. Press Enter to store the last sample scan, paired with its zero scan. To obtain information about a stored spectrum: Press SCAN (Shift Chart-mark) to view the Scan choice list. Press 3, Get scan info.

  • Page 88: Replaying A Spectrum

    Note: If your flow cell does not require a cuvette, you can obtain an analytical flow cell to perform the validation procedures. See the Waters Quality Parts Locator on the Services and Support page of the Waters’ Web site. Using the cuvette option facilitates sample handling and verifying and qualifying the instrument.
  • Page 89 The detector uses a standard, 10-mm path length, spectrophotometric cell (quartz cuvette). You insert the cuvette with one of the two frosted sides facing up in the cuvette holder and place it in the detector flow cell assembly. Figure 3–33: Detector cuvette holder with the cuvette inserted Aperture Frosted sides of cuvette facing up and down...

  • Page 90: Scanning Using A Flow Cell And A Syringe

    Recommendations: • Ensure that you have enough liquid (3 mL) in the cuvette so that when it is inserted into the holder, you can see liquid through the holder’s aperture. That is, the liquid must completely cover the aperture. • Because the cuvette holder is angled, use your thumb or forefinger to ensure the cuvette is secure in the slot and does not slide forward.
  • Page 91 Recommendation: Program the lamp to shut off or turn the lamp off manually only if the value of the “Lamp off” parameter is more than 4 hours. Without powering off the system, you can conserve lamp life by, • turning power to the lamp off and on again manually; •...

  • Page 92: Shutting Down The Detector

    Result: The absorbance screen appears with the message “Igniting.” The lamp can take as much as 1 minute to illuminate. Once the lamp illuminates, the display returns to the absorbance screen, and the X is removed from the lamp icon. You can conserve lamp life by programatically igniting and extinguishing it: for example, overnight, using a timed event method.

  • Page 93: Maintaining The Detector

    For complete information on reporting shipping damages and submitting claims, see Waters Licenses, Warranties, and Support Services. 4.2 Maintenance considerations Perform the procedures in this chapter when you discover a problem with a 2489 UV/Vis detector component or during preventive maintenance. June 11, 2015, 715004752 Rev. A...

  • Page 94: Safety And Handling

    Wait 10 seconds after discontinuing power before attempting to disconnect the assembly. 4.2.2 Spare parts Replace only the parts mentioned in this document. For information about spare parts, see the Waters Quality Parts Locator on the Waters Web site’s Services & Support page. 4.3 Proper operating procedures Note: •...

  • Page 95: Maintaining The Flow Cell

    • 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. This process prevents, – plugging of the solvent lines and flow cell. – damage to the components. –...
  • Page 96 Notice: If the flow cell will not be used for several days, flush it with clean mobile phase, such as a water/acetonitrile or water/methanol, and then cap the flow ports or dry the flow cell with pure nitrogen or pure helium. Notice: To prevent flow cell failure, do not connect any tubing or device that can create backpressure exceeding the flow cell’s maximum rating of 1000 psi.

  • Page 97: Removing And Cleaning The Flow Cell

    Pump a strong cleaning solvent, such as isopropanol, through the flow cell at the recommended flow rate (optional). If the flow cell is dirty, remove any other active detectors from the system, and then pump 1% weak concentration acid solution (such as formic acid solution) through the flow cell at the recommended flow rate.

  • Page 98: Disassembling And Reassembling The Flow Cell

    4.4.3 Disassembling and reassembling the flow cell 4.4.3.1 Before you begin Take the following precautions when disassembling and reassembling the flow cell: • To prevent contamination, wear clean, chemical-resistant, powder-free gloves when touching the flow cell lenses or window. • Take care to prevent scratching of the flow cell parts.
  • Page 99 If the detector has an analytical flow cell (standard in the Alliance HPLC 2489 UV-Vis), once you remove the flow cell assembly from the detector chassis, remove the cuvette subassembly from the flow cell assembly. Figure 4–1: Analytical flow cell assembly with the cuvette...
  • Page 100 • Entrance lens mount • Two gaskets Figure 4–2: Waters TaperSlit flow cell Cell exit Cell entrance For replacement parts for the TaperSlit flow cell, use the Flow Cell Rebuild Kit. Tip: Use nitrogen to clean the flow cell. Use ethanol or methanol to clean the lenses and window.
  • Page 101 To prevent contamination, wear clean, chemical-resistant, powder-free 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 lint-free cloth or similar surface.
  • Page 102 To rebuild the flow cell: Using the plastic tweezers, remove the new clear plastic gaskets from the Flow Cell Rebuild Kit, and inspect them for dirt. Drop one clear, plastic gasket into the groove at the bottom of the entrance lens end of the flow cell body.

  • Page 103: Replacing The Lamp

    To prevent contamination, wear clean, chemical-resistant, powder-free 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 104: Lamp Characteristics

    This section describes the procedure for removing and replacing the detector deuterium lamp. 4.5.1 Lamp characteristics The intensity of the deuterium source lamp varies by wavelength, as shown in the figure below. Figure 4–4: Deuterium lamp sample beam intensity profile Wavelength (nm) 4.5.2 Lamp energy and performance As the lamps used in traditional detectors age, the signal-to-noise performance of the...

  • Page 105: Removing The Lamp

    “Recording sample and reference beam energies” on page 68 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. 4.5.3 Removing the lamp Warning: The lamp housing gets extremely hot during operation.
  • Page 106 Figure 4–5: Lamp assembly and power connector Lamp assembly Power connector Loosen the two captive screws in the base lamp. Figure 4–6: Lamp assembly captive screws Captive screws Withdraw the lamp assembly from the lamp housing. Notice: To prevent shattering the glass, use care when disposing of the lamp as the lamp gas is under slight negative pressure.

  • Page 107: Installing The New Lamp

    Figure 4–7: Removing the lamp assembly Lamp assembly 4.5.4 Installing the new lamp Warning: To avoid exposing your eyes to harmful ultraviolet radiation, never ignite a lamp while it is outside the instrument or if it is not properly secured in place.
  • Page 108 To install the new lamp: Position the lamp so the cut-out located on the lamp base is at the 1 o’clock position, in line with the alignment pin on the lamp housing. Tip: There is no additional alignment required. Figure 4–8: Aligning the lamp Cut-out on lamp base at 1 o’clock...

  • Page 109: Recording The New Lamp Serial Number

    Reconnect the lamp’s power connector. Figure 4–10: Lamp assembly and power connector Lamp assembly Power connector When ready to resume operation of the detector, reconnect the power cord and power-on the unit. Tip: The instrument firmware automatically delays operation for 5 minutes to allow the lamp to warm after it has been reignited.

  • Page 110: Setting The Lamp Threshold

    Figure 4–11: Change lamp screen Press Enter to store the serial number and to move to the “Install date” field. Select the month from the choice list, and press Enter twice to update the month and to select the next field, which specifies the day. Specify the day of the month that the lamp is installed, and then press Enter to enter it and move to the next field, which specifies the year.

  • Page 111: Replacing Fuses

    “Electrical specifications” on page 138. Figure 4–13: 2489 UV/Vis detector rear panel fuse holder Fuse holder Warning: To avoid electric shock, power-off and unplug the instrument before inspecting the fuses.
  • Page 112 Figure 4–14: Power-entry module Power-entry module Pinch the sides of the spring-loaded fuse holder, which is below the power entry module on the rear panel of the detector. Figure 4–15: Sides of spring-loaded fuse holder Sides of fuse holder With minimum pressure, withdraw the spring-loaded fuse holder. Figure 4–16: Removing fuse holder Power-entry module Fuse (2)

  • Page 113: Error Messages, Diagnostic Tests, And Troubleshooting

    Error Messages, Diagnostic Tests, and Troubleshooting The detector provides both user and service diagnostic tests to troubleshoot system problems. 5.1 Error messages 5.1.1 Startup error messages Startup diagnostic tests run automatically when you power-on the detector. They verify the proper operation of the detector electronics. If one or more of the tests fail, the detector beeps and displays an error message.
  • Page 114 Figure 5–1: Diagnostic tests choice list To access a specific diagnostic test, press the up or down arrow key to select a test, and then press Enter. Alternative: Press a number on the keypad corresponding to the test number. Choices that display other choices are indicated by >>. Sticky diagnostic tests remain in effect until you disable them.

  • Page 115: Using The Diagnostic Tests

    • Previous choices << 5.2.2 Using 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. 5.2.2.1 Using the sample and reference energy diagnostic tests You use the sample and reference energy diagnostic tests to plot the output of the analog channels to examine noise fluctuations and to compare with the AU time trace.
  • Page 116 Figure 5–3: 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. Result: When the new wavelength shifts to the left, the corresponding sample and reference energies appear.
  • Page 117 Figure 5–5: Auto-zero offsets diagnostic display 5.2.2.4 Setting a fixed absorbance value From the Input & Output choice list, press 2 (Fix absorbance) to set a fixed absorbance value for channel A or channel B. The allowable range is from –4.0000 AU to +4.0000 AU.
  • Page 118 Figure 5–8: Contact closures & events diagnostic display The Input & Output diagnostic test allows real-time monitoring of the state of the contact closure inputs. A solid (filled in) circle indicates the contact closure is closed (ON = High). An open (empty) circle indicates the contact closure is open (OFF = Low).
  • Page 119 If the keypad is operating properly, each key location is filled in and then cleared with another press of the key. If any key does not respond when pressed, contact your Waters service representative.

  • Page 120: Service Diagnostic Tests

    To deactivate this diagnostic test, press DIAG, and then press 1, or select Automatic from the choice list. 5.2.3 Service diagnostic tests Only qualified Waters service personnel can access the detector service diagnostic tests. June 11, 2015, 715004752 Rev. A...

  • Page 121: Troubleshooting

    Keep in mind that the source of apparent detector problems may lie within the chromatography or your other instruments, as well as the detector. Most detector problems are relatively easy to correct. If you are unable to correct a problem or a failed condition, contact Waters Technical Service (see page 93).
  • Page 122 Keypad not Keypad defective 1. Cycle power to the detector and functioning run the keypad diagnostic test. 2. Contact Waters Technical Service. No sample and Lamp burned out Attempt to reignite using the lamp reference energy key. Replace the lamp.

  • Page 123: Lamp Troubleshooting

    5.3.4 Lamp troubleshooting An apparent lamp-related problem could be caused by air bubbles in the flow cell or incorrect installation of the lamp. Before replacing the lamp, ensure the detector cell is filled with solvent and free of air bubbles, and the current lamp is installed correctly. Flushing the flow cell can sometimes correct the problem of apparently low lamp intensity, which can occur when you have not operated the detector for 2000 hours or more.
  • Page 124 June 11, 2015, 715004752 Rev. A Page 124...

  • Page 125: A Safety Advisories

    The following symbols warn of risks that can arise when you operate or maintain a Waters instrument or device or component of an instrument or device. When one of these symbols appear in a manual’s narrative sections or procedures, an accompanying statement identifies the applicable risk and explains how to avoid it.

  • Page 126: Specific Warnings

    A.1.1.1 Burst warning This warning applies to Waters instruments and devices fitted with nonmetallic tubing. Warning: To avoid injury from bursting, nonmetallic tubing, heed these precautions when working in the vicinity of such tubing when it is pressurized: •...

  • Page 127: Notices

    A.1.1.2 Biohazard warning The following warning applies to Waters instruments and devices that can process material containing biohazards, which are substances that contain biological agents capable of producing harmful effects in humans. Warning: To avoid infection with potentially infectious, human-sourced products, inactivated microorganisms, and other biological materials, assume that all biological fluids that you handle are infectious.

  • Page 128: Bottles Prohibited Symbol

    Use eye protection when refilling or replacing solvent bottles. Requirement: Wear clean, chemical-resistant, powder-free gloves when handling samples. A.5 Warnings that apply to all Waters instruments and devices When operating this device, follow standard quality-control procedures and the equipment guidelines in this section.
  • Page 129 Atencion: cualquier cambio o modificación efectuado en esta unidad que no haya sido expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equipo. 注意：未經有關法規認證部門允許對本設備進行的改變或修改,可能會使使用者喪失操作該設 備的權利。 注意：未经有关法规认证部门明确允许对本设备进行的改变或改装，可能会使使用者丧失操 作该设备的合法性。 주의: 규정 준수를 책임지는 당사자의 명백한 승인 없이 이 장치를 개조 또는 변경할 경우, 이...
  • Page 130 Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione: • Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati. • Spegnere tutte le fiamme vive nell'ambiente circostante. • Non utilizzare tubi eccessivamente logorati o piegati. •...

  • Page 131: Warnings That Address The Replacing Of Fuses

    警告：圧力のかかったポリマーチューブを扱うときは、注意してください。 • 加圧されたポリマーチューブの付近では、必ず保護メガネを着用してください。 • 近くにある火を消してください。 • 著しく変形した、または折れ曲がったチューブは使用しないでください。 • 非金属チューブには、テトラヒドロフラン(THF)や高濃度の硝酸または硫酸などを流さ ないでください。 塩化メチレンやジメチルスルホキシドは、非金属チューブの膨張を引き起こす場合があ り、その場合、チューブは極めて低い圧力で破裂します。 Warning: The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. Attention: L’utilisateur doit être informé...
  • Page 132 Finding fuse types and ratings when that information appears on the instrument or device Warning: To protect against fire, replace fuses with those of the type and rating printed on panels adjacent to instrument fuse covers. Attention: pour éviter tout risque d'incendie, remplacez toujours les fusibles par d'autres du type et de la puissance indiqués sur le panneau à...

  • Page 133: Electrical And Handling Symbols

    Attenzione: per garantire protezione contro gli incendi, sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate nel paragrafo "Sostituzione dei fusibili" del capitolo "Procedure di manutenzione". Advertencia: Para evitar incendios, sustituir los fusibles por aquellos del tipo y características indicados en la sección "Sustituir fusibles".

  • Page 134: Handling Symbols

    Symbol Description Fuse Functional ground Input Output A.7.2 Handling symbols The following handling symbols and their associated statements can appear on labels affixed to the packaging in which instruments, devices, and component parts are shipped. Symbol Description Keep upright! Keep dry! Fragile! Use no hooks! Upper limit of temperature...
  • Page 135 Symbol Description Lower limit of temperature Temperature limitation June 11, 2015, 715004752 Rev. A Page 135...
  • Page 136 June 11, 2015, 715004752 Rev. A Page 136...

  • Page 137: B Specifications

    This appendix lists individual operating specifications for the 2489 UV/Vis detector. Note: All performance specifications are measured following a warm-up period of one hour with ΔΤ ≤ ±2 °C/hr. B.1 Physical specifications The following table lists the physical specifications for the 2489 UV/Vis detector. Table B–1: Physical specifications Attribute Specification Width 34.3 cm (13.5 inches)

  • Page 138: Electrical Specifications

    B.3 Electrical specifications The following table lists the electrical specifications for the 2489 UV/Vis detector. Table B–3: Electrical specifications Attribute Specification Class I Protection class Overvoltage category Pollution degree Normal (IPXO) Moisture protection Grounded AC Line voltages, nominal Power requirements...

  • Page 139: Performance Specifications

    B.4 Performance specifications The following table lists the performance specifications for the 2489 UV/Vis detector. Table B–4: Performance specifications Attribute Specification Wavelength range 190 to 700 nm Bandwidth ≤5 nm Wavelength accuracy ±1.0 nm (via patented erbium filter) Wavelength repeatability ±0.1 nm...

  • Page 140: Flow Cell Specifications

    316 stainless steel, fused silica, PEEK, Tefzel a. US Patent Numbers: US 5,883,721 B.6 Flow cell specifications The following table lists the Waters flow cell specifications for the 2489 UV/Vis detector. Important: The Low Dispersion Analytical flow cell is only compatible with the 2489 UV/Vis detector as a component in an ACQUITY Arc system.

  • Page 141: C Solvent Considerations

    C.1.1 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. C.1.2 Clean solvents Clean solvents provide reproducible results and permit you to operate with minimal instrument maintenance.

  • Page 142: Water

    C.1.5 Water Use water only from a high-quality water purification system. If the water system does not deliver filtered water, filter it through a 0.45-µm membrane filter before use. C.1.6 Using buffers When you use buffers, dissolve salts first, adjust the pH, then filter to remove insoluble material.

  • Page 143: How To Use Miscibility Numbers

    Table C–1: Solvent miscibility: (continued) Viscosit Boiling Miscibilit λ Polarity Solvent y CP, 20 point °C y number Cutoff index °C (1 atm) (nm) Isopropyl ether 0.33 68.3 –– Toluene 0.59 100.6 P-xylene 0.70 138.0 Benzene 0.65 80.1 Benzyl ether 5.33 288.3 ––...

  • Page 144: Buffered Solvents

    To predict the miscibility of two liquids, subtract the smaller M-number value from the larger M-number value. • If the difference between the two M-numbers is 15 or less, the two liquids are miscible in all proportions at 15 °C (59 °F). •...

  • Page 145: Mobile Phase Solvent Degassing

    that occur as the solvents are mixed in different proportions can result in pressure changes during the run. For example, a 1:1 mixture of water and methanol produces twice the pressure of either water or methanol alone. If the extent to which the pressure changes will affect the analysis is not known, monitor the pressure during the run using the Chart Out terminal.

  • Page 146: Solvent Degassing Methods

    C.6.1.3 Effects of partial pressure The mass of gas dissolved in a given volume of solvent is proportional to the partial pressure of the gas in the vapor phase of the solvent. If you decrease the partial pressure of the gas, the amount of that gas in solution also decreases. C.7 Solvent degassing methods This section describes the solvent degassing techniques that will help you to attain a stable baseline.

  • Page 147: 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.

  • Page 148: Mixed Mobile Phases

    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 149: Wavelength Selection For Chromophore Detection

    C.8.3 Wavelength selection for chromophore detection Certain functional groups found in most compounds absorb light selectively. These groups, known as chromophores, and their behavior can be used to categorize the detection of sample molecules. The table below lists some common chromophores, and their detection wavelengths ), as well as the molar absorptivity (ε...
  • Page 150 Table C–4: Electronic absorption bands of representative chromophores*: ∈ ∈ Chemical λ λ Chromophore (L/m/cm (L/m/c configuration (nm) (nm) 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 —...

Table of Contents

Save PDF