Monitor UV-1 User Manual 59-7797-01 Edition AG
Important user information Reading this entire manual is recommended for full understanding and use of this product. ● The exclamation mark within an equilateral triangle is intended to alert the user to the presence of important operating and maintenance instructions in the literature accompanying the instrument.
Contents 1. 2. 3. 4. 5. 6. 7. 8. Introduction ........................................................................................ 4 General Description .......................................................................... 5 2.1 Basic principle ............................................................................. 5 2.2 Optical unit ................................................................................. 7 2.3 Control unit .......................................................
1. Introduction 1. Introduction The UV-1 is a fixed wavelength UV monitor, consisting of a control unit and an optical unit that can be positioned up to 10 meters apart. A mercury lamp is the stable light source, furthermore a built-in reference cell eliminates baseline drift. The output signal can be recorded in either AU or %T. Sensitivity range between 0.01 and 2 AUFS, or (0-100%T). The UV-1 offers a range of three detection wavelengths: 254, 280 and 405 nm.
2. General Description 2. General Description 2.1 Basic principle GE Healthcare LKB Monitor UV-1 consists of an optical unit containing the flow cell, lamp, filter assemblies and preamplifiers, and a control unit containing the signal processing circuits. The two units are connected via a multi-core cable hardwired from the control unit to the optical unit. Connection to the recorder and the mains supply is made via the control unit.
2. General Description Fig. 2 Block diagram The photocurrent from each detector is amplified in a pre-amplifier before passing to the signal processing circuitry in the control unit. If transmission is to be monitored, the signal passes directly to the low pass filter. If AU is to be monitored, the reference cell light intensity (IR) is compared with the sample cell intensity (IS) to form log (IR/IS) in a logarithmic circuit before passing to the low-pass filter.
2. General Description 2.2 Optical unit Front panel controls 5 4 1 2 3 Fig. 3. Optical unit. Front panel. No. Item Description 1 Cell holder The complete cell holder is removed by turning the locking (Fig. 4:6) knob on the rear panel 2 Sample inlet Inlet for sample flow 3 Sample outlet Outlet for sample flow 4 Reference inlet Inlet for flowing reference liquid.
2. General Description Rear panel controls 6 11 9 Guide pin Screw 7 10 8 Fig. 4. Optical unit. Rear panel. No. Item Description 6 Locking knob The cell holder is in locked position when the locking knob is turned fully in the direction of the arrow 7 Filter inlet 8 Converter or Filters, converters or apertures are inserted in the positions indicated. When inserting them, align the arrowhead on the end of the filter or converter Aperturewith the arrowhead next to the appropriate opening.
2. General Description 2.3 Control unit Front panel controls 15 12 16 13 14 Fig. 5. Control unit. Front panel. No. Item Description 12 Absorbance range selector Selector for the desired absorbance range 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1 or 2 AU full scale deflection. In the position SHORT, the signal output terminals are disconnected from the rest of the control circuitry and short-circuited. This position is useful when setting zero on the recorder.
2. General Description Rear panel controls Fig. 6. Control unit. Rear panel. No. Item Description 17 Mains voltage selector Selects mains voltage 110, 130, 220 and 240 V 18 Fuse holder Mains fuses: 19 Signal outputs The output signal is a 10 mV DC signal. The terminals are connected to a potentiometric recorder via shielded cables supplied.
3. Installation 3. Installation 3.1 Site requirements UV-1 should be installed on a stable, flat surface away from all sources of vibration. The atmosphere should be free of both excess humidity and corrosive or contaminated vapours which may form deposits on the component in the optical path. UV-1 can be installed either in a coldroom or at ambient temperature in the laboratory. To minimise drift, the temperature should be kept constant.
3. Installation Mains voltage Voltage selector setting Fuse 110 V 130 V 220-230 V 240 V 110 130 220 240 250 mA 250 mA 125 mA 125 mA 4. 3.4 Installation of the filter Select the mains cable corresponding to your mains outlet. Discard unwanted mains cable immediately. Connect the instrument to a grounded mains outlet. Note: Do NOT switch on. Note: Special care must be taken when handling interference filters. DO NOT touch the filter surface.
3. Installation 3.5 Installation of the flow cell Monitor UV-1 accepts flow cells for Standard Chromatography, FPLC and industrial applications. Product Code No. Material of wetted parts Path length Total dead volume Illuminated Pressure volume limit Application area S-2 19-4840-02 Fluoro-plastic, optical quartz 2 mm 80 µl 2 µl 0.3 MPa (3 bar) Standard Chromatography HR-10 19-6254-02 Fluoro-plastic, optical quartz,titanium 10 mm 24 µl 8.7 µl 1.
3. Installation 3.6 Connecting the optical unit The optical unit may be placed on the bench or mounted on laboratory scaffolding. For scaffolding mounting, mount the support rod on the optical unit. The rod may be mounted horizontally or vertically (Fig. 4:9). Tighten the the Allen screw firmly. The optical unit should be placed as close as possible to the column outlet. Connect the cable from the optical unit to the 1 l-pin socket on the back of the control unit (Fig. 6:21). The plug has a snap lock.
4. Operation 4. Operation 4.1 Choice of wavelength The UV-1 can be operated at either 254 nm, 280 nm or 405 nm. The choice of wavelength will depend on the spectral properties of both the eluent and the substances to be detected. Proteins and polypep-tides containing aromatic amino acids are usually best detected at 280 and 254 nm. Nucleic acids and poly-nucleotides are usually best detected at 254 nm and ferroproteins i.e. hemoglobins, cytochrome and porphyrin derivatives at 405 nm. 4.
4. Operation 4.3 Conversion table T% to AU and OD 4.4 Start-up 4.5 Stabilization time T% AU OD (3 mm cell) OD (10 mm cell) 1 5 10 15 20 25 30 32 35 40 45 50 55 60 63 65 70 75 79 80 85 89 90 95 95.5 97.7 2.000 1.301 1.000 0.824 0.699 0.602 0.523 0.500 0.456 0.398 0.347 0.301 0.260 0.222 0.200 0.186 0.155 0.125 0.100 0.097 0.070 0.050 0.046 0.022 0.020 0.010 6.667 4.337 3.333 2.747 2.330 2.007 1.743 1.667 1.520 1.327 1.157 1.003 0.867 0.740 0.667 0.620 0.517 0.417 0.333 0.323 0.233 0.167 0.153 0.
4. Operation 4.6 Basic operating procedure AU 4.7 Basic operating procedure Transmission (%T) 4.8 Shut down 1. Switch AU/ %T to AU (Fig. 5:13). 2. Set the range selector (Fig. 5:12) to SHORT and zero the recorder with the recorder zero control. 3. Set the range selector to 2 and adjust the recorder baseline with the baseline adjust (Fig. 5:14). 4. Set the range selector to the appropriate range and readjust the recorder baseline with the baseline adjust (Fig. 5:14).
5. Maintenance 5. Maintenance 5.1 General precautions To ensure trouble free running, users are advised to observe the following precautions: All liquids passing through the flow cell should be free of suspended particles. All liquids should be degassed to prevent air bubble formation in the flow cell. Never allow buffer solutions to dry out in the cell. Always rinse the flow cell thoroughly with distilled water after use. Handle interference filters with care.
5. Maintenance Cleaning with detergent: 1. Remove the cell holder from the optical unit. 2. Pump undiluted cleaning detergent through the cell for at least 2 hours. 3. Rinse the cell with a) distilled water (100 ml) b) ethanol/distilled water (50% v/v, 100 ml) c) distilled water (100 ml) 4. Replace the cell in the optical unit and reconnect the system to be monitored. Cleaning with chromic acid: 1.
5. Maintenance 8. 9. Insert the new cell, being careful not to touch either of the optical surfaces. See that the inlet and outlet ports are correctly aligned. Retighten the screw washers with the tool provided. Do not use excessive force. 10. Check for leakage. 11. Replace the black cover. Reference out Sample out Reference in Sample in Fig. 13. Flow cell interior showing tubing connections. 5.
5. Maintenance 4. Set the range selector to SHORT and zero the recorder. 5. Set the range selector as follows: 254 nm, range 1 280 nm, range 0.2 6. Turn Zero fully clockwise. Response 8 mV or greater: the lamp is operating properly and the optical system is clean. Response 0 mV or very close to 0 mV: Contact a GE Healthcare representative. Response less than 8 mV but not very close to zero: check that the cell and filter is clean.
5. Maintenance 6. Use an Allen key (2.5 mm) to unscrew the upper of the two screws securing the lamp holder and the lamp. Note carefully the positions of the insulation sleeve and washer. 7. Disconnect the lamp from the PC-board. 8. Bend the upper part of the lamp holder slightly (2-3 mm) upwards and withdraw the mercury lamp from it by gently pulling the metal socket. 9. Insert the new mercury lamp and connect it to the PC-board. 10.
6. Trouble-shooting 6. Trouble-shooting The UV-1 Monitor has been designed for trouble-free use. If good chromatographic practice is followed, very little difficulty should be experienced. Clean optical surfaces are essential if low noise levels are to be maintained. The following check list of the most frequent problems is meant to be a guide in trouble-shooting. If the checks in this section are executed and the UV-1 still does not work properly, consult your local GE Healthcare representative.
7. Technical Specifications 7. Technical Specifications 254 nm, 280 nm and 405 nm Hg lamp: for 254, 280, and 405 nm Life time lamp: 8000 hours converter: 2000 hours Interference. Stray light maximum 0,1 % Filters at 254 nm, maximum 0.8% at 280 nm AU or Transmission Operating modes 0.01, 0.02, 0.05, 0.1, 0.2, 0.
8. Accessories and Spare Parts 8. Accessories and Spare Parts Please order accessories and spare parts according to the designation and code numbers given below. Designation Code No.
Printed in Sweden by SNITS & DESIGN AB/Västra Aros Tryckeri, Nov 1996
