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P H A R M A C I A B I O T E C H
Monitor UV-1
User Manual
59-7797-01
Edition AG
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Important user information Reading this entire manual is Warranty and Liability GE Healthcare Bio-Sciences AB guarantees that the recommended for full product delivered has been thoroughly tested to understanding and use of ensure that it meets its published specifications. this product. The warranty included in the conditions of delivery is valid only if the product has been installed and used according to the instructions supplied by GE Healthcare Bio-Sciences AB. GE Healthcare Bio-Sciences A
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Contents 1. Introduction........................................................................................ 4 2. General Description ..........................................................................5 2.1 Basic principle.............................................................................5 2.2 Optical unit .................................................................................7 2.3 Control unit .......................................................................
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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 recor- ded 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. In addition, five flow cells ar
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2. General Description 2. General Description GE Healthcare LKB Monitor UV-1 consists of an optical unit 2.1 Basic principle 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. The single path Monitor UV-1 is a dual beam instrument with
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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. The signal finally passes to
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2. General Description Front panel controls 2.2 Optical unit 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. The reference cell may be operated dry, with static reference liquid 5 Reference outlet Outlet for flowing reference liquid 7
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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 Filters, converters or apertures are inserted in the positions indicated. When inserting them, align 8 Converter or the arrowhead on the end of the filter or converter Aperturewith the arrowhead next to the appropriate opening. Filters
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2. General Description Front panel controls 2.3 Control unit 15 12 16 13 14 Fig. 5. Control unit. Front panel. No. Item Description 12 Absorbance range Selector for the desired absorbance range selector 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. 13 Mode switch The AU / % T switch, enables
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2. General Description Rear panel controls Fig. 6. Control unit. Rear panel. Description No. Item 17 Mains voltage selector Selects mains voltage 110, 130, 220 and 240 V 1 x 250 mA for 110-130 V 60 Hz 18 Fuse holder Mains fuses: 1 x 125 mA for 220-240 V 50 Hz The output signal is a 10 mV DC signal. The 19 Signal outputs terminals are connected to a potentiometric recorder via shielded cables supplied. The UV-1 is normally earthed via the mains ground 20 Mains i
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3. Installation 3. Installation UV-1 should be installed on a stable, flat surface away from all sour- 3.1 Site ces of vibration. The atmosphere should be free of both excess humi- requirements dity 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. UV-1 optical unit should be positioned away from all
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3. Installation Mains voltage Voltage selector setting Fuse 250 mA 110 V 110 250 mA 130 V 130 220 125 mA 220-230 V 240 125 mA 240 V 4. 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 mus
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3. Installation Monitor UV-1 accepts flow cells for Standard Chromatography, FPLC 3.5 Installation of and industrial applications. the flow cell Product Material of Path Total dead Illuminated Pressure Application Code No. wetted parts length volume volume limit area S-2 Fluoro-plastic, 2 mm 80 μl 2 μl 0.3 MPa Standard 19-4840-02 optical quartz (3 bar) Chromatography HR-10 Fluoro-plastic, 10 mm 24 μl 8.7 μl 1.0 Mpa FPLC, Standard 19-6254-02 optical quartz,titanium (10 bar) Chromatography
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3. Installation The optical unit may be placed on the bench or mounted on 3.6 Connecting laboratory scaffolding. For scaffolding mounting, mount the support the optical unit 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
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4. Operation 4. Operation The UV-1 can be operated at either 254 nm, 280 nm or 405 nm. 4.1 Choice of The choice of wavelength will depend on the spectral properties of wavelength both the eluent and the substances to be detected. Proteins and poly- pep-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. The UV-1
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4. Operation 4.3 Conversion T% AU OD OD (3 mm cell) (10 mm cell) table T% to AU 1 2.000 6.667 2.000 and OD 1.301 5 1.301 4.337 1.000 1.000 10 3.333 15 2.747 0.824 0.824 20 0.699 0.699 2.330 2.007 25 0.602 0.602 1.743 30 0.523 0.523 1.667 32 0.500 0.500 1.520 35 0.456 0.456 40 1.327 0.398 0.398 45 1.157 0.347 0.347 50 0.301 1.003 0.301 55 0.260 0.867 0.260 60 0.222 0.740 0.222 0.200 63 0.200 0.667 0.186 0.620 0.186 65 70 0.155 0.517 0.155 75 0.125 0.417 0.125 79 0.100 0.333 0.100 80 0.097 0.32
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4. Operation 1. Switch AU/ %T to AU (Fig. 5:13). 4.6 Basic operating 2. Set the range selector (Fig. 5:12) to SHORT and zero the recorder with the recorder zero control. procedure - AU 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). Only minor adjustment should be necessary. 1. Switch AU to %T (Fig. 5:13). 4.7 Basic 2. S
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5. Maintenance 5. Maintenance To ensure trouble free running, users are advised to observe the follo- 5.1 General wing precautions: 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. Never touch the optical surfaces
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5. Maintenance Cleaning with detergent: Remove the cell holder from the optical unit. 1. 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) Replace the cell in the optical unit and reconnect the system to 4. be monitored. Cleaning with chromic acid: 1. Prepare fresh chromic acid by adding concentrated sulphuric acid (100 ml) to a saturated solutio
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5. Maintenance 8. Insert the new cell, being careful not to touch either of the optical surfaces. See that the inlet and outlet ports are correctly aligned. 9. Retighten the screw washers with the tool provided. Do not use excessive force. 10. Check for leakage. 11. Replace the black cover. Sample out Reference out Sample in Reference in Fig. 13. Flow cell interior showing tubing connections. For optimum performance, it is essential that the interference filters 5.4 Inter
