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SpectraSYSTEM UV/Vis Detectors Reference

1. Door of Power Entry Module Fuse Holders si Fuse 8 Figure A 3 Fuses 3 Assuming that you have the proper fuses on hand reinsert the fuses and fuse holders making sure that the arrows on the holders are oriented in the same direction as the arrow inside the cover panel Fig A 3 4 Close the cover panel by svvinging it upvvard and pressing it in until it snaps closed The correct voltage should appear in the cut out panel NOTE To avoid damaging the instrument verify that the new voltage setting displayed in the cut out window is correct before you turn it on Power Cord Attach the power cord at the lower left of the detector s rear panel 82 Thermo Electron 01 2004 MAKING Locate the two connectors 8 pin and 12 pin in your accessory kit REAR PANEL and insert them in the appropriate sockets on the detector s rear panel CONNECTIONS Fig A 4 Note that the connectors are both labeled and keyed to the sockets making it impossible to insert them incorrectly TERMINAL CONFIGURATION UV1 OOO 12 STOP Input 11 GROUND 10 RUN Input D UNRANGED 9 GROUN 8 ZERO Input OUTPUT 1AU V 7 GROUND L E GROUND 4 READY Output D ED C B 1 0 VF S zl GROUND m THIS PRODUCT IS COVERED BY ONE OR MORE OF THE FOLLOWING PATENTS 498 812 TERMINAL CONFIGURATION idi UV2000 12 STOP Input
2. Er ENE E e E aE d tn a xix List of Spare Parts Consumables and 6 2 2222 xxi Chapter 1 Getting Started Introduction esse one dares ete b 1 Learning Your Way Around u eerren a e entearen e CEEE a EEEE E EE lenkte 1 Instrument Control errin a R R A eh n A 2 Manual Conventions a a e ETE T E E E E l 6 Whats NeXt Ae s n s a 8 Chapter 2 A Quick Example ntrod etioh AYAR en te en es E 9 UV1000 An Example cci letaceiastettsislstsceetelisedioleldccttalelasebledscebhdesedbsbalasstsshelectedebaevel Hess 10 UV2000 An Example a ID rel 14 WHat G2 aaa yas alaya Ea IS need 18 Chapter 3 Basic Operations Introduction ea rn Sed de ne 19 Before You Begin nennen RU en der E ln 19 UV1000 Single wavelength Operation ss 20 UV2000 Single and Dual wavelength Operation ss 26 More about Files s s 0 ran iR RB is HEAD s n 32 Analog Outputs un mais RE K l 35 Chapter 4 Advanced Operations Introduction u a ai Ari uate ee cc abe Band cn ceca aaa aie 37 Wavel neth Programming 37 Pr srammed A tozero 2 2 2 2 ni E MM ne na een etre ele re eat Pad dees cd 40 Automatic Lamp Operations sisi 41 Other Festures aa ee es eee 43 U V 2000 Scanners satin
3. Display AU 1 66681 0 28831 Figure 4 35 The Display AU screen Ratioing only occurs when the absorbance value for each wavelength exceeds 12 5 of the corresponding range value So in our example if Ranges 1 and 2 were set to 1 0 in the FILES Edit Options Menu the 250 and 280 nm wavelengths could be ratioed Twelve and a half percent of 1 0 the range is 0 125 Absorbance values less than 0 125 are too low for ratioing No ratio output is produced when the absorbance values fall below 7 5 of the range values Generally good wavelengths to choose are 1 the lambda max of the main peak AU1 2 a wavelength with an absorbance value less than the lambda max but greater than 12 5 of the corresponding range AU2 HINT A good rule of thumb is to select a second wavelength that is either half the height of the lambda max or more than ten nanometers removed from the lambda max Whichever wavelengths you choose don t select a wavelength that has a low absorbance value Low absorbance values decrease the signal to noise ratio thus making the absorbance ratios less meaningful Similarly a small fluctuation in AU2 results in a big difference in the absorbance ratio if AU2 is very small Fortunately by relying on the preset range values the UV 2000 has a built in safeguard that prevents the ratioing of low absorbance values Thermo Electron 01 2004 5 Required Maintenance Introduction Finnigan SpectraS Y
4. Zero on A Change This field toggles between Yes where the detector baseline automatically zeroes at each timed event during a programmed run and No Default is Yes 104 Thermo Electron 01 2004 C Troubleshooting Introduction This Appendix provides you with helpful information for troubleshooting possible detector and chromatographic system problems We have divided it into four sections a brief theory of operation a troubleshooting guide that lists symptoms possible problems and remedies e error messages you might see on the detector s display a description of internal and external diagnostic tests Theory of Operation This brief Theory of operation is included to aid you in troubleshooting problems and performing maintenance for your detector For more detailed information you should contact your Thermo Electron representative 01 2004 Thermo Electron 105 OVERVIEW Figure C 1 shows the optical system used in both the UV1000 and UV2000 detectors The detector operates in a double beam mode using a fiber optic beam splitter that creates sample and reference beams The reference beam is directed to a reference photodiode The sample beam is lens focused prior to passing through the flowcell to a sample photodiode An analog PCB processes the signals from the photodiodes and provides analog output signals through an 8 pin external connector The digital PCB contains the EPROM the built in software p
5. 01 2004 As lamps age there is a reduction in light output that results in increased baseline noise If the noise level on your detector s output signal is increasing and cleaning the flowcell doesn t help you should change the appropriate lamp using the procedures in this section Remove the front panel of the detector The deuterium and tungsten lamps are located in the upper housing Fig 5 4 Both lamps are supplied prealigned in their individual assemblies to make them easy to install and align NOTE Never loosen the screws that hold the lamp to its assembly or attempt to rotate or move the lamp up or down in the assembly Either of these actions can cause a loss of alignment and degrade the system s performance Lamp Housing Flowcell Housing Display RUN STATUS 4 MENU stop lt _ lt Keypad gt ENTER SpectraSYSTEM UV2000 I Figure 5 4 Location of lamp housing Thermo Electron 73 THE DEUTERIUM LAMP D2 Lamp Removal NA 74 The deuterium D2 lamp typically requires a warm up time of twenty to thirty minutes However for applications that demand great sensitivity you may want to allow a warm up period of up to an hour The deuterium lamp s lifetime is usually at least 1000 hours Each D2 lamp assembly is equipped with a chronomet
6. Flowcell Orientation 88 The flowcells listed above are configured for use with any SpectraSYSTEM detector These detectors are vertically oriented detectors that have the tubing clamp located above the flowcell as shown in Figure A 7 If you plan to use any of these flowcells on non SpectraS YSTEM detectors primarily horizontally oriented detectors that have their tubing clamps located to the left side of the flowcell as shown in Figure A 8 you must realign the cell holder as described in the following instructions Figure A 7 shows the flowcell as it is shipped NOTE Figures A 7 and A 8 show the tubing clamp as an aid to the proper positioning of the inlet and outlet tubes The tubing clamp however is actually mounted onto the detector and is not part of the flowcell itself HINT To ensure proper alignment always hold the cell holder and flowcell in the orientation shown in the illustrations Thermo Electron 01 2004 Inlet Tube Tubing Clamp on Outlet Tube DET Z014 FM vertically oriented detectors Photodiode Standoffs Mounting Screws Figure A 7 The alignment of UV1000 and UV2000 flowcells as they are shipped Remove the mounting screws and set them aside 2 Rotate the cell holder 90 counterclockwise Do not rotate the cell body Part B of Figure A 8 shows the cell holder in its new position Note the new position of the photodiode standoffs Photodiode
7. We have also standardized the meanings of two words select and enter select The word select is used when you need to choose from among available options For example to select a particular menu choice you would move the cursor to the appropriate choice and press ENTER To select a field entry move the cursor to the appropriate field and use the and keys to scroll to the desired preset value enter The word enter is used when you need to specify individual alphanumeric digits To enter a particular value move the cursor to the desired field and use the and keys to increment or decrement each digit in the field until the desired value or letter appears Thermo Electron 7 ICONS gt gt p 2 l This manual uses the follovving icons to alert you to various situations Each is called out by an icon in the left margin Caution A caution alerts you to situations that could result in personal injury It also tells you how to avoid them High Voltage This icon alerts you to the presence of high voltage and to the potential injury that could occur from electrical shock were you to come in contact with a specific instrument area or component It also tells you how to avoid contact with the high voltage areas in your instrument Hot Surface This icon alerts you to potential injury that could occur from coming in contact with a heated surface or area on or in an instrument It al
8. an no s nd no Bojeuy s s l H s SI8SJO epolq s ox97 1461 15 m 8 S iz z ebuey 018Z0 ny Sul si suondO 002 99 000 a LY Sul 002 996 Vieng we oid 00 99 v enq weibold 08c 092 000 Ou LY Sul 0SF 061 Vieng wei6olg 097 061 210 weu old 092 0070 yuaja M euuI L x l5ulS HO iy HO J HO Z HO J98J01d awen al d IH nd no Aqvau HO 4007 snye S wn poyW p ds sno SA eBueu Y 4 0127 000 LO Jope M nv z Bojeuy L 96 SHO z Boleuy 18SH0 Bojeuy 00 1 Apeoy wo 00 0 eu UMOPNUyS jenuey uxop nus 00 0 awl jenuen dnyeyS 00 0 Sul Juan 0 s noH due M 0 s noH dwe 20 98 06 za dwe z ses dojanaq 0 814 z s UEOS S allg x l ulS wesbold suondo sdwe 7 91531 s nd no Bojeuy sdwe7 SNOILdO SONVININOO 91531 20300 5313 4 lli 979180 SNOILdO 1 01 OL L 114 402 1098 2 uxopinus s nd no ous HEIN 44943 Adoo b lli peo SONV N NO peo y r q Ado peo weN all weu ord yua M rl salld b 913 HPA
9. 7 Sch den k nnen auch eintreten wenn das Ger t l ngere Zeit unter ung nstigen Umst nden gelagert wurde z B unter der Einwirkung von Hitze oder Wasser 8 Ziehen Sie vor allen Wartungsma nahmen immer zuerst den Netzstecker aus der Steckdose 9 Auch wenn das Ger t abgeschaltet ist k nnen die im Inneren befindlichen Kondensatoren nach wie vor unter Spannung stehen 10 Versuchen Sie niemals Ger tekomponenten zu reparieren oder auszutauschen die nicht in diesem Handbuch beschrieben sind ohne einen Kundendiensttechniker zu Rate zu ziehen XV Thermo Electron 01 2004 GLP VORSCHRIFTEN GOOD LABORATORY PRACTICES 01 2004 Ordnungsgem e Aufzeichnungen Damit Probleme mit Ger ten oder Methoden erkannt und eingegrenzt werden k nnen empfehlen wir Ihnen ordnungsgem e Aufzeichnungen s mtlicher Ger tezust nde z B RSDs zu Retentionszeiten Kurvenfl chen Kurvenformen und Aufl sung Archivieren Sie als Minimum ein Chromatogramm einer typischen Probe und einer Standardmixtur mit umfassender Dokumentation der Systembedingungen zum sp teren Vergleich Ein sorgf ltiger Vergleich von Retentionszeiten Kurvenformen Empfindlichkeitswerten und Hintergrundrauschen liefert wertvolle Hinweise f r den Fall da zu einem sp teren Zeitpunkt Probleme auftreten und eingegrenzt und behoben werden m ssen Chemische Toxizit t Die gro en Mengen an toxischen oder brennbaren L sungsmitteln die im Labor verwendet
10. file G K gradient elution ground terminal K factor keypad M Q menu miscible parameter peak broadening peak suppression photodiode queue 118 A copy of the file used for editing Once loaded the parameters set in the edit file are transferred to the runfile A displayed message that notifies you of a problem The area in a display screen or menu where an entry is required or a choice must be made A list of detector parameters that contains the desired settings for an analysis A liquid chromatographic technique where the mobile phase composition changes over time changes may be continuous or in steps A terminal used to connect the ground or earth lead of a signal or contact closure cable A factor used to calculate a response of zero for one of two coeluting or poorly resolved peaks also known as peak suppression All of the keys that you use to communicate with your instrument A list of choices Two solvents are miscible if they combine with each other to form a single phase A value or set of values used to define the characteristics of behavior of an instrument or system The dilution of a peak as it moves through the chromatographic system A technique that uses a factor the K factor to calculate a response of zero for one of two coeluting or poorly resolved peaks The detector component that measures light intensity A set of items i e samples files in a prearranged or
11. ANALOG 1 11 GROUND OUTPUT F R 10 RUN Input H CET 9 GROUND G 8 ZERO Input F 1 0 VF S 00 1 2 e 5 GROUND 4 READY Output B 1 0 VF S A GROUND THIS PRODUCT IS COVERED BY ONE OR MORE OF THE FOLLOWING PATENTS 498 812 DT Z170E DT Figure A 4 Analog output and remote communications connectors The upper connector holds the detector s analog output terminals The lower connector allows the detector to communicate with other devices in your liquid chromatographic system There is also a communications port labeled COMM Use the cables supplied with your detector to make the connections described in this section For each connection insert the cable s bare wire into the appropriate detector terminal Hold the wire in place while you tighten the small setscrew located next to each opening 01 2004 Thermo Electron 83 UV2000 Analog Output Connections UV1000 Analog Output Connections The terminals on the UV2000 s analog output connector are labeled ANLG 1 Output and ANLG 2 Output Fig A 4 Each output has four terminals labeled H through E for Output 1 and D through A for Output 2 These terminals correspond to 0 01 V full scale terminals H and 0 10 V full scale terminals G and C 1 0 V
12. Each step represents approximately 0 25 nm so if you choose 2 for the number of steps you will have offset the wavelength by 0 5 nm You can offset the wavelength by as much as 2 5 nm NOTE The offset value is not cleared upon resetting the RAM memory It EN can only be changed from the lambda offset screen Self Tests The detector automatically runs eight internal diagnostic tests when it is powered up To run the tests at any other time simply select Self Tests If any test other than the two lamp tests fails you ll see a message to that effect on the display Clear the message and run the remainder of the self tests by pressing ENTER Repeat this process as many times as necessary until all self tests are completed and the Status Screen appears If any test has failed the Status Screen will read NRDY Not Ready Although you can frequently get back to the Ready state on your own e g you can manually turn on the lamps from the Options Menu or load a file the detector may not function properly and your results may be affected For this reason and to help you troubleshoot the detector on your ovvn vve have listed the MLF most likely failure for each test Problems that are not readily resolved should be referred to your Thermo Electron representative 114 Thermo Electron 01 2004 01 2004 The eight self tests are 1 RAM This test checks both non volatile and volatile RAM with a read write test
13. If you are using a stop line in your wavelength program you will start and stop the run with each injection 01 2004 Thermo Electron 29 30 Status Screen Inject your Sample Starting a Run Stopping a Run You can check the detector s status wavelength setting s and absorbance reading s from the Status Screen To access the Status Screen press STATUS The Status Screen for the UV2000 in dual wavelength mode appears below Fig 3 18 Note that in the single wavelength mode the third line does not appear Status AU Figure 3 18 The UV2000 s Status Screen for dual wavelength operation If the Status reads READY the detector is stabilized and ready to run If NRDY appears the detector s lamps may need additional time to warm up or a wavelength outside the selected lamp s range may have been chosen When the detector is stabilized and you are ready to inject your sample first manually zero the detector by pressing the ZERO key If you are not using a stop line in the wavelength program the detector remains in the READY state throughout your chromatographic runs If you are using a stop line you must start and stop the run with each injection following the procedures below If you are using a stop line in your wavelength program you need to start the run with each injection There are two ways to start a run using the UV2000 1 Manually by pressing RUN each time you make an injection 2 Automatic
14. Standoffs Tubing Clamp on horizontally oriented detectors Cell Holder Mounting Screw B Holes C Figure A 8 Changing the alignment of a flowcell so that it can be used on other detectors 01 2004 Turn the cell holder as shown in Part B Align the inlet and outlet tubes with the tubing clamp as shown in Part C Thermo Electron 89 3 Reattach the cell body by replacing and securing the mounting Screws 4 Gently bend the inlet and outlet tubes as shown in Part C of Figure A 8 The inlet tube wound around the cell body should always enter at the bottom of the flowcell the outlet tube should always exit at the top of the flowcell Specifications 90 Wavelength Lamp s Bandwidth Wavelength Accuracy Wavelength Precision Range Selections Absorbance Range Absorbance Linearity 254 nm Analog Outputs UV2000 Outputs 1 and 2 Communications Remote Inputs Remote Outputs Thermo Electron D2 lamp 190 to 380 nm UV1000 190 to 365 nm UV2000 W lamp 366 to 800 nm UV1000 D2 standard W optional UV2000 D2 and W standard 6 nm 1 0nm 0 1 nm 3 0 2 0 1 0 0 5 0 2 0 1 0 05 0 02 0 01 0 005 0 002 0 001 0 0005 AUFS 0 0005 to 3 0 AUFS Better than 1 to 2 0 AU Range selectable over entire absorbance range Run Stop and Zero Ready 01 2004 Specifications cont Noise Single wavelength Mode 254 nm 1 0 sec rise time lt 1
15. UV2000 only This field allows you to identify the file for set up or edit Allowable designations are 1 to 4 S for the Scan file and D for the Develop file Default is 1 UV2000 only n the Scan File Setup this field defines the wavelength at which the detector should finish the scan Allowable values are 191 to 800 nm Default is 350 nm UV2000 only In the Develop File Setup this field defines the wavelength at which the detector should run its last set of injections Allowable values are 191 to 800 nm Default is 350 nm UV2000 only The Event Mark field applies a 15 of full scale spike on the detector s output signals The UV1000 s FILES Menu and the UV2000 s FILES Menu allow you to edit load and delete files On the UV2000 the FILES Menu also lets you copy files Thermo Electron 01 2004 File Name K Factor A1 22 Calibration Interval AMax AMaxAU AMin Offset Lamp Lamps Light Levels 01 2004 This field allows you to enter a file name for a designated file numbered 1 to 4 The name can contain up to eight characters from the following list A to Z 0 to 9 and blank Default is blank UV2000 only For Files S and D the file names are automatically designated as SCAN and DEVELOP respectively No editing of these file names is allowed UV2000 only This field is used in the technique Allowable values are 99 999 to 99 999 Default is 1 000 UV2000 only
16. UV2000 s Analog Output 1 and Analog Output 2 terminals Allowable full scale ranges are 3 0 2 0 1 0 0 5 0 2 0 1 0 05 0 02 0 01 0 005 0 002 0 001 and 0 0005 AUFS Default is 1 0 AUFS UV2000 only This field is used to communicate with other devices through the detector s READY Output terminal This TTL terminal switches the transistor between high and low states whenever the detector starts a run Select Active Hi or Active Lo for the high or low state respectively Default is Active Hi The Replay command sends you to the Replay Menu from which you can set up the parameters for replaying stored spectra UV2000 only This command is used to initiate replay of the designated spectrum UV2000 only This field designates the rate at which the detector replays a stored spectrum Allowable values are 1 2 5 10 and 20 nm sec Default is 5 nm sec UV2000 only This field controls the detector s response time Rise time is inversely proportional to the amount of baseline noise Allowable values are 0 0 0 1 0 2 0 5 1 0 2 0 3 0 4 0 and 5 0 seconds Default is 1 0 second When this field appears in the Status Screen the current run and the total number of injections to be made at the displayed wavelength appear directly below it The field is updated at the beginning of each injection UV2000 only When this field appears in a Queue setup it displays the number of times each file runs in a queue UV2000 only Th
17. Wird ein Verl ngerungskabel verwendet mu auch hier der Schutzleiter durchgef hrt sein 4 Ver ndern Sie nichts an den externen oder internen Schutz bzw Erdungskontakten Wenn Sie sich an diesen zu schaffen machen oder sie unterbrechen k nnen Sie sich selbst und andere gef hrden oder das Ger t k nnte besch digt werden VORSICHT Das Ger t ist bei der Auslieferung vorschriftsm ig geerdet Es brauchen keine Ver nderungen an der elektrischen Verkabelung oder am Ger techassis vorgenommen werden um einen sicheren Betrieb zu gew hrleisten VORSICHT Setzen Sie niemals den Sicherheitsschalter der Lampenabdeckung ausser Kraft Der Sicherheitsschalter schaltet die Lampen aus wenn die Abdeckung entfernt wird Die berbr ckung des Sicherheitsschalters kann zu Gesundheitssch den oder Verletzungen f hren gt gt 5 Nehmen Sie das Ger t nie mit ge ffnetem Geh use in Betrieb da dies zu irreparablen Sch den f hren Kann 6 Schalten Sie das Ger t nicht ein wenn Sie den Verdacht haben da an der Elektrik m glicherweise Sch den eingetreten sind Ziehen Sie in diesem Fall den Netzstecker heraus und lassen Sie das Ger t von einem Kundendiensttechniker untersuchen Versuchen Sie bis zu dieser Untersuchung keinesfalls das Ger t in Betrieb zu nehmen Eine Besch digung der Elektrik kann z B eingetreten sein wenn das Ger t u ere Sch den aufweist oder unter problematischen Umst nden transportiert wurde
18. absorbance contribution from butyl paraben Using the K factor in this way we can quantitate toluene in the presence of butyl paraben without altering the chromatography 25 s Absorbance Ratios Ratioing the detector s outputs from two different wavelengths can be a useful vvay of confirming peak purity VVhen a peak is pure the ratio of the absorbances should remain constant Thus the ratio for a pure compound produces a relatively square wave while the ratio for an impure compound produces a distorted wave see the plots at 1 57 and 0 97 minutes respectively in Figure 4 34 64 Thermo Electron 01 2004 Chromatogram AU1 AU2 Plot Mixture of Two Compounds Pure Compound Mixture of Pure Two Compounds Compound Time in Minutes DET Z17V FM Figure 4 34 Using absorbance ratios to determine the purity of two peaks in a chromatogram Thermo Electron 65 l To use absorbance ratioing you need to select AU1 AU2 for the Analog 2 Output field in the Analog Outputs Menu You also need to select the two wavelengths you want to ratio To select the most appropriate wavelengths use the UV2000 s Scan File to collect a spectrum across a range of wavelengths Then select Display AU from the Replay Menu and examine the collected data The data shown in Figure 4 35 are typical
19. an no SNOILdO SANVNNOD 91531 ANAND 531713 p peo7 enend on OL z abuey OL abuey 0010 lili l zolny 01 si 00701 aul uny IBAeqUI X 096 X pug 022 X HEIS fejdey dnes dnjas SNUSMN 0002 INNIN Yd d suny dojened q alld z 40 z Bd 01 2004 Thermo Electron 96 Menu Reference Analog 1 Offset mV Analog 1 Offset Analog 2 Analog 2 Offset Analog Outputs AU Autozero Time COMMANDS Copy Copy File Current Time Cursor Speed 01 2004 For quick reference we have included this alphabetical list of each field including a short definition and allowable and default values For a more detailed explanation of the functions of your detector you should refer to Chapter 3 Basic Operations and Chapter 4 Advanced Operations Some fields are common to both the UV1000 and the UV2000 so we have indicated the detector name for fields that appear in only one detector This field offsets the Analog 1 output signal by 0 1 2 5 10 20 or 50 mV The default setting of 10 mV is appropriate in most cases UV1000 only This field offsets the Analog 1 output signal by a positive or negative 50 20 10 5 2 1 or 0 percent of the full scale range Default is 0 UV2000 only This field allows you to select the output signal from the Analog Output 2 terminal The selections are AU the absorbance signal for wa
20. brings up Status Screen 5 functions of 3 Status Menu accessing the 24 31 general description of 5 illustration of 24 31 setting parameters 24 31 when Develop File is loaded 56 when Scan File is loaded 50 Status Screen accessing the 23 30 general description of 5 illustration of 23 30 locking the 43 103 when a queue is loaded 58 when Develop File is loaded 55 when Scan File is loaded 48 STOP key example of use 13 18 functions of 3 to stop replaying spectra 53 used to stop arun 23 30 STOP Input terminal 85 Stop line adding a 13 17 affects hovv you start a run 23 30 in a wavelength program 21 27 38 used to stop a run 23 30 Stop time see Stop line Stopping a queue 59 arun from an external device 85 techniques for 23 30 vvith a stop line 21 27 a scan 50 dual wavelength files 30 System error messages 110 T Temperature fluctuations results of 108 Terminals see External contacts Tests Menu accessing the 112 general description of 5 103 illustration of 112 Tests see Diagnostic tests Text conventions of 7 Theory of operation 105 Time current time field 42 field in VVavelength Program 103 setting the 42 stop times 38 Time from READY 103 accessing 42 description of 42 example of 42 illustration of 42 Time lines see VVavelength programming Timed events 85 autozero 40 defined 119 lamp startup and shutdown 42 on wavelength change 40 scanning 51 stop time 38 Tools needed for
21. 3 Remove the two thumbscrews that secure the photodiode assembly to the front of the flowcell Fig E 4 and then reposition the photodiode assembly out of the way to provide access to the flowcell 4 Remove the two thumbscrews top left lower right that secure the flowcell mount Fig E 5 to the front of the detector and then remove reposition the flowcell NOTE You needn t disconnect the flowcell s tubing connections to EN your LC system if the cuvette holder is only going to be used long enough to conduct a calibration Simply reposition the flowcell and omit Step 7 of this procedure 5 Position the cuvette holder and secure the two thumbscrews at top left and lower right that secure the holder to the threaded holes on the detector s front panel 6 Replace the photodiode assembly and secure it to the cuvette holder s standoffs with its two thumbscrews at top right and lower left 7 Snap the detector s front panel back in place 122 Thermo Electron 01 2004 RUN STATUS lt A gt MENU stop m y ENTER Z RO SpectraSYSTEM UV2000 Removable Front Panel Grip and Pull Here DT Z046 FM 01 2004 Figure E 2 Detector front panel Thermo Electron 123 Flowcell Housing DT Z047 FM 124 Figure E 3 Detector with front panel removed to expose flowcell housing Thermo E
22. The Testing RAM message only appears during self initiated testing On power up the test occurs without any special message A failure during either type of testing is indicated by the messages Bad DRAM or Bad NOVRAM MLE Digital PCB Voltages This test checks the circuitry supply voltages MLF Motherboard Analog Outputs This test checks the scale and linearity of the output signal recorder integrator Failure is indicated by a Fail or a Bad Analog Linearity message MLF Analog PCB Diode Offsets This test checks the diodes photodiodes with the lamp s off dark current Either a Bad Sample Diode or an Intense Light Detected message indicates failure You should verify that the sample photodiode is fastened securely to the flowcell and that light is actually passing through the flowcell If Fail or a Bad Ref Diode Detected appear call your Thermo Electron representative MLF Photodiode or Analog PCB Motor The Motor Test checks the monochromator motor and its voltages MLF Motor Deuterium Lamp This test checks the D2 lamp and its voltages when the lamp is on and when it is off If the message D2 Not Detected appears the lamp voltages are good but the lamp is either not present or not functioning properly Try replacing the deuterium lamp and retrying the test If the word Fail appears call your Thermo Electron representative Lamp or Motherboard Tungsten Lamp This test c
23. The wavelength field A in the UV1000 41 A2 in the UV2000 located in the Status Screen shows the current detector wavelength setting s For the UV2000 only these fields also appear in the Wavelength Program for dual wavelength operation The wavelength calibration screen located in the Tests Menu shows the current detector wavelength setting s In the Scan File Setup this field defines the wavelength interval at which the detector should perform the scan Allowable values are 1 2 3 4 5 and 10 nm Default is 5 nm UV2000 only In the Develop File Setup this field defines the wavelength increment the detector monochromator should use for wavelength changes between each set of injections Allowable values are 1 2 3 4 5 10 and 20 nm Default is 5nm UV2000 only This field is the wavelength maximum in a spectral scan UV2000 only This field is the absorbance value for the corresponding wavelength maximum in a spectral scan UV2000 only This field is the wavelength minimum in a spectral scan UV2000 only The lambda offset screen lets you choose a number of steps each representing 0 25 nm by which you want to offset the wavelength This field is used to check the detector s wavelength accuracy Allowable entries are 0 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 and 10 The default is 0 The Lamp field allows you to choose from among several selections D2 190 380 for the UV1000 s deuteriu
24. an undesirable condition exists that prevents the instrument from carrying out an operation Error messages remain on the display until you press a key Figure 1 7 An example of an error message Manual Conventions This manual uses several conventions Among them are menu displays text conventions brackets slashes etc standard words and several different icons DISPLAYS Figure 1 8 shows how we depict the two line display Note that in menu illustrations the triangular cursor location is indicated by a caret gt Q COMMANDS 1 OPTIONS Q TESTS Figure 1 8 A two line menu display Frequently the two lines shown on the display are only part of a longer menu In this manual menus having more than two lines are represented as in Figure 1 9 6 Thermo Electron 01 2004 TEXT STANDARD WORDS 01 2004 Zero on A Change Cursor Speed Status Lock READY Output Active Hi Figure 1 9 A menu longer than two lines Three typographic conventions are used to differentiate betvveen keys menus and fields Brackets Brackets indicate instrument keys For example Press MENU Slashes Slashes are used around menu choices For example From the Main Menu select FILES Capitalization Capitalization is used to make field and menu names appear just as they do on the display Generally the first letters of field names are capitalized For example Select FILES Copy Copy File
25. appears in Figure 3 27 gt Lamps Q Analog Outputs Figure 3 27 The UV2000 s Options Menu 3 Select More The More Menu appears in Figure 3 28 Zero on A Change Cursor Speed Status Lock READY Output Active Hi File Name Protect ds Off 2 Off 3 off 4 off Figure 3 28 The UV2000 s More Menu 4 Scroll down to the Table containing the fields File Name and Protect To protect a file select On in the Protect field corresponding to the appropriate file number To remove the file protection select Off 34 Thermo Electron 01 2004 Analog Outputs There are two analog outputs on the UV2000 Analog Output 1 and Analog Output 2 On the detector s rear panel they appear as ANLG 1 Output and ANLG 2 Output Rear panel connections for both outputs are discussed on page 84 ANALOG OUTPUT 1 By default Analog Output 1 is either the absorbance reading for single wavelength operation or the absorbance reading of wavelength one 21 for dual wavelength operation ANALOG OUTPUT 2 Analog Output 2 is selectable AU AUI K AU2 and AU1 AU2 and so can be used to monitor several different outputs To access these options 1 Press MENU 2 Select OPTIONS 3 Select Analog Outputs The Analog Outputs Menu shovvn in Figure 3 29 appears Analog 1 Offset 5 Analog 2 Offset Figure 3 29 The UV2000 s Analog Outputs Menu 4 Scroll down to Analog 2 The selections are e AU which is either the same ab
26. are automatically returned to the Status Screen and are ready to run your detector A PRACTICE RUN Now you re ready for a practice run Note that the Status Screen Fig 2 1 now displays your wavelength setting the detector s status and the absorbance reading If the Status reads READY the required lamp is lit if it reads NRDY Not Ready there is an error or the lamp isn t lit and if it reads UVW the ultraviolet D2 lamp is still warming up When the baseline is stabilized 1 Press the ZERO key to zero the detector s analog output signal 2 Inject your sample During setup you may have noticed that there was no stop time entered in the detector s parameters In this case the detector stays in the READY state and continually monitors the column eluant You do not need to manually start or stop a run with this set up 12 Thermo Electron 01 2004 ADDING A To add a stop time you need to modify the detector s operating STOP TIME parameters as follows We will then show you how to start and stop a run using the new setting 1 From the Status Screen press the v key to move down to the Status Menu Fig 2 9 which is the programming area below the Status Screen The cursor appears on the tens digit of the wavelength value Wavelength Rise Time Autozero Time Range Figure 2 9 The UV1000 s Status Menu 2 Using the v key move the cursor to the blank line below the 0 00 time line and press Thi
27. autozero 40 protecting files 34 quick example 14 running the detector 29 sample queues 56 scanning ability 46 setting detector parameters 26 31 single wavelength operation 26 software version 113 starting arun 30 stopping arun 30 troubleshooting 107 wavelength programming 37 UVW message 103 V Validation of linearity 116 127 of wavelength 114 127 VDE see Safety Information Voltage AC line problems 107 clipping 84 preset 80 selector barrel illustration of 81 location of 80 setting the 80 setting 80 testing the 115 W W Lamp Hours 103 VVARNINGSI what they signify 8 Wavelength fields defined 99 calibration test 99 114 127 ending in Develop file 55 01 2004 Thermo Electron index xi example of setting the 11 15 X Z field in the Status Screen 99 field in Wavelength Program 103 Zero interval 55 Input terminal 85 offset field described 99 example of use 12 17 recalibration using Cuvette Holder 127 fields described 104 starting 55 from an external device 85 testing accuracy 114 127 key functions of 3 Wavelength Program 103 programming an autozero 40 accessing from the Edit Menu 21 27 used for baseline scans 48 accessing from the Status Menu 24 31 command described 103 End Index description of 21 27 37 example of a stop line 21 28 illustration of 21 27 single line 21 27 tvvo line 21 27 using a stop line 21 27 38 VVavelength programming addin
28. full scale terminals F and B Ground terminals E and A NOTE Analog outputs are driven to twice their range In other words their maximum output is twice the selected range To avoid clipping the voltage be sure to connect integrators and data systems to the 1 0 V terminal and to use caution when connecting recorders to the 0 01 or 0 10 V terminals The terminals on the UV1000 s analog output connector are labeled UNRANGED Output and RANGED Output Fig A 4 The UNRANGED Output is a 1 AU VOLT unrangeable output primarily used for integrators and workstations The UNRANGED Output can be connected to terminal F with a ground at terminal E or A The RANGED Output has four terminals 0 01 V full scale terminals H and D 0 10 V full scale terminals G and C 1 0 V full scale terminals F and B Ground terminals E and A Integrators Workstations For the UV2000 connect your integrator workstation to the 1 0VF S F or B and corresponding ground E or A terminals For the UV 1000 connector your integrator workstation to the 1AU V output terminal F and the ground E terminal Recorders Connect the positive input from your recorder to the full scale voltage 0 01 0 10 or 1 0 V appropriate for your recorder Connect the recorder s floating ground input to the corresponding GROUND terminal NOTE Do not connect the detector s GROUND to any earth ground on your recorder This would lead to an increased noise level a
29. installation 80 Troubleshooting clicking sound 109 flowcell contamination 71 MLF most likely failure 114 index x Thermo Electron 01 2004 Table of common problems 107 term defined 119 Tungsten lamp alignment of 76 changing the 77 illustration of 75 lamp life 103 Low Light Detected message 111 testing the 115 W lamp hours field 103 warm up time 77 T V GS certification see Safety Information U Unpacking your detector 79 Unshort outputs 44 UPS uninterrupTable power supply 107 User messages 5 UV1000 allowable and default values 97 analog outputs 25 84 building a wavelength program 21 checking the status 23 deleting a file 25 98 diagnostic tests 112 features 9 43 installation 79 lamp operations 41 locking the Status Screen 43 maintenance 67 periodic validation 67 programmed autozero 40 quick example 10 running the detector 23 setting detector parameters 20 24 single wavelength operation 20 software version 113 starting arun 23 stopping arun 23 troubleshooting 107 wavelength programming 37 UV2000 allowable and default values 97 analog outputs 35 84 building a wavelength program 27 checking the status 30 copying files 32 97 deleting a file 98 Develop File 54 diagnostic tests 112 dual wavelength operation 26 features 9 43 file identification 27 99 installation 79 K factor 60 lamp operations 41 loading a file 29 locking the Status Screen 43 maintenance 67 periodic validation 67 programmed
30. la dur e de vie et la s curit du syst me 1 Avant de brancher un module d instruments ou de le mettre sous tension toujours s assurer que la tension et les fusibles sont r gl s de fa on correspondre la tension locale du secteur 2 N utiliser que des fusibles du type et du courant nominal sp cifi s Ne pas utiliser de fusibles r par s et ne pas court circuiter le porte fusible Thermo Electron Xi BONNES PRATIQUES DE LABORATOIRE xii 3 Le cordon d alimentation accompagnant l instrument doit tre branch une prise de courant avec mise la terre En cas d utilisation d une rallonge lectrique s assurer que celle ci comporte galement une mise la terre 4 Ne pas modifier les connexions de mise la terre internes ou externes La modification ou le d branchement de ces connexions repr sente un danger pour op rateur et ou risque d endommager le syst me MISE EN GARDE Cet instrument est mis la terre conform ment aux r glements applicables lors de son exp dition Ne pas modifier les branchements lectriques ou le ch ssis de l instrument afin d assurer un fonctionnement en toute s curit MISE EN GARDE Ne pas abroger le contact de s curit de couverture de lampe qui teignez la lampe quand le couverture est enlev ou les blessures pourraient r sulter 5 Ne jamais faire fonctionner le syst me sans son bo tier Des dommages permanents pourraient en r
31. load any other file Because queues are not available on the UV 1000 this message only appears on the UV2000 Thermo Electron 111 Run In Progress Testing Not Allowed You cannot run the detector s built in diagnostics while a run is in progress Run Not In Progress No Scanning Allowed A spectral scan can only be performed when a run is in progress Because there is no scanning feature on the UV 1000 this message only appears on the UV2000 Detector Shutdown This message occurs when you use the Shutdown Detector field to turn off the detector See Shutdown Detector in Chapter 4 Press any key on the keypad to turn on the detector Scan Memory Full This message occurs when the Scan File is loaded and the scan data memory storage is full Because there is no scanning feature on the UV 1000 this message only appears on the UV2000 Run In Progress No Replay Allowed The UV2000 does not allow you to replay stored spectral scans when the Scan file is loaded and a run is active Because there is no scanning feature on the UV1000 this message only appears on the UV2000 Diagnostic Tests 112 INTERNAL DIAGNOSTIC TESTS This section describes the internal diagnostic tests supplied with your detector It also references two external tests that you can run Use these tests if you suspect that your detector is not working properly You can access the detector s internal diagnostic tests by following these steps 1 Pre
32. messages conventions of 6 definition of 118 see also Messages Event mark 44 98 accessing 44 placement of 49 Exclamation points use of 6 01 2004 Thermo Electron index iii External contacts 83 analog outputs connections to 84 scanning 49 shorting 43 ANLG 84 connectors 83 illustration of 83 making connections 83 READY Output 85 accessing command 44 RUN Input 42 85 STOP Input 85 ZERO Input 85 F FCC see Safety Information Fields alphabetical listing of 97 changing entries in 1 4 description of all 97 term defined 118 File Menu UV1000 only accessing the 20 general description of 5 98 illustration of 20 loading the edit file 22 setting parameters 20 Files copying UV2000 only 32 97 103 deleting 25 33 edit file 20 editing from File s Menu 20 26 editing from Status Menu 24 31 escaping without saving changes 25 32 identification of UV2000 only 27 99 loading an example 12 16 fields described 100 technique described 22 29 while a queue is running 59 managing UV2000 only 32 messages Already Running 111 Copied 33 Deleted 25 34 Loaded 22 29 Protected Cannot Be Copied To 33 Protected Cannot Be Deleted 34 Saved 24 32 name field UV2000 only 99 protecting UV2000 only 34 100 run file 20 saving 24 32 term defined 118 Files Menu UV2000 only accessing the 26 general description of 5 98 illustration of 14 26 loading the file 29 setting parameters 26 Floating g
33. reservoir will significantly prolong the life and effectiveness of the inlet filters check valves and seals injector and column Typically HPLC grade solvents do not require filtration Choose a mobile phase that s compatible with the sample and column you ve selected for your separation Remember that some solvents are corrosive to stainless steel Inert biocompatible instrument versions are also available from Thermo Electron Degas the Eluents Degas your eluents using either the vacuum degassing or the helium sparging technique Complete information for using either of these techniques is found in separate documentation provided with degas accessories Solvent Disposal Make sure you have a solvent waste container or other kind of drain system available at or below the benchtop level Most solvents have special disposal requirements and should not be disposed of directly down a drain Follow all governmental regulations when disposing of any chemical High pressure Systems and Leaks LC systems operate at high pressures but since liquids aren t highly compressible they do not store much energy Thus little immediate danger arises from the high pressure in an LC system However if a leak occurs it should be corrected as soon as possible Finally we recommend that you always wear eye and skin protection when working on an LC system and that you always shut down the system and return it to atmospheric pressure before attempt
34. the Scan File is loaded you will notice the fields Zero and Scan in the Status Screen Fig 4 11 Status READY Figure 4 11 The Status Screen with the Scan File loaded Zero Zero is used to perform baseline scans of the solvent s background absorbance With the detector s baseline stabilized and the cursor on the Zero field press ENTER The UV2000 performs and stores a baseline scan using the parameters you set in the Scan File While the detector is performing a baseline scan the Status field displays SCAN 0 Baseline scans may be taken at any time during the run as long as no peak is eluting at that time Subsequent sample scans are corrected using the last baseline scan taken This is especially advantageous for gradient elution where the background absorbance of the eluant may be constantly changing Thermo Electron 01 2004 For example let s say you perform a baseline scan before you initiate a run and then again at 5 00 minutes into the run You also perform sample scans of your eluting peaks at 2 4 and 5 6 minutes into the run The sample scan taken at 2 4 minutes will be corrected using the baseline scan taken before the run began The sample scan taken at 5 6 minutes will be corrected using the baseline scan taken at 5 0 minutes Scan Once you begin the run the cursor will move from Zero to Scan in the Status Screen Each time you wish to perform a sample scan press ENTER NOTE There is a one sec
35. 0 the Scan File 47 analog output 49 background defined 117 baseline scan automatic 48 defined 117 index viii Thermo Electron 01 2004 manual 48 subtracting 48 choosing scan times 49 data storage 53 Display AU 1 screen 53 illustration of display 53 Display AU 2 screen K factor calculation 63 Edit Menu for 47 fields described 101 file name 47 how the detector scans 46 an example 49 identification of stored spectra 52 102 initiating a scan 49 K factor 61 Memory Full message 112 min max wavelength fields 99 no copying 33 No Scans Stored message 53 quantitative analysis 49 Replay Menu accessing the 52 illustration of 52 replay fields described 101 replay rate 52 replay spectra command 52 setting parameters 52 replay spectra command illustration of display 53 stopping replay 53 running replay 52 running the 48 setting the wavelength end of scan 48 98 interval for scan 48 99 monitoring the run 48 102 start of scan 48 102 setting up the 47 Setup Menu for 47 102 spectral scan defined 119 Status Screen for 48 stopping a scanning run 50 subtracting baseline scans 48 summary data screen illustration of 50 wavelength min max fields 99 zero command 48 zero time field 48 SCAN see Scan File Scan Summary Data Screen accessing the 50 illustration of 50 Scan zero time 48 101 Scanning automatically 51 how it works 51 number of spectra stored 46 replaying spectra corrected for baseline absorbance 47 see also Sca
36. 00 Dreieich Germany Phone 49 06103 408 0 Fax 49 06103 408 1222 Milano Italy Phone 39 02 95 059 226 Fax 39 02 95 320 370 Breda Netherlands Phone 31 076 587 8722 Fax 31 076 571 4171 Madrid Spain Phone 34 091 657 4930 Fax 34 091 657 4937 Barcelona Spain Phone 34 093 223 0918 Fax 34 093 223 0982 Stockholm Sweden Phone 46 08 556 468 00 Fax 46 08 556 468 08 Hemel Hempstead United Kingdom Phone 44 01442 233 555 Fax 44 01442 233 667 For all other countries contact your local Thermo Electron dealer Thermo Electron 01 2004 IN AUSTRALASIA AND ASIA 01 2004 In Australasia and Asia customer support replaceable parts and technical support are available from each of the Thermo Electron offices as follows Rydalmere N S W Australia Phone 61 02 9898 9000 Fax 61 02 9898 9800 Yokohama Japan Phone 81 45 453 9100 Fax 81 06 453 9110 Osaka Japan Phone 81 06 6387 6681 Fax 81 06 6387 6641 Beijing P R China Phone 86 010 6621 0839 Fax 86 010 6621 0851 For all other countries contact your local Thermo Electron dealer Thermo Electron IDENTIFYING SAFETY INFORMATION Safety and EMC Information In accordance with Thermo Electron s commitment to customer service and safety these instruments have satisfied the requirements for the FCC and the European CE Mark including the Low Voltage Directive Designed man
37. 0x 1075 AU Dual wavelength Mode 254 280 nm 1 0 sec rise time UV2000 only lt 2 5 x 10 5 AU Drift after warm up 254nm 2 10 4 AU hour Display 2 x 24 character high contrast LCD Dimensions 14 5 37 cm x 6 15 cm x 18 5 47 cm Weight 40 Ib 18 kg Power Requirements 100 120 220 240 VAC 50 60 Hz 1 5 1 6 8 0 0 8A 225VA Environmental 10 40 C 5 95 RH noncondensing 01 2004 Thermo Electron 91 B Menu Reference Introduction This Appendix provides you with two Menu Trees one each for the UV 1000 page 94 and the UV2000 page 95 It also provides you with an alphabetical description of all the instrument s display fields Fortunately it is not necessary to read this Appendix in order to learn how to use your detector It is included in the manual simply as a quick reference and aid to using your instrument The Menu Trees are a representation of the detector s overall menu structure They show the location and interrelation of all the menus for your detector and as such they are a good reference to keep on hand while you work through the operating instructions in Chapters 3 and 4 The menu trees will also help if you become lost while moving through the detector s menus Separate removable copies can be found in the pocket at the front of this manual The Menu Reference is an alphabetical listing of each menu field and command Included in each listing is the field s definition and where appro
38. 26 35 ANLG see Analog outputs Arrow keys 1 4 Asterisks use of 5 6 ASTM Procedure E 925 127 AU analog output choice 35 entries for analog 2 35 field described 97 AU entries for analog 2 97 AUFS acronym defined 117 Auto Scan illustration of 51 Autozero example of 40 programming 39 40 time field described 97 time 22 29 Back pressure causes of 71 detectors in series 87 increasing 71 specifications for flowcells 88 Background scan defined 117 zeroing 48 Baseline problems baseline defined 117 correcting for background absorbance 48 Thermo Electron index 1 drifting causes of 71 108 correcting with autozero 40 correcting with offsets 25 36 erratic causes of 71 noise causes of 71 107 spikes causes of 71 107 Blank key 3 Brackets reason for 7 C Calibration 114 116 127 128 Capitalization standard use of 7 CAUTIONS what they signify 8 Changing the fuses 82 Chemical hazard icon defined 8 Chronometer defined 117 illustration of 74 tracking deuterium lamp life 74 Cleaning procedures deuterium lamp surface 74 flowcell 71 optical surfaces 69 Clicking sound UV2000 only 109 Clock internal 42 Column bleed 108 COMM port 83 Commands Menu accessing the 43 general description of 5 97 illustration of 43 Communications to external devices 83 85 Confirmation messages 6 Connectors see External contacts Conventions used in manual 6 brackets 7 capitalization 7 icons 8 slashes 7 st
39. 4 Thermo ELECTRON CORPORATION The products of Thermo Electron San Jose are produced under ISO 9001 accredited quality management systems Australia P O Box 239 Rydalmere Unit 14 38 46 South Street Rydalmere N S W 2116 e 61 02 9898 9000 Austria Wehlistrasse 27b e A 1200 Wien e 43 01 333 50 34 0 Belgium Technologiestraat 47 B 1082 Brussels 32 02 482 30 30 Canada 5716 Coopers Avenue Unit 1 e Mississauga Ontario e L4Z 2E8 e 1 905 712 2258 France 16 Avenue du Qu bec Silic 765 Z A de Courtaboeuf F 91963 Les Ulis C dex e 33 01 60 92 48 00 Germany Im Steingrund 4 6 D 63303 Dreieich 49 06103 408 Italy Strada Rivoltana 1 20090 Rodano Milano 39 02 95059 226 Japan C 2F e 3 9 Moriya cho Kanagawa ku Yokohama Kanagawa 221 0022 81 45 453 9100 Japan Esaka Grand Building 2 3 1 Esaka cho Suita City e Osaka 564 0063 e 81 06 6387 6681 Netherlands Takkebijsters 1 NL 4817 BL Breda 31 076 5878 722 P R China Room 901 Ping an Mansion e No 23 Jin Rong Street Xi Cheng District e Beijing 100032 e 86 010 6621 0839 Spain Sepulveda 7 A ES 28108 Alcobendas Madrid e 34 091 657 4930 Spain Acer 30 32 Edificio Sertram Planta 2 Modulo 3 ES 08038 Barcelona 1341 093 223 0918 Sweden Pyramidbacken 3 e S 141 75 Kungens Kurva Stockholm e 46 08 556 468 00 United Kingdom Stafford House 1 Boundary Park Boun
40. 4 until the cuvette is completely clean and dry NOTE In cases of serious contamination that resists removal it may be easier to simply replace a dirty cuvette than to spend a lot of time cleaning it Thermo Electron 129 INDEX The asterisks in this index refer to topics that are common to both the UV1000 and the UV2000 detectors For each asterisked item there are two page numbers The first refers to the appropriate page number for the UV1000 detector the second refers to the page number for the UV2000 KEYS and keys 1 4 lt key 4 gt key 4 ENTER key 3 MENU key 3 RUN key 3 STATUS key 3 STOP key 3 ZERO key 3 A key 4 v key 4 A A File Is Already Running message 111 A D defined 117 Absorbance defined 117 linearity 116 127 ratioing UV2000 only 35 64 Accessory kit 79 Alphabetical entries changing 1 Alphanumeric entries changing 4 Amperage requirements see Fuses Analog offset see Analog outputs Analog Output Connections UV1000 84 UV2000 84 Analog outputs adjusting for different devices 25 36 connections to 84 recorders 84 description of 25 35 event mark 44 fields defined 97 illustration of connectors 83 K factor 63 offsets 25 36 term defined 117 range 22 29 scanning 52 shorting the 43 testing the 115 use of negative and positive offsets 25 36 Analog Outputs Menu accessing the 26 35 general description of 97 illustration of
41. 9 illustration of apertures 70 motor testing the 115 movement during a scan 49 More Menu accessing the 34 40 general description of 100 illustration of 34 Motor test 115 Moving within between menus 1 4 N Negative signals 25 36 Nitric acid safety precautions 72 used to clean the flowcell 72 No More Available Memory message 111 No Queue Available message 58 111 No Scans Stored message 53 No Spectra Available message 111 Notes what they signify 8 NRDY not ready message 23 30 41 103 Numerical entries changing 1 4 O Offsets see Analog outputs Optical system 106 index vi Thermo Electron 01 2004 Options Menu chosen from the Edit Menu 100 accessing the 22 28 description of 22 28 illustration of 22 28 chosen from the Main Menu 100 accessing the 34 40 illustration of 40 illustration of UV2000 only 34 general description of 5 Order 1 defined 100 editing 59 97 in a queue 58 Order field 57 Organic solvents used to clean flowcells 71 Outputs see Analog outputs P Parameters allovvable and default values 97 defined 118 setting 20 27 Pausing a queue 59 pause command 100 pause queue command 100 resuming 59 Peak problems broadening 109 118 size 109 tailing 109 purity 35 64 suppression analog output for 35 defined 118 see also K factor Photodiode cleaning the surface 69 defined 118 mount illustration of 69 87 removing the 69 Positioning the detector 80 Powe
42. A brief explanation of the keys and the main menus and screens follows RUN STATUS MENU STOP 4 ZERO SpectraSYSTEMUV2000 DET Z008 FM Figure 1 1 The detector s command center 2 Thermo Electron 01 2004 THE KEYPAD 01 2004 The keypad of each SpectraSYSTEM instrument consists of twelve keys Four keys directly control the instrument s operation RUN STOP STATUS and on the detector a blank key called ZERO The remaining keys either access commands MENU and ENTER or are used to set parameters and move around the display TAL v lt gt FD The function of each is explained below RUN Pressing RUN starts a run The detector must be in the READY state or QREADY if a queue is loaded indicating that the detector is stabilized and waiting to begin a run STOP Pressing STOP halts a run stops the internal clock and returns the detector to a READY state If a wavelength program is operating pressing STOP halts the program and returns the detector to its initial conditions STATUS Pressing STATUS displays the Status Screen Fig 1 1 From the Status Screen you can monitor the run in progress You can also access the Status Menu See page 5 for more information ZERO The unlabeled key is the only variable key in the whole SpectraS YSTEM family On the detector the blank key is the ZERO key The key s name appears on the na
43. Finnigan SpectraSYSTEM UV Vis Detectors Reference Manual A0099 540 Revision G Thermo ELECTRON CORPORATION Finnigan and SpectraSYSTEM are trademarks of Thermo Electron Corporation This manual and the instruments to which it applies have satisfied the requirements for CSA FCC the EMC and Low Voltage Directives Use of non approved components and repair methods may reduce or invalidate the built in protection that is required to meet the above certifications Technical information contained in this publication is for reference purposes only and is subject to change without notice Every effort has been made to supply complete and accurate information however Thermo Electron Corporation assumes no responsibility and will not be liable for any errors omissions damage or loss that might result from any use of this manual or the information contained therein even if this information is properly followed and problems still arise This publication is not part of the Agreement of Sale between Thermo Electron Corporation and the purchaser of an LC system In the event of any conflict between the provisions of this document and those contained in Thermo Electron Corporation s Terms and Conditions the provisions of the Terms and Conditions shall govern System Configurations and Specifications supersede all previous information and are subject to change without notice Printing History Revision G printed in January 200
44. From the Status Screen press the v key to move down to the Status Menu Fig 2 19 which is the programming area below the Status Screen Wavelength 250 v Rise Time Autozero Time Range 1 Range 2 Figure 2 19 The UV2000 s Status Menu 2 Using the v key move the cursor to the blank line below the 0 00 time line and press This adds a second line with a time of 1 00 and the same wavelength setting as the first Change 1 00 to the desired stop time for the run and leave the wavelength unchanged 3 To save your edits scroll down to the words Save File which now appear below Range 2 and press ENTER The confirmation message shown in Figure 2 20 appears and you are automatically returned to the Status Screen File Saved Figure 2 20 The file saved message 01 2004 Thermo Electron 17 RUNNING WITH Now that you have entered a stop time you will need to start the run A STOP TIME with each injection 1 Zero the detector s analog output signal by pressing the ZERO key 2 When the detector is stabilized inject your sample and press RUN Notice that Status now shows the run time If you wish to stop your run before the set stop time simply press STOP What s Next Once you have completed this example and are comfortable with the keypad and display proceed to Chapter 3 Basic Operations to learn more about your detector 18 Thermo Electron 01 2004 3 Basic Operations Introduct
45. HPs 9311 nnan Bd SDU H 0002 95 Thermo Electron 01 2004 1d 3 07 1Q L z abuey L 00 Swi 2 L Owl S M 092 0010 yBuajneM OUI L dojsA9g q 2114 L 10000 092 ACVAY suny NV 7 smes ooo pepeo aij dol A q ul NYXENY XEY LUI L ueos g all 0187 100070 092 4209 nv Y sn p pEo7 lli 4805 L z ebuey L ebuey 070 awl 2 L oul SIY 082 092 0010 A LY Sully TL lli V00000 082 AGVzM V0000 092 nv X smes ooo x 1210 L z ebuey L uey 00 aul 2 L Owl siM osz 000 udusjsnem OWL 11 alld L000 0 osz nv u smes ooo n no esned esned anand e8Je18q esned PPI p peo7 s x l5uls L eweuel L suny OUEN ll i p peo7 all dojenaq xpa papeo7 UEOS Y eng Yy a us SNLYLS peo7 peo nv x Ae ds q x nv Ae ds q e pads Aejday x nv felds q 00 0 eu eoeds G oes uu ayey Ae dayy OL z abuey nv X Ae doy 01 abuey eno ds ejd y 01 2 ebuey 01 ebuey 0070 eUIL l zolny 01 w L 092 xy uny 9 lenau X 096 X pug 022 Y HES peo pa
46. IONS to access the Options Menu Fig 4 3 gt Lamps Q Analog Outputs Figure 4 3 The Options Menu 2 Select More to display the More Menu 3 Place the cursor on the Zero on 2 Change field This field appears on the first line of the More Menu for both the UV1000 and the UV2000 4 Select Yes to automatically zero the detector response with each vvavelength change during a run or No to turn this feature off You can also use this automatic zero feature to add autozeros into your wavelength program without changing the detector s wavelength settings To do this simply add additional time lines Adding autozeros in this way is convenient in cases such as solvent programming where the detector s baseline may drift due to changes in solvent background An example program is shown in Figure 4 4 Wavelength Figure 4 4 An example of a wavelength program with automatic autozeros With Zero on A Change set to Yes the detector will autozero at 2 00 5 00 and 7 00 minutes into the run even though the wavelength will only change once at 5 00 minutes into the run 40 Thermo Electron 01 2004 Automatic Lamp Operations THE LAMPS MENU 01 2004 The Lamps Menu Fig 4 5 allows you to select lamps track lamp life and turn the lamps on and off automatically Field descriptions for this menu follow To access the Lamps Menu 1 Press MENU and select OPTIONS 2 Select Lamps Lamp 190 365 D2 La
47. Manual English Standard Fittings Kit Kit includes stainless steel fittings and tubing used in a Finnigan SpectraSYSTEM LC system Inert Biocompatible Fittings Kit Kit includes PEEK fittings and tubing used in an inert biocompatible Finnigan SpectraSYSTEM LC system Upgrade kits are available for the Finnigan SpectraSYSTEM UV1000 detector Contact your local Thermo Electron Representative for details 01 2004 Thermo Electron XXI Getting Started Introduction AS EASY AS 1 2 3 01 2004 riz This Chapter provides you with the three basic rules you ll need for using your Thermo Electron Finnigan SpectraSYSTEM UV Vis detector It also introduces you to the instrument s command center and describes the conventions we ll use in this manual Before you start this chapter be sure to read the Safety Information section beginning on page v of this manual and to install your detector as described in Appendix A Throughout our explanations we encourage you to explore the general architecture of the instrument s menus and screens Use the Menu Tree in Appendix B as your guide if you wish Learning Your Way Around It s easy to learn your way around a SpectraS YSTEM detector Just remember these three rules 1 arrow keys A v lt gt D move the cursor in the direction printed on the key HINT Press MENU to jump quickly to the top of the menu structure 2 The shape of the cursor dete
48. Offsets Display AU Edit Edit File End Event Mark FILE S 98 Under the top level menu FILE S this field accesses the Delete File command Under the top level menu QUEUE this field accesses the Delete Queue command UV2000 only This field deletes the designated file setting all fields to their default values After pressing ENTER the confirmation message File Deleted appears for one second This field deletes the queue After pressing ENTER the confirmation message Queue Deleted appears for one second UV2000 only This field tracks the total number of hours the detector s deuterium lamp has been in operation up to 9999 When a new lamp is installed you must set this parameter to zero This field displays the analog to digital A D conversion frequencies of the sample and reference diodes when both lamps are turned off These values can be used to measure the detector s digital noise level This command calls up the Display AU X screen a screen that shows the incremental wavelength versus absorbance data for the selected spectral scan Under the top level FILE S menu the Edit Menu allows you to set up or edit files The edits do not change the current settings of the detector until the file is loaded Under the top level QUEUE menu the Edit Menu allows you to set up or edit a Queue Edits may not be made to Order 1 while a queue is loaded or running unless you pause the queue first
49. Once you ve selected the Develop File as described above press either the ENTER or the v key to access the Develop File s Edit Menu Fig 4 18 Edit File D File Name DEVELOP Start End A A Interval Run Time Runs per A Rise Time Autozero Time Range 1 Range 2 Figure 4 18 The Develop Files Edit Menu 54 Thermo Electron 01 2004 2 In the Start A field enter the wavelength at which the first chromatogram is to be monitored 3 Inthe End 2 field enter the wavelength at which the last chromatogram is to be monitored 4 m 2 Interval enter the wavelength increment that the detector s monochromator should use for each wavelength change In Run Time enter how long each run should last In Runs per A enter the number of injections to be made at each wavelength setting 7 Enter Rise Time Autozero Time Range 1 and Range 2 as you would for a typical run Note that Range 1 and Range 2 are the corresponding ranges for Analog Outputs 1 and 2 respectively As an example we will use the Develop File shown in Figure 4 18 The UV2000 would make its first two ten minute runs at 220 nm The monochromator would then change to 225 nm and the detector would make two runs at this wavelength This pattern would continue in five nanometer increments until the detector has made two runs at the last wavelength 350 nm After setting up your Develop File you are ready to load it and run RUNNING THE When the Devel
50. Press STATUS When zeroing a readout device such as an integrator or recorder it s convenient to be able to short the detector outputs You can do this using the Short Outputs feature To access Short Outputs 1 Press MENU 2 Select COMMANDS The Commands Menu Fig 4 7 appears gt Event Mark 14 Short Outputs Q Shutdown Detector Figure 4 7 The Commands Menu Thermo Electron 43 EVENT MARK READY OUTPUT When you select Short Outputs the detector s analog outputs are shorted together zero volts and the field name changes to Unshort Outputs To remove the short and return the outputs to their normal operating state select Unshort Outputs and the field changes back now reading Short Outputs When you leave this screen the field returns automatically to Short Outputs Using the event mark feature you can place an event mark on your chromatogram to note various occurrences such as the turning of a sampling valve The event mark is a spike 15 of full scale for one second in both detector output signals To access Event Mark Press MENU 2 Select COMMANDS The Commands Menu Fig 4 7 appears 3 Place the cursor on Event Mark Press ENTER each time you wish to place an event mark on your chromatogram NOTE You may not want to use event marks if your data will be analyzed by an integrator Integrators can misinterpret event marks as peaks Using the READY Outp
51. STEM detectors are finely tuned scientific instruments that we at Thermo Electron are proud to stand behind Even so routine maintenance is necessary to ensure peak performance so we can only guarantee our detectors performance if you follow proper care and maintenance procedures This Chapter shows you how to replace and clean your detector s flowcell and lamps Also included in this Chapter is a procedure for testing the detector s absorbance linearity This characteristic is particularly useful if your laboratory s standard operating procedures require periodic detector validation You will need the optional cuvette holder to perform the test If you have any questions on proper maintenance or would like to arrange for a preventive maintenance program please contact your local Thermo Electron representative Flowcells NI CHANGING THE FLOWCELL 01 2004 This section describes the changing and general cleaning of your detector s flowcell For other flowcell problems such as a cracked window or leaks that occur in locations other than at the inlet outlet fittings contact your Thermo Electron representative NOTE Flowcells are factory assembled units that should not be disassembled under any circumstance The flowcell needs to be removed whenever you replace a broken cell change specialized applications or clean the cell with nitric acid For a list of available flowcells see Specifications Flovvcells in Appendi
52. To add a second time line scroll down to the first blank line and press The second line will automatically have a time setting of 1 00 and the same wavelength setting as the first Change the Time and corresponding Wavelength fields to the desired values Subsequent lines are added in the same fashion A wavelength program may contain as many as ten lines for a single run On the UV1000 all of the lines wavelengths must be in the same range either UV or visible On the UV2000 however you can cross between the UV and visible ranges in single wavelength mode only If you enter time lines out of sequence the detector will automatically sort the lines and place them all in chronological order The last line of the program the stop line lists the time at which the detector will automatically end the run and return to initial conditions Since wavelength is not important in the stop line it can be set to any value NOTE Remember the last line of the program is always the detector s signal to end a run it is not a programmed wavelength change To delete an entire time line place the cursor in the Time field and press repeatedly until the value goes blank When you leave the line it will be deleted Thermo Electron 01 2004 An Example Figure 4 2 shows a completed wavelength program for single wavelength operation Wavelength Figure 4 2 An example of a completed wavelength program In our example the initia
53. a r solution des pics Il est recommand tout au moins de conserver Thermo Electron 01 2004 01 2004 pour r f rence future un chromatogramme d un chantillon type et d un m lange standard bien document et accompagn des conditions du systeme Une comparaison precise des temps de r tention des formes et de la sensibilit des pics ainsi que des bruits de r f rence peuvent fournir des indices pr cieux pour identification et la r solution de probl mes futurs Toxicit chimique Bien que l utilisation et l entreposage dans les laboratoires de grandes quantit s de solvants inflammables et toxiques puissent repr senter un danger ne pas n gliger les dangers potentiels pos s par les echantillons Veiller particulierement a lire et a suivre toutes les pr cautions indiqu es pour assurer la ventilation le stockage la manutention et l limination des solvants et des chantillons Se familiariser avec les donn es sur la toxicit et les dangers potentiels associ s a tous les produits chimiques en consultant les fiches techniques sur la s curit des substances FTSS du fabricant Pr paration des chantillons Toujours consid rer la solubilit de l chantillon dans la phase mobile La pr cipitation des chantillons peut boucher la colonne les tubes et ou la cellule de dilution et en limiter le d bit Cette obstruction peut endommager le systeme de fa on irr parable L accumulation de particul
54. ad a different file you will get the message shown in Figure 4 28 You are then returned to the Files Menu Queue Loaded Cannot Load File Figure 4 28 The message that appears when you attempt to load a file when a queue is already loaded or running To edit an existing queue follow the procedures outlined in Setting Up a Queue on page 57 You are allowed to edit the Queue while it is running but if you want to edit anything in Order 1 you ll have to pause the queue first To pause a queue 1 Select Pause from the Queue Menu 2 When the words Pause Queue appear press ENTER Ifa file is running the run continues until it is completed at which point the detector returns to its READY state The letter Q no longer appears in the Status Menu When you wish to continue you must reload the queue When the detector receives a start signal the queue will resume operation at the point where it left off Use the following steps to delete an existing queue or to stop a running queue Display the Queue Menu Select Delete When the words Delete Queue appear press ENTER Ifa file is running the run continues until it is completed The confirmation message shown in Figure 4 29 appears for one second and you are returned to the Queue Menu Thermo Electron 59 Queue Deleted Figure 4 29 The queue deleted message You may delete or stop a queue at any time but remember that the queue will be sub
55. al operating state select Unshort Outputs and the field changes back to Short Outputs When you leave this screen the field returns automatically to Short Outputs This field toggles between Manual you turn off the lamp manually Time the lamp turns off automatically at a preset time Time from READY as explained under the Time from READY field and End of Queue the lamp turns off when the queue is finished UV2000 only Default is Manual This command shuts down the detector s lamps and motors leaving the electronics on to preserve memory Press any key to return the detector to the same settings as when this field was activated This field displays the local time ranging from 0 00 to 23 59 at which you have programmed the lamp to turn off automatically Default is 0 00 This field displays the EPROM version of your detector s software This field displays a list of the scans that are currently stored in memory Each scan is identified by the runtime at which it was initiated UV2000 only In the Scan File Setup this field defines the wavelength at which the detector should begin the scan Allowable values are 190 to 799 nm Default is 220 nm UV2000 only In the Develop File Setup this field defines the wavelength at which the detector should run its first set of injections Allowable values are 190 to 799 nm Default is 220 nm UV2000 only The Startup field toggles between Manual where you manually turn on the lamp
56. ally by interfacing the detector with a remote run signal from the injector see Appendix A for details In this scenario a signal that is equivalent to pressing RUN is automatically sent from the injector to the detector with each injection During the run you can monitor the run time from the Status Screen There are two ways to stop a run 1 Manually by pressing STOP before the programmed stop time 2 Automatically by allowing the run to finish at the programmed stop time If you re conducting a dual wavelength run you can also stop the run by loading a single wavelength file Regardless of how you stop a run the detector returns to READY Thermo Electron 01 2004 CHANGING RUN If you wish to change the detector s parameters PARAMETERS 1 You can use the Files Menu and follow the procedures outlined under Setting Parameters on page 26 2 Or you can use the Status Menu which is the programming area below the Status Screen Each method has a distinct advantage Programming in the Status Menu allows you to change the detector s current operating parameters even vvhile the detector is running Programming in the Files Menu allovvs you to prepare an edit file containing the changes vvithout altering the current detector settings The file may then be loaded later Status Menu From the Status Screen scroll down to the Status Menu Fig 3 19 The Status Menu contains the loaded file identification its numb
57. and Time where the lamp automatically powers up at a preset time Default is Manual This field displays the local time ranging from 0 00 to 23 59 at which you have programmed the lamp to start up automatically Default is 0 00 Thermo Electron 01 2004 Status Status Lock TESTS Time Wavelength Time from READY To File W Lamp Hours Wavelength Program 01 2004 This field in the Status Screen shows the current condition of the detector The possible conditions are READY the selected lamp is lit and ready for initiation of a run NRDY the detector is set to the wrong lamp for the run wavelength requested is performing internal tests or has a possible internal problem or UVW the deuterium lamp is warming up The run time is displayed when the running file has a programmed stop time In the UV2000 the letter Q appears at the beginning of this field when a queue is loaded The Status Lock field limits accessibility to the Status Menu the programming section below the Status Screen When set to On only the Status Screen appears on the display and the down arrow icon is not seen Default is Off The allows you to perform the detector s internal diagnostic light level and diode offset tests The Wavelength Program contains the Time and Wavelength fields It allows you to program changes in the detector s wavelength as a function of time Time refers to the run time in minutes when a timed event wave
58. and butyl paraben where the two compound peaks overlap Toluene Peak A is the compound of interest Butyl paraben Peak B is the peak we want to suppress We will use this example throughout the following steps for determining and using the K factor 1 35 1 51 Time in Minutes DT Z023E FM Figure 4 30 A chromatogram of two unresolved peaks toluene A and butyl paraben B Choosing a Pair of Wavelengths The first step in determining the K factor is to choose a pair of wavelengths for your analysis 1 Take an absorbance spectrum of each compound You can do this by injecting samples of compound A and compound B alone separately under the same chromatographic conditions as your analysis and using the UV2000 s scanning feature See Scanning on page 46 For the compounds in our example we get the spectra shown in Figure 4 31 01 2004 Thermo Electron 61 DT Z024E FM Toluene 200 380 Butyl Paraben 200 380 Wavelength nm 62 Figure 4 31 Spectra of individual compounds 2 Label the wavelength maximum for your peak of interest as Al 3 From the spectra pick a wavelength for which compound B absorbs and compound A does not This wavelength is labeled 22 For our example we have chosen 254 nm as Al and 280 nm as 22 Thermo Electron 01 2004 01 2004 u Calculating the K factor Use the UV2000 s Display AU A screen page 53 to obtain the indi
59. andard words 7 two line display 6 Copy Menu accessing the 32 illustration of 33 Copying files 32 103 copy field described 97 Copy File command 97 103 Develop File 33 protected files 33 Scan File 33 CSA see Safety Information Current time field 42 97 Cursor blinking square 1 movement 1 speed setting the 45 97 triangular 1 Curves smoothing spectral replays 53 Cuvette holder cleaning a cuvette 129 cuvette size defined 121 function of 121 inspecting a cuvette 128 installation of 122 maintenance of 128 testing absorbance linearity with 127 wavelength calibration using 127 D D see Develop File D2 Lamp Hours 98 Dark current test for 115 Defaults see Appendix B term defined 117 Degassing 117 Delete commands Delete File 98 Delete Queue 98 fields defined 98 Deleting a queue 59 files 25 33 protected files 34 time lines 38 Detector calibration 114 128 command center 2 front panel illustration of 2 modes described 3 rules for operation 1 Shutdown message 45 Deuterium lamp alignment of 76 changing the 74 chronometer 74 cleaning the surface 74 index ii Thermo Electron 01 2004 D2 lamp hours field 97 illustration of 75 lamp life 74 Low Light Detected message 110 testing the 115 tracking D2 hours 74 warm up time 74 Develop File UV2000 only Runs field 55 101 accessing the 54 advantages of 54 defined 54 117 Edit Menu 54 editing parameters 54 editing while running 56 example of 55 no cop
60. approximately 0 25 nm so if you choose 2 for the number of steps you will have offset the wavelength by 0 5 nm You can offset the wavelength by as much as 2 5 nm NOTE The offset value isn t cleared upon resetting the RAM memory It EN can only be changed from the lambda offset screen Maintenance The cuvette holder contains no user serviceable components however cleanliness of the cuvettes is critical to obtaining accurate analyses Therefore these instructions are provided for inspecting and cleaning the cuvettes Inspecting a Cuvette Cuvettes whether previously used or new should always be visually inspected before use Use the following steps to inspect a cuvette 1 Grasp the cuvette by its two frosted sides and hold it up in front of a bright light source such as a fluorescent fixture incandescent bulb or sunny window 2 Carefully observe the cuvette s two transparent glass sides Look for physical damage such as chips cracks scratches etc Also look for dirt smudges fingerprints and so forth 3 Based on the results of your inspection you can do one of the following three things a If no optical surface damage or contamination is noted you can fill the cuvette with sample and use it for your analysis b If you see physical damage or severe contamination on the optical surfaces you may wish to replace the cuvette with a new one If no physical damage is noted and only light to moderat
61. bleeding from column Leaks in system or flowcell Tiny bubble trapped in flowcell Large temperature fluctuations Thermo Electron Remedy e Allow a 30 minute warm up for normal operation one hour for maximum sensitivity Replace lamp Check for proper cable connections for detector output do not ground at both ends of cable Clean dirty component as described in Chapter 5 Connect integrator to appropriate output connectors on detector see Appendix A Check attenuation setting on integrator Flush flowcell with cleaning solvents as described in Chapter 5 Check for leaks Replace with fresh mobile phase made with high purity solvents Clean or replace column Check all fittings for leaks Replace flowcell Increase flow rate until bubble is removed Connect backpressure device to flowcell outlet check back pressure rating to avoid rupturing flowcell Remove system from drafts Thermo statically control column temperature 01 2004 Troubleshooting Table cont Symptom 4 No peaks or peaks much smaller than expected 5 Broad tailing peaks 6 Clicking sound when UV2000 is in dual vvavelength mode 7 Detector won t power up 01 2004 Cause a Incorrect wavelength setting Lamp not on or defective Integrator input voltage does not match detector output voltage Insufficient sample reaching the detector Rise time is too large too
62. ce Engineers with customer support available in North America in Europe and in Australasia and Asia In North America Thermo Electron Customer Service Engineers are available from each of the Thermo Electron field offices as follows Northeastern Region Phone 111 732 627 0220 Fax 1 732 627 0260 Southern Region Phone 1 770 516 5589 Fax 1 770 516 6916 Central Region Phone 1 847 310 0140 Fax 1 847 310 0145 Western Region Phone 1 408 965 6000 Fax 1 408 965 6123 Canada Phone 111 905 712 2258 Fax 1 905 712 4203 In the Americas use the following telephone number or fax number to order parts for all instruments Thermo Electron Customer Service Operations 1400 Northpoint Parkway Suite 10 West Palm Beach FL 33407 Phone 1 800 532 4752 Fax 1 661 688 8731 Thermo Electron Technical Support is available at the following location Thermo Electron Technical Support Operations 1400 Northpoint Parkway Suite 10 West Palm Beach FL 33407 Phone 1 800 685 9535 Fax 1 561 688 8736 Thermo Electron ili IN EUROPE In Europe customer support replaceable parts and technical support are available from each of the Thermo Electron offices as follows Wien Vienna Austria Phone 43 01 333 50 34 0 Fax 43 01 333 50 34 26 Brussels Belgium Phone 32 02 482 30 30 Fax 32 02 482 30 31 Les Ulis C dex France Phone 33 01 60 92 48 00 Fax 33 01 60 92 49
63. d run For these occasions the UV 2000 offers a queuing feature Using a queue you can program the detector to load and run a specified file for your first group of samples and then automatically load a second file to run your next group of samples The queue feature allows you to run as many as ten groups in a single queue To access the Queue Menu follow these steps 1 Press MENU 2 Select QUEUE When no queue is loaded the Queue Menu appears as shown in Figure 4 21 On page 58 we ll see how the menu appears when a queue is loaded Thermo Electron 01 2004 SETTING UP A QUEUE Entering a Line N Adding More Lines Deleting a Line An Example 01 2004 O Delete Figure 4 21 The Queue Menu with no queue loaded To set up a queue select Edit from the Queue Menu For an empty queue the display appears as shown in Figure 4 22 File Name Figure 4 22 An empty queue A 1 is automatically placed in the Order field of the first file to be run You can t change that so the cursor appears under the first editable field File Name Scroll through the available files and press ENTER when your choice appears NOTE You may only select numbered files The Scan and Develop files are not available in the Queue mode Enter the number of injections to be made in the RUNS field and press ENTER You can have as many as 999 injections per file After completing the first line a second line appears a
64. dary Way Hemel Hempstead Hertfordshire HP2 7GE e 44 01442 233 555 U S A 355 River Oaks Parkway e San Jose CA 95134 1991 1 408 965 6000 Notes The country code is enclosed in square brackets The city code or area code is enclosed in parenthesis For countries other than the U S A when you are dialing from within the specified country dial the 0 of the city code For countries other than Italy when you are dialing from outside the country do not dial the 0 of the city code Published by Technical Publications Thermo Electron Corporation San Jose California Copyright 2004 Thermo Electron Corporation All rights reserved Printed in the United States of America Thermo ELECTRON CORPORATION Customer Registration Register now and receive all the privileges associated with being a Thermo Electron Finnigan product user including application reports and technical reports Name Title Company Address City State Postal Code Country Telephone Ext SpectraSYSTEM UV Vis Detector Serial Date Purchased Tell us more Let us know more about how you use this product My Organization Is Check one only My Primary Application Is Check one only Q Commercial for profit lab Q Analytical Q Government lab Q Biomedical Q Hospital Clinic Q Clinical Toxicology 1 Research Institute Q Energy Q University College Q Food Agriculture Q Veterinary 1 Forensic To
65. der Thermo Electron 01 2004 R S RAM range replay rise time runfile run time signal to noise solvent programming spectral scan status T Z timed events troubleshooting wavelength programming 01 2004 Random Access Memory A detector parameter that controls the full scale range for the output signal Retrieves a stored spectrum that can then be played back as either individual data points or a smoothed spectrum A detector parameter that controls its response time rise time is inversely proportional to the amount of baseline noise The runfile is the file that contains the current detector parameter settings The duration of a sample run from injection to detection A measurement of the sensitivity of a detector the ability to measure a very small sample response over the baseline noise See gradient elution A sample spectrum The current condition An instrument action triggered to occur at a specific preset time during a run e g autozero wavelength change stop time Locating the cause of problems with equipment or procedures and solving these problems Programming the detector to change the monitoring wavelength as a function of time during a run Thermo Electron 119 E Cuvette Holder Accessory Introduction 01 2004 This Appendix provides information on the installation use and maintenance of the Cuvette Holder Accessory Fig E 1 This accessory is available to si
66. e contamination clean the cuvette using the procedure that follows in the next section of this appendix 128 Thermo Electron 01 2004 Cleaning a Cuvette 01 2004 If the visual inspection reveals contamination of or damage to the cuvette s optical surfaces the inner and or outer surfaces of the cuvette s two transparent faces then the cuvette should be cleaned before use Use the following steps to clean a cuvette 1 Immerse the cuvette in a small beaker filled with an appropriate cleaning solution Use detergent and water to clean cuvettes that are contaminated with residue from water based solutions Use an appropriate organic solvent e g methanol ethanol isopropanol etc for cuvettes contaminated with residue from organic solvent based samples Place the beaker containing the cuvette s and cleaning solution in an ultrasonic bath and set the timer Use a time setting that s appropriate for the amount of contamination that has to be removed Remove the cuvette from the beaker handling it by its frosted sides only Rinse it thoroughly with clean deionized water until all traces of detergent and dirt have been flushed away Dry the cuvette thoroughly with a lint free wiper exercising care to handle the cuvette only by its two frosted non optical sides Carefully inspect the cuvette for residual contamination using the steps detailed in the preceding section of this appendix If any is noted repeat Steps 1 through
67. e UV1000 s Status Menu The parameters are set using the same instructions given under Wavelength Program and Options Menu starting on page 21 When you use the Status Menu to change the UV 1000 settings each change is effective immediately upon leaving the field Notice the words Save File below the Range field Press ENTER when the cursor is in the Save File field to save the new settings to the run file The confirmation message shown in Figure 3 9 will appear briefly File Saved Figure 3 9 The File Saved message NOTE When you change the detector settings from the Status Menu the contents of the edit file do not change Only the run file values are modified Thermo Electron 01 2004 DELETING THE FILE To return to your previous setting without saving the new ones do not press ENTER Instead you may reenter the unaltered file as follows 1 Press MENU 2 Select FILE 3 Select Load 4 The words Load File will appear on the screen Press ENTER A confirmation message Fig 3 6 will appear for one second You are then returned to the Status Screen and all settings will contain their original values To delete the edit file select Delete in the File Menu The words Delete File will appear on the screen When you press ENTER the confirmation message shown in Figure 3 10 appears briefly and the display returns to the File Menu The edit file parameters return to the
68. e a queue is running 59 a queue UV2000 only 58 Locking the status screen 43 Low Light Detected messages from deuterium lamp 110 from tungsten lamp 111 Main Menu accessing the 20 26 general description of 4 illustration of 10 14 Maintenance see Chapter 5 Manual conventions used in 6 MENU key function of 3 used to access Main Menu 20 26 Menu Reference 97 Menu Tree 93 Menus display conventions of 4 general description of upper level 5 illustration of longer 6 moving between within 1 4 term defined 118 Messages A File Is Already Running 111 Detector Shutdown 45 112 File Copied 33 File Deleted 25 34 File Loaded 22 29 File Saved 24 32 Invalid Parameters Spectrum Not Allowed 114 Lamp Cover Open 110 Low Light Detected 110 111 No More Available Memory 111 No Queue Available 58 111 No Scans Stored 53 No Spectra Available 111 NRDY for one of the lamps 41 Protected File Cannot Be Copied To 33 111 Cannot Be Deleted 34 111 Cannot Be Edited 111 Queue Deleted 60 98 Queue Loaded 58 100 Queue Loaded Cannot Load File 59 111 real time errors 110 Run In Progress No Replay Allowed 112 Run In Progress Testing Not Allowed 112 Run Not In Progress Spectrum Not Allowed 112 Scan Memory Full 112 system errors 109 types of messages 5 110 user input errors 111 Method development see Develop File Method transfer see Develop File Miscible 118 MLF most likely failure 114 Monochromator cleaning the lens 6
69. e the lamp assemblies Fig 5 6 4 Unplug the deuterium lamp lead from the detector taking care not to twist the connector as you gently pull it out 5 Loosen the two thumbscrews that hold the lamp assembly in place and pull the assembly straight out 01 2004 Thermo Electron 75 D2 Lamp Installation Follow these steps to install a new D2 lamp 1 Hold the deuterium lamp assembly so that the leads are at the top Slide the assembly onto the alignment pin shown in Figure 5 7 The alignment pin is located directly below the detector s monochromator aperture 2 Securely fasten the assembly in place with the two thumbscrews and aluminum standoffs 3 Connect the lamp lead to the right hand terminal in the lamp compartment 4 Replace the lamp housing and secure it with the knurled thumbscrew Replace the detector s front cover 5 Connect the power cord to the rear detector panel Tungsten Lamp Alignment Pins Deuterium Lamp Alignment Pin DT Z021E M 76 Figure 5 7 Deuterium and tungsten lamp alignment pins Thermo Electron 01 2004 THE TUNGSTEN The tungsten W lamp typically requires only fifteen minutes of LAMP warm up time Its lifetime is approximately 2500 hours You can track lamp life automatically See Automatic Lamp Operations on page 41 for details W Lamp Removal Follow the steps below to remove the tungsten lamp 1 Disconnect the power cord from the de
70. ect COMMANDS 3 Scroll down to the Shutdown Detector field 4 Press ENTER The confirmation message shown in Figure 4 8 appears on the display Figure 4 8 Shutdown confirmation message To start the detector up again press any key on the keypad The detector will come up under the same conditions present when the Shutdown Detector command was activated 01 2004 Thermo Electron 45 Scanning HOW IT WORKS 46 UV2000 only The UV2000 can perform a spectral scan on eluting peaks without stopping the eluant flow This unique feature greatly simplifies the determination of wavelength maxima for individual compounds in your sample during method development work When a scan is initiated the monochromator moves from the run wavelength to the scan s start wavelength The detector scans by stepping through a defined spectral range at specified wavelength increments Individual absorbance values are read at each increment until the monochromator has reached the last wavelength The UV2000 can collect and store as many as ten spectra from a single chromatographic run in its memory The actual number of spectra is determined by the number of data points in each scan Since the number of data points varies with the wavelength interval and the scanning range first calculate the number of data points using Equation 1 then use either Equation 2 or Equation 3 to determine the number of spectra you will be able to collect Equation 1 U
71. eep the absorbance values on scale There is no programmed stop in the Scan mode The run will continue until it reaches 99 99 minutes or until you press STOP Thermo Electron 01 2004 Automatic Scanning Model UV2000 only 01 2004 If you ve set the Auto Scan field in the Setup Menu to On your detector will perform an automatic scan whenever there are at least three consecutive data points with positive slopes followed by three consecutive data points with negative slopes The absorbance values for all these data points must exceed 5 percent of the value set in the Range 2 field In our example chromatogram Fig 4 13 a scan would occur automatically for Peak A since it has at least three data points with positive slopes followed by at least three data points with negative slopes all of which exceed 5 of the value set in Range 2 Conversely no scan would occur for peak B since none of its absorbance values exceeds the 5 threshold even though it may satisfy the consecutive slope criteria Peak A 5 of Range 2 Peak B Time DT Z155E DT Figure 4 13 An example of how automatic scanning works An automatic baseline scan will occur at the time specified in the Spectra Menu Scan Zero Time field NOTE Make sure that no peaks are eluting at the specified scan zero time or your baseline scan will be zeroed erroneously for the eluting peak s value at the moment when the scan zero occurs This will
72. eference R1 R2 numbers may vary considerably between instruments A three or four digit number is typical As with the Light Levels test check the photodiodes and the analog PCB the components most likely to affect light intensity if you get an unusual reading If any of the numbers are zero call Thermo Electron To recalculate the diode offsets select C The offsets may need to be recalculated if the light levels are less than the diode offsets This situation normally occurs after slight diode offset drift or while working with extremely low light Selecting A Calibration brings up the screen shown in Figure C 6 You can use this screen in combination with the optional Cuvette Thermo Electron 113 Holder Accessory to offset the factory calibrated wavelength to more closely match an FDA industry or in house calibration standard NOTE If you wish to conduct your calibration using the Cuvette Holder the following procedure has also been detailed in Appendix E for your convenience A Offset steps 0 Figure C 6 The lambda offset screen NOTE The UV1000 and UV2000 detector is calibrated using a mercury NI lamp fixture This provides a very narrow emission line at 254 nm Broad band calibration standards such as holmium oxide and didymium filters make calibration more difficult and less accurate To offset the factory calibrated wavelength select the number of steps by which you want the wavelength to be offset
73. electropherogram of a typical sample and standard mixture well documented with system conditions for future reference Careful comparison of migration times peak shapes peak sensitivity and baseline noise can provide valuable clues to identifying and solving future problems Chemical Toxicity Although the large volume of toxic and flammable solvents used and stored in laboratories can be quite dangerous don t ignore the potential hazards posed by your samples Take special care to read and follow all precautions that ensure proper ventilation storage handling and disposal of both solvents and samples Become familiar with the toxicity data and potential hazards associated with all chemicals by referring to the manufacturers Material Safety Data Sheets MSDS Sample Preparation Always consider the solubility of your sample in the electrolyte buffer Sample precipitation can plug the system by obstructing the flow through the capillary This obstruction may result in irreparable damage to parts of the system Particulate matter can be avoided by filtering the samples through 0 45 or 0 2 micron or less filters Thermo Electron 01 2004 01 2004 Solvent Requirements Many chemical manufacturers provide a line of high purity or spectro quality reagents that are free of chemical impurities Routine filtration of all solvents or eluents through a 0 45 or 0 2 micron or less fluorocarbon filter before placing them in the solvent
74. end stellt der hohe Druck in einem Fl ssigchromatographen auch kaum eine unmittelbare Gefahr dar Jedoch sollten auftretende Undichtigkeiten umgehend beseitigt werden Schlie lich ist noch zu empfehlen bei der Arbeit mit einem Fl ssigchromatographen stets Augen und Haut zu sch tzen und vor allen Wartungsarbeiten darauf zu achten da das Ger t abgeschaltet und druckfrei gemacht wurde Thermo Electron 01 2004 Startup Checklist Use this checklist to ensure that you have completed all the steps necessary for the proper installation of your Finnigan SpectraS YSTEM UV Vis detector Complete installation information can be found in Appendix A UNPACKING POSITIONING DETECTOR POWER CHECKOUT g g REAR PANEL CONNECTIONS g m 01 2004 Unpack and inspect your instrument Check for damage Check your accessory kit and manual Are they O present O complete Read the Safety Information Card Place on benchtop as close as possible to the column and at least 5 inches 13 cm from the wall Set voltage for local requirements Check that the correct fuses are installed Insert the 8 and 12 pin green connectors into their corresponding sockets Connect the Analog Output and the corresponding ground terminals to your data system recorder Connect the desired remote communications terminals to external devices O STOP Input O RUN Input O ZERO Input O READY Output Connect the power cord Th
75. er Fig 5 5 that tracks the total hours of lamp operation To read the chronometer note the position of the gap in the mercury tube against the graduated background You can also track lamp life automatically See Automatic Lamp Operations on page 41 for details NOTE The lamp surface must be kept free of fingerprints and smudges If the surface needs cleaning use a lint free lens paper moistened with methanol or isopropanol Plug In Connector Lamp Leads DT Z038E DG Figure 5 5 Deuterium lamp chronometer These four steps explain how to remove the deuterium lamp 1 Disconnect the power cord from the detector s rear panel and make sure that the instrument is turned off CAUTION Intense UV light can damage your eyes Always disconnect the power cord before exposing the lamp and always allow sufficient time for the lamp to cool before removing it as it gets quite hot when lit 2 If you have not already done so remove the detector s front panel by grasping the bottom of the panel firmly with one hand and pulling back Thermo Electron 01 2004 Tungsten Lamp Assembly Deuterium Lamp Assembly Flowcell Outlet Flowcell lt Inlet Flowcell Assembly DT Z004E FM Figure 5 6 Deuterium and tungsten lamp assemblies 3 Remove the lamp housing by loosening the thumbscrew and pulling the cover straight back to expos
76. er and name Wavelength Program Rise Time Autozero Time and Ranges Rise Time Autozero Time Range 1 Range 2 Figure 3 19 The UV2000 s Status Menu for dual wavelength operation The Status Menu shown in Figure 3 19 is typical for dual wavelength operation In the single wavelength mode only one wavelength field appears in the wavelength program The detector s parameters are set following the same instructions given under Wavelength Program and Options Menu starting on page 27 However you cannot modify either the file identification or the wavelength mode dual or single from the Status Menu NOTE When you modify a file s parameters from the Status Menu you do NI not change the contents of the same file number stored in the detector s memory Only the copy of the active file is modified 01 2004 Thermo Electron 31 Saving the File When you change the UV2000 s settings from the Status Menu each change is effective as soon as you leave the field You ll also see that the File identification on the first line of the Status Menu Fig 3 19 now reads File N xxxx changed where N xxxx is the file number and name and that the words Save File now appear below Range 2 To save the changed file press ENTER The confirmation message shown in Figure 3 20 will appear briefly File Saved Figure 3 20 The File Saved message To keep the original file without saving the changes don t press ENTER In
77. eration you first enter the desired detector parameters into an edit file You then load the edit file into the run file which contains the detector s current operating parameters These instructions will show you how to start and stop a run and how to modify the detector s operating parameters You set up the UV1000 s parameters by using the File Menu to prepare an edit file You then load the edit file into the run file To access the File Menu first press MENU The Main Menu appears on the screen From the Main Menu select FILE The menu shown in Figure 3 1 will appear gt Edit Q Delete Figure 3 1 The UV1000 s Files Menu From the File Menu select Edit to display the Edit Menu The Edit Menu Fig 3 2 selections are Wavelength Program which contains time and wavelength fields and Options which contains the Rise Time Autozero Time and Range fields Qo Wavelength Program 1 Options Figure 3 2 The UV1000 s Edit Menu Thermo Electron 01 2004 Wavelength Select Wavelength Program from the Edit Menu The Wavelength Program Program is a Table containing the Time and Wavelength fields Fig 3 3 Wavelength 254 Figure 3 3 The UV1000 s Wavelength Program In the single wavelength mode you can operate with either a one line or a two line wavelength program Using a one line program the detector is always in the READY state and you can continually monitor the baseline Using a two line pro
78. ermo Electron xix FLOWCELL CONNECTIONS INSTRUMENT POWER UP REGISTRATION CARD XX uu auu Q I I Remove the detector s front panel Remove the flowcell assembly from the detector Connect the flowcell inlet directly to your LC column outlet Connect the flowcell outlet to waste tubing and a waste container Replace the detector s front panel Install the power cord and turn on the instrument Check that self tests are running and that no error messages appear Check that the Status Screen appears on display Complete and return the registration card Thermo Electron 01 2004 Flowcells 9550 0100 9550 0234 9550 0197 9550 0053 9550 0265 9550 0101 9550 0263 Options And Accessories 2103 9119 A4095 010 9551 0022 9551 0023 9051 0143 Manuals A0099 540 Maintenance Parts A4051 010 A4061 010 Upgrade Kits List of Spare Parts Consumables and Kits Shown below is a list of spare parts and consumables available from Thermo Electron for use with your Finnigan SpectraSYSTEM UV Vis detector Contact your local Thermo Electron representative for current prices Analytical LC 6 mm Analytical LC 10 mm Biocompatible LC 6 mm Microbore 3 mm Microbore 6 mm Semi preparative Open Column 3 mm Cuvette Cell Holder External Events Connector Remote Interface Cable Tungsten Lamp prealigned Deuterium Lamp prealigned Regulated Backpressure Accessory UV Vis Detectors Reference
79. ertigungsst tte entwickelt hergestellt und getestet und hat unser Werk in sicherem Zustand verlassen VORSICHT Dieses Ger t darf nur nach den Vorschriften Ger t auf andere Weise als hier beschrieben benutzt wird kann dies zu Sch den am Ger t oder zur Verletzung des Bedieners f hren dieser Bedienungsanleitung benutzt vverden VVenn dieses Dieses Handbuch enth lt Warnhinweise deren genaue Befolgung Personensch den Sch den am Ger t oder Datenverluste verhindern kann Auf alle derartigen Warnhinweise wird durch Fettschrift und durch Verwendung der nachfolgenden Symbole gesondert aufmerksam gemacht N A VORSICHT OBERFL CHE HOCHSPAN HEISS NUNG Jedes Ger t kann unter bestimmten Umst nden gef hrlich sein Lesen Sie daher in jedem Fall die nachstehenden Sicherheitshinweise und ergreifen Sie die entsprechenden Ma nahmen Auf dieses Weise sorgen Sie f r einen sicheren Betrieb und eine lange Lebensdauer des Ger ts 1 Bevor Sie eines der Ger temodule einstecken oder das Ger t einschalten berpr fen Sie in jedem Fall ob die Nennspannung und die Sicherungen der Netzspannung der rtlichen Stromversorgung entsprechen 2 Verwenden Sie nur Sicherungen des angegebenen Typs und der angegebenen Amperezahl Verwenden Sie keine reparierten Sicherungen und berbr cken Sie die Sicherung nicht Thermo Electron XV 3 Das mitgelieferte Netzkabel mu in eine Steckdose mit Schutzleiter eingesteckt werden
80. es peut tre vit e par la filtration des echantillons 3 travers des filtres de 0 45 ou 0 2 um ou moins Caract ristiques des solvants Un grand nombre de fabricants de produits chimiques fournissent des r actifs de puret lev e ou de qualit spectrographique d pourvue de toute impuret chimique La filtration syst matique de tous les solvants ou luants 3 travers un filtre fluorocarbon de 0 45 ou 0 2 um ou moins avant de les placer dans le r servoir de solvants prolonge de fa on significative la dur e de vie et l efficacit des filtres d entr e des clapets et des joints d tanch it de l injecteur et de la colonne De fa on g n rale les solvants pour chromatographie liquide sous haute pression ne n cessitent pas de filtration Choisir une phase mobile qui est compatible avec l chantillon et la colonne s lectionn s pour la s paration Noter que certains solvants sont corrosifs pour l acier inoxydable Des versions inertes et biocompatibles des instruments sont disponibles aupr s de Thermo Electron Degazage des luants Effectuer le d gazage des luants selon la m thode de d gazage par le vide ou a l h lium Une description complete de ces m thodes est disponible dans la documentation fournie s par ment avec les accessoires de d gazage Thermo Electron xiii X V Elimination des solvants S assurer qu il existe un conteneur pour solvants a liminer ou tout autre syste
81. etector The chapter s first section covers the capabilities offered by both the UV1000 and the UV2000 detectors the second section beginning on page 46 contains those functions specific to the UV2000 You should be familiar with the instructions presented in Chapter 3 Basic Operations before you begin UV1000 and UV2000 Wavelength Programming BUILDING THE PROGRAM 01 2004 N Your detector can change wavelength as a function of time a feature vve call VVavelength Programming This feature gives you maximum detection sensitivity for each component of a mixture vvithout making multiple injections of the sample NOTE A wavelength program can be built in either the Status Menu or the File s Menu In wavelength programming you enter time lines into a Wavelength Program Each time line specifies the time at which you want a wavelength change to occur The following instructions are for single wavelength operation but if you have a UV2000 you can build a dual wavelength program using the same procedure Thermo Electron 37 Initial Conditions Adding Lines The Stop line Deleting a Line Access the Wavelength Program Fig 4 1 through either the Status Menu or the Files Menu Wavelength 250 Figure 4 1 The wavelength program for single wavelength operation The initial time entry is 0 00 Move the cursor to the corresponding Wavelength field and enter the initial wavelength for your analysis
82. fer EN solution be sure to pump 25 40 mL of HPLC grade water or equivalent through the system to remove any salts before flushing with the cleaning solvent s 2 Flush the flowcell with 40 50 milliliters of solvent HPLC grade water methanol or isopropanol You can either pump the solvent through the flowcell with the chromatographic pump or you can draw the solvent through the flowcell using a large volume syringe If you use an LC pump to flush the flowcell first remove the column from your chromatographic system to avoid column degradation Replace the column with an appropriate length of tubing ensuring that all connections are snug and leak free If you use a syringe always draw the solution through the flowcell flowcell Pressurizing the syringe could cause a leak or rupture that would result in an extremely dangerous uncontrolled spraying of solvent CAUTION Never use a syringe to force solvent through a 01 2004 Thermo Electron 71 Cleaning with Methanol or isopropanol is generally sufficient for cleaning a Nitric Acid flowcell However if the flowcell is still contaminated after flushing with organic solvents follow this nitric acid procedure with alcohols especially methanol Be sure to adhere to your company s safety procedures for handling and disposal of corrosive acids Flush the flowcell with water to remove all traces of alcohol prior to flushing with nitric acid CAUTION Nitric acid is ex
83. g time lines 38 automatic ordering 38 autozeroing with or without a wavelength change 40 building a program 37 deleting a line 38 described 37 editing while running 39 example of 39 illustration of 39 last line 38 sorting time lines 38 term defined 119 using a stop line 38 zeroing the baseline 39 index xii Thermo Electron 01 2004
84. gram you can use a stop line and you can start and stop the detector during a chromatographic run Stop lines are useful for example in an automated series of runs where you want to autozero the detector s baseline after each injection For a one line program enter the wavelength s for your analysis in the Wavelength field that corresponds to the time of 0 00 For a two line program add an additional line the stop line by scrolling down to the blank line below the time 0 00 line and pressing The second line will automatically have a time setting of 1 00 and the same wavelength setting s as the first Change 1 00 to the desired stop time for the run and leave the wavelength value unchanged An example of a two line wavelength program for a nine minute run at 283 nm is shown in Figure 3 4 Wavelength Figure 3 4 An example of a two line wavelength program with a programmed stop time 01 2004 Thermo Electron 21 Options Menu Loading the Edit File 22 Q fl 4 Select Options from the Edit Menu to display the Options Menu Fig 3 5 Use this menu to set the detector s rise time autozero time and range Rise Time Autozero Time Figure 3 5 The UV1000 s Options Menu Rise Time This field controls the detectors response time Rise time is inversely proportional to the amount of baseline noise For example the longer the rise time the less noise detected The one second defa
85. have several detectors fluorescence refractive index electrochemical etc hooked up in series place your UV1000 or UV2000 detector closest to the column outlet as its flowcell can withstand the greatest backpressure Replace the front panel of the detector making sure that the tubing passes through the slots without being pinched 01 2004 Flovvcell _ Flovvcell nlet Outlet Tubing Clamp Photodiode Mount Flovvcell Assembly Flowcell Assembly Thumbscrews Figure A 6 The flowcell assembly showing thumbscrews photodiode mount and flowcell inlet Thermo Electron 87 DT Z019E FM OPTIONAL FLOWCELLS Thermo Electron offers several different flowcells for use in different applications Each flowcell possesses distinct design characteristics and performance specifications These characteristics are compiled in Table A 1 Contact your Thermo Electron representative for details Table A 1 Design and performance specifications for Finnigan SpectraSYSTEM flovvcells Path Max Path Length Tubing I Max Flow Press Flowcell mm Volume uL Diam in Material mL min psi Analytical LC 6 9 OI 551 50 1000 Analytical LC 10 15 01 551 50 1000 Microbore 3 1 2 005 551 10 1000 Microbore 6 7 0 007 551 20 1000 Semi Prep Open Column 3 4 5 02 551 100 1000 cells use sapphire for windows All but the preparative flowcells have a heat exchanger SS Stainless Steel with TFE Gaskets
86. hecks the W lamp and its voltages when the lamp is on and when it is off If the message W Not Detected appears the lamp voltages are good but the lamp is not present or is not functioning properly Try replacing the tungsten lamp and retrying the test Ifthe word Fail appears call your Thermo Electron representative MLF Lamp or Motherboard Lamp and Shutter UV2000 only This test actually has several parts each of which checks a different part of the lamps and shutter s operation If either of the lamps fails an appropriate message will be displayed Try replacing the lamp and retrying the test If a Bad Shutter message appears call your Thermo Electron representative MLF part listed on display Thermo Electron 115 EXTERNAL DIAGNOSTIC TESTS LC Test Mix l Absorbance Linearity fl a 4 l 116 This section describes tvvo external diagnostic tests that can be used to verify that your detector is vvorking properly An ampule of prepared LC Test Mix is included as part of your detector s accessory kit An instruction sheet describing the parameters for running the test mix and showing the resulting chromatogram is also enclosed This is a good test to run when you first set up your LC system HINT Keep the chromatogram that you generate with the LC Test Mix It can be a useful baseline for troubleshooting problems later on Use the optional cuvette holder see Appendix E and certified
87. hermo Electron representative SYSTEM RESET e RAM ERROR e ADDRESS ERROR BUS ERROR e DIVIDE BY ZERO LOW LO ERROR e LOW LI ERROR e DISTANT QUEUE ERROR REAL TIME Real time error messages indicate that you need to correct a certain ERRORS hardware condition Possible messages are Lamp Cover Open Check that the detector s lamp housing is in place and properly installed Low Light Detected From Deuterium Lamp This message indicates that the deuterium lamp may not be on may be improperly installed or needs to be replaced due to low light energy It can also appear if the lamp cover is replaced while the lamp is on Using the Lamps Menu see Automatic Lamp Operations on page 41 turn the lamp state to off wait five seconds and then switch the lamp on If the error message recurs check for proper lamp installation according to the procedure outlined in Appendix A If the lamp is installed correctly its surface is clean and the message still appears replace the lamp 110 Thermo Electron 01 2004 INPUT ERRORS 01 2004 Low Light Detected From Tungsten Lamp This message indicates that the tungsten lamp may not be on may be improperly installed or needs to be replaced due to low light energy Using the Lamps Menu see Automatic Lamp Operations on page 41 turn the lamp state to off wait five seconds and then switch the lamp on If the error message recurs check for proper lamp installation accordi
88. ile in the Edit File you will see file choices of S and D that represent the Scan and Develop files respectively These files are some of the UV2000 s advanced features that you will learn about in Chapter 4 From the Edit Menu select Wavelength Program The Wavelength Program designates dual or single wavelength operation and also contains a Table of time and wavelength A wavelength program for dual wavelength operation appears in Figure 3 14 Program Dual A 190 450 Figure 3 14 The UV2000 s Wavelength Program in dual wavelength mode Select Single A Dual 2 190 450 or Dual 2 366 700 in the Program field The Table for time and vvavelength s vvill appear For single wavelength operation there is only one wavelength field You can operate with either a one line or a two line wavelength program Using a one line program the detector is always in the READY state and you can continually monitor the chromatographic eluant Using a two line program you can use a stop line and you can start and stop the detector during a chromatographic run Stop lines are useful for example in an automated series of runs where you want to autozero the detector s baseline after each injection Thermo Electron 27 Options v r For a one line program enter the wavelength s for your analysis in the Al and 22 or Wavelength fields that correspond to the time of 0 00 For a tvvo line program add an additional line the s
89. ing any maintenance Thermo Electron ix Information sur la s curit et la compatibilite electromagnetique CEM Selon notre engagement 3 assurer a nos clients service et s curit ces instruments sont d clar s conformes aux normes de la FCC et a la r glementation europ enne CE y compris a la directive sur les basses tensions Con u fabriqu et test dans une installation homologu e 1509001 cet instrument a t livr a partir de notre usine de fabrication dans le respect des r gles de s curit MISE EN GARDE Cet instrument doit tre utilis selon les d utilisation de cet instrument d crites dans le pr sent manuel risque instructions figurant dans ce manuel Le non respect des consignes IDENTIFICATION DES INFORMATIONS SUR LA SECURITE 01 2004 d endommager l instrument et ou d infliger des blessures a l operateur Ce manuel de r f rence contient des pr cautions d usage afin de pr venir tout dommage corporel ou mat riel ainsi que toute perte de donn es lorsque I op rateur se conforme aux instructions indiqu es Ces instructions sont accompagnees des ic nes suivantes et sont affich es en caract res gras pour attirer I attention de op rateur N A A MISE EN SURFACE HAUTE GARDE 1 BRULANTE TENSION Chaque instrument pr sentant des dangers sp cifiques il incombe a l op rateur de lire les pr cautions suivantes et de s y conformer afin de maintenir
90. ion a narrovv tip screwdriver 2 mm wide a 2 Phillips screwdriver Place the detector on the benchtop as close as possible to the chromatographic column outlet thus minimizing the length of tubing necessary for connection to the flowcell inlet Allow at least five inches 13 cm of clear space between the detector s rear panel and any wall or obstruction This provides both access to the rear panel connectors and a free flow of cooling air The detector is shipped with the voltage and fuses preset for your location To verify the correct setting look through the cut out window on the voltage selector cover Figure A 1 The cover is located on the detector s rear panel but if your instrument is new it may be hidden behind a precautionary sticker If the voltage setting satisfies you local site requirements skip to Fuses on page 82 If not proceed to the next section Voltage Selection NOTE Do not plug in the instrument without first verifying that the voltage is properly set for your location And never run the detector at more than 10 below the nominal line voltage If the preset voltage does not satisfy your local requirements select the correct voltage by following these steps 1 Insert a small flat blade screwdriver into the slot at the top of the voltage selector cover Fig A 1 PRY OPEN HERE DT Z033E FM CT il Figure A 1 Opening the voltage selector c
91. ion This Chapter provides step by step instructions for the most frequently used detector operations including setup and run procedures for single and dual wavelength modes detector file management and protection and analog output operations To keep the instructions easy to follow we have divided the Chapter into two sections Instructions for the UV1000 begin on page 20 Instructions for the UV2000 begin on page 26 You may wish to keep the Menu Tree and the Menu Reference from Appendix B on hand as you work through this chapter NOTE You should be aware that your display s values may differ from those presented in this manual especially if the detector has been previously programmed Before You Begin 01 2004 Before you begin this chapter your detector should be installed in a chromatographic system see Appendix A and you should have completed the Startup Checklist located at the front of this manual We also recommend that you review Chapter 1 Getting Started which includes general instructions for using the detector keypad and which lists the conventions used throughout this manual Thermo Electron 19 UV1000 Single wavelength Operation SETTING PARAMETERS 20 The UV1000 uses a standard deuterium lamp to operate in a single wavelength mode in the ultraviolet UV range Adding an optional tungsten lamp increases the detector s capabilities to the visible Vis range To perform a single wavelength op
92. ir default settings File Deleted Figure 3 10 The File Deleted message Analog Outputs ANALOG OFFSETS 01 2004 There are two analog outputs for the UV1000 Analog Output land Analog Output 2 On the detector s rear panel they appear as Unranged output and Ranged Output Analog Output 1 is set at 1 V AU and is intended for an integrator interface Analog Output 2 is range selectable and is used for recorders and other devices Rear panel connections for both outputs are discussed on page 84 Analog offsets may be used when there is a high background absorbance reading or when there is considerable baseline drift from your chromatographic system and you are unable to keep your integrator s recorder s signal on scale Because integrators have very limited capacity for handling negative signals you may wish to set a small positive offset 1 when using an integrator Use negative offsets with recorders where you may wish to set the pen at either side of the strip chart Thermo Electron 25 riz l Offset options are selectable from the Analog Outputs Menu To access these options 1 Press MENU 2 Select OPTTONS 3 Select Analog Outputs The Analog Outputs Menu is shown in Figure 3 11 Analog 1 Offset mv Analog 1 Offset Figure 3 11 The UV1000 s Analog Outputs Menu HINT Although the default for the Analog 1 offset is set at zero we recommend a 1 mV setting fo
93. is field designates the monitoring wavelength to be used when running the Scan file Allowable values are 190 to 800 nm Default is 254 nm UV2000 only Located in the Develop file this field is the amount of time designated for each chromatographic run Allowable values are 0 01 to 999 99 minutes Default is 10 00 minutes UV2000 only Located in the Develop file this field designates the number of injections to be performed at each wavelength increment Allowable values are to 9 Default is 1 UV2000 only This field appears in the Status Screen when the Scan file is loaded To initiate a scan move the cursor to this field and press ENTER UV2000 only This field allows you to set a runtime at which the detector will perform a baseline scan automatically Allowable values are 0 00 to 99 99 minutes Default is 0 00 UV2000 only Thermo Electron 101 Self Tests Setup Short Outputs Shutdown Shutdown Detector Shutdown Time Software Version Specira Time Start Startup Startup Time 102 This command tells the detector to run through its internal diagnostic tests The Setup Menu allows you to set up the parameters in the Scan file UV2000 only This command allows you to short the detector s outputs together When you select Short Outputs the detector s analog outputs are shorted together zero volts and the field changes to Unshort Outputs To remove the short and return the outputs to their norm
94. is particularly useful with autosamplers where the detector can signal that it is ready for the next injection in an automated series of runs To hook up the READY Output terminal connect the input from the other instrument to the detector s READY Output and GROUND terminals terminals 4 and 5 For more information on accessing this feature through the detector s keypad see page 44 01 2004 Thermo Electron 85 CONNECTING TO Use the following steps to connect the flowcell THE FLOWCELL 1 Remove the front panel of the detector Although the flowcell assembly is located behind the lower housing Fig A 5 the housing does not need to be removed to connect your inlet and outlet lines Display Keypad Lamp Housing Flowcell Housing Figure A 5 The flowcell assembly is located behind the flowcell housing Thermo Electron 01 2004 Z fl a 4 l Use the finger tight fitting and ferrule sets included vvith the installation kit to connect the column outlet directly to the detector s flovvcell fluid inlet Figure A 6 shows how the inlet line enters the detector from the left side and winds around the flowcell before entering the flowcell from the bottom NOTE If additional tubing is required to reach the inlet use a zero dead volume union Connect the detector s fluid outlet to the low pressure union and waste tubing supplied in the installation kit HINT If you
95. kinds of information menus screens and messages Menus require you to make selections or enter specific values Screens display information that cannot be edited Messages confirm actions and point out errors The Menu Tree in Appendix B outlines the structure and content of the detectors menus and screens Main Menu The Main Menu is the top level of the menu structure In the UV 1000 Fig 1 2 the Main Menu gives you access to four other menus FILE COMMANDS OPTIONS and TESTS In the UV2000 there is and additional menu choice QUEUE Fig 1 3 To see the Main Menu press the MENU key at any time gt FILES 14 COMMANDS Q OPTIONS Q TESTS Figure 1 2 The UV1000 s Main Menu gt FILES 141 QUEUE Q TESTS Q COMMANDS Q OPTIONS Figure 1 3 The UV2000 s Main Menu Thermo Electron 01 2004 MESSAGES 01 2004 From the UV1000 s and the UV2000 s File s Menu you can edit load delete or copy files The UV2000 also lets you copy files The Commands Menu lets you insert an event mark onto your chromatogram short outputs or shut down the detector The Tests Menu lets you run built in instrument tests and diagnostics In the Options Menu you can set up or change your instrument s configuration From the Queue Menu you can edit or change the order of files in the sample queue Refer to Chapters 3 4 5 and Appendix B for more information on any of the instrument s menus Status Screen The Status Screen Fig 1 4 disp
96. l detection wavelength is 254 nm At 5 00 minutes into the run the wavelength changes to 280 nm At 7 00 minutes it changes to 265 nm The run ends at 10 00 minutes and the detector returns to its initial wavelength of 254 nm and to its READY state RUNNING THE After you set the rest of your parameters the detector is ready to run PROGRAM It is good practice to zero the detector at the beginning of every run and at each wavelength change See the next section titled Programmed Autozero for details Once you start the run you may edit any timed event wavelength change autozero or stop time that has not yet taken place These edits can only be made from the Status Menu however Each edit is entered immediately into the detector s operating wavelength program For example for the program displayed in Figure 4 2 the stop time is 10 0 minutes If at 7 00 minutes into the run you determine that the run should be 9 00 minutes long you can edit the last line of the program such that the current run will stop at 9 00 minutes 01 2004 Thermo Electron 39 EE Programmed Autozero The detector can be programmed to perform an automatic zero with each wavelength change during a run using the Zero on A Change field To access this feature 1 Press MENU and select OPT
97. lays the detector status wavelength setting s and the absorbance reading It automatically appears whenever the instrument is powered on or the STATUS key is pressed No entries are made on the Status Screen Status AU READY 0 00001 Figure 1 4 The Status Screen Status Menu Just below the Status Screen is the Status Menu To access the Status Menu press the down arrow key from the Status Screen The Status Menu lets you review and edit run parameters during a run Chapter 3 discusses the Status Menu in more detail There are three different kinds of messages that can appear on your detector s display user messages confirmation messages and error messages User Messages User messages indicated on the display by double asterisks tell you about an existing instrument condition or ask for further actions Some of these will only appear on the display for three seconds An example of a message requiring further action is shown in Figure 1 5 Protected File No Editing Allowed Figure 1 5 An example of a user message Thermo Electron 5 Confirmation Messages Confirmation messages Fig 1 6 also indicated on the display by asterisks appear for one second after an operation has been carried out successfully File Loaded Figure 1 6 An example of a confirmation message Error Messages Error messages Fig 1 7 indicated on the display with capital letters and exclamation points are shown whenever
98. lectron 01 2004 Flowcell lt Flowcell Inlet Outlet Tubing Clamp Rear of i Mounting Photodiode Assembly Thumbscrews DT Z048 FM 01 2004 Figure E 4 Detector with flowcell assembly exposed to show photodiode assembly mounting thumbscrews Thermo Electron 125 Flovvcell lt Flovvcell nlet Outlet Tubing Clamp Photodiode Assembly Mounting Thumbscrews DT Z049 FM Figure E 5 Detector with photodiode assembly repositioned to expose flowcell 126 Thermo Electron 01 2004 Using the Cuvette Holder The two procedures that follow allow you to use the Cuvette Holder to test the linearity of your detector s absorbance and to recalibrate the detector if necessary NOTE Be sure to insert the cuvette inside the holder so that its transparent EN sides rather than the frosted ones are in line with the beam of light from the detector s lamp Failure to insert the cuvette properly may result in insufficient light levels for accurate analyses ABSORBANCE You can use the optional Cuvette Holder and certified standards to LINEARITY test the absorbance linearity of your detector in the UV range approximately 235 to 350 nm HINT This procedure is particularly useful for laboratories that require periodic detector validation r To perform the test you ll need proced
99. length change autozero or run stop is to occur Allowable values range from 0 00 to 999 99 minutes Default is 0 00 minutes Wavelength refers to the wavelength that will be set at a specified time Allowable values for the UV1000 are 190 to 380 nm with the deuterium lamp and 366 to 800 nm with the tungsten lamp Allowable values for the UV2000 are 190 to 365 nm with the deuterium lamp and either 366 to 700 nm or 366 to 800 nm with the tungsten lamp depending on whether the detector is operating in the dual wavelength or the single wavelength mode respectively Default is 250 nm A preset time interval from the Ready state of the detector after which the detector lamp will turn off if a start signal has not been received from the keypad or external Run Input terminal Allowable values range from 0 30 to 9 59 hours Default is 1 00 This field along with the Copy File field allows you to copy a file to the specified file identification UV2000 only This field tracks the total number of hours the detector s tungsten lamp has been in operation up to 9999 When a new lamp is installed you must set this parameter to zero This command allows you to access the Wavelength Program See the Time Wavelength description above for details Thermo Electron 103 Zero This field appears in the Status Screen when the Scan file is loaded To initiate a background scan move the cursor to this selection and press ENTER UV2000 only
100. loads and runs the file designated in Order 1 It will continue to run this file each time it receives a start signal until the file has run the number of times specified in the Runs field The detector will then load and run the file designated in Order 2 and run it the number of times specified in that line and so on until the entire queue has run To view a queue s progress while it is running Press MENU 2 Select QUEUE Note that when a queue is loaded the Queue Menu Fig 4 27 looks different The Load field has been replaced by Pause which we will discuss on page 59 1 Delete Figure 4 27 The Queue Menu with a queue loaded Thermo Electron 01 2004 Loading other Files EDITING A QUEUE PAUSING A QUEUE DELETING STOPPING A QUEUE 01 2004 3 Select Edit to display the queue Refer to Figure 4 23 for an example queue While the queue is running the Runs field automatically decreases by one with each injection When a particular file s last injection is made the queue is automatically resorted In other words the information for Order 2 is now moved up to Order 1 the information for Order 3 is moved up to Order 2 and so forth This process continues until the queue becomes empty is paused or is deleted When a queue is loaded or running you may not load any other file from the Files Menu without first pausing or deleting the queue If you forget to pause or delete the queue and attempt to lo
101. m lamp D2 190 365 for the UV2000 s deuterium lamp W 366 800 for the tungsten lamp D2 W 190 800 for dual lamp operation UV2000 only or Off to shut the lamp s off Default is D2 190 380 or D2 190 365 for the UV 1000 and UV2000 respectively The Lamps Menu allows you to control the detector s lamp operations This field displays the analog to digital A D conversion frequencies of the light detected by the sample and reference diodes when the D2 lamp is on Thermo Electron 99 Load Load File Load Queue More OPTIONS Options Order Pause Pause Queue Program Protect QUEUE Range 100 Under the top level menu FILE S the Load selection accesses the Load File command Under the top level menu QUEUE the Load selection accesses the Load Queue field UV2000 only The Load File field loads the designated file settings into the active runfile After pressing ENTER the confirmation message File Loaded appears for one second The Load Queue field loads the queue After pressing ENTER the confirmation message Queue Loaded appears for one second UV2000 only This menu allows you to access the Zero on A Change Cursor Speed Status Lock and READY Output fields In the UV2000 you can also protect files from this menu Found in the Main Menu the Options Menu allows you to perform lamp and analog output operations The Options selection in the Edit Menu of FILE S allows you t
102. me Figure 4 9 The Scan File s Edit Menu 4 Select Setup to set up your spectral scanning parameters SETTING UP The Scan File s Setup Menu is shown in Figure 4 10 THE SCAN FILE Start A A Interval Run A Rise Time Scan Zero Time Range 1 Range 2 Figure 4 10 The Scan Files Setup Menu 01 2004 Thermo Electron 47 48 RUNNING THE SCAN FILE l Use these steps to set the parameters for scanning 1 In the Start field enter the wavelength at which each scan should start 2 Inthe End 2 field enter the wavelength at which each scan should end 3 In Interval enter the wavelength interval to be used To perform a scan the UV2000 takes individual absorbance readings at wavelengths incremented by the interval you specify HINT Five nanometers is an excellent wavelength interval for most applications At this interval you get very rapid scans and you can still display the to I nm accuracy 4 In Run 2 enter the wavelength at which the chromatographic run will be monitored 5 In Scan Zero Time enter the runtime at which you wish the detector to perform an automatic baseline scan If you use an automatic baseline scan make sure no peaks are eluting during the designated scan time 6 Fill in entries for Rise Time Range 1 and Range 2 as you vvould for any chromatographic run VVhen you are finished setting up the Scan File you are ready to load it and run When
103. me de vidange au niveau de la table de travail ou au dessous de celle ci La plupart des solvants doivent tre limin s dans des conditions particuli res et ne doivent pas tre vacu s directement par les canalisations Respecter la r glementation en vigueur concernant l vacuation des produits chimiques Systemes a haute pression et fuites Les systemes de chromatographie liquide CL fonctionnent a des pressions lev es Les liquides accumulent pas de grandes quantit s d nergie car ils ne sont pas hautement compressibles Par cons quent le risque d un danger imm diat caus par les pressions lev es dans un syst me CL est faible En revanche si une fuite survient il est n cessaire de la r parer le plus rapidement possible Enfin il est recommand l op rateur de se prot ger en permanence les yeux et la peau lorsqu il travaille sur un systeme CL De plus il doit toujours mettre le syst me hors tension et le ramener la pression atmosph rique avant de proc der tout entretien Thermo Electron 01 2004 ERKENNEN VON SICHERHEITS INFORMATIONEN 01 2004 Informationen zu Sicherheit und Funkentstorung Wir sind dem Dienst am Kunden und der Sicherheit des Kunden verpflichtet Diese Ger te entsprechen den Anforderungen f r die FCC Zulassung und f r das CE Zeichen sowie den Bestimmungen der Richtlinie f r Niederspannungsger te Dieses Ger t wurde in einer nach ISO 9001 zertifizierten F
104. meplate below the key Pressing ZERO resets the detector output to zero volts plus or minus any offset MENU Pressing MENU displays the Main Menu Figs 1 2 and 1 1 See page 4 for more information ENTER Pressing ENTER accepts a selected choice or menu entry The ENTER key also advances the cursor to a new field either on the same line of the display or in the line below Thermo Electron 3 MENUS SCREENS AND MESSAGES N NI lt and gt Pressing any arrow key up down left or right moves the cursor in the direction indicated on the key The up and down arrow keys also move the cursor between menus and displays and Pressing the and keys scrolls you through a field s available choices or changes the value of alphanumeric entries Holding down either key will continuously scroll the list of choices forward or backward until you release the key In fields that require numerical entries the value of each digit is increased or decreased by one unit each time you press the or key In fields that accept either numeric or character entries such as the File Name field the and keys scroll through the alphabet from A to Z then through the numbers 0 to 9 and finally to a slash hyphen and blank space In other fields the key advances you through a preset list of choices while the key takes you back through the list Your detector s display can show you three
105. meters you are ready to send the spectral data to your chart using the Replay Spectra command To initiate the Replay Spectra command in the Replay Menu press ENTER While the replay is occurring the screen shown in Figure 4 15 appears on the display Thermo Electron 01 2004 Stopping Replay SPECTRAL DATA STORAGE Viewing Data 01 2004 AU 0 00001 Figure 4 15 The display as it appears while spectra are being replayed The screen s Replay field displays the start time of the spectrum being replayed The A and AU fields display the individual data points being plotted The UV2000 uses advanced curve fitting algorithms to present a smooth continuous plotted spectrum The spectrum is replayed in 1 nm steps regardless of the wavelength interval selected To change the appearance of replayed spectra from 1 nm stepped curves to smooth curves or vice versa vary the recording device s replay rate and response time If no spectra are stored in memory when you activate the Replay Spectra command the message shown in Figure 4 16 will appear on the display When the replay is finished the display returns to the Replay Menu No Scans Stored Figure 4 16 The message that appears when no spectra are stored in memory You may stop a replay at any time by pressing STOP Spectral data are stored in the UV2000 s memory until a new file or queue is loaded or the detector is turned off You can display the individ
106. mp Hours W Lamp Hours Current Time Manual Startup Time 0 00 Shutdown Shutdown Time 0 00 Startup Manual Time from READY 1 00 Figure 4 5 The UV2000 s Lamps Menu Lamp The Lamp field allows you to select from the following D2 190 365 for deuterium the UV1000 reads D2 190 380 W 366 800 for tungsten D2 W 190 800 for dual lamp operation e or Off to shut the lamp s off Actually though the wavelength setting in the loaded file automatically selects the appropriate lamp for you In fact the wavelength setting you choose in your file has priority over any selection you make here in the Lamp field For example if the loaded file designates a wavelength in the UV range but you manually selected W 366 800 in the Lamp field the detector s display will read NRDY not ready for the deuterium lamp Thermo Electron 41 Lamp Hours VV and D2 fields These fields automatically track the number of hours each lamp has been in operation For the value to be accurafe set the appropriate Lamp Hours field to zero each time you install a new lamp HINT If you switch lamps before they are burned out with the intention of using them again at a later date keep a record of how many hours they have been in operation Startup and Shutdown When you set the Startup and Shutdown fields to Manual the lamp designated in the Lamp field turns on and off when the detector power is switched o
107. mplify calibration standardization of your UV2000 UV Vis detector using FDA industry or in house calibration standards The cuvette holder is a modular accessory that installs in place of the detector flowcell It allows analysis of calibration standards for example potassium dichromate to ensure your detector s compliance with FDA industry and or in house regulations To use the cuvette holder prepare your calibration standard according to the instructions provided with the sample Then place the sample in a standard 10 0 mm LD 12 5 mm O D quartz cuvette Analyze the sample and compare its measured maxima to its certified maxima If there s a discrepancy in the measured wavelength the detector can be recalibrated using the procedure described on page 114 DT Z045 DG Cuvette Photodiode Standoffs Mounting Screws Figure 1 Cuvette Holder accessory Thermo Electron 121 Installation The cuvette holder attaches to your detector using the standard flowcell mounting hardware Use the following steps to remove the flowcell and install the cuvette holder 1 Remove the front panel of the detector Fig E 2 to gain access to the flowcell mounting area Note that the front panel is a friction grip mount and will snap free if you pull outward on its lower edge 2 Remove the thumbscrew that secures the flowcell cover to the front of the detector Fig E 3 Remove the flowcell cover and set it aside
108. n File 47 Scrolling through choices 4 Select as a standard word 7 Self Tests 116 analog outputs 115 command described 102 deuterium lamp 115 diode offsets 115 lamp and shutter UV2000 only 115 motor 115 RAM 115 tungsten lamp 115 voltages 115 Setting parameters current time 42 Develop File 54 example of 55 File s Menu 20 26 replaying spectra 52 sample queue 57 scanning 47 Status Menu 24 31 Setup Menu general description of 102 illustration of 47 Short outputs accessing 43 command described 44 102 Shortcuts icon for 8 Shutdown detector accessing the command 45 command described 102 message described 112 restarting the detector 45 Shutdown lamps how 102 when 41 102 Signal to noise defined 119 Single wavelength operation analog output 2 UV2000 only 35 selecting 100 using 20 26 Slashes reason for 7 01 2004 Thermo Electron index ix Software version 102 113 Solvents disposal of wastes see Safety Information programming 40 119 used to clean the flowcell 71 Specifications flowcells 88 instrument 90 Spectra changing the appearance of 53 corrected with baseline scan 48 getting ready to replay 50 identification of 52 replaying 52 Spectra Time field defined 102 storage of 53 Spectral scanning see Scan File Spiking 71 Standard words 7 Start 102 Starting a run 23 30 from an external device 85 Startup fields defined 102 Status field defined 103 lock feature 43 103 term defined 119 STATUS key
109. n and off respectively Startup and Shutdown Times When you set the Startup and Shutdown fields to Time see above the designated lamp will automatically turn on and off at the local time set in the Startup Time and Shutdown Time fields respectively NOTE For the detector to perform automatic lamp startup and shutdown correctly the detector s 24 hour clock must be set to your local time Set the clock in the Current Time field Since the clock resets to zero each time the detector is turned off it will have to be reset prior to performing automatic lamp startup and shutdown unless the detector has been left on continuously Time from READY If you prefer you can program the detector to shut the lamp off after a series of automated runs by using the Time from READY feature Time from READY is a preset time interval that automatically begins each time the detector returns to its READY state If the Time from READY interval elapses without a run signal being received from either the keypad or the detector s Run Input terminal the detector s lamp turns itself off To use the Time from READY feature Select Time from READY in the Shutdown field 2 Inthe Time from READY field enter the length of time during which a run signal must be received by the detector before the lamp turns off For example let s say your chromatographic system is set up for an automated run and the autosampler signals the detector to run after each injecti
110. nd 44 connecting to terminal 85 field described 101 use of 44 state 23 30 time from ready field 42 Real time error messages 110 Recorders connecting to analog outputs 84 Remote communications 85 Replay fields described 101 rate 52 term defined 119 Replay Menu UV2000 only accessing the 52 replay fields described 101 replay spectra command 52 illustration of display 53 stopping replay 53 setting parameters 52 Reviewing parameters during arun 5 Rise time defined 119 field described 101 incorrect setting results of 109 suggestions for use 22 28 Run fields fields described 101 used in a queue 57 used in the Develop File 55 input terminal 85 shutting down lamps 42 parameters 20 27 programming during a 24 31 progress messages No Replay Allowed 112 No Scanning Allowed 112 Testing Not Allowed 112 ready signal for external devices 85 starting a example of 13 18 from an external device 85 technique described 23 30 stopping a example of 13 18 from an external device 85 technique described 23 30 the run file 20 the run file defined 119 time defined 119 RUN key example of use 13 18 functions of 3 Runfile defined 119 S S see Scan File 47 Safety certification see Safety Information definition of icons 8 general precautions see Safety Information nitric acid precautions 72 Sample Queue see Queue Save File command 24 32 Saving anentry 1 Scan File UV2000 only 46 accessing summary data 5
111. nd a subsequent decrease in sensitivity Thermo Electron 01 2004 Remote Your detector can accept inputs from and send inputs to remote Communications devices through the remote communications connector Fig A 4 Connections For example if your chromatographic system has programmable timed events contact closures or TTL you can use one to automatically zero the detector signal during a run The terminals available on the detector s remote communications connector are labeled STOP Input RUN Input ZERO Input and READY Output each with an associated ground terminal The terminals are labeled 12 through 1 STOP Input You can use a timed event from your chromatographic system to take the detector out of run by connecting the system s event to the detector s STOP Input and GROUND terminals terminals 12 and 11 RUN Input You can use the remote start event on your injector or autosampler to automatically put the detector into run whenever an injection occurs by connecting the event to the detector s RUN Input and GROUND terminals terminals 10 and 9 ZERO Input You can zero the detector signal automatically by connecting a timed event on your chromatograph to the detector s ZERO Input and GROUND terminals terminals 8 and 7 READY Output The detector is capable of driving one TTL load each time it goes to its READY state through the READY Output terminal This ability to signal other instruments
112. ng to the procedure outlined in Appendix A If the lamp is installed correctly its surface is clean and the message still appears replace the lamp The following error messages indicate improper use of the detector s menu system A File Is Already Running You cannot start a different file while a file is already running Invalid Parameters Spectrum Not Allowed Invalid scanning setup parameters have been entered so the detector cannot perform a spectral scan No More Available Memory All available system memory is full No Queue Available You cannot load a queue if none has been set up first Because queues are not available on the UV 1000 this message only appears on the UV2000 No Spectra Available You cannot run Replay Spectra when no spectra are available in memory Because there is no scanning feature on the UV 1000 this message only appears on the UV2000 Protected File Cannot Be Copied To You cannot copy to a protected file File protection is not offered on the UV 1000 so this message can only appear on the UV2000 Protected File Cannot Be Deleted You cannot delete a protected file File protection is not offered on the UV 1000 so this message can only appear on the UV2000 Protected File Cannot Be Edited You cannot modify a protected file File protection is not offered on the UV 1000 so this message can only appear on the UV2000 Queue Loaded Cannot Load File When a queue is loaded you cannot
113. ns in voltage on AC line 2 Random noisy baseline a Contaminated flowcell b Leak in sample inlet line c Bubble trapped in flowcell d Leaking flowcell 01 2004 Thermo Electron Remedy a Degas mobile phase Supply backpressure device to flowcell check back pressure rating Check for leaks at high pressure fittings Flush flowcell with 2 propanol then with mobile phase Check electrical lines for good connections and or interference from broadcast radiation Check for ground loops Remove systems e g ovens that cause voltage fluctuations isolate the detector to quiet circuit or use UPS UPS uninterruptible power supply Flush flowcell with cleaning solvents as described in Chapter 5 Check for leaks Check all fittings from column outlet to flowcell inlet for leaks Increase flow rate until bubble is removed Supply backpressure device to flowcell check back pressure rating to avoid rupturing flowcell Replace flowcell 107 Troubleshooting Table cont Symptom 2 Random noisy baseline cont d 3 Excessive baseline drift See Baseline problems 108 Cause e a Insufficient lamp warm up Lamp aging or defective Ground loop problem between integrator and detector Flowcell lamp lenses or photodiode dirty Integrator input voltage does not match detector output voltage Flowcell contaminated Mobile phase contamination Material
114. o 84 Invalid Parameters Spectrum Not Allowed message 111 J K K factor 60 algorithm for 60 allowable values of 98 calculation of 63 choosing wavelengths for 61 defined 118 operating in the K factor mode 63 setting analog output for 35 uses for 60 using Display AU A screen 63 using the Scan File 61 Keypad 3 118 illustration of 2 moving around 1 Keys description of 3 L Laboratory safety see Safety Information Lambda A fields defined 50 max IMax 50 maxAU IMaxAU 50 min IMin 50 Lambda A fields defined 99 Lamps aging 73 alignment illustrated 76 and shutter tests 115 changing the 73 77 Cover Open message 110 deuterium see Deuterium lamp 74 illustration of assemblies 75 installation of 76 77 lamp hours fields 42 98 lamp life 74 77 location of 73 removal of 74 77 selecting the 41 99 priority over 41 shutting down the at end of queue 43 automatically 42 manually 42 shutdown fields described 102 Time from READY 42 103 tungsten see Tungsten lamp 77 turning on the 102 automatically 42 manually 42 Lamps Menu accessing the 41 general description of 99 illustration of 41 LC test mix 116 LCD see Display Light Levels test 99 113 Linearity of response 116 127 Load fields described 100 used illustrated 01 2004 Thermo Electron index v Load command 12 16 Load File command 12 16 Load Queue command UV2000 only 58 Loading a file an example 12 16 technique described 22 29 whil
115. o edit Rise Time Autozero Time and Range This field designates the order in which the selected files in a queue will be run UV2000 only This field accesses the Pause Queue command UV2000 only This field pauses an active queue If a file is running it continues until completed and the detector returns to a READY state UV2000 only This field allows you to select single or dual wavelength operation The selection toggles between Single Dual 2 190 450 and Dual 366 700 Default is Single 2 This field along with the File Name field protects a specified file from being edited copied to or deleted The field toggles between On allowing no changes to the file and Off where changes may be made Default is Off UV2000 only The Queue Menu allows you to edit load delete or pause a queue A queue is a series of files that are run in a specific order and is typically used for automated runs UV2000 only The Range field controls the full scale output range for the UV1000 s Analog Output 2 terminal Allowable full scale ranges are 3 0 2 0 1 0 0 5 0 2 0 1 0 05 0 02 0 01 0 005 0 002 0 001 and 0 0005 AUFS Default is 1 0 AUFS UV1000 only Thermo Electron 01 2004 Range 1 Range 2 READY Output Replay Replay Spectra Replay Rate Rise Time Runs Run Run Time Runs per Scan Scan Zero Time 01 2004 The Range 1 and Range 2 fields control the full scale output ranges for the
116. o the chromatographic column and the outlet tubing to the waste reservoir 5 Taking care not to pinch the cable or tubing replace the flowcell housing and secure it with the knurled thumbscrew Replace the detector s front cover 6 Connect the power cord to the rear detector panel Thermo Electron 01 2004 DT Z020E FM CLEANING THE The exterior and or interior surfaces of the flowcell can become FLOWCELL contaminated When flowcell contamination occurs it is usually caused by precipitation or solubility problems such as when the quality of your mobile phase solvent components and the cleanliness of your samples are variable Signs of a contaminated flowcell are increased baseline noise signal spiking erratic or drifting baselines and increased backpressure Cleaning with If you suspect that your flowcell needs to be cleaned start with the Organic Solvents following procedure using organic solvents NOTE Flowcells are factory assembled units that should not be EN disassembled under any circumstance If you encounter contamination problems that are not remedied by this cleaning procedure contact your local Thermo Electron representative to arrange for repair or replacement 1 Make certain that the cleaning solvent s you plan to use is are miscible with the solvent already present in the flowcell and pump Isopropanol is a good choice for most applications NOTE If the last solvent in the pump was an aqueous buf
117. on With the detector settings shown in Figure 4 6 the lamp will turn off ten hours after the last run is completed Shutdown Time from READY Shutdown Time Time from READY Figure 4 6 An example of the Time from Ready feature Thermo Electron 01 2004 You can also program the UV2000 s lamps to turn off at the end of a queue by selecting End of Queue in the Shutdown field For more information on the Queue feature see page 57 Other Features STATUS LOCK SHORT OUTPUTS 01 2004 Additional features offered by the UV1000 and UV2000 include the abilities to lock the Status Screen to short the detector outputs to place an event mark on the chromatogram and to send a ready signal to external devices You can also control the display s contrast and cursor speed and do a quick shutdown of the detector s lamps and motors You can lock the detector s display using the Status Lock field This feature lets you prevent accidental changes to a file that is currently being run With the lock on in the UV 1000 only the Status Screen appears In the UV2000 you can scroll down from the Status Screen as far as the Status Menu s File Name field You will still be able to access the Main Menu and the rest of the menu structure using the MENU key however To access Status Lock 1 Press MENU 2 Select OPTIONS 3 Select More 4 Scroll down to Status Lock Select On or Off to turn the lock on or off respectively 5
118. ond delay from the time the detector takes its EN absorbance readings to the time you see the same reading on the analog readout Keep this in mind when choosing your scan times Each time you perform a sample scan the detector s monochromator moves from the run wavelength to the start wavelength The detector performs each scan from the start wavelength to the end wavelength by taking individual absorbance readings at wavelengths incremented by the interval you set in the Scan File When the scan is finished the monochromator returns to the run wavelength For example using the default Scan File Setup Menu shown in Figure 4 10 the detector would monitor the run at 250 nm Each scan would include absorbance readings for wavelength settings of 220 225 230 235 and so on up to 350 nm NOTE If you chose starting and ending wavelengths that were not an exact EN multiple of your wavelength interval the ending spike event mark on your chromatogram would be placed at the last multiple of the wavelength interval that falls within the scanning range For example with a starting wavelength of 200 nm an ending wavelength of 365 nm and a wavelength interval of ten the end spike on your chromatogram would be at 360 nm the last full wavelength multiple within the range While the detector is scanning the Status field displays SCAN NOTE During scanning the output signal will hold at the last absorbance EN value taken before
119. op File is loaded you will notice an additional field DEVELOP FILE in the Status Screen Runs Fig 4 19 Status READY Figure 4 19 The Status Screen with the Develop File loaded Runs The Runs field in the Status Screen shows the current run number followed by a forward slash and the total number of injections for the wavelength specified in the field For example if the file is set up to make three injections per wavelength and the detector is in the second run for the 250 nm setting the Runs field would appear as 2 3 The field is updated with each injection 01 2004 Thermo Electron 55 Status Menu N REPEATING THE DEVELOP FILE The Status Menu looks the same for a Develop File as it does for a typical chromatographic file Fig 4 20 DEVELOP Wavelength 250 250 Rise Time Autozero Time Range 1 Range 2 Figure 4 20 The Status Menu with the Develop File NOTE You can change any of the parameters in the Status Menu while the detector is running but the changes will be effective only until the next wavelength is loaded After the last wavelength is run the detector is reset automatically to the starting wavelength in the Develop File The file can be run as many additional times as you wish as long as the detector continues to receive run signals Sample Queue 56 QUEUE MENU Sometimes it s convenient to group samples together under different detector conditions in an automate
120. ou ve set your detector parameters in the edit file and have DETECTOR loaded the parameters into the run file you re ready to run your analysis First check the detector s status by viewing the Status Screen If you re using a stop line in your wavelength program you will start and stop the run with each injection Status Screen You can check the detector s status wavelength setting and absorbance reading from the Status Screen Fig 3 7 To access the Status Screen press STATUS Status AU READY 254 0 00001 W Figure 3 7 The UV1000 s Status Screen If the Status reads READY the detector is stabilized and ready to run If NRDY appears the detector s lamps may need additional time to warm up or a wavelength outside the selected lamp s range may have been chosen Inject your Sample When the detector is stabilized and you are ready to inject your sample first manually zero the detector by pressing the ZERO key If you are not using a stop line in the wavelength program the detector remains in the READY state throughout your chromatographic runs If you are using a stop line you must start and stop the run with each injection following the procedures below Starting a Run If you are using a stop line in your wavelength program you need to start the run with each injection There are two ways to start a run using the UV 1000 1 Manually by pressing RUN each time you make an injection 2 Automatically by inte
121. over Thermo Electron 01 2004 01 2004 Current Voltage Selection Drum Fuse Holder Q Voltage Selector DT Z026E DG Figure A 2 Voltage selector barrel and fuse holders Gently pry open the cover Once unlatched the cover will swing downward to reveal the voltage selector barrel and the fuses Remove the voltage selector barrel from the detector The selector resembles a wheel with four settings 100 120 220 and 240 V Fig A2 Rotate the barrel such that the desired voltage setting will be visible through the cut out in the cover when it is replaced Replace the barrel in the detector Before closing the cover check the fuses according to the procedure on page 82 Thermo Electron 81 Fuses To verify that your detector is fitted with the correct fuses follow these steps If you haven t already done so first open the voltage selector cover according to step 1 in the Voltage Selection procedure listed above 1 Pull each fuse holder straight towards you The fuse holders are the black squares with arrows located directly beneath the voltage selector Fig A 2 2 Remove each fuse from its holder Check the fuse amperage voltage and type according to the following description You should have either e two T2A 250V fuses for 100 120 VAC operation or e two T1A 250V fuses for 220 240 VAC operation
122. priate all allowable and default values Menu Trees The UV1000 and UV2000 Menu Trees are useful tools for learning your way around your detector You may wish to keep one handy while you learn where each display is located in the overall menu structure 01 2004 Thermo Electron 93 S S L 4 S IH indino AqvaM sesyo bo jf HO 3907 sn e S S A 7 44617 EN UNIPON p ds Josin9 UOISI9 ACMYOS g a ueyg y uo 0107 Sissi OJON OL 95 si z Boleuy OL AW s40 Boleuy s nd no Bojeuy 8 SJON s nd no Bojeu 00 1 Apeoy wol l ndino uni Ol abuey 00 0 aw L 000 eu 2 jenueyy uMOP NYS SNOI LdO eWIL S M 00 0 aul dnyels 092 00 0 jenueyy dnueis ufuejeaen eu 00 0 aw L 14 V00000 092 AQV3M 0 s noy d le7 M nv Y smes 0 s noy dwe 20 086 061 za dwe sn v s sdwe Sisal SNOIL4O 3713 J lli nam 2 99 40 99 9q UMOPInyS ndino ous HEIN 1 01 abuey SOANVWWOI 0000 ls zomny _ OL Sul siM ll i peo suondo peo Saas peo xpa 3714 suondo 092 0070 U 6U l ABAA uIdusjsnem Suu YBU M ii S n u IN 0 0 0 L A N 01 2004 Thermo Electron 94 alo z abed s
123. produce a baseline scan that is heightened artificially Thermo Electron 51 52 REPLAYING YOUR SPECTRA Setting Replay Parameters Running Replay When you have completed your run you can retrieve your stored sample spectra using the Replay Menu Fig 4 14 To access the Replay Menu 1 Press MENU Select FILES 2 Select Edit to display the Scan File s Edit Menu Fig 4 9 3 Select Replay Range 1 Range 2 Replay Rate nm sec Spectra Time Replay Spectra Display AU A Figure 4 14 The Replay Menu To set the parameters for replay 1 Set Range 1 and Range 2 for Analog Output 1 and Analog Output 2 If you are using only one output disregard the appropriate range 2 Enter the Replay Rate nm sec This is the rate at which the detector will read out the spectral data to your chart You will use this value and an appropriate chart speed to calculate wavelength increments on your printed sample spectrum For example if your sample scan were taken between 190 and 340 nm a span of 150 nm a replay rate of 5 nm sec would print the spectrum in 30 seconds A chart speed of 30 cm min would give you a scan of 15 centimeters in increments of 10 nm cm 3 Select the spectrum you want to replay by selecting its start time in the Spectra Time field Each spectrum taken during the run is individually identified by the run time at which it was initiated When you finish setting your replay para
124. r cord 82 requirements 80 switch location of 10 14 Preset choices selecting from 4 Preventative maintenance 67 Program field 27 100 Programmed autozero 39 40 example of 40 illustration of 40 Protected files 34 100 copying to 33 deletion of 34 illustration of 34 messages Cannot Be Copied To 33 111 Cannot Be Deleted 34 111 Cannot Be Edited 111 Pumps used to clean flowcells 71 72 Purity determinations 35 64 Q Q see Queue Queue UV2000 only 56 a Q in the display 58 103 defined 118 deleting a line 57 deleting a queue 59 98 description of 56 editing a 59 example of 57 field described 100 file name 57 illustration of an empty queue 57 lamp shutdovvn at end of 43 Loaded message 59 loading a 58 99 messages No Queue Available 58 111 Queue Deleted 60 Queue Loaded 58 100 Queue Loaded Cannot Load File 111 monitoring while running 58 number of files per queue 57 number of injections per file 57 order 1 58 order field 57 100 pausing a 59 100 resuming after a pause 59 running a 58 setting up 57 Status Screen 58 stopping a 59 01 2004 Thermo Electron index vii Queue Menu accessing the 56 general description of 5 illustrations with no queue loaded 57 with queue loaded 58 pause field 58 100 R RAM defined 119 testing 115 Range example of use 11 15 fields defined 100 setting the 22 29 term defined 119 Ratioing absorbance values UV2000 only 35 64 READY output accessing the comma
125. r analysis Remember that each digit must be edited individually 4 Press ENTER to accept the new wavelength setting Range Range is an example of a field that gives you a preset list of choices Note that Range 1 and 2 correspond to Analog Outputs 1 and 2 on the rear panel of your detector To set the range 1 Select Options from the Edit Menu Fig 2 14 to display the Options Menu Fig 2 16 Rise Time Autozero Time Range 2 Figure 2 16 The UV2000 s Options Menu 01 2004 Thermo Electron 15 16 Loading the File A PRACTICE RUN 2 Scroll down in the Options Menu and move the cursor to Range 1 using the v key 3 Using the or key select the desired setting from the list of choices 4 Press ENTER to accept the new Range 1 setting We will use the rise time autozero time and range 2 default settings for this example You will learn more about setting these parameters in Chapter 3 You are now ready to load the settings from File 1 into the detector s operating parameters To load the file 1 Return to the Files Menu Fig 2 13 by pressing either ENTER or the v key 2 Select Load The screen in Figure 2 17 appears gt Load File 1 filename Figure 2 17 The Load File command 3 You will be able to select from among several files in the Load File field Depending on whether or not your detector has ever been used before these files will either contain previously stored set
126. r use with your data system or integrator UV2000 Single and Dual wavelength Operation SETTING PARAMETERS 26 You can operate the UV2000 in either a single or a dual wavelength mode In the dual wavelength mode the detector simultaneously monitors two wavelengths in either the UV range or the visible range in a single run To perform a single or dual wavelength operation you need to be able to identify and enter a file load that file into the detector s current operating parameters and start and stop a run This section will also show you how to modify the detector s current operating parameters Before you set any detector parameters you need to access the Files Menu to identify the file you wish to edit To access the Files Menu first press MENU The Main Menu appears on the screen From the Main Menu select FILES The menu shown in Figure 3 12 will appear QO Load O Delete Figure 3 12 The UV2000 s Files Menu Thermo Electron 01 2004 File Identification Wavelength Program 01 2004 Select Edit from the Files Menu to display the Edit Menu Fig 3 13 Edit File File Name Q Wavelength Program Q Options Figure 3 13 The UV2000 s Edit Menu Enter the number of the file you wish to edit in the Edit File field The UV2000 can store up to four files in memory so file numbers from 1 to 4 are allowed You may also enter a name of up to eight characters in the File Name field Wh
127. raphic instrumentation As you go through unpacking and installation you may want to use the Start up Checklist located at the beginning of this manual The checklist is an abbreviated version of this Appendix and is supplied as a quick reference of how to conduct a successful installation After installation verify that the detector is working properly by running the two tests described on page 116 Also included in this Chapter is a list of your detector s specifications m Installation UNPACKING Carefully remove the detector from the shipping container and inspect both the detector and packing for any signs of damage If you find any damage immediately contact the shipping company The shipping container should contain the detector an accessory kit any options you ordered for your detector and this manual The accessory kit should contain the following items 8 pin connector 12 pin connector e Nut and ferrule tubing set e Teflon tubing 1 16 nut fitting 1 4 28 thread 1 16 ferrule fitting e Union e External run autozero cable e Analog cable e Extra cap screws 2 e LC test mix vial 3 foot 4 conductor cable 01 2004 Thermo Electron 79 CHECKING THE POWER Voltage Selection 80 Carefully check to make sure you received all the items listed on the packing list If any items are missing contact your Thermo Electron representative immediately You will need the following tools for installat
128. rfacing the detector with a remote run signal from the injector see Appendix A for details In this scenario a signal that is equivalent to pressing RUN is automatically sent from the injector to the detector with each injection During the run you can monitor the run time from the Status Screen Stopping a Run There are two ways to stop a run 1 Manually by pressing STOP before the programmed stop time 2 Automatically by allowing the run to finish at the programmed stop time In either case the detector returns to its READY state 01 2004 Thermo Electron 23 24 CHANGING RUN PARAMETERS Status Menu There are two ways to change the detector s run parameters 1 You can use the Files Menu and follow the procedures outlined under Setting Parameters on page 20 2 Or you can use the Status Menu which is the programming area belovv the Status Screen Each method has a distinct advantage Programming in the Status Menu allows you to change the detector s current operating parameters even while the detector is running Programming in the Files Menu allows you to prepare an edit file containing the changes without altering the current detector settings The file may then be loaded later From the Status Screen scroll down to the Status Menu Fig 3 8 The Status Menu contains the Wavelength Program Rise Time Autozero Time and Range Wavelength Rise Time Autozero Time Range Figure 3 8 Th
129. rmines how you make a selection e Ifa triangular Cursor appears press ENTER e If a blinking square cursor appears press the or keys to change values Depending on the field you will scroll up or down through preset choices or change alphanumeric entries one letter or digit at a time 3 There are four ways to accept and automatically save an entry Just move the cursor out of the field by any of the following methods Pressing ENTER e Using the arrow keys Pressing MENU e Pressing STATUS NOTE You won t be able to leave a menu if errors are present or if you haven t filled in all the necessary entries Thermo Electron VISUAL CLUES The following conventions are used on the detector s display 1 Top level menu choices are displayed in all capital letters 2 A field s square cursor changes to an underscore cursor when you re scrolling through preset choices or entering numerical values and characters 3 A solid down arrow on the right side of some displays indicates that the current menu continues on additional screens To access additional menu lines press the down arrow key v 4 The last line of a longer menu is frequently a blank display line without a solid down arrow Instrument Control Take a look at the keypad and two line display located on the front panel Fig 1 1 This is the command center from which you ll access menus and control the instrument s operations
130. rogram to display the Wavelength Program Fig 2 5 Time Wavelength 0 00 254 Figure 2 5 The UV1000 s wavelength program 2 Using the and keys edit the wavelength field to the desired setting for your analysis Remember that each digit must be edited individually 3 Press ENTER to accept the new wavelength setting Range Range is an example of a field that has a preset list of choices To set the range 1 Select Options from the Edit Menu Fig 2 4 to display the Options Menu Fig 2 6 Rise Time Autozero Time Figure 2 6 The UV1000 s Options Menu 2 Scroll down in the Options Menu and move the cursor to Range 1 using the v key 3 Using the or key select the desired setting from the list of choices 4 Press ENTER to accept the new range setting We will use the rise time and autozero time default settings for this example You will learn more about setting these parameters in Chapter 3 01 2004 Thermo Electron 11 Loading the File You are now ready to load the settings from the edit file into the detector s operating parameters its run file To load the file 1 Return to the File Menu Fig 2 3 using the A key 2 Select Load The screen in Figure 2 7 appears gt Load File Figure 2 7 The Load File command 3 Press ENTER to execute The confirmation message shown in Figure 2 8 appears for one second File Loaded Figure 2 8 The file loaded message You
131. round 84 Flowcell bubbles in the 107 108 changing the 67 cleaning the precautions during 71 safety precautions 72 using a pump 71 72 using a syringe 71 72 windows 69 with nitric acid 72 with organic solvents 71 connecting to inlet outlet lines 86 contamination of 71 different types of 88 disassembly of 67 installing the 70 leaks 107 location of 67 maintenance of 67 orientation for non Finnigan SpectraS YSTEM detectors 88 removing flowcell and housing 68 specifications 88 Fluid connections 86 Front panel illustration of 2 Fuses changing the 82 illustration of fuses 82 illustration of holders 81 requirements 82 G Glossary 117 Good laboratory practices see Safety Information Gradient elution 48 118 index iv Thermo Electron 01 2004 Ground loop problems causes of 108 Ground terminal 118 hooking to recorders 84 Grounding 1 H I et High voltage warning 8 Hints defined 8 Icons how they re used 8 Identification of files 27 99 Injecting a sample 23 30 Input errors 111 Installation accessory kit 79 analog outputs 84 connecting to recorders 84 connecting to inlet outlet lines 86 flowcell connections 86 fluid connections 86 fuses 82 illustration of 82 illustration of holders 81 requirements 82 positioning the detector 80 power cord 82 power requirements 80 rear panel connections 83 tools for 80 unpacking 79 voltage selection 80 Instrument control 2 Integrators Workstations rear panel connections t
132. rovides digital processing circuitry and interfaces with the keyboard display and the remote communications devices Additional software is held on an EPROM PCB The Motherboard provides all the necessary interconnections and power supplies Lamps N Shutter N Baffles B Reference eam Photo Splitter y diode amp Lens Sample Cell Sample Photodiode DT Z001E FM 106 Figure 1 The optical system for the UV1000 and UV2000 detectors Only the UV2000 has a shutter The deuterium and tungsten lamps are continuum light sources that provide high light intensity over the UV and visible wavelength ranges Two sets of baffles minimize stray light A concave holographic grating actuated by a microprocessor controlled stepper motor provides wavelength selection Thermo Electron 01 2004 Common Problems This next section contains a table of symptoms possible causes and remedies for some common problems you may observe in detector response Many of the problems attributed to the detector may actually be due to other components in the chromatographic system so we have included references to these types of problems and solutions as well Troubleshooting Table Symptom Cause 1 Spikes on baseline a Gas bubbles in the flowcell b Immiscible solvent bubbles following mobile phase changeover c Electrical interference d Extremely large fluctuatio
133. s adds a second line with a time of 1 00 and the same wavelength setting as the first Change 1 00 to the desired stop time for the run and leave the wavelength unchanged 3 To save your edits scroll down to the words Save File which now appear below Range and press ENTER The confirmation message shown in Figure 2 10 appears and you are automatically returned to the Status Screen File Saved Figure 2 10 The file saved message RUNNING WITH Now that you have entered a stop time you will need to start the run A STOP TIME with each injection 1 Zero the detector s analog output signal by pressing the ZERO key 2 When the detector is stabilized inject your sample and press RUN Notice that Status now shows the run time If you wish to stop your run before the set stop time simply press STOP 01 2004 Thermo Electron 13 UV2000 An Example In this example specifically designed for the UV2000 we will show you how to prepare a file and how to load it into the detector s operating parameters After a practice run we will add a stop time To keep the instructions simple we will use the single wavelength mode HINT You may wish to keep the Menu Tree in Appendix B on hand as you work through this example If you lose your place at any time you can 1 Press the A key to move back to a previous screen 2 Or press STATUS to return to the Status Screen and retrace your steps STARTUP Se
134. se this equation to calculate the number of data points for any scan between Al the lower wavelength and 22 the higher wavelength 2 of data points 1 interval Equation 2 Use this equation to calculate the number of spectra you can collect when using wavelength intervals of 2 nm or greater Round the resulting number down to the nearest integer 5000 of data points 12 of spectra of data points 4 14 Equation 3 Use this equation to calculate the number of spectra you can collect when using wavelength intervals of 1 nm Round the resulting number down to the nearest integer 5000 of data points 4 of spectra of data points 4 14 Thermo Electron 01 2004 For example if you want to scan from 190 to 564 nm in 2 nm steps there would be 188 data points and the UV2000 would be able to store up to 3 spectra 5000 188 12 2744 of spectra 3 58 3 188 4 14 766 Each scan is corrected for baseline absorbance before being played back either as individual data points or as a smoothed continuous spectrum SELECTING To select spectral scanning follow these step by step instructions THE SCAN FILE Press MENU Select FILES Select Edit Use the key to increment the Edit File field until an S is displayed Fig 4 9 The File Name field is automatically named SCAN You cannot edit the Scan File s name Edit File File Na
135. sequently erased from the detector s memory It is good practice to delete an existing queue prior to designing a new one K Factor The K factor calculates a factored response that can be used to eliminate add or subtract absorbances This technique is useful for suppressing peaks when there are two co eluting or poorly resolved peaks in your chromatogram It is also useful in applications where you want to add or subtract absorbances at two different wavelengths in real time For example if you want to quantitate a peak without interference from another peak you would use the K factor to calculate a response of zero More specifically let s say you want to analyze for Compound A in the presence of Compound B If both absorb at the monitoring wavelength 41 but only Compound B absorbs at a second wavelength 2 2 you can calculate a K factor for Compound B using its absorbances at and A2 You can then use the K factor to calculate the absorbance due only to Compound A at the monitoring wavelength 2 1 by subtracting Compound B s contribution from the total absorbance The UV2000 uses the algorithm Absorbance due to A at Al TAbs A1 K x TAbs A2 where TAbs A1 is the sum of the absorbances of A and B at the monitoring wavelength K is the K factor and TAbs 22 is the total absorbance obtained at 22 Thermo Electron 01 2004 AN EXAMPLE Figure 4 30 shows a chromatogram of a mixture of toluene
136. slow Flowcell volume too large Noise comes from grating motor and is normal Tripped circuit breaker at AC wall outlet Blown detector fuse Incorrect voltage selected Power cord not connected Thermo Electron Remedy a Check wavelength setting Make sure the correct file is selected Make sure lamp is lit Run detector s diagnostic tests to check lamp Replace lamp if necessary Connect integrator to appropriate output connectors on detector see Appendix A Check attenuation setting on integrator Check entire chromatographic system for leaks Verify sample injection volume Lower the rise time selection Change to a flowcell with smaller volume No action necessary Resolve problem reset circuit breaker Resolve problem replace fuse Reset detector for correct incoming line voltage see Appendix A Connect power cord 109 Error Messages There are three types of error messages that you may see on your detector s display e System errors e Real time errors User input errors Each type of error is explained below in further detail SYSTEM ERRORS System errors are indicated on the display by a double set of exclamation points 11 11 They occur whenever an undesirable condition exists that prevents the detector from operating If one of these messages appears try turning the detector s power switch off and on If the message recurs contact your T
137. so tells you how to avoid contact with the heated surfaces in your instrument Note Notes alert you to the correct operating or maintenance procedures needed to prevent equipment or data damage They also alert you to important exceptions side effects or unexpected occurrences that may result from certain action s Hint Hints call out general rules or shortcuts They specify ways to obtain the best performance and results from your instrument What s Next Now youre ready to try the practice example in Chapter 2 A Quick Example Thermo Electron 01 2004 2 A Quick Example Introduction THE UV1000 THE UV2000 BEFORE YOU BEGIN 01 2004 In Chapter 1 you read about the three easy rules for using your detector s command center and some of its menus and screens In this chapter you will find an example procedure that shows you how the rules and keys actually work as you move through the various menus Instructions begin on page 10 for the UV1000 and on page 14 for the UV2000 This quick example uses only a fraction of the features available on your detector and is included only as a first step in becoming familiar with your new instrument After experimenting with this example you ll want to turn to Chapters 3 and 4 which cover the detector s basic and more advanced operations It is in those chapters that you ll learn about the full capabilities of your detector First though to give you a general unders
138. sorbance reading you got from Analog Output 1 in single wavelength operation or the absorbance reading of wavelength two 2 2 for dual wavelength operation e AUI1 K AU2 which is the readout of the suppressed signal using the K Factor technique See page 60 for more details e AU1 AU2 which is the ratio of the dual wavelength absorbance values This ratio is sometimes used to check peak purity See page 64 for more details 01 2004 Thermo Electron 35 ANALOG OFFSETS Both analog outputs 1 and 2 can be offset on the UV2000 Analog offsets may be used in cases where there is a high background absorbance reading or when there is considerable baseline drift from your chromatographic system and you are unable to keep your integrator s recorder s signal on scale Because integrators have very limited capacity for handling negative signals you may wish to set a small positive offset 1 when using an integrator Negative offsets are available for use with recorders where you may wish to set the pen at either side of the strip chart The offset options are selectable from the Analog Outputs Menu shown in Figure 3 29 HINT Although the offset for each output is set at 0 of full scale readout by default we recommend a 1 setting for use with your data system or integrator 36 Thermo Electron 01 2004 4 Advanced Operations Introduction In this chapter you will learn to use the more advanced capabilities of your d
139. ss MENU 2 Select TESTS 3 The Tests menu appears in Figure C 2 Software Version Light Levels Diode Offsets A Calibration Self Tests Figure C 2 Detector s Tests Menu Thermo Electron 01 2004 Software Version Light Levels Diode Offsets Calibration 01 2004 Select this field to display the EPROM version of your detector s software Fig C 3 Version 1 01 Figure C 3 The software version The Light Levels test displays numbers related to the level of light intensity seen by the sample and reference photodiodes When you select Light Levels the screen in Figure C 4 appears SI nnnnn n R1 S2 R2 Figure C 4 The Light Levels screen The sample S1 S2 and reference R1 R2 numbers may differ considerably between instruments A five or six digit number is typical If you get an unusual reading check the photodiodes and the analog PCB These components are the ones that are the most likely to affect light intensity If any of the numbers is zero call Thermo Electron The Diode Offsets test presents numbers related to the level of background signal received from the sample and reference photodiodes when the lamps are off dark current When you select Diode Offsets the screen in Figure C 5 appears gt C 81 1 592 2 Figure C 5 The diode offsets screen The sample S1 S2 and r
140. standards to test the absorbance linearity of your detector in the UV range approximately 235 nm to 350 nm For your convenience the following procedure is also detailed in Appendix E HINT This procedure is particularly useful for laboratories that require periodic detector validation To perform the test you will need procedure E 925 from the American Society for Testing and Materials ASTM and standard potassium dichromate SRM 930 from the National Institute of Standards and Technology NIST formerly the National Bureau of Standards NBS The test involves the preparation of acidic solutions of potassium dichromate at four concentrations and the absorbance measurement of each solution at four wavelengths between 235 and 350 nm After correcting for an absorbance blank the linearity deviation of a plot of absorbance versus concentration should be less than 1 If you want more information on this test or find that your instrument does not conform to these specifications and requires service contact your local Thermo Electron representative Thermo Electron 01 2004 D Glossary Introduction A C A D AUFS absorbance analog offset background scan baseline chronometer D F defaults develop file degassing diagnostics display 01 2004 We have included a glossary to define certain technical terms used throughout the manual s text These terms are consistent with standard definitions used througho
141. stead reload the unaltered file using the Files Menu as follows 1 Press MENU 2 Select FILES 3 Select Load 4 The words Load File will appear on the screen Enter the desired file number and press ENTER A confirmation message Fig 3 17 will appear for one second You are then returned to the Status Screen and all settings will contain their original values More about Files COPYING FILES 32 On page 26 you learned how to edit and load files from the Files Menu The UV2000 also allows you to copy and delete files and to protect files from being edited copied to or deleted in a few easy steps To copy a file 1 Press MENU 2 Select FILES to display the Files Menu Fig 3 21 4 Load Q Delete Figure 3 21 The UV2000 s Files Menu Thermo Electron 01 2004 DELETING FILES 01 2004 3 Select Copy The Copy Menu will appear on the screen Fig 3 22 gt Copy File 1 filename 4 To File 2 filename2 Figure 3 22 The UV2000 s Copy Menu Enter the identification number for the file you wish to copy in the Copy File field Enter the number of the file to which you wish to copy in the To File field Press ENTER The confirmation message shown in Figure 3 23 appears briefly and you are returned to the Files Menu File Copied Figure 3 23 The message that s displayed when a file is copied If you attempt to copy to a protected file see the section belo
142. stop time HINT You may wish to keep the Menu Tree in Appendix B on hand as you work through this example If you lose your place at any time you can 1 Press the A key to move back to a previous screen 2 Or press STATUS to return to the Status Screen and retrace your steps STARTUP Set the povver svvitch located on the detector s rear panel to On After a series of power up tests the Status Screen Fig 2 1 appears on the display We will discuss the Status Screen after you have set up your operating parameters Status AU READY 250 0 00001 Figure 2 1 The UV1000 s Status Screen SETTING To set your parameters you need to prepare an edit file The PARAMETERS following steps will show you how to access the Edit Menu and prepare the file 1 Press the MENU key The detector s Main Menu appears on the screen Fig 2 2 gt FILE 14 COMMANDS Q OPTIONS Q TESTS Figure 2 2 The UV1000 s Main Menu 2 Now select FILES to display the Files Menu Fig 2 3 gt Edit 34 Load O Delete Figure 2 3 The UV1000 s Files Menu 10 Thermo Electron 01 2004 3 Select Edit to display the Edit Menu Fig 2 4 gt Wavelength Program Q Options Figure 2 4 The UV1000 s Edit Menu Wavelength You use the Wavelength program to set the monitoring wavelength Wavelength is an example of a field that requires a numeric entry To set the wavelength 1 From the Edit Menu Fig 2 4 select Wavelength P
143. sulter 6 Ne pas mettre l instrument sous tension si celui ci a subi des dommages lectriques D brancher le cordon d alimentation de l appareil et consulter un repr sentant du service technique pour proceder un examen du produit Ne pas essayer d utiliser instrument avant qu il n ait t examin Des dommages lectriques peuvent s tre produits si le systeme montre des signes visibles d endommagement ou si les conditions de transport ont t extr mement difficiles 7 L instrument peut galement tre endommag s il est entrepos pendant une p riode de temps prolong e dans de mauvaises conditions par exemple s il est expos a la chaleur a Vhumidit etc 8 Toujours d brancher le cordon d alimentation avant d effectuer n importe quel type d entretien 9 Les condensateurs pr sents l int rieur de I instrument peuvent toujours tre charg s meme si l instrument est hors tension 10 Ne jamais tenter de r parer ou de remplacer un composant du systeme non d crit dans ce manuel sans obtenir de l aide aupres d un repr sentant du service technique Bonne tenue des dossiers Pour permettre d identifier et d isoler les probl amp mes pouvant survenir avec l quipement ou la m thodologie utilis s il est recommand de tenir correctement des dossiers de toutes les conditions du syst me p ex CV sur les temps de r tention et les zones de pics la forme et l
144. sure safe operation N CAUTION Do not override the lamp cover safety interlock which 01 2004 turns the lamps off when the cover is removed or personal injury could result 5 Never run the system without the top cover on Permanent damage can occur Thermo Electron Vil GOOD LABORATORY PRACTICES viii 6 Do not turn the instrument on if you suspect that it has incurred any kind of electrical damage Instead disconnect the power cord and contact a Thermo Electron Service Representative for a product evaluation Do not attempt to use the instrument until it has been evaluated Electrical damage may have occurred if the system shows visible signs of damage or has been transported under severe stress 7 Damage can also result if the instrument is stored for prolonged periods under unfavorable conditions e g subjected to heat water etc 8 Always disconnect the power cord before attempting any type of maintenance 9 Capacitors inside the instrument may still be charged even if the instrument is turned off 10 Never try to repair or replace any component of the system that is not described in this manual without the assistance of Thermo Electron Keep Good Records To help identify and isolate problems with either your equipment or your methodology we recommend that you keep good records of all system conditions e g RSDs on migration times and peak areas peak shape and resolution At a minimum keep an
145. t the power switch located on the detector s rear panel to On After a series of power up tests the Status Screen Fig 2 11 appears on the display We will discuss the Status Screen after you have set up your operating parameters Status AU READY 0 00001 VW Figure 2 11 The UV2000 s Status Screen SETTING To set your parameters you need to prepare an edit file The PARAMETERS following steps will show you how to access the Edit Menu and prepare the file 1 Press the MENU key The detector s Main Menu appears on the screen Fig 2 12 Q QUEUE Q TESTS Q COMMANDS Q OPTIONS Figure 2 12 The UV2000 s Main Menu 2 Now select FILES to display the Files Menu Fig 2 13 QO Load O Delete Figure 2 13 The UV2000 s Files Menu 14 Thermo Electron 01 2004 3 Select Edit to display the Edit Menu Fig 2 14 Edit File File Name gt Wavelength Program Q Options Figure 2 14 The UV2000 s Edit Menu For this example we will use a file designation of 1 and leave the File Name field blank Wavelength Wavelength is an example of a field that requires a numeric entry To set the wavelength 1 From the Edit Menu Fig 2 14 select Wavelength Program to display the Wavelength Program Fig 2 15 Program Wavelength 254 Figure 2 15 The UV2000 s wavelength program Scroll down to the wavelength field Using the and keys edit the wavelength field to the desired setting for you
146. tanding of the detectors capabilities and design we will briefly describe the features and benefits of the UV1000 and UV2000 here The UV1000 detector is a time programmable variable wavelength UV Vis ultraviolet visible absorbance detector It operates in single wavelength mode in either the UV range using a deuterium lamp or in the visible range with an optional tungsten lamp The UV1000 s optical system has a novel high light throughput design that provides high sensitivity detection along with maximal application versatility The UV 1000 detector can be upgraded to a UV2000 The UV2000 detector is a full featured time programmable dual wavelength UV Vis absorbance detector It operates in both single and dual wavelength modes in the UV and visible ranges The UV2000 offers the same optical system design as the UV1000 In addition to the features of the UV 1000 UV2000 also offers spectral scanning a Develop File for method development multiple file storage a Queue feature that allows you to link files and more Once the detector is installed in your chromatographic system according to the procedures described in Appendix A and you have completed the Startup Checklist you are ready to begin your quick example Thermo Electron UV1000 An Example In this example we will show you how to prepare an edit file and how to load the edit file into the detector s run file After a practice run we will add a
147. tector automatically at the start of each run This will keep the detector output in range throughout an automated series of runs r Range 1 and 2 These parameters range the signal from Analog Output 1 and Analog Output 2 shown as ANLG 1 Output and ANLG 2 Output on the detector s rear panel Set each range to an appropriate full scale absorbance for your sample For more information on the use of ranges and analog outputs see pages 35 and 84 HINT We recommend a range of 1 0 when you are using an integrator or data system Loading a File When you are ready to load a file into the detector settings select Load from the Files Menu The screen vvill display the vvords Load File 1 filename Use the keys to view the number and name of available files When the desired file number appears press ENTER The confirmation message shown in Figure 3 17 will appear for one second You are then returned to the Status Screen File Loaded Figure 3 17 The message that s displayed when a file is loaded NOTE When a dual vvavelength program is loaded you ll hear the motor NI start to operate in dual wavelength mode even though you didn t press RUN RUNNING YOUR Once you ve set your detector parameters in the designated file and DETECTOR have loaded the file into the detector s operating parameters you are ready to run your analysis First check the detector s status by viewing the Status Screen
148. tector s rear panel and make sure that the instrument is turned off CAUTION Hot Surface Avoid burns Always allow sufficient time for the lamp to cool before removing it If you have not already done so remove the detector s front panel by grasping the bottom of the panel firmly with one hand and pulling back Remove the lamp housing by loosening the thumbscrew and pulling the cover straight back to expose the lamp assembly Fig 5 7 Unplug the tungsten lamp lead from the detector taking care not to twist the connector as you gently pull it out Loosen the thumbscrew and the aluminum standoff that hold the lamp assembly in place and pull the assembly straight out W Lamp Installation These five steps explain how to replace the tungsten lamp 1 01 2004 Hold the lamp assembly so that the leads are at the top Slide the assembly onto the two alignment pins shown in Figure 5 7 The alignment pins are located on either side of the detector s monochromator aperture Securely fasten the assembly in place with the thumbscrew and aluminum standoff Connect the lamp lead to the left hand terminal in the lamp compartment Replace the lamp housing and fasten securely with the thumbscrew Connect the power cord to the rear detector panel Thermo Electron 77 A Installation and Specifications Introduction This Appendix covers the initial installation of your UV Vis detector including hookup to other chromatog
149. the scan was initiated until the scan is finished For this reason quantitative analysis should never be performed when scanning 01 2004 Thermo Electron 49 Scan Summary Data Screen STOPPING THE SCAN FILE When the Scan File is loaded the normal Status Menu no longer appears below the Status Screen Instead several new lines that we call the Scan Summary Data Screen appear The Scan Summary Data Screen is useful in setting up the parameters to replay your stored spectra An example of the Scan Summary Data screen as it appears after two sample scans is shown in Figure 4 12 File S SCAN Figure 4 12 An example of the Scan Summary Data Screen The Scan Summary Data Screen has four fields e Time which is the run time at which the scan was initiated e Max which is the scan wavelength where the maximum absorbance occurred AMaxAU which is the maximum absorbance e Min which is the scan wavelength where the minimum absorbance occurred If no maximum was found the AMax and AMaxAU fields read 0 zero The summary information is updated as each sample scan is completed NOTE The UV2000 uses a second derivative to find the local XMax In our example Fig 4 12 scans were taken at 10 50 and 11 66 minutes into the run The scan taken at 10 50 minutes has a maximum absorbance of 1 6668 AU at 280 nm The minimum absorbance occurred at 230 nm To replay your 10 50 minute scan you would use a range of 2 0 AUFS to k
150. then aline y s s d l m 46 UV2000 AutomatiC Scanning 2 2a reine een 51 Le Developer Zentren aaa a a 54 Sample QUEUE kerinan teesina oe neei NE a EEA EEE AE E EEEE ls meta 56 K Facto neinon ninani e a a ein a a a e ese tee ve 60 Absorbance RaUOS en ara E TEET E TE TAE E ta 64 01 2004 Thermo Electron i Chapter 5 Required Maintenance Introduction 22 ee ae b as Floweells 2 82 22 rar un an ee Lamps urn asien lien Appendix A Installation and Specifications Introduction rain a r na b Tastallat on nantes AA A io one tie Appendix B Menu Reference Introduction ars els de Mita ees Appendix C Troubleshooting Introduction s n vets eaten See de eee Theory 0f Operation 2 4 nu ee Eee Common Problems Troubleshooting Table Error Messages og wheter Diagnostic Tests nun einen Appendix D Glossary Introduction ae a Appendix E Cuvette Holder Accessory as no s s Installationy cc s mt ie lets BS Beads Using the Cuvette Holder Maintenance aa Index 1 Thermo Electron 01 2004 IN NORTH AMERICA 01 2004 Customer Support Thermo Electron San Jose products are supported by Thermo Electron Customer Servi
151. tings or default settings Use the and keys to scroll through available choices When the file you wish to load appears we re using the default settings for this example press ENTER to execute the load command 4 The confirmation message shown in Figure 2 18 appears for one second after which you are automatically returned to the Status Screen File Loaded Figure 2 18 The file loaded message Now you re ready for a practice run Note that the Status Screen Fig 2 11 now displays your wavelength setting the detector s status and the absorbance reading If the Status reads READY the required lamp is lit if it reads NRDY Not Ready there is an error for example you may have chosen a wavelength outside the selected lamp s range or the lamp isn t lit and if it reads UVW the ultraviolet D2 lamp is still warming up Thermo Electron 01 2004 When the detector is stabilized 1 Press the ZERO key to zero the detector s analog output signal 2 Inject your sample During setup you may have noticed that there was no stop time entered in the detector s parameters In this case the detector stays in the READY state and continually monitors the column eluant You do not need to manually start or stop a run with this set up ADDING A To add a stop time you need to modify the detector s operating STOP TIME parameters as follows We will then show you how to start and stop a run using the new setting 1
152. todiode surface or monochromator lens all of which are exposed during these procedures If dirty the surfaces should be cleaned with spectroscopic grade methanol or isopropanol and lint free lens paper only Flowcell Flowcell Inlet Outlet Tubing Clamp Photodiode Mount Flowcell Assembly Flowcell Assembly Thumbscrews DT Z019E FM 01 2004 Figure 5 2 Removing the cell cover to expose the flowcell and the photodiode mount 6 T Loosen the thumbscrew that holds the tubing clamp in place Gently pull the clamp toward you just far enough to disengage the tubing Loosen the two thumbscrews that hold the flowcell assembly Carefully pull the assembly toward you to remove it from the detector Thermo Electron 69 Flowcell Installation Too install a flowcell follow these steps 1 With the inlet tube on the bottom slide the flowcell assembly onto the alignment pins Fig 5 3 and securely fasten it in place with the two thumbscrews Tubing Clamp Flowcell Alignment Flowcell Pins Assembly Photodiode Mount Figure 5 3 Installing the flowcell assembly 2 Slip the flowcell s inlet and outlet tubes into the slots of the tubing clamp and tighten the thumbscrew that holds the clamp in place 3 Replace the photodiode mount and fasten it securely with the two thumbscrews 4 Connect the inlet tubing t
153. top line by scrolling down to the blank line below the time 0 00 line and pressing The second line will automatically have a time setting of 1 00 and the same wavelength setting s as the first Change 1 00 to the desired stop time for the run and leave the wavelength value s unchanged An example of a dual wavelength nine minute run at 254 and 283 nm is shown in Figure 3 15 Figure 3 15 A wavelength program with a programmed stop time Select Options from the Edit Menu to display the Options Menu Fig 3 16 Use this menu to set the detector s rise time autozero time and ranges Rise Time Autozero Time Range 2 Figure 3 16 The UV2000 s Options Menu Rise Time This field controls the detectors response time Rise time is inversely proportional to the amount of baseline noise For example the longer the rise time the less noise detected The one second default value is appropriate for most applications HINT To minimize baseline noise while retaining maximum resolution select a rise time that is at least one tenth of the peak width at the base of the narrowest peak of interest Thermo Electron 01 2004 Autozero Time This parameter tells the detector when to perform an automatic zero of the baseline If you do not wish to set an autozero and you are using a stop line in your wavelength program simply set the autozero time to a value greater than your stop time HINT It is good practice to zero the de
154. tremely corrosive and react explosively 1 Remove the flowcell assembly from the detector housing following the procedure on page 68 before cleaning with a nitric acid solution This will prevent possible leaks from harming the mechanical or electronic components of the detector 2 Flush the flowcell with water before proceeding This step is very important 3 Prepare a 20 v v solution of nitric acid in HPLC grade water 4 Pump the nitric acid solution through the flowcell with the chromatographic pump or draw it through with a large volume syringe If you use an LC pump replace your column with tubing and make sure water was the last solvent in the pump and solvent reservoir If you use a syringe always draw the solution through the flowcell flowcell Pressurizing the syringe could cause a leak or rupture that would result in an extremely dangerous uncontrolled spraying of nitric acid CAUTION Never use a syringe to force nitric acid through a 5 After you have finished the cleaning procedure and before returning to the buffer solution pump another 25 40 mL of water through the flowcell to remove all traces of nitric acid before returning to your chromatographic solvents Reinstall the flowcell assembly NOTE Flush the pump with water immediately after the nitric acid EN flush Leaving nitric acid solution in the pump for prolonged periods can damage pump seals 72 Thermo Electron 01 2004 Lamps
155. ual data points of your stored spectra by selecting the Display AU field in the Replay Menu Fig 4 14 A screen similar to that shown in Figure 4 17 will appear on the display Display AU 10 50 0 00001 Figure 4 17 The Display AU screen Thermo Electron 53 N NOTE Only actual data points separated by the proper wavelength interval can be displayed The Display AU screen shows the time at which the scan was initiated along with each wavelength and absorbance reading collected You can scroll through the data using the and keys To return to the Replay Menu press A The Develop File The Develop File is unique to the UV2000 It allows you to make sequential sample injections at different wavelengths automatically This automation makes method development much easier because you can use an automated run to determine the optimum wavelength of detection for each component in your sample You can also use the Develop File to troubleshoot chromatographic problems or to confirm method transfer from laboratory to laboratory SELECTING THE Use the following instructions to select the Develop File DEVELOP FILE Press MENU Select FILES Select Edit 3 Use the key to increment the Edit File field until a D is displayed The File Name field will read DEVELOP You cannot edit the Develop File s name EDITING THE Follow these instructions to edit Develop File parameters DEVELOP FILE 1
156. ufactured and tested in an ISO9001 Registered facility this system has been shipped to you from our manufacturing facility in a safe condition This reference manual contains precautionary statements that can prevent personal injury instrument damage and loss of data if properly followed All statements of this nature are called to your attention through the use of bold type and the following icons A A A HOT HIGH CAUTION SURFACE VOLTAGE Every instrument has specific hazards so be sure to read and comply with the following precautions They will help ensure the safe long term use of your system 1 Before plugging in any of the instrument modules or turning on the power always make sure that the voltage and fuses are set appropriately for your local power supply 2 Only use fuses of the type and current rating specified Do not use repaired fuses and do not short circuit the fuse holder 3 The supplied power cord must be inserted into a power outlet with a protective earth contact ground When using an extension cord make sure that the cord also has an earth contact 4 Do not change the external or internal grounding connections Tampering with or disconnecting these connections could endanger you and or damage the system CAUTION The instrument is properly grounded in accordance with these regulations when shipped You do not need to make any changes to the electrical connections or to the instrument s chassis to en
157. ult value is appropriate for most applications HINT To minimize baseline noise while retaining maximum resolution select a rise time that is at least one tenth of the peak width at the base of the narrowest peak of interest Autozero Time This parameter tells the detector when to perform an automatic zero of the baseline If you do not wish to set an autozero and you are using a stop line in your wavelength program simply set the autozero time to a value greater than your stop time HINT It is good practice to zero the detector automatically at the start of each run This will keep the detector output in range throughout an automated series of runs Range This parameter ranges the signal from the Ranged Output labeled as Ranged Output on the detector s rear panel Set the range to an appropriate full scale absorbance for your sample For more information on the use of ranges and analog outputs see pages 25 and 84 HINT We recommend a range of 1 0 when you are using an integrator or data system When you are ready to load the settings from the edit file into the detector s run file select Load from the File Menu The screen will display the words Load File Press ENTER to accept the settings The confirmation message shown in Figure 3 6 will appear for one second You are then returned to the Status Screen File Loaded Figure 3 6 The file loaded message Thermo Electron 01 2004 RUNNING YOUR Once y
158. und aufbewahrt werden K nnen ein erhebliches Gefahrenpotential darstellen doch darf man hier ber nicht die m gliche Gef hrdung durch die Proben selbst vergessen Achten Sie insbesondere darauf s mtliche Warnhinweise hinsichtlich ausreichender Bel ftung Lagerung Handhabung und Entsorgung von L sungsmitteln ebenso wie von Proben sorgf ltig zu lesen und zu befolgen Machen Sie sich mit den Toxizit tsdaten und den m glichen Gefahren s mtlicher verwendeter Chemikalien anhand der betreffenden Sicherheitsdatenbl tter vertraut die von den Produktherstellern zur Verf gung gestellt werden Probenvorbereitung berpr fen Sie stets die L slichkeit der Probe in der mobilen Phase Durch das Ausf llen von Feststoffen k nnen die S ule die Leitungen oder die Durchflu zelle verstopfen und damit den Durchflu hemmen Durch eine solche Verstopfung k nnen irreparable Sch den am System entstehen Die Ablagerung von Partikeln l t sich durch Filtrieren der Proben durch ein Filter mit einer Porengr e von 0 45 oder 0 2 um oder weniger vermeiden Anforderungen an das L sungsmittel Viele chemische Hersteller bieten eine Produktserie hochreiner Reagenzien in spektroskopisch reiner Qualit t an die frei von chemischen Unreinheiten sind Die routinem ige Filtrierung aller L sungs und Extraktionsmittel durch ein Fluorkohlenwasserstoff Filter mit einer Porengr e von 0 45 oder 0 2 um oder weniger vor dem Einf llen in den L s
159. ungsmittelbeh lter verl ngert die Lebensdauer der Einla filter der Ventile und Dichtungen des Injektors und der S ule betr chtlich Spezielle HPLC L sungsmittel brauchen normalerweise nicht filtriert zu werden Thermo Electron xvii xviii W hlen Sie eine mobile Phase die zur Probe und zur f r die Separation verwendete S ule kompatibel ist Dabei ist darauf zu achten da Edelstahl durch bestimmte L sungsmittel korrodiert wird Reaktionstr ge biokompatible Ger teausf hrungen werden ebenfalls von Thermo Separation Instruments angeboten Entgasen des L sungsmittels L sungs und Extraktionsmittel sollten entgast werden und zwar entweder durch Vakuum oder Heliumdurchperlung Eine umfassende Beschreibung dieser Techniken finden Sie in dem separaten Handbuch das dem Entgasungszubeh r beiliegt Entsorgung von L sungsmitteln Sorgen Sie daf r da ein Auffangbeh lter f r L sungsmittel oder eine andere Auffangvorrichtung in H he des Arbeitstisches oder darunter zur Verf gung steht F r die meisten L sungsmittel gelten besondere Entsorgungsvorschriften eine Entsorgung ber die Abwasserleitung ist hier nicht zul ssig Bei der Entsorgung von Chemikalien gleich welcher Art sind die einschl gigen Vorschriften streng zu beachten Hochdrucksysteme und Undichtigkeiten Fl ssigchromatographen arbeiten unter hohem Druck Da Fl ssigkeiten kaum komprimierbar sind k nnen sie nicht viel Energie speichern Dementsprech
160. ure E 925 from the American Society for Testing and Materials ASTM as well as standard potassium dichromate SRM 930 from the National Institute of Standards and Technology NIST formerly the National Bureau of Standards NBS The test involves the preparation of acidic solutions of potassium dichromate at four concentrations and the absorbance measurement of each solution at four wavelengths between 235 and 350 nm After correcting for an absorbance blank the linearity deviation of a plot of absorbance versus concentration should be less than 1 If you want more information on this test or find that your instrument does not conform to these specifications and requires service contact your local Thermo Electron representative CALIBRATION Selecting A Calibration brings up the screen shown in Figure E 6 You can use this screen to offset the factory calibrated wavelength to more closely match FDA industry or in house standards A Offset steps 0 Figure E 6 The lambda vvavelength offset screen 01 2004 Thermo Electron 127 N NOTE The UV2000 detector is calibrated using a mercury lamp fixture AN This provides a very narrovv emission line at 254 nm Broad band calibration standards such as holmium oxide and didymium filters make calibration more difficult and less accurate To offset the factory calibrated vvavelength select the number of steps by which you want the wavelength to be offset Each step represents
161. ut terminal on the detector s back panel the detector can send a signal to other devices each time it goes to its READY state This feature is frequently used with autosamplers to signal that the detector is ready for the next injection To access the READY Output field 1 Press MENU 2 Select OPTIONS 3 Select More 4 Scroll down to the READY Output field Select Active Hi or Active Lo depending on which signal you wish to send HINT All SpectraSYSTEM instruments are set to receive high signals so select Active Hi if you are hooking up to this type of chromatograph For any other type of instrument refer to the appropriate reference manual Thermo Electron 01 2004 DISPLAY You can vary the display s contrast to make it easier to read CONTRAST To change the display s contrast first press STATUS to access the Status Screen Then simultaneously press the gt key and the key to increase the contrast or the gt key and the key to reduce the contrast CURSOR SPEED You can control the display s cursor speed to make it easier to use To access Cursor Speed 1 Press MENU 2 Select OPTIONS 3 Select More 4 Scroll down to Cursor Speed and select Fast Medium or Slow SHUTDOWN This feature offers a quick shutdown and subsequent startup of the DETECTOR detector s lamps and motors The electronics stay on to maintain the detector s memory To shut down the detector 1 Press MENU 2 Sel
162. ut the analytical industry and are added here as a quick reference only Analog to digital Converts a detector s analog signal to a digital signal Absorbance units full scale a measure of sensitivity A process where the intensity of light shining through a sample is decreased the transmitted light is measured in absorbance units which are directly proportional to the concentration of the absorbing sample A voltage applied to the output signal in order to keep the signal on scale throughout a run The reference spectrum of the mobile phase It is subtracted from the sample spectral scans to correct for baseline absorbances Also called baseline scan The reference line at the bottom of a chromatogram from which measurements are made A baseline represents the chromatogram that would be drawn if only the mobile phase with no sample were run through the column A gauge for measuring the total amount of time something has been in operation The values or choices built in to a system If no specific choice is made the detector will run using the default settings A feature that allows you to make multiple injections of a sample at different wavelengths automatically The practice of removing air from the mobile phase usually by sparging or applying a vacuum Methods used to detect and isolate problems The two line screen on all SpectraS YSTEM instruments Thermo Electron 117 edit file error message field
163. utomatically The Order field reads 2 and the rest of the line is blank Select the proper file and the number of injections to be made for that file You can have as many as ten groups in the queue To delete a line use the key in the File Name field until it goes blank When you leave the line it is deleted and the queue is resorted automatically An example of a queue appears in Figure 4 23 File Name 2 THEOPHYL 3 ABCD 1 BARBITUA Figure 4 23 An example of a queue In our example we have programmed the detector to run File 2 for the first five injections File 3 for the next 25 injections and File 1 for the last ten injections Thermo Electron 57 58 LOADING A QUEUE RUNNING A QUEUE Viewing its Progress To load a queue select Load in the Queue Menu When the words Load Queue appear press ENTER The confirmation message in Figure 4 24 appears for one second Queue Loaded Figure 4 24 The confirmation message when a queue is loaded When a queue is loaded the letter Q appears at the extreme left of the Status Screen Fig 4 25 Status AU Q READY 40 00001V Figure 4 25 The Status Screen when a queue is loaded If you attempt to load a queue when no queue exists the message shown in Figure 4 26 will appear on the display No Queue Available Figure 4 26 The message that is displayed when no queue is available When the detector receives its first start signal it
164. velength one in single wavelength operation or from wavelength two in dual wavelength operation AU1 KxAU2 a calculated peak response using the K Factor technique and AU1 AU2 the absorbance ratio of wavelength 1 to wavelength 2 Default is AU UV 2000 only This field offsets the Analog 2 output signal by a positive or negative 50 20 10 5 2 1 or 0 percent of the full scale range Default is 0 This menu allows you to offset the signals from the analog output terminals located on the back panel of the instrument For the UV2000 you can also select the output signal for Analog Output 2 and input a K factor This field located in the Status Screen shows the detector s current absorbance reading It is a six digit number ranging from 3 00000 to 3 00000 AUFS This field tells the detector when to perform an automatic zero Allowable values are 0 00 to 999 99 minutes Default is 0 00 minutes The Commands Menu lets you put an event mark into your chromatogram short detector outputs and shut down the detector This field accesses the Copy File field UV2000 only This field along with the To File field allows you to copy from the specified file to another file designation UV2000 only This field displays local time ranging from 0 00 to 23 59 This field may be set to Slow Medium or Fast according to your need Default is Medium Thermo Electron 97 Delete Delete File Delete Queue D2 Lamp Hours Diode
165. vidual absorbance value data from your scan of compound B Calculate the K factor using the following equation K AUI AU2 where AU and AU2 are the absorbance values for compound B at A1 and 222 respectively For our example the absorbance values are 0 0144 and 0 0032 for 254 and 280 nm respectively so our K factor is 4 5 calculated as follovvs K 0 0144 0 0032 4 5 Using the K Factor To use the K factor set the parameters in the Analog Outputs Menu inject your sample and monitor the results as follows 1 2 3 Press MENU Select OPTIONS Select Analog Outputs The menu shown in Figure 4 32 will appear Analog 1 Offset Analog 2 Offset Analog 2 K Factor Figure 4 32 UV2000 s Analog Outputs Menu Scroll down to Analog 2 and select AU1 K AU2 Scroll down to K factor and enter your calculated value 4 5 for our example Inject your sample HINT Make sure your file was set to dual wavelength mode as described in Chapter 3 Also remember that in this example AUl 1 is 254 nm and AU2 X2 is 280 nm Use Analog Output 2 on the detector s rear panel to monitor the chromatograms for your peak of interest Thermo Electron 63 DT Z025E FM 1 51 Time in Minutes Figure 4 33 Chromatogram of toluene with butyl paraben suppressed Our example chromatogram would now appear as shown in Figure 4 33 with a slightly lowered response for toluene and no
166. w titled Protecting Files you will get the message shown in Figure 3 24 If a file is not protected make sure it s empty or unwanted before you copy to it as it will be overwritten Protected File Cannot Be Copied To Figure 3 24 The message that is displayed when you attempt to copy to a protected file You cannot use Copy for the Scan or Develop files You will learn more about these files in Chapter 4 To delete a file 1 2 3 4 Press MENU Select FILES to display the Files Menu Fig 3 21 Select Delete The Delete File field will appear on the screen Enter the identification number of the file you wish to delete When you press ENTER the confirmation message shown in Figure 3 25 appears briefly and the display returns to the Files Menu The parameters in the file you have just deleted return to their default values Thermo Electron 33 File Deleted Figure 3 25 The message that s displayed when a file is deleted If you attempt to delete a protected file see the next section Protecting Files you will get the message shown in Figure 3 26 Protected File Cannot Be Deleted Figure 3 26 The message that s displayed when you try to delete a protected file PROTECTING FILES The UV2000 allows you to protect files from being edited copied to or deleted To access the file protection operation follow these steps 1 Press MENU 2 Select OPTIONS The Options Menu
167. x A All flowcells are shipped premounted in a holder for easier installation and alignment To access the flowcell remove the front panel of the detector The flowcell assembly is located behind the lower housing Fig 5 1 Once the housing is removed the flowcell is easily identified by the tubing that extends from the fittings on either side of the cell body Fig 5 2 Thermo Electron 67 Display Keypad Lamp Housing Flowcell Housing Figure 5 1 Detector flowcell and lamp housings Flowcell Removal Use the following steps to remove the flowcell 1 Disconnect the power cord from the rear panel of the detector and make sure that the instrument is turned off 2 If you have not already done so remove the detector s front panel by grasping the bottom of the panel firmly with one hand and pulling back 68 Thermo Electron 01 2004 Loosen the knurled thumbscrew that holds the flowcell housing in place and remove and set aside both the thumbscrew and the housing Disconnect the flowcell inlet tube from the chromatograph and free the flowcell outlet tubing from the waste reservoir Loosen the two thumbscrews on the photodiode mount and carefully pull the mount straight back Fig 5 2 The cable that connects the photodiode mount to the detector is sufficiently long to allow the mount to rest on the bench top NOTE Avoid putting fingerprints or scratches on the flowcell windows pho
168. xicology Q Other Q Pharmaceutical 1 Research Education Job Function Check one only Q Administration 1 Lab Management 1 Operator Q Other Reader Survey Help us to improve the quality of our documentation by answering a few questions Finnigan SpectraSYSTEM Revision G UV Vis Detector Reference Manual A0099 540 Strongly Strongly Agree Agree Disagree Disagree The manual is well organized 1 2 3 4 The manual is clearly written 1 2 3 4 The manual contains all of the information need 1 2 3 4 The instructions are easy to follow 1 2 3 4 The instructions are complete 1 2 3 4 The technical information is easy to understand 1 2 3 4 The figures are helpful 1 2 3 4 Additional Comments Attach additional sheets if necessary Tear this sheet from the manual fold it closed stamp it and drop it in the mail From fold Place Stamp Here EDITOR TECHNICAL PUBLICATIONS THERMO ELECTRON SAN JOSE 355 RIVER OAKS PARKWAY SAN JOSE CA 95134 1991 UNITED STATES OF AMERICA fold Contents Back Pocket Quick Reference Card Menu Trees EUSTOMET SUP POM t sisiecedsccccesaisasdenedivocccossvonsstaeseoecetesacoscsdseeseeseoessetevesecesostdoaessescoessuseceosavedsceteucesssesavets iii Safety Informati n vecsscekissecccssscescesisseecdcenscscadcssponccevanecseecegrnnnssacevessetsevencsacecsdecnedsenssvecvonsesevssendeosroveess vii
169. ying 33 repeating the file 56 setting parameters 54 Status Menu 56 Status Screen 55 DEVELOP see Develop File Diagnostic tests analog outputs 115 deuterium lamp 115 diagnostics defined 118 external absorbance linearity 116 127 LC test mix 116 internal diode offsets self test 115 user selected 113 light levels 113 software version 113 wavelength calibration test 114 127 lamp and shutter UV2000 only 115 light levels 98 motor 115 RAM 115 self tests 114 software version 102 Tests Menu illustration of 112 tungsten lamp 115 voltages 115 Diode Offsets field described 98 self test 115 user selected test 113 Display changing the contrast 45 changing the cursor speed 45 illustration of the 2 term defined 118 the Display AU I screen 53 98 Down arrow t key right side of display 2 Drift see Baseline problems Dual wavelength operation 26 analog output 2 35 selecting 100 Status Screen 30 stopping arun 30 wavelength program 27 E Edit File 20 98 118 Edit Menu accessing the 20 27 general description of 98 illustration of 20 27 while in the Develop File 54 while in the Scan File 47 Editing a file from the File s Menu 20 26 from the Status Menu 24 31 a queue 59 parameters during a run 5 the Develop File 54 time lines 39 Electrical interference causes of 107 End 98 End of queue 43 Enter as a standard vvord 7 ENTER key 3 Equipment damage avoiding 8 see also Safety Information Error

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