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PerkinElmer Perkin Elmer - lambda 20 - Perkin Elmer Uvvis-spectrophotometer Lam

1. XXX Xnm Xxx nm Wavelength X X min X XXX A Xxx A Measured value ordinate as selected Xxx min Time units as selected C xx Repeat measurement cycles still to be performed This appears on the top right when cycles gt 1 Printout The result is printed out at the end of each sample analysis 84 Using Methods Scanning a Spectrum Select a SCAN method to scan and record a spectrum of the sample Procedure l Select the desired SCAN method The following table lists typical SCAN parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter 85 Lambda 20 Lambda 40 UV Vis Spectrometers No Parameter Value No Parameter Value 17 SLIT 2 0 nm 1 ORDINATEMODE A 3 WAV MAX 1100 00 nm 4 WAV MIN 190 0 nm 13 SPEED 960 nm min 14 SMOOTH 2 nm 15 LAMP UV Vis 16 AUTOZERO YES 38 ACCESSORY MANUAL 18 SAMPLES BATCH 0 Only available with Lambda 40 When a particular parameter is set to YES the extra parameters required automatically appear in their correct order For example when GRAPHICS PLOT is YES the extra parameters ORD MAX ORD MIN SCALE and GRID appear When GRAPHICS PLOT is NO the extra parameters ORD MAX ORD MIN SCALE and GRID no longer appear 2 Ifnecessary modify the method parameters 3 Press START to start the method 86 Using Methods 4 Check the display If the dis
2. XXX nm XXX nm Wavelength XXX min XXX A XXX min Time XXX A Measured value The display shown when using a cell changer x C n X C Temperature XXX min XXX A n Cell location XXX min Interval time XXX A Measured value 112 NOTE NOTE Using Methods Printout The results are printed out at the end of the analysis Determining the Blank Value Determine the blank value of the reaction as follows 1 Select the appropriate SUBSTRATE method 2 Set parameter BLANK 0 0 in the method 3 Carry out a measurement according to your procedure using a cell filled with redistilled water in place of the sample 4 Enter the result of the measurement in the parameter BLANK Recalculation of Results with POSTRUN KIN You can recalculate the results from ENZYME and SUBSTRATE methods using the POSTRUN KIN parameter When you set the POSTRUN KIN parameter to YES the following parameters in the corresponding method can be identified ENZYME method LAG time and TOTAL TIME SUBSTRATE method END TIME manual operation only The time set for the LAG TIME must be smaller than the TOTAL TIME If a cell changer is being used make sure that both LAG TIME and TOTAL TIME are whole multiples of the INTERVAL time Proceed as follows to calculate the results 1 Create a method with POSTRUN KIN YES 113 Lambda 20 Lambda 40 UV Vis Spectrometers 2 114 Carry out a measurement see Enz
3. 87 Printout cess 5 Reli deerant im een ient 87 Measurement at Several Wavelengths essere 88 Proceduren fost ica rb n eos UH toe nctucesay saver TA 88 Example of the Display Shown During the Measurement 90 PrititOUL co eere seen oe ee ref rette een 90 Concentration Determination sess 91 CONCENTRATION 1 Method Peak heights ene 91 CONCENTRATION 2 Method Peak Areas 2 Derivative 96 Processing the Calibration Curve CONCENTRATION Methods 103 Enzyme Kinetics ee tege to s deci eda ee bide iv eg a eps 105 Juni on 105 Determining the Blank Value essere 108 Substrate Kinetics iie Dire et T ri 109 Contents Man al Procedure mn ee e nte atten ees 109 Procedure with a Cell Changer 112 Determining the Blank Value esses 113 Recalculation of Results with POSTRUN KIN eee 113 Methods for Quantitative Analysis of Oligonucleotides 115 POC Uli rice eden te t dn ene e thd Rr Re RE TERRE 116 Date Time o oa ee nnt ce ek Or e ee eR ees 121 Makeup iet ettet bent re erbe IIR sehen 123 Self Check a bre ie air ene eb NE M esa ire EDS 125 Operating with Accessories eee e eee ee eee e esee eene een neenon nest tn aseo seta 127 Operating with Accessories ccccceesseesseeesseetseeeeesecseceseceseceseeeeeceseenseeses 129 SICHERER 129 ACCESSOHEs u
4. Default values Port Enable No ETXT Character 015 Respond Prompt Prompt Character 021 Baud Rate 5800 Break Character 043 Bits Character 8 Erase Character 010 Stopbit 1 Kill Character 010 Parity none Range 2 Terminator CRLF Usage of the RS 232 interface marked Serial Port on the spectrometer and the centronics interface marked Parallel Port on the spectrometer PRINTER PORT Parallel Port you can choose to have this interface active ON or inactive OFF Select the printer using PRINTER CONFIG RS232 PORT Serial Port you can choose to connect a personal computer or printer to this port COMPUTER PC default setting PRINTER serial printer After a full reset the value is set to COMPUTER To configure the printer output Parameters PRINTER ON Switches output to the printer on and off PRINTER Selects the printer NO no printer connected EPS EX FX EPSON LQ 219 Lambda 20 Lambda 40 UV Vis Spectrometers Mood 220 Function BIDIRECTIONAL INTERNAL internal printer EPSON INTERNAL HP PCL GAP Sets the gap between two pages 4 6 8 FF PERFORATION Skip over Perforation Appears when GAP is set to FF form feed YES NO COLOR ON Switches color printing on and off only for color printers YES NO PLOT HEADER Header printed at the start of each plot YES NO PAPER When the PRINTER parameter is set to EPSON INTERNAL you can select Z fold or single sheet paper Z FOLD SINGLE S
5. Acetone Tetrachloroethylene m Xylene Toluene Eum Benzene N N Dimethylformamide Ethyl Propionate Carbon Tetrachloride Methyl Form Chloroform 1 2 Dichloroethane Dichloromethane G lycerol XIII IE UU MUT oa Dioxan committe E Hexane EXE A AAA i Octane minim mn me 2 2 i 4 Tri methyl pe ntane EXIIT UIS UU UI MU UU UU TIU UI MUI I UIT EEUU TTE Acetonitrile 9 p22 2iIiInt o ee T iUt Cyclohexane zum METUUNT eer eee Methanol uui Me AE TT Ethanol STE TESTS ESCHE UN T ENT UU EUTP NIU U NU UM UU TT eU NEEDED eee Methylcyclohexane i Propanol Water 190 Figure 7 Lower Wavelength Limits of Solvents 154 Error Messages 1 0 Error Messages Error Messages If an error occurs during the operation of the spectrometer an error message is shown on the display or is printed out if a printer is connected Error Messages Shown on the Display Errors remain displayed until they are deleted To delete press PARAM RANGE ERROR The value entered is outside the displayed range XXXX X XXXX X 2 Press PARAM and enter a value within the range shown PARAMETER XX The parameter XX does not exist DOES NOT EXIST i Press PARAM to continue PARAMETER XX The parameter XX is not used or is not active in this NOT USED method Press PARAM to continue HINT The Interval time is greater than the total time INT GREAT
6. Number of reference solutions used for the calibration Range 1 to 20 WAVELENGTHS Number of wavelengths at which measurements are made Range 1 to 20 If ordinate modes RAT and DIF are used the number must be divisible by 2 If ordinate mode COR is used the number must be divisible by 3 See also parameter Ordinate Mode ABS COEF dA 71 To set the molar absorption coefficient of the Adenine nucleotide Deoxyadenosine monophosphate dAMP Range 0 to 99999 Appears with the Oligoquant applications 169 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter To set the molar absorption coefficient of the Cytosine nucleotide Deoxycytidine Monophosphate dCMP Range 0 to 99999 ABS COEFF dC 72 Appears with the Oligoquant applications ABS COEFF dG 73 To set the molar absorption coefficient of the Guanine nucleotide Deoxyguanosine monophosphate dGMP Range 0 to 99999 Appears with the Oligoquant applications 38 To set the molar absorption coefficient of the Thymine nucleotide Deoxythymidine monophosphate dTMP Range 0 to 99999 Appears with the Oligoquant applications ABS COEFF dT ABS COEFF dN To set the molar absorption coefficient of any nucleotide Deoxynucleoside monophosphate dNMP N is used to represent any base Range 0 to 9999 Appears with the Oligoquant applications ACCESSORY Lets the spectrometer recognize the accessory type connected MANUAL is for operations without usin
7. Range 9999 9 to 9999 9 The measured value is multiplied by the factor and the result displayed Thus concentration can be read off directly or a dilution taken into account If only the true measured value is to be shown then choose FACTOR or FACTOR n 1 If concentration units are to be read off directly calculate the factor according to the Beer Lambert law ecd Where A is the absorbance e is the molar absorption coefficient c is the concentration of the sample d is the pathlength of the cell Thus the concentration c A amp d And the factor f 1 d Number of the first sample in the batch All subsequent samples are automatically numbered consecutively The first sample tube location of the batch to be analysed n is any location number up to the maximum location for the tray type 183 Lambda 20 Lambda 40 UV Vis Spectrometers Range 1 to the maximum location number of the tray being used Appears with the autosampler accessory GRAPHICS PLOT Graphics printout Option YES NO Graphics printout with grid valid only when GRAPHICS PLOT YES Option YES NO INTERVAL 21 Interval time units as selected for TIME UNIT Range 0 1 to 999 9 When using CALCULATE REGRESSION the change in absorbance dA dt is printed out When using CALCULATE INTERVAL the slope is calculated for each interval The mean of all slopes is then used for the calculation of the enzyme activity see also CALCULAT
8. Insert a cell containing the sample solution and press START 3 Continue to insert samples when asked until they have all been measured Example of the Display Shown During the Measurement CONC 2 SMPL 1 xxx nm Wavelength XXX X nm XXX c XXX C Result units as selected CYCLES XX CYCLES XX Repeat measurement cycles still to XXX X nm DIS be performed This appears on the top right when cycles gt 1 Printout If PLOT REFERENCES and PRINT DATA are set to YES the calibration curve and results are printed out 102 Using Methods Processing the Calibration Curve CONCENTRATION Methods Changing the type of Curve Fit The type of calibration curve fit linear or quadratic can be altered without having to carry out additional measurements The procedure is as follows l Select REFS OLD OR CUR FIT as required This modifies the method Press START The new curve fit is calculated on already available data Remeasuring the Reference Solution Should a measuring point lie outside the calibration curve and need to be remeasured proceed as follows l 2 Select REFS NEW Press START You will be asked for first reference solution Enter the number of the reference to be remeasured and press ENTER REF n XXX START STOP AUTOZERO Place the reference solution in the sample cell holder Press START 103 Lambda 20 Lambda 40 UV Vis Spectrometers
9. Method 9 Factory Contig 2 n n 225 Lambda 20 Lambda 40 UV Vis Spectrometers 226 Enzyme Kinetics Enzymatic Analytical Procedures Enzyme analysis makes use of enzyme controlled reactions to determine a substance These procedures are used especially in clinical chemistry and food chemistry There are two methods of enzymatic analysis e The determination of enzyme concentration or enzyme activity enzyme kinetic measurement For example the determination of enzymes in blood serum e The determination of the concentration of a substrate substrate kinetic measurement For example the determination of components in food alcohol in wine The basis of both methods is the conversion of a substrate into a product whereby the enzyme acts as a catalyst The reaction can be followed photometrically an added coenzyme for example NADH is oxidized or reduced in the course of the reaction and the resulting change in absorbance measured Or the substrate or product may be photoactive and the absorbance will change with the concentration In enzyme kinetics the substrate is added in excess and the reaction rate as dA dr measured With excess substrate it is constant and directly proportional to enzyme activity In substrate kinetics the substrate reacts completely The substrate concentration can then be calculated from the change in absorbance AA Enzyme Kinetics In enzyme kinetics the enzyme activity of a samp
10. NEW OLIGO1 PARAM gt Y Param y nnn OLIGOQUANT 1 lt gt PARAM START Procedure 1 Select the appropriate OLIGOQUANT method see Oligoquant Parameter Tables on page 118 2 Ifnecessary modify the method parameters 116 Using Methods 3 Press START This starts the measurement 4 Check the display If the display is AUTOZERO START STOP OR If the display is OLIGO n SMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START 5 Continue to insert samples when asked until they have all been measured Example of the Display Shown During the Measurement OLIGO n SMPL 1 XXX nm Wavelength XXX nm XXX A XXX A Measured value ordinate as selected Printout Graphics are printed out during the measurement process numerical data follow at the end of the analysis 117 Lambda 20 Lambda 40 UV Vis Spectrometers 118 Oligoquant Parameter Tables The following table lists typical OLIGOQUANT 1 parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter No Parameter Value No Parameter Value SLIT 20nm 1 ORDINATE MODE At WAVELENGTHS WAV 1 260 0 nm FACTOR 1 PATHLENGTH 1 0 cm 61 SEQ 2 65 CHANGE CONSTANTS NO Dem o SAMPLE ID Only available with Lambda 40
11. 145 Lambda 20 Lambda 40 UV Vis Spectrometers Sample Compartment Windows e Generally the windows should be installed at all times e The windows are optical components and require the same care and handling as cells e You can remove windows to clean them They are held in place by a magnetic frame Windows are most suitably cleaned by wiping them with a soft lint free cloth moistened with ethanol 146 Analytical Notes 9 Analytical Notes Autozero The type of autozero depends on the method type selected In methods with a fixed wavelength as TIME DRIVE WAVELENGTH PROGRAM CONCENTRATION 1 the displayed measurement value for absorbance is set to 0 for transmission to 10096 at the measurement wavelength this is called an autozero You can use the AUTOZERO key to perform a manual autozero in fixed wavelength methods see Manual Autozero on page 42 In methods with measurement over a wavelength range such as SCAN CONCENTRATION 2 a background correction is performed over the selected wavelength range A background correction can only be performed in a method The ordinate mode of the last used method always appears on the display To change from absorbance to transmittance or vice versa select a TIME DRIVE method and then select the desired ordinate mode An autozero or background correction must be performed e Atthe start of a new method e When the wavelength is changed e When the wavelength range is ext
12. Only data above the given threshold value is printed out Concentration2 Threshold value for MODE DERIV PEAK 2 Only values above the given threshold value will be recognized as peaks Units for all subsequent time parameters min minutes s seconds Calculates the theoretical melting point Ty Refer to the Biochemical Application Manual Total time from the start of the method or end of DELAY TIME to the end of the measurement units as selected for TIME UNIT Range 0 1 to 999 9 Select the measuring time so that the end of the measurement is still within the linear portion of the curve Manual operation For CALCULATE REGRESSION the time is that between the start of the method and end of the measurement For CALCULATE INTERVAL the time is that between the end of the DELAY TIME and end of the measurement Operation with Cell Changer The time from the end of the DELAY TIME to the end of 201 Lambda 20 Lambda 40 UV Vis Spectrometers the measurement Within this measuring period all samples are measured n times consecutively The number of cycles is determined by the measuring time and the INTERVAL time Number of cycles TOTAL TIME INTERVAL 1 If the result proves to be a decimal fraction the next higher whole number is taken For example 100 30 1 4 33 5 cycles The cycle time is identical to the INTERVAL time The final cycle begins at the end of the measuring time Measurement e
13. START to start the measurement 2 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is REF n XXX START STOP AUTOZERO Insert a cell containing the sample solution and press START 94 Using Methods 3 Insert the references in sequence when asked When they have all been measured the instrument prints out the calibration curve and the results You can now amend the calibration curve see Processing the Calibration Curve CONCENTRATION Methods on page 103 if required You can use previously established calibration curves or reference values see REFS in Parameter Numbers and Descriptions page 169 Measuring the Sample 1 Press START to start the measurement 2 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is CONC 1 SMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START 3 Continue to insert samples when asked until they have all been measured 95 Lambda 20 Lambda 40 UV Vis Spectrometers Example of the Display Shown During the Measurement CONC 1 SMPL 1 XXX nm Wavelength XXX X nm XXX c XXX C Result units as selected CYCLES XX CYCLES XX Repeat measurement cycles still to XXX X nm ANO be performed This
14. Value 2 0 nm 3 3 WAV 1 459 9 nm 4185nm 3 WAV 3 360 0 nm 1 0 11 FACTOR 2 1 0 1 0 14 RESPONSE 0 55 UV Vis 16 AUTOZERO YES 18 SAMPLES BATCH 0 21 CYCLES 1 MANUAL 1 0 10 min 25 GRAPHICS PLOT YES 27 ORD MIN 0 000 A 1 000 A 29 GRID YES Using Methods 32 PRINT DATA YES 35 AUTO METHOD NO 36 OPER ID 37 SAMPLE ID Only available with Lambda 40 When a particular parameter is set to YES the extra parameters required automatically appear in their correct order When GRAPHICS PLOT is YES the extra parameters ORD MAX ORD MIN SCALE and GRID appear When GRAPHICS PLOT is NO the extra parameters ORD MAX ORD MIN SCALE and GRID no longer appear 2 Ifnecessary modify the method parameters 3 Press START to start the measurement 4 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is WAVPROG SAMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START 5 Continue to insert samples when asked until they have all been measured For additional information see Replot on page 79 89 Lambda 20 Lambda 40 UV Vis Spectrometers Example of the Display Shown During the Measurement WAVPROG SMPL 1 xxx nm Wavelength Xxx nm X XXx A Xxx A Measured value ordinate as selected CYC xx Repeat measurement cycles still to be perform
15. background correction irrespective of the locations selected as described above If AUTOZERO YES has been selected place the blank at location 1 and the sample solutions from location 2 If AUTOZERO NO has been selected all locations can be used for sample measurement 136 Operating with Accessories Cell changer 13 Cell Changer CELL 1 7 The locations at which measurements are to take place CELL 8 13 When CELL 1 7 shows the numbers 1 to 7 represent sample locations 1 to 7 When CELL 8 13 shows the numbers 1 to 6 represent sample locations 8 to 13 If none of the locations is to be used enter 0 Press ENTER to confirm the location numbers If measurement is to be carried out at locations 2 and 5 only enter 25 and then press ENTER CELL 1 7 and enter 0 and then press ENTER CELL 8 13 If AUTOZERO YES has been selected place the blank solution at location 1 and the sample solutions from location 2 If AUTOZERO NO has been selected all locations can be used for sample measurement Example 1 CELL 1 7 246 Measurement will take place at CELL 8 14 0 locations 2 4 and 6 the rest will not be used Example 2 Measurement will take place at locations 5 to 13 CELL 1 7 567 CELL 8 13 123456 137 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter Description STIRRER Switches the magnetic stirrer on and off Option YES NO Select with arrow key If the magnetic
16. offered by PerkinElmer The minimum sample volume required is a function of the cell internal width or volume and is specified below Cell Type Cell athlength Minimum Part Internal Volume Number Width Required Height of 2mm 200 uL B0631071 liquid slightly pair Cell athlength Minimum Part Volume Volume Number Required completely liquid 30 uL 50 uL B0631079 NOTE You should align microcells very carefully in the radiation beam by following the procedures in Aligning the Single Cell Holder on page 33 When aligning microcells fill each cell with the minimum volume of liquid specified in the above table to make sure that the liquid meniscus is not in the radiation beam P more than height of 4 mm lcm 400 uL B0631064 beam pair P 37 Lambda 20 Lambda40 Operation and Parameter Description 38 Operating without 4 Methods Operating without Methods Operating without Methods Overview Measurements are usually carried out using methods containing all the necessary parameters see Using Methods on page 71 The following functions can be carried out via the keyboard Setting the wavelength Manual autozero Quick sample measurement Reset Print Help Setting the Wavelength Manually The wavelength can be set manually in standby using the GOTO key as follows l 2 Set the spectrometer to the standby display Press GOTO Enter the desired wavelength for example 325 5 Press ENTER The
17. 104 6 Waituntil the measurement is complete 7 Press STOP 8 Set REFS OLD 9 Press START The new calibration curve is calculated with the new value 10 Ifthe new curve is satisfactory measure the sample OR If it is not acceptable repeat the procedure Deleting a Point from the Calibration Curve To delete such a point proceed as follows Linear Curve through Zero 1 In the method parameters set REF n 0 000 and VALUE n 0 000 n is the number of reference solutions to be deleted 2 Press ENTER to confirm the changes Non linear curves and curves with intercept 1 In the method parameters note the ordinate value and the concentration value of the last point 2 Replace the ordinate value and the concentration value of the point to be deleted REF n with the values noted in step 1 above 3 Reduce the value for OF REFS by 1 4 Press ENTER to confirm the changes Using Methods Enzyme Kinetics Select an ENZYME method for enzyme kinetic measurements NOTE Enzyme activity is strongly dependent on temperature Thus the following should be taken into account All measurements should be carried out at a constant temperature You can use the temperature sensor B0185227 for monitoring the temperature All solutions and essential instrument accessories especially cells and cell holders should be thermostatted prior to use Procedure l Select the appropriate ENZYME method The follo
18. 193 Lambda 20 Lambda 40 UV Vis Spectrometers PRINT DATA Prints out a table containing analytical data when activated Option YES NO Substrate Kinetics Enzyme Kinetics Option ALL FINAL NO ALL All readings and the final calculated results are printed FINAL Only the final calculated results are printed NO No data printed PRINT REFS Prints out data from the references when activated Option YES NO READER Read mode for the linear transporter Option RANDOM CONT RANDOM Measurements at user defined positions CONT Continuous measurement over the entire 100 mm measurement range of the linear transporter Reference solutions Choice as to whether a calibration curve should be established or not at the start of a method Option OLD NEW OLD means you wish to use the OLD stored calibration curve NEW means you wish to make a NEW calibration curve If a calibration curve is to be used again or if the values of the curve are to he entered directlv select 194 Parameter Numbers and Descriptions REFERENCES OLD see also VALUE It is often useful to tag REFERENCES For example To tag a reference as a CALL parameter select REFERENCES NEW when selecting the first method and generate a calibration curve At the next call up select REFERENCES OLD The available calibration curve is used and measurement can start immediately LEE Concentration of reference solution n 14 Response time Time constan
19. DIALOG software or after a full reset FULL RESET DONE Make a note of the steps you made leading up to this message Press PARAM to continue If you cannot continue call your PerkinElmer office and inform them of the error and the steps you made leading up to the error BUS ERROR a This error plus a message is shown when the instrument has an address error message Make a note of this message and the steps you made leading up to the error Press PARAM to continue If you cannot continue call your PerkinElmer office and inform them of the error the error message and the steps you made leading up to the error After a full reset all methods are erased 162 Error Messages Error Reports on the Printer Error Meaning Baseline correction data do not fit Start baseline correction The background correction last carried out did not correlate to the method used Carry out a new background correction with the proper method Cannot approximate calibration curve Check references or change curve fit algorithm The calibration data deviate strongly from the curve form selected or not enough points were measured for the curve form selected Check the references select another curve form or carry out more measurements Cannot calculate delta absorbance Because too few points read A kinetic method was interrupted with STOP Too few points available to calculate delta
20. HELP for additional information about a parameter Enter the desired value with the numeric keys Press ENTER to confirm the value entered Appears If several values have to be entered Use the arrow key to select the desired parameter Enter the required value Press ENTER to confirm the value entered For example Several reference values have to be entered Enter the value for REF 1 Confirm with ENTER Use the arrow key to move to REF 2 Continue until values have been entered for all the references When a particular parameter is set to YES the extra parameters required automatically appear in their correct order For example when GRAPHICS PLOT is YES the extra parameters ORD MAX ORD MIN SCALE and GRID appear When GRAPHICS PLOT is NO the extra parameters ORD MAX ORD MIN SCALE and GRID no longer appear 53 Lambda 20 Lambda 40 UV Vis Spectrometers Tagging a Parameter You tag a parameter to change it at appropriate times during the analysis for example prior to the start of each sample measurement Untagged parameters can only be changed prior to the start of a method The following table shows the type of tagging and when it appears during the analysis Tag a parameter as follows 1 Select the parameter to be tagged 2 Press PARAM 3 Select the appropriate tagging with the arrow keys 54 Methods 4 Press ENTER to confirm the tag Every parameter can be tagged For par
21. HIGH HAUTE VOLTAGE TENSION WARNING UV RADIATION HARMFUL TO THE EYES HOT COMPONENTS RISK OF BURNS ACHTUNG UV STRAHLUNG GEFAHRDUNG DER AUGEN HEISSE BAUTEILE VERBRENNUNGSGEFAHR ATTENTION RADIATION UV DOMMAGEABLE POUR LES YEUX PARTIES CHAUDES RISQUE DE BRULURES 18 Introduction 2 Introduction Introduction The Lambda 20 and Lambda 40 are versatile spectrometers operating in the ultraviolet UV and visible Vis spectral ranges The spectrometers have some common features Keyboard and Display Cover Display Power Switch Keyboard Space for Optional Printer onnector Panel Figure3 Features common to Lambda 20 and 40 Spectrometers Keys f JF dl A Help Param lt I F4 8 9 CE i i 4 5 6 BE Method Print Stop 11213 B NE T Goto Autozero Start 0 s Enter j wes 21 Key HELP PARAM 3 gt METHOD PRINT STOP GOTO AUTOZERO START 0 to 9 E ENTER CE Lambda 20 Lambda 40 UV Vis Spectrometers Provides additional parameter information on the display Selects next parameter Switches to next lower level Selects previous or next element in a particular level Selects methods Use with numerical keys see Selecting a Method on page 48 Prints out the top line of the current standby or method header display Stops a method Switch
22. Rare lesben 9 TEC 1010 Compliance eese 10 ESA Compliance inerenti ntes 10 UL Compliance eie te pee erect m tke dp ted 10 Electrical Protect sedeo tnr a ae 10 Electrical Safety et nr nern su 10 Electromagnetic Compatibility EMC sse 12 Environment rere terere iate ete dea 14 Symbols Used on the Instrument sese 17 Introduction coc R SSS 19 InthOdUCHON a A ed e Pi da 21 Keys EE 21 Startup and Shutdown 4 erae eee eee esee esee eene eene tn seen sesta ss ennu 27 Startup and Shutdown uuessessessnssnesnnennnennnennnnennennnennnnnsn esse enne nnne 29 SL rena ni i o EED S 29 SN E en 30 Single Cell Holder ete ee teet aereo ee hen 31 D SCrIDtiOT a aaa PE eH RUE 31 Installing the Single Cell Holder eene 32 Aligning the Single Cell Holder sese 33 Minimum Volume Applications 37 Operating without Methods ceres eee ee eren ener eene tn neto nest tn se tnun 39 Operating without Methods sss ene ener 41 OYEIVIEW UMEN 41 Setting the Wavelength Manually esee 41 Manual AUtoZ6eto eerie tete ceat lite coe i ata 42 Quick Sample Measurement sese 42 A 43 Proa Rei Wet atenta Heredia 44 Help ee mp et ei ona ha e ident 44 ji unn see 45 What are Methods nice
23. The Print key can be used on its own to print the top line of the standby or method header displays or in combination with other keys to provide other functions as listed in the following table PRINT Prints out the current values shown on the top line of the standby and method header displays Calls up the graphics plot parameters after each sample analysis see Replot on page 79 1 PRINT Prints out the method parameters 2 PRINT Prints out a directory of methods available in the branch 3 PRINT Prints out the additional method or parameter information 6 PRINT Prints out the Peltier cell holder temperature shown on the display Functions only when Peltier accessory is installed 69 Lambda 20 Lambda 40 UV Vis Spectrometers Help Key To view the additional information text for a particular method or parameter proceed as follows 1 Select the desired method or parameter 2 Press HELP 3 Continue to press HELP to view all the text OR Press STOP to interrupt the help function NOTE Help text is available in German refer to the print configuration in SuperUser 70 Using Methods 6 Using Methods Methods Overview The spectrometer incorporates the basic types of methods shown in the table below 1 TIME DRIVE Measurement over a Single Wavelength certain period at one Measurements page 82 wavelength SCAN Scanning spectra and Scanning a Spectrum derivative spectra page 85
24. WAVELENGTH Measurement at several Measurement at Several PROG wavelengths differential Wavelengths page 88 and ratio analysis at several wavelengths 4 CONCENTRATION 1 Determination of CONCENTRATION 1 concentration using peak Method Peak heights height page 91 CONCENTRATION 2 Determination of CONCENTRATION 2 concentration using peak Method Peak Areas 2nd area or 2 derivative Derivative page 96 ENZYME Enzyme kinetics cac Kinetics page 105 KINETICS SUBSTRATE KIN Substrate kinetics Substrate Kinetics page 109 OLIGOQUANT 1 Quantitative analysis of Methods for Quantitative oligonucleotides up to 50 Analysis of bases long Oligonucleotides page 115 73 Lambda 20 Lambda 40 UV Vis Spectrometers OLIGOQUANT 2 Quantitative analysis of Methods for Quantitative oligonucleotides longer Analysis of than 50 bases Oligonucleotides page 115 DATE TIME To enter and change the Date Time page 121 date and time 74 901 WAKEUP To switch on the lamps Wakeup page 123 and allow them to warm up before the start of the working day SELF CHECK Instrument internal Self Check page 125 check of the topics Preprogrammed Methods Using the preprogrammed methods you only need to change a few parameters to quickly create your own individual methods A list of the preprogrammed methods is given in the table below TIME DRIVE Measurement at 500 nm SCAN Absorbance scan from 900 nm to 200 nm CA WAVPRO
25. appears on the top right when cycles gt 1 Printout If PLOT REFERENCES and PRINT DATA are set to YES the calibration curve and results are printed out CONCENTRATION 2 Method Peak Areas 2 Derivative Summary of the procedure for creating a CONCENTRATION 2 method e Determine the measurement wavelength s see Determining the Measurement Wavelengths Peak areas on page 96 e Determine the threshold value 2 derivative see Determining the Measurement Wavelengths 2nd derivative on page 97 e Create a CONCENTRATION 2 method see Creating a Method on page 99 e Establish a calibration curve using references see Establishing the Calibration Curve on page 101 e Measure the sample see Measuring the Sample on page 102 Determining the Measurement Wavelengths Peak areas To determine the wavelengths 96 Using Methods 1 Record the spectrum of the sample see Scanning a Spectrum on page85 2 Selecta strong peak and note the wavelength at its start WAVE MAX and end WAVE MIN WAVE MIN WAVE MAX 2 WAVE MIN WAVE MAX 4 Determining the Measurement Wavelengths 2 derivative To determine the wavelengths 1 Record the spectrum of the sample see Scanning a Spectrum on page 85 2 Select a strong peak and note the wavelength at its start WAV MAX and end WAV MIN 3 Record the spectrum of the same sample using the 2 derivative D2 mode over the wavelength range determined in ste
26. ceteri area 47 Selecting a Method sse eene nnne enne 48 Default Methods x reae 22728238 RES RAR 49 Lambda 20 Lambda 40 UV Vis Spectrometers Editing Methods ooh ite a etait ets 5 Modifying a Method sese 52 Deleting a Method sse 56 Creating a New Method sss eee 57 New Method Namoe eese ener a 60 Checking a Method ee ie ect rhe sten 62 Copying Method Parameters into a New Method File 63 Printing Out a Method ssssesssesseeeeeen eee 66 Printing Out a Directory eese enne enne nennen 67 Spectrometer Directory cnans aa a a a ennt 67 Branch Directory 2 esce ren e reri end 67 Print Key ie stet al eee eese tee itt 69 Help Key tede nio decade esie tis 70 Using MethOdS 2 iere rret eerta erro reb renta ae sra e C Pro ter pe eta eco PE orcos se be Paga 71 Methods Overview neenon er niert nes 73 Method Procede PER REUS T Analysis Procedure 4 eerie ete RI Ce REO ts 78 iP fees 79 Single Wavelength Measurement essere eene een 82 PLOCE LT 82 Example of the Display Shown During the Measurement 84 Printout oesie hae dre aid 84 Scanning a Spectrum isr eein ie anaE E e R E E EN 85 Proceduren seco tem else r etate ede S 85 Example of the Display Shown During the Measurement
27. gel plate reader Vacuum Sipper or Peristaltic Sipper 129 Lambda 20 Lambda 40 UV Vis Spectrometers 130 Requirements for Operation with Accessories The following preconditions must be fulfilled in order to operate with accessories e The accessory in use must be properly selected in the accessory parameter on the appropriate method e The connector panel for the accessory in question must be installed in the spectrometer Some accessories such as Peltier or Temperature sensor also require an accessory board The various circuit boards and connector panels are described in the Installation and Maintenance Guide NOTE Operating with Accessories Using Methods with Accessories You must set the parameter ACCESSORY in the method to the appropriate accessory being used see Parameter Numbers and Descriptions on page 169 You can simulate operation with an accessory by setting the ACCESSORY parameter in the method to the accessory required However with cell changers only one sample can be measured Simulation of Accessories You can simulate operation with an accessory without the accessory being connected Proceed as follows to set up a method to simulate operation with an accessory 1 Switch on the spectrometer in SuperUser mode see SuperUser on page 207 2 Select the CONFIGURATION branch 3 Select method 7 ACCESSORY CONFIG 4 Select ACCESSORY YES This modifies the method 5 Press START This st
28. mL of solution in a cell of 1 cm pathlength 176 Parameter Numbers and Descriptions CREEPING CYCLE 21 color number optical density absorbance Number of cycles after the reaction end time Range 0 no further measurements after end point 2 to 99 This parameter is used to compensate for creeping reactions The spectrometer calculates the slope for each interval If it remains constant the substrate reaction is complete the spectrometer will then determine the difference in absorbance for the substrate reaction If the slope does not remain constant two further measuring intervals are added 177 Lambda 20 Lambda 40 UV Vis Spectrometers Absorbance Difference CREEP TIME Duration of the measuring interval units as selected for TIME UNIT Appears only if CREEP CYCLES is 2 or more Range 0 1 to 999 9 A CREEPING CYCLES s aon al bo me CREEP TIME fe A Creeping Reaction AA END TIME P IME gt Time e ee pStart ofthe Method Enzyme Added 3 Start of Measurement CUR FIT 10 Different types of calibration Curve Fit can be calculated by the spectrometer software LINEAR Used when the measured values vary linearly with the concentration the curve passes through the origin LIN INTERC Used when the measured values vary linearly with the concentration the curve has an intercept on the measured value axis to compensate for background interferences 178 Parameter Numb
29. sample compartment cover 9 Press AUTOZERO and wait until the autozero is completed 10 Open the sample compartment cover 11 Remove the blank and insert the cell with sample solution in the sample cell holder 12 Close the sample compartment cover 13 The absorbance A or transmittance T reading and wavelength are shown on the display The ordinate mode of the last used method always appears on the display Use a TIME DRIVE method to change from absorbance A to transmittance T 14 Press PRINT to print out the reading Reset By a full reset the spectrometer and its program are returned to the default condition You can carry out a full reset at any time 43 Lambda 20 Lambda 40 Operation and Parameter Description 44 NOTE In carrying out a full reset all methods will be erased Before carrying out a full reset make sure that all important methods are printed out To carry out a full reset l 2 Switch off the spectrometer Press 7 9 seven nine point simultaneously Keep the keys pressed and switch on the spectrometer Keep the keys pressed until the display appears The instrument requests the printer type Select the printer type and press START After the full reset is completed a status report is printed out when a printer is connected NOTE There are default methods stored in the internal memory of the spectrometer These methods are not deleted after a full reset and can be
30. stirrer has been switched on place a small magnetic stirring bar in each of the cells The arrangement is such that whilst measurement is taking place in the one cell the following cell will be stirred SAMPL TIME Sample aspiration time in seconds for the sipper Range 0 1 to 99 9 Enter value and confirm by pressing ENTER DELAY TIME Delay between the end of the aspiration process and the start of the measurement Range 0 0 to 99 9 Enter value and confirm by pressing ENTER AUTO PURGE Switches the autopurge function on and off Option YES NO Select with arrow keys RETURN Pump in reverse direction after sample measurement peristaltic sipper only Option YES NO Select with arrow keys 138 Operating with Accessories Parameter Description RETURN TIME To enter the reversed flow time for the optimization peristaltic sipper only Range 0 0 to 99 9 Enter value and confirm by pressing ENTER To select the sipper accessory Further information about operation is described in the user documentation provided with the sipper AS 90 91 To select the autosampler accessory Further information about operation is described in the user documentation provided with the autosampler LINTRANS To select the linear transporter accessory Further information about operation is described in the user documentation provided with the linear transporter 139 Lambda 20 Lambda 40 UV Vis Spectrometers
31. surfaces such as the matt finish surfaces Always wipe the optical surfaces of cells dry and free of fingermarks using a soft cloth or cleaning tissue just before placing them in the cell holder Protect cells from scratches and never permit them to rub against one another or against other hard surfaces Avoid abrasive corrosive or stain producing cleaning agents and make certain that the exposed surfaces of cells are optically clean When measuring cold solutions always bear in mind that condensation can form on the optical surfaces Make certain no bubbles cling to the inner surfaces of the cell particularly when handling cold solutions For maximum precision and accuracy calibrate and test with cells of the same type and always insert cells into the holders with the same orientation Pressure Buildup in Cells Only fill the cell so full that the liquid meniscus is just above the radiation beam The remaining air space in the cell is then adequate to compensate for any slight increase in pressure in the cell during routine operation If for analytical reasons it is necessary to fill the cell completely insert the stopper only lightly so that the liquid in the cell has a chance to expand Do not insert a stopper forcefully into a completely filled cell since this is likely to cause the cell to burst Care e When working at higher temperatures use a drilled stopper 0 4 mm hole to allow for expansion in the cell
32. t Do not change the value for this parameter otherwise you will get the wrong result Using Methods Once the OLIGOQUANT 1 method is created the method parameters can be edited The base sequence must be entered from base 5 to 3 Use the numeric keys according to the table below to enter the base sequence Lo em INC s qum NC e eme o Lr mmm E o 119 Lambda 20 Lambda 40 UV Vis Spectrometers The following table lists typical OLIGOQUANT 2 parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter No Parameter Value No Parameter Value 17 SLIT 20nm 1 ORDINATE MODE A 2 WAVELENGTHS 1t 3 WAV 1 260 0 nm FACTOR 1 59 PATHLENGTH 1 0 cm 60 NUMBER OF dA 60 NUMBER OF dC 60 NUMBER OF dG 60 NUMBER OF dT 60 NUMBER OF N 0 65 CHNGE CONSTANTS NO pense o 37 SAMPLE ID Only available with Lambda 40 t Do not change the value for this parameter otherwise you will get the wrong result 120 Using Methods Date Time 10 11 12 Select the DATE TIME method 900 Press PARAM Press PARAM again Use the arrow keys to select the realtime or the internal clock Press PARAM again Use the arrow keys to select the day Press PARAM again Type in the date year month day 960429 using the numeric keys Press ENTER to confirm the entry Press PARAM again Type in the time hours minu
33. the entry Press PARAM again Use the arrow keys lt gt to select UV lamp On or Off Press ENTER to confirm the entry Press PARAM again Use the arrow keys lt gt to select Vis lamp On or Off Press START to activate the wakeup method OR Press STOP to cancel When this method is activated the lamps are switched off and are then switched on again at the preselected WAKEUP time 123 Lambda 20 Lambda 40 UV Vis Spectrometers To exit the WAKEUP method press STOP and the display returns to standby Both lamps go on 901 WAKEUP lt gt PARAM Y so MODIFY METHOD y Para DATE 000000 ENTER Y Ente v Para TIME 0000 ENTER Y Ente v Para Y LAMP UV NO Sot Y Ente v Para Y LAMP Vis NO lt Y Star 124 Using Methods Self Check The spectrometer calibrates the signals from the optics and prints a report at the end We recommend you do a self check at regular intervals l 2 8 Make sure that the beam path is not obstructed Select the SELF CHECK method 999 Press START to activate the self test The spectrometer then asks you to select the printer type you are using Use lt gt to select your printer type Press START to confirm the selection and move to the next parameter Depending on the type of printer selected the spectrometer
34. then be selected see Selecting a Method on page 48 Can only be set in SuperUser mode APPLICATION PARAM gt Y lt gt y CONFIGURATION PARAM gt Param 500 0 nm 0 000 A INPUT gt lt About the Various Branches With branches that do not contain analytical methods you must always press START to make the changes effective APPLICATION the Application Branch This branch contains the analytical methods Appendix COMMUNICATION the Communication Branch This branch contains a method for controlling the spectrometer via an external computer Control of the spectrometer via an external computer A precondition is that the computer should be equipped with a program compatible with the method in use including UV WinLab or PECSS version 3 1 or later For example To control the spectrometer using the UV WinLab software proceed as follows 1 Set the RS 232 interface to computer see RS 232 PORT CONF 2 Connect the computer to this interface 3 Start UV WinLab 215 Lambda 20 Lambda 40 UV Vis Spectrometers CALIBRATION the Calibration Branch Only for service personnel or very experienced users This branch contains methods for wavelength calibration of the spectrometer 19 Switches the dark signal compensation on and off CALIBRATION i A residual current dark signal flows through the detector even when there is no beam This s
35. under reduced pressure When you dispose of lamps that are defective or otherwise unusable handle them correctly to minimize the implosion risk UV Radiation You should be aware of the health hazards presented by ultraviolet radiation e When the deuterium UV lamp is illuminated do not open the spectrophotometer covers unless specifically instructed to do so in the manual e Always wear UV absorbing eye protection when the deuterium lamp is exposed e Never gaze into the deuterium lamp Safety Information Symbols Used on the Instrument Warning symbol shown on the spectrometer housing SS x I D ENS CM Pi lc Sede Figure 1 Lambda 20 40 Spectrometers m UV Radiation Risk of Eye Damage 2 The lamp emits intense UV radiation which can damage your eyes A Do not gaze into a lighted lamp Always wear UV absorbing safety glasses when looking at the WARNIN s radiation from the lamp Hot Surfaces Risk of Burns IA Lamps soon become hot and also heat up other components in the lamp compartment Allow the components to cool to room temperature before installing new lamps or making adjustments in the lamp compartment WARNING 17 Lambda 20 Lambda 40 UV Vis Spectrometers Warning Labels on the Instrument Warning labels shown on the inside of the lamp compartment Figure 2 Lambda 20 40 Spectrometers The following warnings are shown on the inside of the lamp compartment DANGER DANGER
36. voltages can still be present within the instrument e When the instrument is connected to line power terminals may be live and opening covers or removing parts except those to which access can be gained by hand is likely to expose live parts e Capacitors inside the instrument may still be charged even if the instrument has been disconnected from all voltage sources Gef hrliche Spannung im Ger t e Auch in ausgeschaitetem Zustand kann an einigen Stellen im Ger t Netzspannung anliegen wenn das Ger t am Stromnetz angeschlossen ist e Auch bei ausgeschaltetem Ger t und getrennter Netzverbindung k nnen Kondensatoren im Ger t noch mit gef hrlicher Spannung geladen sein Farlig sp nding i apparatet fare for kv stelser e Ogs i slukket tilstand kan der v re netspsending nogle steder i apparatet hvis apparatet er tilsluttet til str mnettet e Selv n r apparatet er slukket og str mforbindelsen er afbrudt kan kondensatorerne i apparatet v re ladet med farlig sp nding En el aparato existen voltajes letales e Incluso con el interruptor desconectado puede haber voltaje dentro del equipo e Cuando el instrumento se encuentre conectado a la red el ctrica los terminales pueden estar bajo corriente y stos quedar expuestos al abrir las cubiertas o al extraer componentes exceptuando aquellos a los cuales se puede acceder con la mano e Los condensadores internos del aparato pueden permanecer car
37. when asked until they have all been measured When the graphics plot parameter in the method is set to YES the display is XXX SAMPL n START STOP AZ PRINT This allows you to change the scale of the plot for the sample just analyzed and replot the result see Replot on page 79 NOTE Pressing STOP interrupts a method in progress and returns you to the ready display 78 Using Methods Replot You can replot sample data for last analyzed sample in certain methods when the graphics plot parameter in that method is set to YES Replot the data as follows l Set the graphics plot parameter to YES in the method parameters before starting the method Start the method The following ready display appears after the first sample analysis XXXSCAN SMPL 2 START STOP AZ PRINT Press PRINT The following display appears ORD MAX 0 000 A ENTER gt lt Enter a new upper value for the ordinate if required and then press ENTER to confirm the value Press START to move to the next parameter The following display appears ORD MIN 0 000 A ENTER gt lt Enter a new lower value for the ordinate if required and then press ENTER to confirm the value 79 Lambda 20 Lambda 40 UV Vis Spectrometers 7 Press START to move to the next parameter The following display appears SCALE 50 0 nm cm lt gt 8 Select a new abscissa scale value u
38. 140 Care 8 Care Care WARNING Unauthorized Adjustments and Servicing Do not attempt to make adjustments replacements or repairs to this instrument except as described in the accompanying User Documentation Only a PerkinElmer service representative or similarly trained and authorized person should be permitted to service the instrument Daily Care CAUTION Do not leave samples particularly those given to fuming or evaporation in the sample compartment for longer than necessary If any type of sample handling system is installed and portions of it are left in the sample compartment such as a sipper and flowcell make certain that the system is cleaned at the end of the working day Generally such systems should be filled with deionized water when left overnight Immediately clean all spilled materials from the affected area and wipe it dry with lintless paper or cloth If you have to wipe the sample compartment windows make sure you do not introduce scratches Sample windows are optical components and you should handle them in the sampe way as high quality cells Risk of damage to Spectrometer Take care not to spill liquids onto the spectrometer Expensive damage can result to the optics or electronics if liquids are spilled and run inside the instrument or onto the keyboard 143 Lambda 20 Lambda 40 UV Vis Spectrometers 144 Use and Care of Cells Cell Handling Only hold cells by non optical
39. C WAV2 and CALC WAV1 and calculates the difference in height between these two points Only values that have exceeded the set THRESHOLD are taken into account This function can avoid errors that may occur through displacement of the spectrum CALC WAV2 CALC WAV1 MOL MASS dA Sets the relative molecular mass of the Adenine nucleotide Range 0 to 99999 Appears with the Oligoquant methods 188 MOL MASS N NUMBER OF dA r NUMBER of dC Parameter Numbers and Descriptions MOL MASS dC MOL MASS dG i MOL MASS dT i Sets the realitve molecular mass of the Cytosine nucleotide Range 0 to 99999 Appears with the Oligoquant methods Sets the relative molecular mass of the Guanine nucleotide Range 0 to 99999 Appears with the Oligoquant methods Sets the relative molecular mass of the Thymine nucleotide Range 0 to 99999 Appears with the Oligoquant methods Sets the relative molecular mass of the nucleotide N is used to represent any base Range 0 to 99999 Appears with the Oligoquant methods Sets the number of Adenine nucleotides Range 0 to 999 Appears with the Oligoquant methods Sets the number of Cytosine nucleotides Range 0 to 999 Appears with the Oligoquant methods 189 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter Description NUMBER of dG NUMBER of dC NUMBER OF N OPER ID Sets the number of Guanine nucleotides Range 0 to 999 Appears with the Oligoquan
40. E Operating with the Cell Changer During the interval time the spectrometer measures once at each location To do this you need to know the minimum measuring time t min tmn N x 3 x RESPONSE x 0 1 seconds where N the number of cells 3 x RESPONSE measuring time per cell x relocation time from cell to cell The interval time should always be greater than the required minimum time 184 Parameter Numbers and Descriptions LAG TIME Lag time This is the time from the start of the method to the start of calculation units as selected for TIME UNIT After this time a constant reaction rate should have been reached Range 0 0 to 999 9 Measurement begins with the start of the method However enzyme activity is only calculated from the end of the lag time Only when using POSTRUN KIN YES or manual operation together with CALCULATE REGRESSION LAMP 15 Switched on lamps UV 190 nm to 326 nm VIS 326 nm to 1100 nm UV VIS 190 nm to 1100 nm NOTE n order to preserve the UV lamp switch off the lamp only at the end of the working day and allow the lamp to cool off for at least 2 minutes before switching on again LAST TUBE n The last sample tube location of the batch to be analysed n is any location number up to the maximum location for the tray type Range 1 to the maximum location number of the tray being used Appears with the autosampler accessory 185 Lambda 20 Lambda 40 UV Vis Spectrometers LINE
41. ER TR TOT TIM This message appears with kinetic measurements Select a shorter interval time HINT Appears with enzyme methods with cell changers if the SELECT interval time does not correlate with the measurement INTERVAL TIME time Select the INTERVAL time so that the TOTAL TIME can be divided by the INTERVAL time evenly 157 Lambda 20 Lambda 40 UV Vis Spectrometers 158 PROBLEM MARK NOT SET PROBLEM METHOD NOT FOUND PROBLEM ACCESSORY NOT INITIALIZED PROBLEM METHOD NO LIMITS 1 999 ERROR LAST METHOD PROBLEM METHOD PROTECTED DON T PROTECT ALL METHODS COPY FROM MARK was selected without first initiating MARK FOR COPY First use MARK FOR COPY and then select COPY FROM MARK A method has been tagged MARK FOR COPY although the method in question is no longer available This error is shown when the instrument gets no response from an attached accessory during startup Check that the accessory is correctly connected switched on functioning normally The method number entered is outside the displayed range Enter a number within the range 1 to 999 An attempt was made to delete all methods Retain at least one method in the memory otherwise the spectrometer cannot work An attempt was made to select a fully protected method The protection has to be modified if the method is to be used An attempt was made to ALL protect all the methods Retain at least one meth
42. G Wavelength program at 260 nm FACTOR CONCENTR Quantitative analysis without calibration Absorbance at a specific wavelength is multiplied by a factor to give concentration of analyte Using Methods ES Type of Method CALIB CONC Quantitative DEED measurement after calibration using 3 references 805 ONE REF CONC Quantitative analysis using 1 reference for calibration 806 TWO REF CONC Quantitative analysis using 2 references for calibration 807 THREE REF CONC Quantitative analysis using 3 references for calibration CONC VARIABLE Quantitative analysis measurement after calibration using 3 references SURVEY SCAN Scans a survey spectrum fast scan speed a survey spectrum fast scan Scans a survey spectrum fast scan speed ABSORBANCE SCAN Scans absorbance spectra T SCAN Scans transmittance spectra 000 transmittance Scans transmittance spectra 000 812 UV SCAN Scans absorbance spectra between 400 nm and 200 nm 813 VIS SCAN Scans absorbance spectra between 900 nm and 400 nm REPETITIVE SCAN Scans absorbance spectra between 400 nm and 200 nm CYCLE WAVPROG Wavelength program at 60mm 000 program at Wavelength program at 260mm nm 816 RATIO Ratio of absorbance readings at different wavelengths 75 Lambda 20 Lambda 40 UV Vis Spectrometers Ne Type of Method 817 DIFFERENCE Difference of absorbance N at different wavelengths 76 Using Methods Method Procedure Wh
43. HEET Appendix ACCESSORY CONFIG USER CONFIG Accessory mode Parameters ACCESSORY Switches accessory mode on and off MANUAL To select operation with the standard cell holder CELL To select the type of cell changer NO is no cell changer CALL5 CELL6 CELL8 CELL9 CELL 13 is for cell changer type SIPPER To select the type of sipper VASI PESI SI sipper VA vacuum PE peristaltic AS90 91 To select the autosampler AS 90 91 LINTRANS To select the linear transporter Switches from single to double beam mode switches background correction on and off Parameters BEAM To set the beam DB Double beam SBR Single reference beam SBS Single sample beam BASELINE CORR YES NO Background correction on off LAMP UV YES NO Switch UV lamp on off LAMP Vis YES NO Switch UV lamp on off NOTE When using single beam mode operate only with ordinate mode T 221 Lambda 20 Lambda 40 UV Vis Spectrometers FACTORY Calibration peaks offsets and filter change points CONFIG Parameters ONM OFFSET Only for service personnel Default 0 0 nm ABS FACT Only for service personnel Default 1 0 D2 OFFSET Only for service personnel Default 0 0 nm FILTER n Wavelengths for filter change filter 2 7 Default filter 1 980 0 nm filter 2 794 0 nm filter 3 683 0 nm filter 4 558 0 nm filter 5 420 0 nm filter 6 383 0 nm filter 7 326 0 nm lamp changeover Wavelengths for lamp chan
44. Lambda 20 Lambda 40 Operation and Parameter Description Release History Part Number Publication Date 09935056 November 1996 B June 2000 User Assistance PerkinElmer Ltd Post Office Lane Beaconsfield Buckinghamshire HP9 1QA Printed in the United Kingdom Notices The information contained in this document is subject to change without notice PerkinElmer makes no warranty of any kind with regard to the material including but not limited to the implied warranties of merchantability and fitness for a particular purpose PerkinElmer shall not be liable for errors contained herein for incidental consequential damages in connection with furnishing performance or use of this material Copyright Information This document contains proprietary information that is protected by copyright All rights are reserved No part of this publication may be reproduced in any form whatsoever or translated into any language without the prior written permission of PerkinElmer Inc Copyright O 2000 PerkinElmer Inc Trademarks Registered names trademarks etc used in this document even when not specifically marked as such are protected by law UV WinLab is a trademark of PerkinElmer Inc PerkinElmer is a registered trademark of PerkinElmer Inc Contents Contents Contes edema esu 3 Safety InformatiOn cree eee ee eere eese enne een neta seen osse ense etes etos a seen ose eo see 7 Safety Informations nhe a
45. Lambda 40 UV Vis Spectrometers 108 Printout The results are printed out at the end of the analysis Determining the Blank Value Determine the blank value of the reaction as follows l 2 Select the appropriate ENZYME method Set parameter BLANK 0 0 in the method Carry out a measurement according to your procedure using a cell filled with redistilled water in place of the sample Enter the result of the measurement in the parameter BLANK Using Methods Substrate Kinetics Select a SUBSTRATE KIN method for substrate kinetic measurements Manual Procedure 1 Select the appropriate SUBSTRATE KIN method The following table lists typical SUBSTRATE KIN parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169for a detailed description of each parameter 109 Lambda 20 Lambda 40 UV Vis Spectrometers No Parameter Value No Parameter Value 17 SLIT 20nm 3 WAVELENGTH 340 0 nm 14 20 22 11 12 6 18 25 27 29 34 36 110 RESPONSE 0 55 15 LAMP UV Vis TIME UNIT min 23 DELAY TIME 0 0 min CONC FACTOR 1 0 10 DIL FACTOR DIVISOR 1 0 7 BLANK CONC UNIT C 16 AUTOZERO SAMPLES BATCH 0 19 FIRST SAMPLE 1 GRAPHICS PLOT YES 26 ORD MAX 1 000 A ORD MIN 0 000A 28 SCALE 20 mm min GRID YES 32 PRINT DATA POSTRUN KIN YES 35 AUTO METHOD OPER ID 37 SAMPLEID Only available with Lambda 40 Modify the method as requir
46. Samples that are polarizing in nature or have a double index of refraction are often difficult to measure accurately The emerging monochromatic radiation is slightly polarized due to having been refracted 151 Lambda 20 Lambda 40 UV Vis Spectrometers Thin film samples also pose a problem since optical interferences may develop causing a regular interference pattern to be superimposed on the spectral curve 152 Analytical Notes Solvent Properties The solvent should meet the following requirements e Jt should dissolve the sample without reacting with it e The radiation absorption in the scanning region should be low High absorption by the blank reduces the reference energy thus increasing noise e Evaporation should be fairly low at ambient temperature In general aromatic compounds exhibit high absorption in the UV region and hence are not suitable as solvents for measurements in this region Water is virtually the only useful solvent below 195 nm but it must be freed from oxygen to attain best transmission Whenever you are going to use a solvent with unknown absorption characteristics scan its spectrum first to determine whether it is suitable The lower wavelength limits of a number of commonly used solvents are presented in the following table The lower limit has been defined as that wavelength at which 10 mm of pure solvent has a transmission of 1095 153 Lambda 20 Lambda 40 UV Vis Spectrometers
47. TYPE 31 Type of line used for the graphics printout valid only when GRAPHICS PLOT YES DASH1 AUTO For each curve a different type of line is used in the sequence DASH1 DASH2 DASH3 DASH4 DASH1 1 Concentration 1 Sets the mode in which the measurements are made 1 Wavelength 2 Wavelength 3 Wavelength 1 Wavelength measuring absorbance at one wavelength Measured Value A A1 2 Wavelength correction for a sloping baseline by measuring absorbance at two wavelengths A Measured Value A A1 A A2 186 Parameter Numbers and Descriptions 3 Wavelength correction for a sloping baseline by measuring absorbance at three wavelengths Measured Value 63 Alda A Ay 4 Alda 1 Concentration 2 Sets the mode in which the measurements are made There are four possibilities TOTAL AREA PEAK AREA DERIV 2 FIX DERIV 2 PEAK TOTAL AREA the total peak area between wavelength maximum and wavelength minimum is calculated WAVE MIN WAVE MAX A PEAK AREA the peak area between wavelength maximum and wavelength minimum is calculated with a correction for the baseline drift WAVE MN WAVE MAX x 187 Lambda 20 Lambda 40 UV Vis Spectrometers DERIV 2 FIX height of the 2 order derivative curve at wavelengths CALC WAV2 and CALC WAV is measured CALC WAV1 DERIV 2 PEAK height of the 2 order derivative peak The software locates the extreme values around wavelengths CAL
48. UE 1 VALUE 2 VALUE 3 CUR FIT LINEAR 22 CYCLE TIME 0 1 min Only available with Lambda 40 100 Using Methods 2 Modify the parameters as required using the wavelengths and threshold values determined above Once a method is created you can save it and use it for the same analysis when required without having to redetermine the values Establishing the Calibration Curve 1 Press START to start the measurement 2 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is REF n XXX START STOP AUTOZERO Insert a cell containing the sample solution and press START 3 Insert the references in sequence when asked When they have all been measured the instrument prints out the calibration curve and the results You can now amend the calibration curve see Processing the Calibration Curve CONCENTRATION Methods on page 103 if required You can use previously established calibration curves or reference values see REFS in Parameter Numbers and Descriptions on page 169 101 Lambda 20 Lambda 40 UV Vis Spectrometers Measuring the Sample 1 Press START to start the measurement 2 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is CONC 2 SMPL 1 START STOP AUTOZERO
49. absorbance Cannot calculate slope Because too few points read A kinetic method was interrupted with STOP Too few points available to calculate the slope End time out of limits Change end time In recalculation an out of limits value for END TIME was entered Enter a lower END TIME value and repeat the procedure 163 Lambda 20 Lambda 40 UV Vis Spectrometers 164 Error Meaning Lag or total time is not divisible by interval time 0 0 In recalculation the LAG TIME or TOTAL TIME entered was not divisible by the INTERVAL time Enter a value for LAG TIME or TOTAL TIME that is fully divisible by the INTERVAL time Lag or total time out of limits Change lag and or total time In recalculation a greater value for LAG TIME or TOTAL TIME was entered than the actual measuring time Enter a lower value for LAG TIME or TOTAL TIME Lag time greater than total time Change lag time In recalculation a value for LAG TIME was entered that was greater than the measuring time Enter a LAG TIME less than the TOTAL TIME More than one peak within wavelength limits Change threshold or measurement wavelengths More than one peak identified within the selected measuring range Change WAV MIN and WAV MAX or THRESHOLD so that only one peak is detected No peak detected Change threshold The THRESHOLD is too high to detect peaks Select a lowe
50. alignment Figure 4 Single Cell Holder B0505071 NOTE Depending on the spectrometer the single cell holder can be installed in two different positions in the sample compartment Always install the holder such that the arrow on the cell holder lines up with the center point on the baseplate see Installing the Single Cell Holder on page 32 31 Lambda 20 Lambda40 Operation and Parameter Description 32 Inscription legible Use in Spectrometer on Holder LAMBDA In this position the cell holder can be used with all Lambda Series Spectrometers BIO LAMBDA 2 In this position the cell holder can be used with Lambda 2 Series Spectrometers as Lambda 10 20 40 Bio baseplate with 4 threaded holes The smallest beam diameter is exactly in the middle of the cell This is useful especially for operation with micro and semi micro cells Installing the Single Cell Holder There are two single cell holders provided with the instrument one for the sample beam and one for the reference beam Install the single cell holder in the sample compartment as follows 1 Orientate the holder so that the lifter is toward the rear of the sample compartment 2 Lower the holder so that the two alignment holes slip onto the two studs on the baseplate at the bottom of the sample compartment The arrow on the cell holder must line up with the centre point of the baseplate and BIO LAMBDA 2 must be legible Startup
51. ameters where tagging is less meaningful LAMP GRAPHICS PLOT tagging Is accepted but not carried out The tagged parameter appears at the appropriate time but cannot be changed AUTOZERO NO lt gt Param AUTOZERO FIX lt gt lt gt AUTOZERO CALL lt gt Enter AUTOZERO NO lt gt 55 Lambda 20 Lambda 40 UV Vis Spectrometers Deleting a Method l 2 56 Select a method that can be deleted Press PARAM Use the arrow keys to select DELETE METHOD Press PARAM again to delete the method The method is deleted as soon as PARAM is pressed and the display returns to the next method header in the list OR Press STOP to cancel 2 SCAN lt gt PARAM START Param MODIFY METHOD PARAM gt Y E DELETE METHOD PARAM gt Y Param Methods Creating a New Method You can create a new method in one of the following ways Create a new empty method file Overwrite an existing method file Creating an Empty Method File l 2 Press METHOD Enter a method number nnn not previously used Press ENTER The first method of the NEW METHOD level appears Use the arrow keys to select the required method type Press PARAM This confirms the creation of the new method OR Press STOP to cancel 57 Lambda 20 Lambda 40 UV Vis Spectrometers 6 Modify
52. and Shutdown Center Point Tube Ports 3 Move the milled posts a little to locate the threaded holes in the baseplate and then tighten the milled posts The tube ports located at the front of the sample compartment allow you to lead tubes from for example flowcells water thermsotatted cell holders in and out of the sample compartment When not in use you should always insert the caps into the ports Aligning the Single Cell Holder Coarse alignment of the single cell holder is carried out as follows 1 Open the sample compartment cover 2 Fill matching cells with a low absorbing solvent deionized water or ethanol 3 Insert one cell into the sample cell holder and one into the reference cell holder Make certain that the cell is pushed down fully NOTE The alignment procedure is for a given cell in a given holder After alignment the cell should always be used in the same holder 33 Lambda 20 Lambda40 Operation and Parameter Description 4 Block the sample and reference beam window on the right hand side of the sample compartment with a card to prevent white light from saturating the detector 5 Return to standby display 6 Using the GOTO key slew the monochromator to 0 nm to obtain a beam of visible zero order radiation in the sample compartment 7 By holding a piece of matt white paper behind each cell holder visually examine the light spot to see that the radiation beam is passing through the cel
53. arameters including sample locations upper and lower limits for the sampler arm AS 90 91 autosampler parameters UP OFFSET To set the up offset in mm Range 0 to 145 mm DOWN OFFSET To set the down offset in mm Range 0 to 145 mm OF ROWS To set the number of rows for the tray Range 1 to 20 X ORIG ROWn To set the X coordinate left to right of row n Range 0 to 302 217 Lambda 20 Lambda 40 UV Vis Spectrometers ZORIG ROWn To set the Z coordinate back to front of row n Range 0 to 185 mm COL ROWn Tosetthe number of columns location in each row Range 1 to 20 DISTANCE n To set the distance from the centre of one column location to the next Range 1 to 99 9 mm X ORIGIN FLUSH To set the X coordinate left to right of the flush solution column location Range 1 to 302 nm Z ORIGIN FLUSH To set the z coordinate back to front of the flush solution column location Range 1 to 185 mm For further details see the autosampler documentation LINTRANS To set the number of positions at which CONFIG measurements are to be made and their location from a fixed point Parameters POSITIONS The number of positions 1 to 20 POSITIONn The position in mm from a fixed point 0 00 to 97 00 218 Appendix COMM CONFIG PRINTER CONFIG RS 232 PORT CONF Protocol for the RS 232 interface for use with a PC The default parameter values are for use with PECSS and an Epson PC
54. are 43 Daily Care 143 Sample Compartment Windows 146 Use and Care of Cells 44 Checking a Method 62 CONCENTRATION 2 Method Peak Areas 2nd Derivative 73 96 Concentration Determination 91 CONCENTRATION Methods with Cell Changers 134 CONFIGURATION the Configuration Branch 217 Copying Method Parameters into a New Method File 63 Creating a New Method 57 D Date Time 121 Default Methods 49 Deleting a Method 56 E Editing Methods 51 Enzyme Kinetics 105 226 Index Error Messages 157 Error Messages Shown on the Display 157 Error Reports on the Printer 163 H Help Key 70 I Instrument Branches 213 Introduction 2 M Measurement at Several Wavelengths 88 Method Procedure 77 Methods 47 Methods for Quantitative Analysis of Oligonucleotides 115 Methods Overview 73 Modifying a Method 52 N New Method Name 60 O Operating with Accessories 129 General 29 Operating without Methods 41 Help 44 Manual Autozero 42 Overview 41 Print 44 Quick Sample Measurement 42 245 Lambda 20 Lambda 40 UV Vis Spectrometers 246 Reset 43 Setting the Wavelength Manually 41 P Parameter Numbers and Descriptions 169 Print Key 69 Printing Out a Directory 67 Printing Out a Method 66 Processing the Calibration Curve CONCENTRATION Methods 103 Protect Functions 208 R Recalculation of Results with POSTRUN KIN 113 Replot 79 Requirements f
55. aseline select a second wavelength at the baseline minimum WAV 2 A WAV 1 WAV 2 OR With a sloping baseline select a wavelength at the beginning and at the end of a peak WAV 2 and WAV 3 WAV 3 WAV 1 WAV 2 92 Using Methods Creating a Method l No Create a new CONCENTRATION 1 method see Creating a New Method on page 57 The following table lists typical CONCENTRATION 1 parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter Parameter Value No Parameter Value 17 SLIT 2 0 nm 1 MODE 1 WAVELENGTH 3 WAV 1 500 0 nm 6 CONC UNIT pg mL 7 REF 1 1 0 pg mL 7 REF2 2 0 ug mL 7 REF 3 3 0 pg mL 8 10 12 15 38 19 8 VALUE 1 VALUE 2 0 2 8 VALUE 3 CUR FIT LINEAR 11 FACTOR DIVISOR 1 0 14 RESPONSE LAMP UV Vis 16 AUTOZERO YES ACCESSORY MANUAL 18 SAMPLES BATCH 0 FIRST SAMPLE 1 21 CYCLES 1 22 CYCLE TIME 0 01 min 32 PRINT DATA YES 25 PLOT REFS NO 26 PRINT REFS YES 93 Lambda 20 Lambda 40 UV Vis Spectrometers 35 AUTO METHOD NO 36 OPER ID 37 SAMPLE ID Only available with Lambda 40 2 Modify the parameters as required using the wavelengths determined above Once a method is created you can save it and use it for the same analysis when required without having to redetermine the wavelength Establishing the Calibration Curve 1 Press
56. ay with wavelength and measured 900 0 nm value INPUT gt Starting point appears after switch on following initialization routine Standby display gt set up absorbance manually set wavelength manually select method print out ordinate reading print out method directory of the relevant branch return to branch header Branch header with branch name APPLICATION gt select the branch method change to another branch select default method of branch view help message print out the spectrometer display return to standby display SGAN Method header with method number and type start method select method processing select another method view help message print out method parameters print out help message return to standby display 24 Introduction Method processing with selected processing MODIFY METHOD tunetion PARAM gt start processing function select another processing function return to method header Parameter directory with parameter names and ORDINATE MODE A value lt gt change parameter select next previous parameter start method view help message for current parameter return to method header WAVe MAX 900 0 nm Parameter with parameter names value and value ENTER gt lt entry field gt change parameter value select next previous parameter start method view help messages for current parameter return to method header Disp
57. copied and amended see Selecting a Method on page 48 Print Press PRINT to print out the top line shown on the standby and method header displays Other functions using the print key are described in Printing Out a Directory on page 67 Help Press HELP to view additional information about the current method or parameter on the display Methods 5 Methods What are Methods Methods are a collection of those parameters necessary for a particular analysis using the spectrometer and are stored as method files You can process large numbers of samples efficiently using the methods The parameter values necessary for the analysis in question need only be set once and are then available on request Up to 200 methods can be stored in the spectrometer each method can be allocated a number between 1 and 999 On delivery 10 basic methods are programmed in the spectrometer these are immediately available for use 47 Lambda 20 Lambda 40 UV Vis Spectrometers Selecting a Method l 48 Switch to the standby display using STOP or PARAM 500 0 nm 0 000 A INPUT gt lt Standby display Press METHOD Enter the method number Press ENTER 500 0 nm 0 000 A SELECT METHOD gt lt Entry Field The method is loaded onto the operational memory The method header then appears on the display OR In the standby display enter the method number Press METHOD Method Numb
58. cription of each parameter No Parameter Value 3 WAVELENGTH 500 00 nm 17 SLIT 2 0 nm 14 RESPONSE 0 5s 16 AUTOZERO YES 18 SAMPLES BATCH 0 21 CYCLES 25 GRAPHICS PLOT YES 27 ORD MIN 0 000 A 29 GRID YES No Parameter Value 1 ORDINATE A MODE 11 FACTOR 1 0 15 LAMP UV Vis 38 ACCESSORY MANUAL 19 FIRST SAMPLE 1 22 CYCLE TIME 0 10 min 26 ORD MAX 1 000 A 28 SCALE 20 nm min 32 PRINT DATA NO Using Methods 35 AUTO METHOD NO 36 OPERID 37 SAMPLE ID Only available with Lambda 40 When a particular parameter is set to YES the extra parameters required automatically appear in their correct order For example when GRAPHICS PLOT is YES the extra parameters ORD MAX ORD MIN SCALE and GRID appear When GRAPHICS PLOT is NO the extra parameters ORD MAX ORD MIN SCALE and GRID no longer appear If necessary change the method parameters Press START to start the measurement Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is TIMEDRIVE SAMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START Continue to insert samples when asked until they have all been measured For additional information see Replot on page 79 83 Lambda 20 Lambda 40 UV Vis Spectrometers Example of the Display Shown During the Measurement
59. d Descriptions esee creer eere eee en eee nne 167 Parameter Numbers and DeScriptions ccccccccesscesteceseceeeeeeeeeeeeeeeeeeneeses 169 O c 205 SUDETU SET cec Let ee tet t deett md etr er of tete 207 Protect Functions eo er ero eer ee wee eee n ntes 208 Setting Protect Functions sess 208 Instrument Branches errearen enne enne nennen nnns 213 Overview of the Instrument Branches 213 Selecting a Branch nennen ia ur 213 About the Various Branches ssensenseenneenneennnennnennnne nn 214 APPLICATION the Application Branch sssssses 214 COMMUNICATION the Communication Branch 215 CALIBRATION the Calibration Branch sss 216 CONFIGURATION the Configuration Branch 217 VALIDATION the Validation Branch 223 TEST the Test Branch aae a tr tet t eite eg 223 Enzyme Kinetic Se auis ee D e I ente edet ores nad 226 Enzymatic Analytical Procedures essere 226 Enzyme Kinetcs a ei E Haein adn 226 Translations of Warnings eese crees eee esee neenon eren nettes tn osse tn seen 231 AS M 243 O NN 245 Safety Information 1 Safety Information Safety Information This manual contains information and warnings that must be followed by the user to ensure safe operation and to maintain the instrument in a safe condit
60. dit kan leiden tot verwondingen van de gebruiker Warning Aviso Significa que a n o observ ncia da instrug o referida poder causar um ferimento ao usu rio 233 Lambda 20 Lambda 40 UV Vis Spectrometers 234 Caution Co1 01 We use the term CAUTION to inform you about situations that could result in serious damage to the instrument or other equipment Details about these circumstances are in a box like this one 000006809 Caution Achtung Bedeutet daB die genannte Anleitung genau befolgt werden muB um einen Ger teschaden zu vermeiden Caution Bem rk Dette betyder at den neevnte vejledning skal overholdes neje for at undg en beskadigelse af apparatet Caution Advertencia Utilizamos el t rmino CAUTION ADVERTENCIA para advertir sobre situaciones que pueden provocar aver as graves en este equipo o en otros En recuadros ste se proporciona informaci n sobre este tipo de circunstancias Caution Attention Nous utilisons le terme CAUTION ATTENTION pour signaler les situations susceptibles de provoquer de graves d t riorations de l instrument ou d autre mat riel Les d tails sur ces circonstances figurent dans un encadr semblable celui ci Caution Attenzione Con il termine CAUTION ATTENZIONE vengono segnalate situazioni che potrebbero arrecare gravi danni allo strumento o ad altra apparecchiatura Troverete informazioni su tali circostanze in un riquadro come ques
61. dure is not used enter 0 Range 0 to 999 Abscissa scale in nm cm or mm min for graphics printout valid only when GRAPHICS PLOT YES Options 0 5 1 2 5 10 20 50 100 nm cm 1 2 5 10 20 50 100 mm min SAMPLE FLUSH 51 To flush the flowcell after each sample measurement Option YES NO 196 Parameter Numbers and Descriptions Parameter SEQUENCE Length of the oligonucleotide base sequence EP Range 2 to 50 Sequence of the oligonucleotide bases The sequence must be entered in order from base 5 to 3 Use the numeric keys according to the table below to enter the base sequence Option Base Adenine Cytosine Guinine Thymine any base Required slit width in nm Options 0 5 1 2 4 SLIT SMOOTH NOTE Only shows with Lambda 40 spectrometer Smoothing according to Savitsky Golay in nm The acceptable level of smoothing is dependent on the scan speed SPEED Up to 1920 nm min 2880 960 nm min nm min 0 6 8 10 nm 197 Lambda 20 Lambda 40 UV Vis Spectrometers Guideline 0 5 x peak width of lowest peak Smooth 2 nm corresponds to Peak width Spectrum h quality not compromised Smooth 6 nm Spectrum quality poor j Smooth value is 10 nm Benzene spectrum not recognizable Smoothing influences resolution and noise an increase in smoothing decreases the noise but resolution suffers A too high degree of smoothing tends to average out the values and t
62. e linear region of the curve is used for calculating the reaction rate Lag Time Linear Region me Substrate Kinetics In substrate kinetics the substrate concentration of a sample solution is determined via enzyme controlled reactions The advantages of such a process are e High specificity that is only one substrate is converted This avoids the necessity of complex sample preparation e A quicker reaction with measuring times of only 3 30 minutes 228 Appendix In the course of the reaction the substrate is converted to product and the reaction can be followed photometrically The reaction is started by the addition of enzyme and proceeds relatively quickly until a state of equilibrium is attained The substrate has been converted by this time and the absorbance does not alter any more The measured difference in absorbance AA is directly proportional to the substrate concentration Cs f AA where Csu is the substrate concentration f is the concentration factor AA isthe measured difference in absorbance Enzyme added yl ime The course of the reaction can deviate from the ideal described above creeping reactions can take place and the absorbance can hence increase even after the substrate reaction has been completed The end point of such a reaction is reached when the slope of the curve remains constant The actual end point can then be determined by extrapolation 229 Lambda 20 La
63. ect functions define the right of access to methods and branches They can only be defined in the SuperUser mode You can prevent access to branches and methods using the protect function You can set the protection so that for routine sample checking only the method dedicated to that task is accessible The table below lists the protect functions available in order of priority WRITE has lowest priority ALL full has highest priority Protect Function Effect Designation Write protection Prevents method parameters from WRITE being written over Read Write protection Prevents methods from being read RD WR and from being overwritten Execute protection Prevents methods from being used EXECUTE Full protection Prevents access to a branch and all of ALL its methods Setting Protect Functions Please observe the following when setting protect functions e Protect functions set for a method are valid for this particular method only e Protect functions set for a branch are automatically valid for all the methods contained in the branch Appendix If a particular method has a higher protect function priority than the branch the method protection is valid If the method has a lower priority than the branch the branch protection is valid For example when a branch has read and write protection all the methods in this branch have the same protection However full protection can be set for individual methods since full pr
64. ed If necessary determine the blank value of the reaction see below and enter the value in the parameter BLANK Press START This starts the measurement Using Methods 5 Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution distilled water and press START OR If the display is SUBSTRATE SMPL 1 START STOP Place solutions with the exception of enzyme solution in a cell and mix Place the cell in the sample cell holder and press START 6 Allow equilibrium time delay time to elapse DELAY TIME XXX nm XXX min XXX A 7 Addthe enzyme solution and mix WAIT SAMPLE 1 START STOP 8 Press START 111 Lambda 20 Lambda 40 UV Vis Spectrometers Procedure with a Cell Changer Analysis is performed analogous to manual operation the essential difference being that instead of one cell several can be used for measurement in the one operation The procedure is as follows 1 Once the equilibrium time has elapsed prior to addition of enzyme the absorbance in each cell is measured automatically 2 After adding enzyme only location 1 is measured to follow the course of the reaction 3 Once the reaction is complete the absorbance in all the remaining cells is measured 4 Continue to insert samples when asked until they have all been measured Example of the Display Shown During the Measurement
65. ed This appears on the top right when cycles gt 1 Printout The graphic result is printed out at the end of each sample analysis 90 Using Methods Concentration Determination You use CONCENTRATION 1 and CONCENTRATION 2 methods to determine the sample concentration Using CONCENTRATION methods you first establish a calibration curve and then measure the sample concentration The instrument calculates the calibration curve from the corrected or uncorrected values at defined wavelengths via the peak heights CONCENTRATION 1 or the peak areas CONCENTRATION 2 or the 2 derivative CONCENTRATION 2 of the spectrum CONCENTRATION 1 Method Peak heights Summary of the procedure for creating a CONCENTRATION 1 method e Determine the measurement wavelength s see Determining the Measurement Wavelength s on page 91 e Create a CONCENTRATION 1 method see Creating a Method on page 93 e Establish a calibration curve using references see Establishing the Calibration Curve on page 94 e Measure the sample see Measuring the Sample on page 95 Determining the Measurement Wavelength s To determine the wavelengths 1 Record the spectrum of the sample see Scanning a Spectrum on page 85 2 Selecta strong peak and note the wavelength at its absorbance maximum WAV 1 91 Lambda 20 Lambda 40 UV Vis Spectrometers WAV 1 A Select the type of baseline correction required 3 With a straight but offset b
66. en a method is selected it can be used for measurements When starting the method the system automatically makes requests via the display AUTOZERO START STOP SCAN SMPL n START STOP AUTOZERO ORDINATE MODE amp A lt gt Autozero Place a cell containing a blank solution in each of the sample and reference cell holders OR Place an empty cell in each of the sample and reference cell holders measurement against air Press START to start the autozero Sample measurement Place the cell containing the sample solution in the sample cell holder n ENTER can be used to switch directly to SAMPLE n n is the sample number Press START to start the measurement Tagged parameter amp is the CALL tag If desired enter a new value and press ENTER OR Select a new value using the arrow keys Press START to proceed with the analysis 77 Lambda 20 Lambda 40 UV Vis Spectrometers Analysis Procedure l Select the appropriate method see Selecting a Method on page 48 If necessary modify the method parameters Press START Check the display If the display is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is XXX SAMPL n START STOP AUTOZERO Insert a cell containing the sample solution and press START XXXX is the method type Continue to insert samples
67. ended e When the scan speed is changed e Fach time the solvent is changed To perform an autozero or background correction place cells with a blank solution or as directed in the method in the sample cell holder and reference cell holder 149 Lambda 20 Lambda 40 UV Vis Spectrometers 150 Unusual Samples If a sample is chemically stable and undergoes no physical or chemical change other than to absorb incident radiation errors in photometric values should not be caused by the sample Many samples are not stable and special consideration must be given to them Volatile Samples Some liquid samples are so volatile that their concentration can change while recording is in progress If this occurs the resulting data will lack reproducibility If you are analyzing volatile samples use stoppered cells to prevent this problem Samples not Governed by the Beer Lambert Law Quantitative analyses utilizing the absorption of spectral radiation are based on the Beer Lambert law which states that the absorption is proportional to the concentration of the analyte The law can be expressed in the form A cd Where A is absorbance e Is molar absorption coefficient c is molar concentration d is thickness through which the radiation is transmitted This law 1s mostly true for dilute solutions but at higher concentrations a plot of absorbance aga inst concentration will be nonlinear for a number of reasons Analytical Note
68. er 2SCAN k Method Name lt gt PARAM START Method Header The method is loaded onto the operational memory Methods The method header then appears on the display The method can now be used for measurement If an unused method number is entered the function NEW METHOD appears A new method can now be created see Creating a New Method on page 57 If you don t know the method number return to the standby display and use PARAM to switch to the first method header Then use the arrow keys to view the available methods in turn Default Methods Default methods are stored in the spectrometer The default methods can be read and copied but not modified The copied default methods can then be modified to suit your own requirements You access the default methods as follows 1 Switch on the spectrometer in SuperUser mode see SuperUser on page 207 2 Press STOP repeatedly until the APPLICATION branch header is displayed APPLICATION PARAM 3 Press PARAM to select the first of the default methods 4 Use the arrow key to select the required method type 5 Press PARAM and then use the arrow key to select MARK FOR COPY 6 Press STOP to return to the APPLICATION branch 7 Create an empty method file see Creating an Empty Method File on page 57 49 Lambda 20 Lambda 40 UV Vis Spectrometers 8 Copy the default method into the empty method
69. ers and Descriptions QUADRATIC Used when the measured values do not vary linearly with the concentration the curve passes through the origin QUAD INTERC Used when the measured values do not vary linearly with the concentration the curve has an intercept on the measured value axis to compensate for background interferences CYCLE TIME The time in minutes between the start of one sample measurement to the start of the next sample measurement Range 0 002 to 999 9 CYCLE TIME CYCLE TIME Measurement1 Measurement2 Duration of the analysis CYCLES x CYCLE TIME NOTE When using accessories set the CYCLE TIME longer than required for scanning the spectrum This avoids time problems CYCLES 21 The number of times one sample is scanned or measured Range 0 to 99 NOTE f CYCLES is set to 0 the sample is scanned or measured continuously until you stop the method DELAY TIME 23 Equilibration time 47 Only in the case of use with the cell holder or in manual operation with CALCULATE INTERVAL This is the time from the start of the method to the start of measurement units as selected for TIME UNIT 179 Lambda 20 Lambda 40 UV Vis Spectrometers Range 0 0 to 999 9 NOTE Measurement begins after equilibration time has elapsed Substrate Once the equilibration time has elapsed add the enzyme solution to the cell and mix Operating with a Sipper Delay between the end of the aspiration process and the start of the measure
70. es and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his own expense Changes or modifications not expressly approved by the manufacturer could void the user s authority to operate the equipment 13 Lambda 20 Lambda 40 UV Vis Spectrometers 14 Environment Operating Conditions Explosive Atmosphere This instrument is not designed for operation in an explosive atmosphere WARNING The instrument will operate correctly under the following conditions e Indoors e Ambient temperature 15 C to 35 C e Ambient relative humidity 20 to 80 without condensation e Altitude in the range 0 m to 2000 m Storage Conditions You can store the instrument safely under the following conditions e Ambient temperature 20 C to 60 C e Ambient relative humidity 20 to 80 without condensation e Altitude in the range 0 m to 2000 m When you remove the instrument from storage before putting it into operation allow it to stand for at least a day under the approved operating conditions Safety Information Chemicals Use store and dispose of chemicals that you require for your analyses in accordance with the manufacturer s recommendations and loca
71. es to next higher level To change the wavelength setting see Setting the Wavelength Manually on page 41 Starts autozero background correction see Manual Autozero on page 42 To start and continue a method Numerical keys Decimal point Minus used to enter negative values Confirms parameter Clears unconfirmed parameter entry Introduction Key Combinations nnn METHOD Selects method nnn nnn n GOTO To change to wavelength nnn n nn PARAM Selects parameter nn when you are in the parameter list level PARAM Selects parameter tagging Selects default methods from Application level l PRINT Prints out a method or branch directory see Printing Out a Directory on page 67 PARAM Selects previous parameter 1 PRINT Prints out method parameters 2 PRINT Prints out a directory of the methods available in the branch 3 PRINT Prints out the additional method information 4 PRINT Line feed 5 PRINT Form feed 6 PRINT Prints out the Peltier cell holder temperature shown on the display 7 PRINT Prints out the spectrometer status 7 9 Full reset see Reset on page 43 Functions only when the Peltier accessory is installed Y Caution All methods are deleted 23 Lambda 20 Lambda 40 UV Vis Spectrometers 1 5 E Switches to SuperUser mode see Activating SuperUser Mode on page 207 Displays Standby displ
72. ese wavelengths In MODE DERIV 2 PEAK a derivative maximum or minimum is located around these wavelengths and the nearest one evaluated see Error Reference source not found CALCULATE 9 Slope calculation mode REGRESSION The slope is calculated using all data points by means of linear regression INTERVAL The slope is calculated for each interval The mean of all the slopes is then used for the calculation of enzyme activity NOTE When using CALCULATE INTERVAL nlease 173 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter 174 CELL 1 7 41 bear in mind instead of the parameter LAG TIME parameter DELAY TIME equilibration time appears Measurement begins only when the DELAY TIME has elapsed The TOTAL TIME must be a whole multiple of the INTERVAL time A ko A A ALCULATE REGRESSION CALCULATE INTERVAL qpstart of the Method aptart of Kiesshreimeht Start of Calculation To enter the location of the cell or cells to be measured in the 5 cell changer To enter the location of the cell or cells to be measured in the 6 cell changer To enter the location of the cell or cells to be measured in the 8 cell changer To enter the location of the cell or cells to be measured in the 9 cell changer To enter the location 1 to 7 of the cell or cells to be measured in the 13 cell changer Example Parameter Numbers and Descriptions Cell 1 7 2246 Measurement will Cel
73. et o e 129 Requirements for Operation with Accessories 130 Using Methods with Accessories 131 Simulation of Accessories ucsesesenseseennennensnennennen nennen 131 Running Methods with an Accessory oooooocoocccoocnnonnoonnononooncconnononocnonos 132 Autozero with Cell Changers sss 133 CONCENTRATION Methods with Cell Changers sessss 134 Accessory Parameters nennen enne nnne nene nnns 135 CAL D OS 141 ar p ne ehren 143 Diath y Care eere idet DE S RR IIIS 143 Use and Care ot Coll erret ert e teen nes 144 Cell Handling eie ieee nran oe ener enne 144 Pressure Buildup in Cells sese 144 Sample Compartment Windows esses 146 Analytical Notes c 147 ANULOZEIO vitara eie inn cidit ve e Dis oie 149 Unusual Samples rs eet etn eet eremi ann 150 Volatile Samples docta aiit te a Ra 150 Samples not Governed by the Beer Lambert Law sss 150 Chemically Reactive Samples eese 151 Photoactive Samples unseessessssnesnesnsennnnnnnennsennnennne i enn nn 151 Other Sample Properties ensaen tet ens 151 Solvent Properties ectetuer reisten 153 Error MESSAGES m E M M 155 Error Messages 2er IRRE HERE 157 Lambda 20 Lambda 40 UV Vis Spectrometers Error Messages Shown on the Display 157 Error Reports on the Printer essen 163 Parameter Numbers an
74. file using the NEW FROM MARK parameter see Copying the Method into Another Method File on page 64 The default method can now be amended as required 50 Methods Editing Methods The following options are available e MODIFY METHOD e DELETE METHOD e NEW METHOD e NEW METHOD NAME e CHECK METHOD e MARK FOR COPY NEW FROM MARK e PRINT METHOD To recreate methods that have been inadvertently erased or written over regularly print out all important methods 51 Lambda 20 Lambda 40 UV Vis Spectrometers 52 Modifying a Method 1 Select the method to be modified 2 Press PARAM 3 Press PARAM again 4 Change the displayed parameter values if required OR Press PARAM to select the next parameter OR Press PARAM to recall the previous parameter OR Enter the appropriate parameter number and press PARAM to select a particular parameter See Parameter Numbers and Descriptions on page 169 for parameter description and parameter numbers OR Press STOP to cancel 2 SCAN lt gt PARAM START Param MODIFY METHOD PARAM gt Param SLIT 1 0nm lt gt Param Methods Changing a Parameter 1 Select the parameter to be changed 2 Depending on the parameter shown change as described in the following table Select option with the arrow keys ENTER gt ENTER 3 Press PARAM to proceed to the next parameter You can press
75. g an accessory or for manual cell changers CELL is for a cell changer SIPPER is for a sipper 170 Parameter Numbers and Descriptions AS 90 91 is for the AS 90 91 autosampler LINTRANS is for the linear transporter NOTE Sipper operation is not possible when using SUBSTRATE or ENZYME methods NOTE Autosampler operation is not possible when using ENZYME methods AUTO METHOD Prints method parameters prior to each method start Options YES NO AUTO PURGE Switches the autopurge function on and off Options YES NO AUTOZERO 16 Autozero Options YES NO Without tagging autozero should be carried out at the start of the method If tagging has been carried out autozero is offered At the start of the method CALL tagging Prior to each sample batch BATCH tagging Prior to each sample START tagging Time Drive Wavelength Prog Autozero or setting the baseline to zero is carried out at every wavelength Scan Background correction is carried out within the range WAV MAX and WAV MIN The baseline is set to zero 171 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter No Description Concentration 1 Concentration 2 If AUTOZERO YES has been selected autozero must be carried out prior to measurement of the first reference solution Autozero should be carried out at wavelengths WAVELENGTH 1 3 That is the baseline signal is set to zero at these points Substrate Enzyme Autozero is carried ou
76. gados incluso cuando el aparato haya s do desconectado del voltaje de la l nea Pr sence de tensions mortelles dans l instrument e M me lorsque l interrupteur de puissance est sur ARRET des tensions de secteur peuvent encore tre pr sentes dans l instrument e Lorsque l instrument est reli au secteur les raccords peuvent tre sous tension et des parties sous tension peuvent tre d couvertes en ouvrant des capots ou en retirant des pieces l exception de celles auxquelles il est possible d acc der manuellement e Les condensateurs contenus dans l instrument peuvent encore tre charg s m me si l instrumeni a t d connect de toutes les sources de tension Translations of Warnings W02 02 e Even with the power switch OFF line power voltages can still be present within the instrument WARNING e When the instrument is connected to line power terminals may be live and opening covers or removing parts except those to which access can be gained by hand is likely to expose live parts e Capacitors inside the instrument may still be charged even ifthe instrument has been disconnected from all voltage sources Lethal voltages are present in the instrument Nello strumento sono presenti tensioni mortali CD e Anche se l interruttore di alimentazione e in posizione OFF la tensione di linea pu essere ancora presente all interno dello strumento e Quando lo strumento collegato alla rete d
77. geover Default 326 0 nm Only for service personnel Default off 222 Appendix VALIDATION the Validation Branch These methods are used to test that the spectrometer s performance lies within specification The methods in this branch are described in detail in the document UV Vis Spectrometer Performance Validation 09935026 from PerkinElmer TEST the Test Branch These methods are for use by PerkinElmer Service only 223 Lambda 20 Lambda 40 UV Vis Spectrometers S Status Display p M Status Display P Method 29 Freerun Status Display P Method 19 O T Calibration A Method 20 One Wavel Calib Method 21 Two Wavel Calib Method 1 Timedrive Method 2 Scan Method 3 Wavprog Method 4 Conc1 Method 5 Conc2 HA A Method 6 Enzyme ek 4 iu Method 7 Substrate A y Method 900 Date Time EL Y API Method 901 Wakeup Method 999 Self Check NE 224 Methodx J Appendix Validation DEE Status Display Status Display Method 1 Help Config Validation Methods Service Methods Method 2 Autosampler Config Method 3 Liintrans Config Method 4 Comm Config Method Enter Param Stop gt Arrow keys Method 5 RS232 Port anuUu 5 Z Method 6 Printer Config Method 7 Acoy Config Method 8 User Config
78. he fine structures of the spectrum disappear If smoothing is too low spectral fine structures may be hidden by noise If an unacceptable value is selected an appropriate indication is displayed If then no other value is substituted the spectrometer will automatically continue to operate at smoothing 0 NOTE Derivative spectra cannot be scanned at smoothing 0 Concentration methods Select a smoothing factor that is as low as possible so that the peak is smoothed as little as possible The value selected should depend on the level of noise 198 STATISTIC Parameter Numbers and Descriptions Scanning speed in nm min Select the scanning speed depending on the type of sample and the desired resolution The following can be used as a general rule for narrow peaks select a low scanning speed and low degree of smoothing in order to limit the noise level for broader peaks higher scanning speeds can be selected For derivative spectra observe the comments given under ORDINATE MODE Guideline values Overview spectra 2880 1920 nm min Broad peaks 960 480 240 nm min Solid and liquid samples 120 60 30 nm min Gaseous samples spectra with higher resolution expanded spectra 30 15 7 5 nm min To select the statistic calculation function To calculate and print out the average the standard deviation and relative standard deviation for each sample repeat measurements or sample group batch measure
79. hromator to your measurement wavelength or to 460 nm Call up a method that uses transmission T as the ordinate If necessary change the ordinate mode to transmission Open the sample compartment cover Insert the cell with a low absorbing solvent into the sample cell holder Leave the reference cell holder empty Make horizontal fine alignment to the sample cell holder locking screws and milled posts loosened to obtain the highest possible transmittance reading on the display Close sample compartment cover while measuring transmittance Make vertical fine adjustment using the vertical adjustment screw again to obtain the highest possible reading Close sample compartment cover while measuring transmittance 35 Lambda 20 Lambda40 Operation and Parameter Description 7 When you are satisfied with the alignment tighten the milled posts and the locking screws on the cell holder 8 Insert the matching cell with the same low absorbing solvent into the reference cell holder The first cell remains in the sample cell holder 9 Repeat steps 5 through 7 with the reference cell holder but this time obtain the lowest possible transmittance reading on the display This completes the fine alignment procedure When the cell holder has been aligned once you can take it out and reinstall it without aligning it again 36 Startup and Shutdown Minimum Volume Applications To measure minimum sample volumes use microcells
80. i alimentazione i terminali possono essere sotto tensione aprendo le calotte di protezione o rimuovendo alcune parti ad eccezione di quelle raggiungibili con la mano possibile esporre altre parti sotto tensione e Anche se lo strumento stato scollegato da tutte le fonti di tensione i condensatori al suo interno possono essere ancora carichi Gevaarlijke spanningen in het instrument gevaar van letsel NL e Ook in uitgeschakelde toestand kan wanneer het instrument op de neivoeding is aangesloten op sommige plaatsen in het instrument netspanning staan e Ook bij een uitgeschakeld instrument en een onderbroken netvoeding kunnen condensatoren in het instrument nog een gevaarlijke lading bevatten Tens o perigosa no aparelho perigo de ferimento e Mesmo desligado o aparelho poder ainda ter tens o de rede em alguns pontos enquanto estiver ligado rede de corrente e Mesmo com o aparelho desligado e a liga o rede de corrente interrompida os condensadores dentro do aparelho ainda poder o ter ums tens o perigosa aplicada 239 Lambda 20 Lambda 40 UV Vis Spectrometers 240 CA WARNING W02 03 Electrical Hazard To prevent potential injury to yourself and damage to the instrument switch OFF all instruments in the system and disconnect them from the line power supply before you alter or make any new electrical connections amp 9 00206098 Gef hrdung durch Elektrizit t Um Ver
81. ignal is taken into account when the dark signal function is switched on Compensation then takes place automatically either at the start of a method or every 10 minutes whichever occurs first ONE WAVEL Wavelength calibration with one peak FANE To check the calibration record the spectrum of a wavelength standard and compare the wavelength values recorded by the spectrometer with the values for the standard If they do not correlate the spectrometer should be recalibrated Parameters 0 nm PEAK Internal calibration at 0 nm D2 PEAK Internal calibration at 656 1 nm SPEC PEAK Calibration using an external wavelength standard for example holmium oxide the standard If they do not correlate the spectrometer should be recalibrated 21 TWO WAVEL Wavelength calibration with two peaks CALIB To check the calibration record the spectrum of a wavelength standard and compare the wavelength values recorded by the spectrometer with the values for 216 Appendix Parameters AUTO PEAK Internal calibration at 0 0 and 656 1 nm SPEC PEAK Calibration using an external wavelength standard for example holmium oxide CONFIGURATION the Configuration Branch This branch contains methods for configuration of the spectrometer to the individual requirements of the user 1 HELP CONFIG Level and language of help messages Parameters LEVEL Extent LANGUAGE Language AUTOSAMPLER CONF To set the autosampler p
82. ing a letter or number for an empty space You can combine letters and numbers Methods 6 Press PARAM to confirm the new name 13 TIME DRIVE lt gt PARAM START Param MODIFY METHOD PARAM gt Y lt gt y NEW METHOD NAME PARAM gt Param lt gt y NEW METHODNAME gt TIME DRI lt Param 13 TIME DRIVE 2 lt gt PARAM START 61 Lambda 20 Lambda 40 UV Vis Spectrometers Checking a Method When using the CHECK METHOD function the parameter values are displayed but cannot be changed 1 Select the method to be checked 2 Press PARAM 3 Use the arrow keys to select CHECK METHOD 4 Press PARAM to check each parameter in turn OR Press STOP to cancel 2 SCAN lt gt PARAM START Y Param y MODIFY METHOD PARAM gt Y lt gt y CHECK METHOD PARAM gt Y Param y SLIT 1 0 nm CHECK ONLY Param Y 62 Methods Copying Method Parameters into a New Method File This is useful when you wish to make a new method with only a few parameters different from the original Two steps are involved in this procedure e Marking the method for copy e Copying the method into another method file Marking for Copy 1 Select the method whose parameters are to be copied 2 Press PARAM 3 Use the arrow keys t
83. ion Possible hazards that could harm the user or result in damage to the instrument are clearly stated at appropriate places throughout this manual The following safety conventions are used throughout this manual We use the term WARNING to inform you about situations that could result in personal injury to yourself or other persons Details about these circumstances are in a box like this one WARNING We use the term CAUTION to inform you about situations that could result in serious damage to the instrument or other equipment CAUTION Details about these circumstances are in a box like this one Translations of the warning messages used in this manual are given in Translations of Warnings on page 231 Before using the instrument it is essential to read the manual carefully and to pay particular attention to any advice concerning potential hazards that may arise from the use of the instrument The advice is intended to supplement not supercede the normal safety code of behavior prevailing in the user s country Lambda 20 Lambda 40 UV Vis Spectrometers 10 IEC 1010 Compliance This instrument has been designed and tested in accordance with IEC 1010 1 Safety requirements for electrical equipment for measurement control and laboratory use and Amendment 1 to this standard CSA Compliance This instrument meets the Canadian Standards Association CSA Standard CAN CSA C22 2 No 1010 1 92 Laboratory Equipment UL Comp
84. irst measurement 133 Lambda 20 Lambda 40 UV Vis Spectrometers 134 CONCENTRATION Methods with Cell Changers Please note the following when using CONCENTRATION methods When AUTOZERO YES has been selected an autozero is carried out once the method has been started irrespective of whether the parameter has been tagged or not e Insert the reference solutions in sequence when asked e Always start at location 1 REFS INTO CELL 1 5 START STOP AUTOZERO Operating with Accessories Accessory Parameters Parameter Description ACCESSORY Select the accessory MANUAL Operation with standard cell holder no accessories CELL For cell changers including 5 Cell Changer 6 Cell Changer 8 Cell Changer 9 Cell Changer 13 Cell Changer SIPPER For Vacuum Sipper or Peristaltic Sipper AS 90 91 Forautosamplers AS 90 and AS 91 LINTRANS For linear transporter t If SUBSTRATE methods are used Sipper operation is not possible Autosampler operation is not possible when using ENZYME or SUBSTRATE methods 135 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter Description Cell Changer 5 Cell 6 Cell 8 Cell 9 Cell CELL 1 n The locations at which measurements are to take place Enter the number of the locations and press ENTER If measurement is to be carried out at locations 2 and 5 only enter 25 Therefore If AUTOZERO YES has been selected location 1 is used for
85. ive of the ordinate mode selected Ratio difference and corrected ratio RAT DIF COR modes are determined according to the equations below QUAQUS DIF A Az As As 3 A E A A T As COR 48 AA o where subscript 1 stands for the first wavelength subscript 2 stands for the second wavelength and so on 192 Parameter Numbers and Descriptions OVERLAY PATHLENGTH E PLOT REFS dl POST RUN KIN Prints spectra from the same batch onto the same graphics printout valid only when GRAPHICS PLOT YES Option YES NO Overlaying graphics printouts functions only if the printer used has an automatic paper reverse function If CYCLES 1 all the results of a particular batch are printed out sequentially This enables spectra to be more easily compared than if they are printed out separately To enter the pathlength cm of the cell The result is corrected for pathlength Range 0 001 to 10 0 Prints out the calibration curve Option YES NO Allows you to recalculate the results Option YES NO Enzyme When POSTRUN KIN YES is used the results for a changed TOTAL TIME and LAG TIME can be recalculated When using a cell changer the values for TOTAL TIME and LAG TIME must be a whole multiple of the set INTERVAL time Substrate When POSTRUN KIN YES is used the results for a changed reaction END TIME can be recalculated When using a cell changer no recalculation is possible
86. l sample area Diffraction patterns become apparent if the radiation beam impinges on the cell wall 8 Ifthe radiation beam is not centered exactly loosen the two locking screws and the two milled posts on the relevant cell holder and shift the cell holder plate to center the radiation beam horizontally Then retighten the two milled posts and the two locking screws 9 Now visually check the vertical alignment of the radiation beam in the cell sample area Alignment is correct when the radiation beam is just above the floor of the cell sample area min 2 mm or covers the cell window NOTE The center of the window for micro flowcells should be ideally approximately 15 mm above the base of the cell Min 2 mm Figure 5 Correct Alignment of the Radiation Beam in the Cell Sample Area 34 10 11 12 13 Startup and Shutdown If vertical alignment is required turn the vertical adjustment screw on the lifter either clockwise to raise the cell or counterclockwise to lower the cell Recheck the horizontal alignment of the radiation beam through the cell and correct if necessary Using the GOTO key slew the monochromator to any value above 200 nm Remove the card blocking the sample beam window and close the sample compartment cover This completes the coarse alignment of the cell holder Fine Alignment If fine alignment is necessary proceed as follows l Using the GOTO key slew the monoc
87. l 8 13 0 take place at locations 2 4 and 6 the rest will not be used CELL 8 13 To enter the location 8 to 13 of the cell or cells to be measured in the 13 cell changer Example Cell 1 7 567 Measurement Cell 8 13 123456 will take place at locations 5 to 13 CHNGE To change the molecular mass and molar absorption CONSTANTS coefficient values of oligonucleotide bases Option YES NO Refer to the Biochemical Application manual CONC FACTOR 11 Concentration factor Range 0 00001 to 9999 9 NOTE The concentration factor is calculated as follows Concentration Factor V x M_ dx vx 1000 where V is the volume of the total solution in the cell in mL M is the molar mass of the substrate in g mol d is the pathlength in cm v is the volume of sample in mL 1000 is the conversion factor for volume units in liters Denendine on the nrocedure used the molar ahsorntion 175 Lambda 20 Lambda 40 UV Vis Spectrometers WE coefficient may need to be taken into account CONC UNIT 6 Concentration Unit defines the concentration unit used for the analysis c any concentration unit g L gram per liter mg L milligrams per milliliter mg dL milligrams per deciliter Substrate methods only Hg microgram g mL micrograms per mililiter mol mole mmol millimole nmol nanomole pmol picomole ppm parts per million mg kg ppb parts per billion ug kg percent A1 absorbance of a solution containing 1 g substance in 100
88. l safety regulations Hazardous Chemicals Some chemicals used with this instrument may be hazardous or may Y become hazardous after completion of an analysis WARNING The responsible body for example Laboratory Manager must take the necessary precautions to ensure that the surrounding workplace and instrument operators are not exposed to hazardous levels of toxic substances chemical or biological as defined in the applicable Material Safety Data Sheets MSDS or OSHA ACGIH or COSHH documents Venting for fumes and disposal of waste must be in accordance with all national state and local health and safety regulations and laws OSHA Occupational Safety and Health Administration U S A ACGIH American Conference of Governmental Industrial Hygienists U S A COSHH Control of Substances Hazardous to Health U K Toxic Fumes If you are working with volatile solvents or toxic substances you must provide an efficient laboratory ventilation system to remove vapors that may be produced when you are performing analyses Waste Disposal Waste containers may contain corrosive or organic solutions and small amounts of the substances that were analyzed If these materials are toxic you may have to treat the collected effluent as hazardous waste Refer to your local safety regulations for proper disposal procedures 15 Lambda 20 Lambda 40 UV Vis Spectrometers 16 Deuterium lamps and other spectral lamps are maintained
89. lays shown during the measurement Display Description AUTOZERO Request to start autozero START STOP Display during autozero AUTOZERO XXX X nm 25 Lambda 20 Lambda 40 UV Vis Spectrometers Request to start sample measurement NNNN SMPL 2 NNNN is the method name START STOP AUTOZERO Request to start sample measurement or replot NNNN SMPL n a START STOP AZ PRINT graphics Appears in methods with replot function when graphics plot set to yes NNNN is the method name Display during sample measurement SCAN method Display of a tagged parameter If necessary the parameter can be changed AUTOZERO IN CELL1 Cell Changer request to insert blank solutions and START STOP start autozero SAMPLES INTO 9 CELL Cell Changer request to insert sample solutions and START STOP AUTOZERO start the measurement AUTOZERO SIPPER Sipper request to press start key on the Sipper ACCESSORY START autozero SAMPLE 1 SIPPER Sipper request to press start key on the Sipper ACCESSORY START sample measurement REF 1 XXX Request to measure a reference solution START STOP AUTOZERO 26 Startup and 3 Shutdown Startup and Shutdown Startup and Shutdown When operating the spectrometer wait until the BUSY display has disappeared before pressing the next key This allows the software to complete the calculations and the motors to move the optics to their required setting Before starting analysis leave
90. le solution is determined the sample solution containing enzyme for example serum is reacted with a high excess of substrate Appendix The substrate is converted to product by the enzyme the rate of the reaction can be followed photometrically and is a direct measure of the enzyme activity Enzyme activity is given as International Units U 1U the enzyme activity required to convert 1 umole of substrate per minute under optimal conditions The following reaction can be assumed for the conversion of substrate S into product P where E enzyme and k reaction rate constant The rate of reaction S E gt P E is governed by the Michaelis Menten equation Cog Cs v k Ku 7 Cs i dE oS dA m i with v di dr di Reaction Rate and ky amp k a Michaelis Constant 1 Due to the large excess of substrate there is a limiting case Cs gt Ky and hence v k C is valid Thus the reaction rate is constant no consumption of enzyme and directly proportional to enzyme concentration or enzyme activity Hence the enzyme activity of a solution can be directly determined by measuring the reaction rate 227 Lambda 20 Lambda 40 UV Vis Spectrometers A d we Pd p d dA Coe rd A dt dt gt Time In practice the course of the curve obtained can deviate from the ideal form it becomes linear only after a certain lag time and flattens out towards the end In such a case only th
91. letzungsgefahr durch elektrische Spannung und Besch digungsge fahr der Ger te zu vermeiden schalten Sie alle Ger te des MeBplatzes AUS und trennen Sie die Netzverbindungen bevor Sie irgendwelche elektrischen Verbindungen herstellen oder ndern Fare p grund af elektricitet For at undg faren for kveestelser fra den elektriske speending og faren for beskadigelse af apparaterne skal alle apparater p m lepladsen vere SLUKKET og stramforbindelserne skal vere skilt inden De opretter eller eendrer noget i de elektriske forbindelser Peligro el ctrico Para evitar posibles da os personales y al equipo apague todos los m dulos del sistema y descon ctelos de la toma de corriente antes de establecer nuevas conexiones el ctricas o de modificar conexiones existentes Risque d lectrocution Afin d viter des blessures ventuelles et des d t riorations de l instrument mettre tous les instruments du syst me hors tension et les d connecter de l alimentation par le secteur avant de modifier des connexions lectriques ou d en r aliser de nouvelles Pericolo elettricit Per evitare potenziali incidenti alle persone e danni allo strumento mettere in posizione OFF l interrutiore di tutti gli strumenti del sistema e scollegarli dalla fonte di alimentazione prima di modificare i collegamenti elettrici o di creare nuovi collegamenti Risico s door elektriciteit Om het gevaar van letsel door elektrische spanning en het gevaar v
92. liance This instrument meets the Underwriter Laboratories UL Standard UL 3101 1 Oct 93 Electrical Equipment for laboratory use part 1 general requirements Electrical Protection Insulation Class I as defined in IEC 1010 1 Installation Category The instruments are able to withstand transient overvoltage according to Installation Category II as defined in IEC 1010 1 and IEC 664 Pollution Degree The equipment will operate safely in environments that contain non conductive foreign matter and condensation up to Pollution Degree 2 as defined in IEC 1010 1 and IEC 664 Electrical Safety To ensure satisfactory and safe operation of the instrument it is essential that the green yellow lead of the line power cord is connected to true electrical earth ground If any part of the instrument is not installed by a PerkinElmer service representative make sure that the line power plug is wired correctly Safety Information Protective Conductor Green Yellow Green earth ground Electrical Hazard Any interruption of the protective conductor inside or outside the instrument or disconnection of the protective conductor earth ground terminal is likely to make the instrument dangerous WARNING Intentional interruption is prohibited p P Lethal voltages are present in the instrument e Even with the power switch OFF line power voltages can still be present within the instrument When the instrument is connected to line power termi
93. may ask you to select the type of paper you are using Z fold or single sheet Use lt gt to select your paper type Press START to confirm the selection and start the checks When the checks are completed the spectrometer display shows SPECTROMETER FULL RESET DONE Press STOP to print the report and return to standby If there are any FAIL results in the report then repeat the test If you have any further enquiries contact your PerkinElmer office 125 Lambda 20 Lambda 40 UV Vis Spectrometers 999 SELF CHECK lt gt PARAM START Y Start y PRINTER ON Y lt gt y Start y PAPER SINGLESHEET lt gt Y lt gt Y Start Y SPECTROMETER FULL RESET DONE y Stop 126 Operating with 7 Accessories Operating with Accessories Operating with Accessories General Accessories are components or instruments that are installed or connected in the sample compartment or otherwise connected to the spectrometer For some of these accessories parameters have to be taken into account in the methods The accessories described below have parameters in the various methods Accessories Samples can be applied either manually or with the help of a number of accessories The following accessories are currently available Changers 5 Cell Changer 6 Cell Changer 8 Cell Changer 9 Cell Changer 13 Cell Changer 8 cell changer or
94. mbda 40 UV Vis Spectrometers 230 A A Enzyme added ss P ME Pd Creeping reaction ES Dime Translations of Warnings Translations of Warnings gt WARNING Wo1 01 Warning We use the term WARNING to inform you about situations that could result in personal injury to yourself or other persons Details about these circumstances are in a box like this one 080 0 086 Warning Warnung Bedeutet daB es bei Nichtbeachten der genannten Anweisung zu einer Verletzung des Benutzers kommen kann Warning Advarsel Betyder at brugeren kan blive kveestet hvis anvisningen ikke overholdes Warning Peligro Utilizamos el t rmino WARNING PELIGRO para informarle sobre situaciones que pueden provocar darios personales a usted o a otras personas En los recuadros como ste se proporciona informaci n sobre este tipo de circunstancias Warning Danger Nous utilisons la formule WARNING DANGER pour avertir des situations pouvant occasionner des dommages corporels l utilisateur ou d autres personnes Les d tails sur ces circonstances sont donn es dans un encadr semblable celui ci Warning Pericolo Con il termine WARNING PERICOLO vengono segnalate situazioni che potrebbero provocare incidenti alle persone Troverete informazioni su tali circostanze in un riquadro come questo Warning Waarschuwing Betekent dat wanneer de genoemde aanwijzing niet in acht wordt genomen
95. ment Range 0 to 99 9 DIL FACTOR 10 Dilution factor The measured value is multiplied by the factor and the result displayed Thus concentration can be read off directly or a dilution taken into account Range 0 00001 to 9999 9 DIVISOR 12 Divisor Range 0 00001 to 999 9 The molar absorption coefficient value can be entered as divisor Values can be obtained from the literature NOTE f the absorption coefficient is already included in the ENZ FACTOR enzyme factor enter DIVISOR 1 Enzyme Enzyme activity is automatically calculated as follows Ag enzyme factor x dilution factor x 1 divisor x dA dt Substrate Substrate concentration cs is calculated as follows 180 END TIME E Parameter Numbers and Descriptions Csu concentration factor x dilution factor x 1 divisor x AA Where AA absorbance difference Concentration 1 Concentration 2 The measured value is multiplied by the factor or divided by the divisor and the resultant value displayed Thus dilution procedures or differing masses can be taken into account If a dedicated correction factor is to be used for each sample select FACTOR or DIVISOR as a START TAG PARAMETER see Tagging a Parameter on page 54 The factor or divisor can be entered immediately prior to each analysis Example A particular component of a powder is to be determined and displayed in mg g The calibration curve is compiled using pure substance in solution made
96. mnts when YES Option YES NO Appears with the autosampler accessory Not available with all applications 199 Lambda 20 Lambda 40 UV Vis Spectrometers STIRRER Switches magnetic stirrer on and off Option ON OFF If magnetic stirring has been switched on place a small magnetic stirring bar in each of the cells The arrangement is such that whilst meaurement is taking place in one cell the following cell is stirred Appears with the 13 cell changer accessory TEMP CHECK Temperature measurement Option YES NO If TEMP CHECK YES has been selected a temperature sensor B0185227 must be installed The temperature measured in the cell is included in the printout If a temperature sensor has not been installed select TEMP CHECK NO TEMPERATURE 51 To enter the required temperature in C only in combination with TEMP CHECK YES Range 0 0 to 150 0 Measurement begins as soon as the temperature in the cell has reached the required temperature 40 1 C When minus is selected the temperature is only registered and printed out together with the anaytical results Operation with a Peltier cell holder To enter the required temperature of the Peltier cell holder Range 15 0 to 45 0 200 Parameter Numbers and Descriptions THRESHOLD TIME UNIT E TIME 76 CALCULATION TOTAL TIME Scan Threshold value for the printout of analytical data valid only when PRINT DATA YES Range 0 0 to 9999
97. monochromator slews to the selected wavelength OR Set the spectrometer to the standby display Enter a wavelength using the numeric keys for example 325 5 41 Lambda 20 Lambda 40 Operation and Parameter Description 42 3 Press GOTO The monochromator slews to the selected wavelength Manual Autozero In this operation the AUTOZERO key is used to set the measured absorbance value to 0 or transmittance value to 10095 for the actual wavelength shown on the display 1 Open the sample compartment cover 2 Place cells with blank solution in the reference and sample cell holders OR Use air as blank 3 Close the sample compartment cover 4 Set the desired wavelength using GOTO 5 Press AUTOZERO Autozero is complete once the standby display reappears The ordinate mode of the last used method always appears on the display To convert absorbance to transmittance or vice versa select a TIME DRIVE method and then select the desired ordinate mode Quick Sample Measurement You can make quick sample measurements as follows l 2 3 Prepare the sample Set the spectrometer to the standby display Press GOTO Operating without Methods 4 Select your desired wavelength 5 Press ENTER The monochromator slews to the selected wavelength 6 Open the sample compartment cover 7 Insert cells with blank solutions in the reference and sample cell holders OR Use air as blank 8 Close the
98. nals may be live and opening covers or removing parts except those to which access can be gained without the use of a tool is likely to expose live parts Capacitors inside the instrument may still be charged even if the instrument has been disconnected from all voltage sources When working with the instrument e Connect the instrument to a correctly installed line power outlet that has a protective conductor earth ground e Do not attempt to make internal adjustments or replacements except as directed in this manual e Do not operate the instrument with any covers or parts removed 11 Lambda 20 Lambda 40 UV Vis Spectrometers 12 Servicing should be carried out only by a PerkinElmer service representative or similarly authorized and trained person Disconnect the instrument from all voltage sources before opening it for any adjustment replacement maintenance or repair If afterwards the opened instrument must be operated for further adjustment maintenance or repair this must only be done by a skilled person who is aware of the hazard involved Use only fuses with the required current rating and of the specified type for replacement Do not use makeshift fuses or short circuit the fuse holders Whenever it is likely that the instrument is no longer electrically safe for use make the instrument inoperative and secure it against any unauthorized or unintentional operation The instrument is likely to be elect
99. nds only on completion of the final cycle TRAY Enter the number of the tray used with the AS90 91 autosampler VALUE n If the calibration curve is to be compiled from known values for example from the literature values or from previous measurements these values should be entered here Make sure that the entering sequence is the same as for the corresponding concentrations parameter REF If the calibration curve is to be recorded do not enter anything here After calibration the absorbance values for the reference solutions are automatically entered The spectrometer calculates the calibration curve from these values WAV MAX Upper limit ofthe wavelength range in nm Value range 190 0 to 1100 0 in steps of 0 1 202 Parameter Numbers and Descriptions WAV MIN WAVE n Lower limit of the wavelength range in nm Range 190 0 to 1100 0 in steps of 0 1 If peak areas are used for the calculation see Error Reference source not found the area is calculated between the upper and lower wavelength values If calculation is made via a 2 order derivative the derivative spectrum is recorded between the upper and lower wavelength values Wavelength program Oligoquant 1 Measuring wavelength in nm Range 190 0 to 1100 0 in steps of 0 1 NOTE The wavelengths is run in the sequence in which they are entered when using the ORDINATE MODE take RAT and DIF into account Enter wavelengths in decrea
100. nt access to branches and methods using the protect functions Example If only one particular method is to be used without access to the parameters set the protect functions as follows e Set full protection for all branches except the application branch e _ Set read write protection for the application branch e Set full protection for all methods in the application branch not to be used For the method to be used the read write protection of the branch is valid 211 Lambda 20 Lambda 40 UV Vis Spectrometers This method can be used that is started outside of the SuperUser mode but it cannot be read or altered Tagged parameters continue to appear when methods are being run and may then be modified 212 Appendix Instrument Branches Overview of the Instrument Branches The basic spectrometer has 6 branches Application Analysis methods Communication Interface methods Calibration Methods to calibrate the spectrometer Configuration To set up basic spectrometer settings Test Test methods Validation Validation methods NOTE can only be accessed as SuperUser NOTE Methods can only be modified You cannot create any new methods Selecting a Branch 1 Switch on the spectrometer in SuperUser mode 2 Press STOP until a branch header is displayed 3 Use the arrow keys to select the required branch 213 Lambda 20 Lambda 40 UV Vis Spectrometers 214 4 Press PARAM The branch methods can
101. ntration see also FACTOR Derivative modes D1 to D4 can be used to resolve overlapping peaks to reduce interference and to enhance the fine structure of a particular peak This facilitates the qualitative evaluation of spectra with overlapping peaks and the quantitative evaluation of spectra with undesired background absorption The derived values obtained are multiplied by 10 for every degree of derivation in order to produce graphics that are easier to interpret Resolution and noise increase with the degree of derivation In general the 2 derivative is more helpful in this respect than the 1 the resolution is better and the characteristic maximum of the signal is easy to recognize as a derivative minimum 191 Lambda 20 Lambda 40 UV Vis Spectrometers Should the 1 or 2 derivatives prove insufficient the 3 or 4 derivatives can be used providing the noise level remains within acceptable limits The parameters SPEED and SMOOTH influence the quality of derived spectra In choosing parameter values take the following into account SPEED guideline value peak width in nm x 10 High scan speeds decrease resolution low scan speeds increase noise SMOOTH in the case of derivation spectra smoothing exerts a greater influence than in absorbance measurement Smoothing should thus be kept to a minimum Guideline value peak width in nm x 0 5 Derivation is not possible with a degree of smoothing of 0 irrespect
102. o una persona de formaci n y autorizaci n similares podr n realizar trabajos de revisi n y mantenimiento delinstrumento Reglages et entretien non autoris s Ne pas essayer d effectuer des r glages des remplacements ou des r parations sur cet instrument d une mani re autre que celle d crite dans la Documentation Utilisateur jointe Seul un repr sentant du service apr s vente Perkin Elmer ou une personne autoris e et de m me formation devraient tre autoris s intervenir sur l instrument Regolazione e manutenzione non autorizzate Non tentare di regolare sostituire o riparare questo strumento se non come descritto nell allegata Documentazione Utente Solo un rappresentante dell assistenza Perkin Elmer o una persona dalla analoga esperienza pu eseguire se autorizzata la manutenzione dello strumento Niet toegestane werkzaamheden aan het instrument Onderhouds reparatie of afstelwerkzaamheden die niet in de gebruikersdocumentatie van het instrument worden omschreven mogen alleen door personeel van de klantenservice of door overeenkomstig geschoolde en geautoriseerde vakmensen worden uitgevoerd Trabalhos n o permitidos no aparelho Trabalhos de manuteng o ou de reparag o ou ajustamentos n o descritos na documentag o do usu rio para o aparelho apenas podem ser executados pelo Servico T cnico da Perkin Elmer ou por pessoal especializado devidamente formado e autorizado 235 Lambda 20 Lambda 40 UV Vis S
103. o select MARK FOR COPY 4 Press PARAM to mark the method The method is now marked for copying in the next step OR Press STOP to cancel 2 SCAN lt gt PARAM START Y Param y MODIFY METHOD PARAM gt Y lt gt y MARK FOR COPY PARAM gt Param 63 Lambda 20 Lambda 40 UV Vis Spectrometers 64 Copying the Method into Another Method File l 2 Create a method see Creating a New Method on page 57 Press PARAM Use the arrow keys to select NEW METHOD Press PARAM again Use the arrow keys to select NEW FROM MARK Press PARAM The parameters from the marked method are copied into the newly created method OR Press STOP to cancel 7 13 SCAN lt gt PARAM START Y Param y MODIFY METHOD PARAM gt y lt gt y NEW METHOD PARAM gt Y Param Y lt gt y NEW FROM MARK PARAM gt y Param 13 SCAN lt gt PARAM START Modify the new method as required Methods 65 Lambda 20 Lambda 40 UV Vis Spectrometers Printing Out a Method Printing out a method provides a list of its parameters and their current values A printer must be installed and configured refer to the installation manual l 2 66 Select the method to be printed out Press PARAM Use the arrow keys t
104. o select PRINT METHOD Press PARAM again to print out the method OR Press STOP to cancel You can also press 1 and then PRINT to print out the method parameters 2 SCAN lt gt PARAM START Param MODIFY METHOD PARAM gt Y lt gt y PRINT METHOD PARAM gt Param Y NOTE Methods Printing Out a Directory Printing out a directory provides a list of its methods A printer must be installed and configured refer to the installation manual There are two directories the spectrometer directory and the branch directory Spectrometer Directory The spectrometer directory is a list of all methods for the spectrometer including the SuperUser methods Print out the directory as follows 1 Select a branch header 2 Press and then PRINT to print out a directory of the methods APPLICATION PARAM Print Branch Directory The branch directory is a list of all the methods in the selected branch Print out all the methods in the selected branch as follows To select SuperUser branches you must first enter as SuperUser see SuperUser on page 207 1 Select a method in the branch to be printed out 67 Lambda 20 Lambda 40 UV Vis Spectrometers 2 Press 2 and then PRINT This prints out the branch directory 2 SCAN lt gt PARAM START Y Print 68 Methods Print Key
105. od otherwise the spectrometer cannot work Error Messages DON T PROTECT ALL BRANCHES PROBLEM BRANCH WRITE PROTECT PROBLEM DIRECTORY FULL PROBLEM MEMORY FULL ERROR NO ENERGY An attempt was made to ALL protect all branches Retain at least one branch otherwise the spectrometer cannot work An attempt was made to modify methods in a write protected branch To modify the method alter the branch protection An attempt was made to store more than 200 methods To create space for the new method delete a method that is no longer required The available memory is insufficient to cope with the new method To create space for the new method delete a method that is no longer required This error is shown when not enough energy is detected Possible causes Beam is blocked in the sample compartment Loose lamp connection Lamp burnt out Lamp s switched off Defective detector 159 Lambda 20 Lambda 40 UV Vis Spectrometers ERROR This error is shown when not enough energy is received NO ENERGY UV from the UV lamp LAMP Possible causes Beam is blocked in the sample compartment UV lamp loose connection UV lamp burnt out UV lamp switched off Defective detector PROBLEM This error is shown when not enough energy is received NO ENERGY VIS from the Vis lamp LAMP Possible causes Beam is blocked in the sample compartment Vis lamp loose connection Vis lamp burnt out Vis lam
106. oor beschadiging van het instrument te voorkomen schakelt u voordat u welke elektrische verbinding dan ook wilt maken of wilt veranderen alle instrumenten van de meetplaats UIT en ontkoppelt u de netvoeding Perigo por electricidade A fim de evitar o perigo de ferimento por tens o el ctrica e o perigo de danificag o dos aparelhos DESLIGUE todos os aparelhos do posto de medi o e interrompa as liga es rede antes de estabelecer ou modificar qualquer ligag o el ctrica Translations of Warnings WO3 02 UV Radiation Risk of Eye Damage The lamp emits intense UV radiation which can damage your eyes Do not gaze into a lighted lamp Always wear UV absorbing safety glasses when looking at the radiation from the lamp 9000000 UV Strahlung Gef hrdung der Augen Die StrahlungsquellejLampe emittiert intensive UV Strahlung und kann dadurch Augenschaden verursachen Tragen Sie eine Schutzbrille die die emittierte UV Strahlung ausreichend absorbiert wenn Sie in die leuchtende Strahlungsquelle Lampe schauen Ultraviolet str ling farligt for jnene Str lingskilden lampen emitterer ultraviolet str ling og kan derfor for rsage gjenskader Beer beskyttelsesbriller som absorberer den emitterede ultraviolette str ling tilstreekkeligt n r De ser ind i den lysende str lingskilde Radiaci n de rayos ultravioleta La l mpara emite una intensa radiaci n de rayos ultravioleta que puede ser pe
107. or Operation with Accessories 130 Running Methods with an Accessory 132 S Safety information Electromagnetic Compatibility EMC 12 Safety Information 9 CSA Compliance 10 Electrical Protection 10 Electrical Safety 10 Environment 14 IEC 1010 Compliance 10 Symbols Used on the Instrument 17 UL Compliance 10 Scanning a Spectrum 85 Selecting a Method 48 Self Check 125 Single Cell Holder 31 Single Wavelength Measurements 82 Solvent Properties 153 Startup and Shutdown 29 Shutdown 30 Startup 29 Substrate Kinetics 109 SuperUser 207 T TEST the Test Branch 223 U Unusual Samples 150 Using Methods CONCENTRATION 1 Method Peak heights 9 Using Methods with Accessories 131 V VALIDATION the Validation Branch 223 W Wakeup 123
108. ores the changes and activates the accessory mode 6 Switch off the spectrometer This deactivates the SuperUser mode 7 Wait about two minutes to allow the lamps to cool down 8 Switch on the spectrometer 9 Setup the method as required 131 Lambda 20 Lambda 40 UV Vis Spectrometers 132 Running Methods with an Accessory When using methods with accessories the spectrometer automatically presents the necessary actions AUTOZERO IN CELL 1 Insert blank at location 1 of the cell changer and START STOP then press START Insert samples in the cell changer and then press SAMPLES INTO 9 CELL START START STOP AUTOZERO Press START on the Sipper AUTOZERO SIPPER Press START on the Sipper sample SAMPLE 1 SIPPER Operating with Accessories Autozero with Cell Changers The locations used for sample measurement depend on the tagging and option chosen for the AUTOZERO parameter Not all locations can be used for sample measurement as shown in the following table AUTOZERO Procedure MARET Blank Sample AUTOZERO solution solution YES AUTOZERO zNO FIX CALL Insert solutions 1 1 n 1 n according to display START When selecting an 1 autozero first insert all solutions and then select YES or NO and press START BATCH If no autozero is 1 selected AUTOZERO is only selectable in the appropriate method Thus on demand to insert the sample first insert all solutions and then start the f
109. otection has a higher priority than read write protection Write protection is not possible for individual methods since the branch has the higher priority Setting Method Protect Function Set the method protect function as follows l 2 Switch on the spectrometer in SuperUser mode Select appropriate method Press PARAM Use the arrow keys to select CHANGE PROTECTION Press PARAM again Use the arrow keys to select the desired protect function Press PARAM to confirm the protection OR Press STOP to cancel 209 Lambda 20 Lambda 40 UV Vis Spectrometers 8 Exit SuperUser mode to activate the protect function 2 SCAN lt gt PARAM START Start Y MODIFY METHOD lt PARAM gt Y lt gt y CHANGE PROTECTION PARAM gt Param lt gt v NO PROTECTION Y Param Setting Branch Protect Function Set the branch protect function as follows 1 Switch on the spectrometer in SuperUser mode 2 Select the appropriate branch 3 Preset e METHOD 210 Appendix 4 Use the arrow keys to select the required protect function 5 Press PARAM to confirm the protection OR Press STOP to cancel 6 Exit SuperUser mode to activate the protect function APPLICATION PARAM gt Y e Method Y NO PROTECTION Y Param Preventing Access to the Methods and Branches You can preve
110. p 2 above 4 From the 2 derivative spectrum determine the wavelength at the peak maximum and peak minimum Use these values for CALC WAV 1 and CALC WAV 2 97 Lambda 20 Lambda 40 UV Vis Spectrometers E D2 Em WAV MIN CALC WAV 2 CALC WAV 2 WAV MAX Peak minimum Peak maximum Determining the Threshold Value 2 derivative To determine the threshold value 1 Record the spectrum of the most dilute reference solution using the 2 derivative D2 mode over the wavelength range determined in step 2 or Determining the measurement wavelengths 2 derivative above 2 Determine the value D2 of this spectrum D2 1s the height from peak maximum to peak maximum D2 D2 WAV MIN CALC WAV 2 CALC WAV 2 WAV MAX Peak minimum Peak maximum 3 Select a value for the threshold parameter somewhat lower than this D2 value 98 Using Methods Creating a Method l Create a new CONCENTRATION 2 method see Creating a New Method on page 57 The following table lists typical CONCENTRATION 2 parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter 99 Lambda 20 Lambda 40 UV Vis Spectrometers No Parameter Value No Parameter Value 17 SLIT 2 0 nm 1 MODE PEAK AREA 3 WAV MAX 600 0 nm 4 WAV MIN 500 0 nm 7 REF 1 1 0C 7 REF2 REF 3 3 0C VAL
111. p switched off Defective detector 160 Error Messages PROBLEM SYSTEM ERROR SPECTROMETER FULL RESET DONE This error is shown when the instrument operating software crashes A full reset is then automatically carried out After the instrument 1s reset one of the following messages is shown on the second line of the display BATTERY LOW TIMER FAIL RS232 IRQ FAIL TIMER IRQ FAIL Make a note of this message Press PARAM to continue If you cannot continue call your PerkinElmer office and inform them of the error message This error is shown after changing the instrument software or after a full reset or when the spectrometer data is defect Make a note of the steps you made leading up to this message Press PARAM to continue If you cannot continue call your PerkinElmer office and inform them of the error message and the steps you made leading up to the error After a full reset all methods are erased 161 Lambda 20 Lambda 40 UV Vis Spectrometers DIALOG This error is shown after changing the instrument FULL RESET software or after a full reset or when the method DONE memory is defect Make a note of the steps you made leading up to this message Press PARAM to continue If you cannot continue call your PerkinElmer office and inform them of the error and the steps you made leading up to the error SPECTROMETER This error is shown after changing the instrument
112. paratet eller i stramledningen Peligro el ctrico Cualquier interrupci n del conductor de protecci n dentro o fuera del aparato o la desconexi n del terminal del mismo toma de tierra podr an ocasionar serios peligros al usar el equipo Prohibida la interrupci n intencionada Risque d lectrocution Toute interruption du conducteur de protection l int rieur ou l ext rieur de l instrument ou d connexion du raccord du conducteur de protection terre peut rendre l instrument dangereux li est interdit d interrompre volontairement ce conducteur Pericolo elettricit Qualsiasi interruzione della protezione del conduttore all interno o all esterno dello strumento o lo scollegamento del terminale di terra massa del conduttore di protezione possono rendere pericoloso lo strumento E vietato provocare volontariamente queste interruzioni Risico s door elektriciteit Het instrument moet voor de werking altijd geaard zijn In geen geval mag de aarding van het instrument of de netvoeding worden onderbroken of worden verwijderd Per go por electricidade Para a operag o o aparelho tem de estar sempre ligado terra De forma alguma separar ou retirar os condutores de protecg o terra no apareiho ou no cabo de alimenta o da rede 237 Lambda 20 Lambda 40 UV Vis Spectrometers 238 A WARNING W02 02 Lethal voltages are present in the instrument e Even with the power switch OFF line power
113. pectrometers 236 Wo01 03 Explosive Atmosphere This instrument is not designed for operation in an explosive atmosphere 9 60008901 Explosionsf hige Atmosph ren Das Gerat darf nicht in explosionsfahigen Atmosph ren betrieben werden Eksplosive omgivelser Apparatet m ikke anvendes i eksplosive omgivelser Atm sfera explosiva Este aparato no ha sido disenado para utilizarlo en atm sferas explosivas Atmosph re explosive Cet instrument n est pas congu pour fonctionner dans une atmosphere explosive Atmosfera esplosiva Questo strumento non adatto per luso in atmosfera esplosiva Explosiegevaarlijke omgevingen Het instrument mag niet in een explosiegevaarlijke omgeving worden gebruikt Atmosferas explod veis O aparelho n o pode ser utilizado em atmosferas explod veis Translations of Warnings WO2 01 Electrical Hazard Any interruption of the protective conductor inside or outside the instrument or disconnection of the protective conductor earth ground terminal is likely to make the instrument dangerous Intentional interruption is prohibited 00 0008 9 i Gef hrdung durch Elektrizit t Das Ger t muB zum Betrieb immer geerdet sein Auf keinen Fall die Schutzleiter im Ger t oder in der Netzzuleitung trennen oder entfernen Fare p grund af elektricitet Apparatet skal altid veere jordet Man m under ingen omsteendigheder skille eller fjerne jordlederen inde i ap
114. play is AUTOZERO START STOP Insert cell s containing a blank solution and press START OR If the display is TIMEDRIVE SAMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START 5 Continue to insert samples when asked until they have all been measured For additional information see Replot on page 79 Example of the Display Shown During the Measurement SCAN SMPL 1 Xxx nm Wavelength XXX nm XXXX A Xxx A Measured value ordinate as selected C xx Repeat measurement cycles still to be performed This appears on the top right when cycles gt 1 Printout The result is printed out at the end of each sample analysis numerical data follow at the end of the analysis 87 Lambda 20 Lambda 40 UV Vis Spectrometers Measurement at Several Wavelengths Select a wavelength program WAVELENGTH PROG method to measure a sample at several different wavelengths Procedure 1 Select the desired WAVELENGTH PROG method The following table lists typical WAVELENGTH PROG parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter No Parameter 17 SLIT 2 WAVELENGTH 3 WAV 2 11 FACTOR 1 11 FACTOR 3 15 LAMP 38 ACCESSORY 19 FIRST SAMPLE 22 CYCLE TIME 26 ORD MAX 28 SCALE 2 88 0 nm min No Parameter Value 1 ORDINATEMODE A
115. r value for THRESHOLD Error Messages Error Meaning No points stored Start measurement The method was interrupted with STOP before data could be stored for example during the equilibrium time of a substrate method Restart the method Two solutions to 1 ABS value Change curve fit algorithm In a non linear calibration curve two reference solutions exhibit the same absorbance value Check the references or change the concentration range or change the curve fit algorithm Value not within valid limits Check references or change curve fit alogrithm The measured sample concentration is outside the calibration range Measure additional calibration solutions within the concentration range or change the curve fit algorithm Wavelength data do not fit Start background correction Appears with TIME DRIVE methods if the wavelength has been modified since the last background correction Perform a new background correction using the proper method 165 Lambda 20 Lambda 40 UV Vis Spectrometers 166 Parameter Numbers 1 1 and Descriptions Parameter Numbers and Descriptions Parameter Numbers and Descriptions OF BATCHES To enter the number of batches to be analyzed Range 1 to 50 Appears with the autosampler accessory MEASUREMENTS To set the number of repeat sample measurements from each sample tube Range 1 to 9 Appears with the autosampler accessory
116. rically unsafe when it Shows visible damage Fails to perform the intended measurement Has been subjected to prolonged storage under unfavorable conditions Has been subjected to severe transport stresses Electromagnetic Compatibility EMC European Union EMC Directives This instrument has been designed and tested to meet the requirements of the EC Directives 89 336 EEC and 92 31 EEC It complies with the generic EMC standards EN 50 081 1 rf emissions and EN 50 082 1 immunity for domestic commercial and light industrial environments This instrument has passed the following EMC tests Safety Information Emission EN 50 081 1 92 Immunity EN 50 082 1 92 Emission of conducted and Electromagnetic Compatibility radiated noise EN55 011 91 IEC 801 2 91 EN 60 555 2 87 IEC 801 3 84 EN 60 555 3 87 IEC 801 4 88 IEC 801 5 90 United States FCC This instrument is classified as a digital device used exclusively as industrial commercial or medical equipment It is exempt from the technical standards specified in Part 15 of the FCC Rules and Regulations based on Section 15 103 c Japan FCC This instrument has been tested and found to comply with the limits of a Class A digital device pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates us
117. rjudicial para los ojos No mire fijamente a la l mpara Al examinar la radiaci n de las l mparas utilice siempre gafas de seguridad que absorban este tipo de rayos Rayonnement UV La lampe met un rayonnement UV intense susceptible de provoquer des l sions oculaires Ne pas regarder fixement une lampe allum e Toujours porter des lunettes de protection absorbant les UV pour regarder le rayonnement des lampes Radiazioni ultraviolette La lampada emette intense radiazioni ultraviolette dannose per gli occhi Non fissare le lampade accese Indossare sempre occhiali protettivi ad assorbimento degli ultra violetti quando si guardano le radiazioni emesse dalla lampade UV straling Risico s voor de ogen De stralingsbron lamp emitteert intensieve UV straling en kan daardoor letsel aan de ogen veroorzaken Draag een veiligheidsbril die de ge amp mitteerde UV straling voldoende absorbeert wanneer u in de brandende stralingsbron moet kijken Radiac o UV perigo para os olhos A fonte de radiag o l mpada emite uma radia o UV intensapodendo assim provocar ferimentos aos olhos Use culos de protec o que absorvam suficientemente a radia o UV emitida ao olhar directamente para a fonte de radiag o luminosa 241 Lambda 20 Lambda 40 UV Vis Spectrometers 242 Index Index A Accessories 129 Accessory Parameters 135 Analysis Procedure 78 Autozero 149 Autozero with Cell Changers 133 C C
118. s The absorption characteristics of a sample can be changed during sample preparation depending on the amount of reagent added for color development and so on For details refer to reference books covering these subjects Temperature has an influence to a greater or lesser degree on the absorption characteristics of a sample You should check this effect 1f non repeatable results are obtained If you are measuring temperature dependent samples either wait until temperature equilibrium has been attained or use a thermostatted cell or cell holder Chemically Reactive Samples If a reaction takes place in the cell between the sample material and the solvent spectral data based on that sample cannot always be expected to have sufficient reliability or repeatability For samples of this type use a quantitative method that takes advantage of the change in transmittance with time at a fixed wavelength For details refer to reference books covering this specific subject Photoactive Samples Some samples are known to be photoactive in that they fluoresce upon absorbing radiation Since a small portion of the fluorescent radiation will be measured by the detector a higher apparent transmittance will often result Samples are also known that undergo photochemical reactions as they absorb radiation With such samples which are mostly biochemical lack of reproducibility will characterize the resultant data Other Sample Properties
119. sing sequence if possible highest first lowest last Concentration 1 Wavelength n in nm Range 190 0 to 1100 0 in steps of 0 1 When using MODE 1 WAVELENGTH only 1 wavelength can be entered When using MODE 2 WAVELENGTH 2 wavelengths must be entered When using MODE 3 WAVELENGTH 3 wavelengths must be entered WAVELENGTH 1 measuring wavelength WAVELENGTH 2 and 3 wavelengths for baseline correction See Error Reference source not found 203 Lambda 20 Lambda 40 UV Vis Spectrometers WAVELENGTH The wavelength nm at which measurements are made Range 0 for adjustment of cell holder 190 0 to 1100 0 in steps of 0 1 204 Appendix Appendix SuperUser Activating SuperUser Mode SuperUser mode gives you access to all the branches in the instrument You activate SuperUser mode as follows 1 Switch off the spectrometer 2 Wait about two minutes to allow the lamps to cool down 3 Press 1 5 one five minus simultaneously 4 Keep the keys pressed and switch on the spectrometer 5 Release the keys when the display appears The SuperUser mode is now active Deactivating SuperUser Mode 1 Switch off the spectrometer 2 Wait about two minutes to allow the lamps to cool down 3 Switch on the spectrometer The SuperUser mode is now deactivated 207 Lambda 20 Lambda 40 UV Vis Spectrometers 208 Protect Functions General Information about Protect Functions Prot
120. sing the arrow keys if required 9 Press START REPLOT SPECTRUM BUSY The data is then replotted using the new ordinate and abscissa values The display returns to the ready display SCAN SAMPL 2 START STOP AZ PRINT You can replot the data as often as required Press STOP to interrupt the replot and return to the ready display NOTE Once you start the analysis for the next sample the data for the previous sample is deleted from the spectrometer memory and replaced with that of the current sample being analyzed 80 NEXT GROUP Using Methods n METHOD lt gt PARAM START CALL PARAMETER START BATCH PARAMETER l START START PARAMETER START MEASUREMENT NEXT SAMPLE Measurement repeated All samples in the group measured Figure 6 Flow diagram of a typical method procedure 81 Lambda 20 Lambda 40 UV Vis Spectrometers 82 Single Wavelength Measurements Select a TIME DRIVE method to measure a sample at one wavelength over a defined period of time For Enzyme Activity methods see Enzyme Kinetics on page 226 Procedure 1 Select the TIME DRIVE method The following table lists typical TIME DRIVE parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed des
121. t at the wavelength selected BLANK 7 Blank value Enzyme Blank value units as selected for ENZ UNIT Range 9999 9 to 9999 9 The enzyme activity is calculated as follows AEnz ATotal 7 ABlank Substrate Blank value units as selected for CONC UNIT Range 9999 9 to 9999 9 The substrate concentration is calculated as follows Csub 7 CrTotal CBlank CALC WAV 1 Wavelength in nm used in CONCENTRATION 2 methods for calculations using the 2 order derivative only Range 190 0 to 1100 0 in steps of 0 1 The values must lie within the WAV MAX and WAV MIN values set Enter the value at peak maximum for CALC WAV 1 see 172 Parameter Numbers and Descriptions Determining the Threshold Value 2nd derivative on page 98 In MODE DERIV 2 FIX the height of the derivative curve is measured at these wavelengths In MODE DERIV 2 PEAK a derivative maximum or minimum is located around these wavelengths and the nearest one evaluated see Error Reference source not found CALC WAV 2 Wavelength in nm used in CONCENTRATION 2 methods for calculations using the 2 order derivative only Range 190 0 to 1100 0 in steps of 0 1 The values must lie within the WAV MAX and WAV MIN values set CALC WAV 2 E cont Enter the value at peak minimum for CALC WAV 2 see Determining the Threshold Value 2nd derivative on page 98 In MODE DERIV 2 FIX the height of the derivative curve is measured at th
122. t in seconds s Option 0 1 0 2 0 5 1 2 5 10 A large value for time constant gives good signal noise ratios but can cause undesirable smoothing of the curve especially with rapidly altering signals Thus the time constant should be kept as low as possible but maintaining an acceptable signal noise ratio NOTE 7n analyses involving creeping reactions the time constant set must be lower than the CREEP TIME set NOTE The time constant set must be lowe than the INTERVAL in order to calculate dA dt see INTERVAL RETURN 79 To operate the pump in the reverse direction to return the sample to the sample tube Option YES NO Appears with the peristaltic sipper accessory RETURN TIME To set the amount of time for pumping in reverse Range 0 0 to 99 9 195 Lambda 20 Lambda 40 UV Vis Spectrometers Parameter Description SAMPL TIME To enter the salt concentration monvalent cation concentration of the oligonucleotide sample Used for the melting point calculation Units in mmol L Range 0 1 to 999 9 Shows only when Ty theoretical melting point is YES Refer to the Biochemical Application manual Aspiration time in seconds for sipper Range 0 0 to 99 9 Appears with the sipper accessory Sample identification up to a maximum of 8 characters SAMPLE ID SAMPLES BATCH Sample identification appears on every printout Number of samples per batch does not appear when using the cell changer If batch proce
123. t methods Sets the number of Thymine nucleotides Range 0 to 999 Appears with the Oligoquant methods Sets the number of bases N is used to represent any base Range 0 to 999 Appears with the Oligoquant methods 78 To enter the concentration of primer DNA in the solution This is not necessarily the concentration in the cell Used for the melting point calculation Units are nmol L Range 0 001 to 99999 9 Shows only when Ty theoretical melting point is YES Refer to the Biochemical Application manual User identification up to a maximum 8 characters User identification appears on each printout Maximum ordinate scale range for graphics printout valid only if GRAPHICS PLOT YES Range 9999 9 to 9999 9 If the analytical value obtained is outside the set values the latter should be changed NOTE Derivative spectra can also have negative values 190 Parameter Numbers and Descriptions al ORDINATE MODE Minimum ordinate scale range for graphics printout valid only if GRAPHICS PLOT YES Range 9999 9 to 9999 9 If the analytical value obtained is outside the set values the latter should be changed NOTE Derivative spectra can also have negative values Ordinate Mode T transmittance in percent A absorbance D1 to D4 Ist to 4 derivatives of the spectrum derivative spectra RAT absorbance ratio DIF absorbance difference COR Corrected Absorbance ratio CONC conce
124. tes 1430 using the numeric keys Press START to activate the clock OR Press STOP to cancel The realtime clock need only be set once and has the following functions day Monday Tuesday date yymmdd and time hhmm and continues working when the instrument is switched off The internal clock is limited to the following functions date yymmdd and time hhmm and counts from the time the spectrometer is switched on The internal clock must be reset to actual time after each switch on 121 Lambda 20 Lambda 40 UV Vis Spectrometers 122 900 DATE TIME lt gt PARAM Y Param MODIFY METHOD PARAM gt Y Para CLOCK INTERNAL Y lt gt Y CLOCK REALTIME S M Y DAY MONDAY Y lt gt y DAY MONDAY Y Y DATE 000000 ENTER m Y Para TIME 0000 ENTER Y Start Using Methods Wakeup You can use WAKEUP to set the spectrometer to switch on the lamps to warm up before the start of the working day l 2 10 11 12 13 14 Select the WAKEUP method 901 Press PARAM Press PARAM again Type in the wakeup date year month day 960429 using the numeric keys Press ENTER to confirm the entry Press PARAM again Type in the wakeup time hours minutes 0655 using the numeric keys Press ENTER to confirm
125. the new method as required 0 000 A 500 0 nm SELECT METHOD gt lt y nnn y Enter y NEW TIMEDRIVE PARAM gt Y lt gt Y NEW SCAN PARAM gt y Param y nnn SCAN lt gt PARAM START Overwriting an Existing Method 1 Select a method that can be overwritten 2 Press PARAM 3 Use the arrow keys to select NEW METHOD 4 Press PARAM again 58 Use the arrow keys to select the method type Press PARAM The existing method is written over OR Press STOP to cancel Modify the new method as required 13WAVELENGTHPROG lt gt PARAM START Param MODIFY METHOD PARAM gt NEW METHOD PARAM gt Y Param Y Es NEW TIME DRIVE PARAM gt Y Param Y 13 TIME DRIVE lt gt PARAM START Methods 59 Lambda 20 Lambda 40 UV Vis Spectrometers New Method Name 60 You can rename methods using the NEW METHOD NAME parameter The method number remains the same when you rename a method l 2 Select the method to be renamed Press PARAM Use the arrow keys to select NEW METHOD NAME Press PARAM again Use the arrow keys to select letters Confirm each letter by pressing ENTER OR Use the numeric keys to enter numbers Confirm each number by pressing ENTER OR Press ENTER without enter
126. the spectrometer switched on for approximately 10 minutes to allow the lamps to warm up and stabilize Startup 1 Open the sample compartment cover 2 Make sure that the beam paths are free that is e No objects for example cables project into the beam paths e No samples are in the sample compartment e Accessories are properly installed NOTE fthe sample compartment is obstructed during the startup procedure the spectrometer will not initialize correctly 3 Close the sample compartment cover 4 Switch on the power switch LAMBDA 20 VERSX XX 5 Wait for the standby display to appear Lambda 40 shows on those Initialization display spectrometers The standby display cud oe INPUT gt lt Other values may be shown Standby display 6 Switch on the accessories 29 Lambda 20 Lambda40 Operation and Parameter Description Shutdown 30 l Return the spectrometer to standby use STOP or PARAM Switch off the accessories Open the sample compartment cover Remove samples and cells from the sample compartment If accessories for example flowcell are installed in the sample compartment clean them thoroughly Close the sample compartment cover Switch off the spectrometer See also Wakeup on page 123 Startup and Shutdown Single Cell Holder Description Locking screw for horizontal alignment Vertical adjustment Screw Milled post Lifter Locking screw for horizontal
127. to Caution Opgelet Betekent dat de genoemde handleiding nauwkeurig moet worden opgevolgd om beschadiging van het instrument te voorkomen Caution Atenc o Significa que a instrug o referida tem de ser respeitada para evitar a danificagao do aparelho E gt z z o 90 00089 Translations of Warnings W01 02 Unauthorized Adjustments and Servicing Do not attempt to make adjustments replacements or repairs to this instrument except as described in the accompanying User Documentation Only a Perkin Elmer service representative or similarly trained and authorized person should be permitted to service the instrument Unzul ssige Arbeiten am Ger t Wartungs oder Reparaturarbeiten oder Justierungen die in der Benutzer dokumentation zum Ger t nicht beschrieben sind d rfen nur vom Perkin Elmer Kundendienst oder von entsprechend ausgebildeten und autorisierten Fachkr ften ausgef hrt werden Ikke tilladte arbejder p apparatet Vedligeholdelses eller reparationsarbejder eller justeringer som ikke er beskrevet i dokumenterne til brugeren m kun udfares af Perkin Elmers kundeservice eller af tilsvarende uddannede og autoriserede fagfolk Ajustes y servicios sin autorizaci n No intente realizar ning n tipo de ajuste sustituci n o reparaci n en este aparato a excepci n de lo descrito en la Documentaci n del Usuario que se adjunta Tan s lo un ingeniero de Servicio T cnico de Perkin Elmer
128. up in mg L In order that the results can be displayed independently of the actual mass of powder used the mass of the powder should be entered as the divisor If the powder were dissolved in 0 25 L instead of in 1 L an additional dilution factor of 0 25 should be entered The results are then automatically calculated as follows 0 25 mass of powder in g x concentration in mg L concentration in mg g Time from the start of the reaction addition of enzyme to the end of the reaction units as selected for TIME UNIT Range 0 1 to 999 9 181 Lambda 20 Lambda 40 UV Vis Spectrometers ENZ FACTOR 11 Enzyme factor Range 0 00001 to 9999 9 Calculate the enzyme factor as follows Enzyme factor V dv Where V is the volume of the total solution in the cell in mL d is the pathlength in cm vis the volume of sample in mL NOTE Depending on the procedure used the molar absorption coefficient may need to be taken into account ENZ UNIT Enzyme Unit U L Units per liter U mL Units per milliliter mU L Millinunits per liter U Units mU Milliunits mg mL Milligrams per milliliter Any unit Enzyme Unit U The amount of enzyme which catalyzes the conversion of 1 umol or micro equivalent of substrate per minute The unit may appear in capitals on the display 182 Parameter Numbers and Descriptions FIRST SAMPLE FIRST TUBE n u Factor Range 0 000001 to 9999 99 Factor for each wavelength n
129. wing table lists typical ENZYME parameters in the order left to right in which they appear See Parameter Numbers and Descriptions on page 169 for a detailed description of each parameter 105 Lambda 20 Lambda 40 UV Vis Spectrometers No Parameter Value 14 RESPONSE 05s 15 LAMP UVevis 22 TOTAL TIME 1 0 min 21 INTERVAL 0 2 min 24 LAG TIME 0 0 min 11 ENZ FACTOR 1 0 10 DIL FACTOR 1 01 12 DIVISOR 1 0 Only available with Lambda 40 2 Modify the method as required 3 Ifnecessary determine the blank value of the reaction see below and enter the value in the parameter BLANK 4 Press START This starts the measurement 106 5 Check the display If the display is AUTOZERO START STOP Using Methods Insert cell s containing a blank solution distilled water and press START OR If the display is ENZYME SMPL 1 START STOP AUTOZERO Insert a cell containing the sample solution and press START 6 Continue to insert samples when asked until they have all been measured Example of the Display Shown During the Measurement XXX nm XXX nm XXX A XXX nm XXX min XXX A Wavelength Time Measured value The display shown when using a cell changer xc n XXX min XXX A C 001 X C n C 001 XXX min XXX A Temperature Cell location Cyclic number Interval time Measured value 107 Lambda 20
130. yme Kinetics on page 105 or Substrate Kinetics on page 109 At the end of the measurement POSTRUN KIN appears on the display Press START to recalculate results OR If the results are not to be recalculated select NO using the arrow keys and press START Enter new values as required for the parameter displayed and press ENTER Press START to continue POSTRUN KIN YES lt gt Start i LAG TIME 0 0 min ENTER gt lt Enter i Start Using Methods Methods for Quantitative Analysis of Oligonucleotides Select an OLIGOQUANT 1 method for quantitative analysis of oligonucleotides up to 50 bases long and to calculate the theoretical melting point Select an OLIGOQUANT 2 method for quantitative analysis of oligonucleotides longer than 50 bases and to calculate the theoretical melting point You create an OLIGOQUANT method as follows l 2 Press METHOD Enter a method number nnn not previously used Press ENTER The first method of the NEW METHOD level appears Use the arrow keys to select the required method type Oligoquant 1 or Oligoquant 2 Press PARAM to confirm the creation of the new method OR Press STOP to cancel 115 Lambda 20 Lambda 40 UV Vis Spectrometers 6 Modify the new method as required Method Y 0 000A 500 0 nm min SELECT METHOD gt lt Y nnn Y Enter y NEW TIMEDRIVE PARAM gt Y lt gt y

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