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1. 128 TT 2 BEES SUBSROUTINIE dev Donee 128 eee 129 Ue ETE Ci uv 130 PIB IAS Melamine pine eet Nagios das ea 131 1724 Curves transformation IN ASCIEIIES oido tme oo i t o caedes EE e 131 ME ITROLOGY ac ree 133 Tod MERIBIGATIONVPES cis Co Nen 5 133 E admi TA Y 133 SCHOTT INSTRUMENTS Uvi Light XTD 15 9 S TAIRTUP dcn etait debi S Ree eto hiuc Mes 133 TO A PERIODICITY itat eu tene tom UO MN ICE tec MES 133 18 5 BACKGROUND NOISE VERIFICATION ON THE COMPLETE WAVELENGTH RANGE 133 18 6 STRAIGHT LIGHT VERIFICATION 340 134 18 7 VERIFICATION OF MEASURE STABILE use E Ee ma 134 S WAVELENGTH VERIFICATION ett dos e ui ppc 135 15 9 ABSORBANCE VERIFICATION
2. 98 TOO IVI H 99 Uvi Light XTD SCHOTT INSTRUMENTS VOR P ao 0 11118 A PTEE E E 100 19 040 SUSU ACO tierce UM E ALME ER M M 100 10054 O COlSIQiE ous ease n A LEM M ME 100 pS tes PUR I POE ROT Oot eR MU ewe oe A SEDE TEC 101 PSS AGE Lu Cm Mate ud Uu E A 102 125276 RT T 102 PAD SOMO IOS TERT 102 TANS AIC Carri setae So Vibe ee 102 103 14 1 PROGRAMMING THE 5 103 105 T4 9 SAVING THE METHOD ici abi a ted 105 144 SAVING THE ASGILPIBES 2 5 2 4 2 dodi isto vob d tede Solicitud 106 14 5S MEASUREME NT eon tout M AA EM EM daca AM ee e UE 106 TA SONIOQOE CM NETT 108 14057
3. 108 T46 ERR EOM TERCER 109 14 0 122 tte su edit ined eme 109 EY 109 109 14 5 1 5 Hard6ODV n et ouai Li oa ea 109 OG Bc na 109 NO INO P 4 110 110 111 AICO 111 112 14602 E M E E 113 Io caue tM D 115 LEN OG 116 14639 ETE ETT T 116 IU 5 9 0 OH EM IM EIDEM LAM SPA D LI ILU AR NRI IU OM IDOL MEUM EA ERE 116 4 6 3 7 Km calculation Michaelis Constant den i recen cres a ecd
4. ac faa 81 1255 ES E LEE E 81 125 2 dz A 82 12 9 D SQVIMg HG alvei eee oh aoe ease EE 83 12 5 4 Recalling ora stored QV SIS tate me dox eoe as se a MEC CIA 84 12 54 T Analysis slo TAG ied asm emu A IM IM LAM MU LAU AM A 84 12042 Analysis using One Sandal suite eti Tot ooo M Auge a mie eee eae 85 12 5 4 9 A DalVSiS HsIng several standards ss ima an et bo EH tedio t Lue 85 LU d uid Pu LE 86 12 5 65 damples TCIM CHING Mau ac 87 1236 1 EE RETE 87 12590 2 Ia io METRE 88 12 5 6 9J408nie aor a NSE 88 129 7 Locked m 90 13 SPECTRUM MODE ERR 91 131 THE PARAMETERS cux iv
5. 2 The software displays the procedure to be used for creating retrieving and running macro command in a rated fashion Macros commands create a macro instruction Go to the menu where you want the macro to start Press Ctrl F1 Go on normally At the end of macro press Ctrl F2 Press a function key to associate to the macro run a macro instruction Qo to the menu where you want the macro to start Press the chosen function key repeat a macro instruction Press Ctr Press the chosen function key Enter the number of repetition Time between two macros in sec Copyright SECOMAM 15 4 2 Listing of macro commands Enter F1 Listing Macros commands Choice of macro command Copyright SECOMAM Select the macro command to be listed Confirm and confirm 123 SCHOTT INSTRUMENTS Uvi Light XTD Press key to go Macros commands macro 1 SPECTRUM MEAS F1 8 F1 F4 Esc F1 0 10 F1 F2 1 5 00 Esc F1 F4 Esc sc Copyright SECOMAM The key s associated to the macro command its name and the corresponding sequence of commands are displayed In our example Key Alt F1 name SPECTRUM MEAS F1 Measure 8 Number of cuvettes F3 Tools F1 F5 Hardcopy Esc Quit F3 Calculation F3 Maxi F1 Peaks 0 10 Difference between the peak and the vall
6. TENER 75 12 o MUI TIEVVAVEDENG Saccus cad ved 75 PZ OI AE cl dete c I cde e tetur soda 5 UAPRITAZOCU NEL cm celts ess 76 the parameters dica Io 78 123 4 ENERO TT 79 t229 Menoa Savin Fee tans educ Me etu 79 3 6 Measure meN Rd 79 MO HO S i as uus LIMINE M C MM MI Pl 79 123 0 VNC TOSUNS 79 80 zi REGIS c c m PI 80 12 4 1 Meaning of the blank 80 80 12 4 3 Explanation or the 81 12 5 PARTICULAR WORKING S
7. EE ERR RR 117 USN OC RINT TREE 120 120 15 MACRO COMMANDYS RR E eM uU REPRE E VER ENS 121 15 1 CREATION OF A 8 121 15 2 EXECUTION OF 121 15 3 RATED EXECUTION OF MACRO COMMAND 122 15 4 RECALL OF APPLICATION AND LISTING OF A 122 19 4 4 MEET 122 DAZE O macro ww 123 16 UTILIZATION AS A SINGLE CELL HOLDER nnne 125 usui EM 127 a 127 SE SEM ATOAN c 127 otc et AR 127 127 TAS KIDS S Macc REO 127 17 1 5 Image tiles in spectrum Tode d dead Rao Dea duda dud 127 uw od TIES 128 I T OO Mn f 128 1771 60 MaCITO MIIBS
8. Tes Tape lv N 0 Del pointe Paint ho Next NN orrelation 0 0 7 m 2 l g m 2 o 2 q DETAILS ON THE CURVE 15 Concentration The values displayed on the absorbance axis are the values of the standards or samples calculated as follows Displ Abs Abs Std or Sam Bl Std or Sam Blank BI Blank Values read as against the air Press F8 when the calibration curve is over Enter the number of samples to be measured Confirm Fe SimuTation riParaneters 2003 12 02 08 02 48 Concentration Mode v1 00 CELL Reference Sam bl Sam 1 Sam bl sam sam bl sam Sam bl HI GCO mmuodou pr Prepare the multicell holder as indicated Start reading by pressing F1 Measure RESULTS Sample Conc Blank b1 Std bl Std 61 Std bl Blank Std 10 0 G L Std 20 0 G L Std 30 0 G L Copyright SECOMAM 71 SCHOTT INSTRUMENTS Uvi Light XTD Entrer number of cell 1 8 2003 12 02 08 04 37 Concentration Mode v1 RESULTS Sample Cone CELL Std bl Reference Blank Sam bl Std 10 0 G L 1 Std 20 0 G L Sam bl Std 30 0 G L Sam 2 Sam Sam 61 Sam 5 5 G L sam 3 sam sam sam sam sam sam E D E F G H I 10 4 G L UVWXYZ S t opyright SECOMAM THE DISPLAYED ABSORBANCES OF THE SAMPLES STANDARDS
9. 7 8 Sam 9 Sam 10 0 5 0 8 1 4 MG L 1 9 2 0 MG L 2 8 3 4 MG L 3 7 3 0 2 4 Abs 0 185 Abs 0 320 Abs 0 575 Abs 0 751 Abs 0 814 Abs 1 102 Abs 1 348 Abs 1 476 Abs 1 204 Abs 0 973 Time 10 32 am Time 10 33 am The sample concentration will be calculated as follow Sam Conc Sam Abs reference F actor Values read against the air Enter number of samples to be measured Confirm Follow indications displayed to prepare a new multicell holder for another series of measurement Press F1 Measure to read the following series Press ESC to leave the measuring mode b With a blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method 48 Uvi Light SCHOTT INSTRUMENTS Concentration Mode v1 00 PARAMETERS of measure Abs wavelength nm Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Factor 10000 Unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute Enter the number of samples to be measured Confirm 2003 11 21 11 14 21 Concentration Mode v1 00 RESULTS conc gt Copyright 5 Prepare the mul
10. Io ME me aiU 47 12 L4 1 Measure WI a Taclor oon t etum alea deba tee acia uude bee 47 12 4 Measurement witha SIBTdSl sss tee L Aet LA MT 50 12 14 3 Measurement with several standards di tul pte eap qtia detulit tt t dr tup ae tet adu qu lend aeu 53 12 1 5 Measurement with a sample 14 044 00000 000000000000005500000 60 PASS Ni PME Torr 61 12 52 WII T oc a dtes osse E ose eut Cube dob EA ebd UU 64 12 1 5 3 Measurement with several standards 68 2 CAEIBIATEB OUSINETIG otia ui a 72 7221 Hrooramming the ad ee 73 12 2 2 1 OF parameter oii 205 M ous 74 RMT OT SAV genii f emerat if cenas cct DD cM Sosa E P 74 1224 Measann trame ohn cm tete uiuit ear peo RPM D LEN bas DU Le 74 210 mE EE 74 22 6 VOCS UES S 74
11. B2 are coefficients which allow for absorbance calculations 51 Abs 2 Absn are the absorbances of a solution at LO LO2 being wavelength 75 SCHOTT INSTRUMENTS Uvi Light XTD 12 3 2 Calculations Example Ratio calculation calculation of the purity index of DNA The DNA purity index is given by a ratio of the absorbance read at 260 nm by the absorbance read at 280 nm with a trouble deletion by a measurement at 320 nm Ha 5 ADS gonm ADS Here we used 3 wavelengths then 3 absorbances The formula will therefore be _ AO A1x Abs1 A2x Abs2 4 A3x Abs3 BO B1x Abs1 B2x Abs2 B3x Abs3 Coefficients AQ 2 2 B3 will be chosen in such a way that the result will be If Abs ADS 560m Abs2 Abs Abs3 8 0 The calculation will be _ AO ALX ADS 6555 2 ADS 35 BO B1x Abs B2x 3 A0 0 Al 1 42 0 Hes 0 0 Bl 0 2 1 B3 1 0 1 ADS 0 X ADS CD 0 0 ADS 1 X ADS ADS 330m Then R ADS 562m APS33 09m ADS gonm ADS Other example Warburg Christian formula for proteins quantification Co oteines 1 55 x ADS 0 757 x ADS x60 nm given in mg mL Here we used 2 wavelengths then 2 absorbances The calculation is Uvi Light XTD SCHOTT INS
12. 000 0 100 0 000000000000 assis seas essa aas 36 9 3 VALIDATION OF ALL THE PARAMETERS 37 9 4 SPECIFIC CASE PARAMETERS FOR CONCENTRATION 37 95 PRINTING THE PARAMETERS x uide t 38 96 SAVING TRE PARAMETER Suc otii eo cM c e eal cata 38 10 HOW USE WITH SINGLE 0 22222 1 39 TO FMEAS URING 39 43 43 UT 43 CONCENTRATION etia Deb DR 45 12 1 CONCENTRATION MEASUREMENT 45 SCHOTT INSTRUMENTS Uvi Light XTD T2 1 T Erogrammalloh uo parameters Pita Eder eU 45 12 d 24 npressml of parameters uccisi ON ri aub ua ictu ae 47 TZ eo AVICHIOO SAVIN RR Ner 47 12 1 4 Measure without sample DIANK ss detiene toe e eed ee abet
13. 1 avs ofthe sample blanks RI Sam Water R2 Sam Abs of the sample s RI R2 RE Sam The coefficient is given by the following formula Std Conc Std Std Bl Blank Factor The sample concentration is given by the formula Sam Conc Sam Abs Sam BI Blank BI Blank actor 12 4 2 Logit axis The LOGIT is given by the formula logx 1 log x With 80 7 Uvi Light XTD SCHOTT INSTRUMENTS EE 4 BO Sample absorbance Reagent blank absorbance X Zero standard absorbance Reagent blank absorbance IMPORTANT If the event of a calibration with a LOGIT axis Ex Immuno enzymology it is necessary to program a zero standard with no concentration but with an absorbance higher than that of all the standards 12 4 3 Explanation of the various types of curve Straight line The calibration line consists of segments of straight line which link the calibration points together Spline To be used mainly for a non linear calibration It is the curve defined by the juxtaposition of several 3 degree curves which meet between each point and the 1 and 27 derivatives of which also meet To calculate all the coefficients of the 3 degree curves the algorithm moves along a sequence of approximations with the following criteria curves must across a rectangle of uncertainty centered on the measuring point The number of points of in
14. EN 9 9 m 2 o 2 q Wavelength nm a With cursor The absorbance of the one or several curves can be read simultaneously depending on the position of a cursor moved by the operator Press F1 Cursor 97 SCHOTT INSTRUMENTS Uvi Light XTD u 9 m 2 o 2 At the top left of the screen wavelength in nanometers with name of each curve and absorbance corresponding to such wavelength below are displayed It is possible to change the position of the cursor by means of the keys With each new position of the cursor new Wavelength Absorbance couples are displayed b Manual Press F2 Manual Enter wavelength in nanometers ANTHELIE Data will display the absorbance value for the curve s visualized on the monitor For both modes with cursor and manual the command F4 Hardcopy allows to print the curve s with the values The command F3 Print only prints out the values L O 220 1 Time 1 50 pm SP1 Abs 0 349 T9o 44 8 SP2 Abs 0 932 T 11 7 The command ESC allows to return to the previous menu 13 5 4 2 Derivatives It is possible to derive a spectrum up to 4 order It allows to eliminate a linear or non linear noise to look for the absorption peaks of solutions in a mixture etc NOTE The derivative of zero order allows to smooth ou
15. 90 Uvi Light XTD SCHOTT INSTRUMENTS 1 3 SPECTRUM MODE The spectrum mode is used for measuring the absorbance or transmission of a sample by reference to the wavelength It is especially useful to look for peaks of absorption in solutions and more generally speaking to know the behavior of a solution at different wavelengths The spectrum mode also offers the opportunity to process curves which have already been saved or acquired by the operator Load the method or select Spectrum Mode Confirm Introduction of datas User s name Batch number Origin Kit number Enter The user s name 20characters signs letters numbers The batch number 8 characters signs letters numbers The origin 30 characters signs letters numbers Kit number 8 characters signs letters numbers These information are necessary for measurement traceability ISO 9000 G L P and will be printed with the results Press ESC to skip this step 13 1 PROGRAMMING THE PARAMETERS For the parameters modification see page 36 F1 Modification F2 Printer Spectrum Mode v1 00 PARAMETERS RESULTS wavelength 2 nm 900 wavelength 1 nm 190 1 320 375 330 Speed 100 2400 1800 Y axis Absorb Parameters locked No Copyright SECOMAM 91 SCHOTT INSTRUMENTS Uvi Light XTD Press F2 Parameters then F1 Modification Wavelength 2 Final wavelength of the scan C
16. 2 1 U g m 2 o 2 o 58 Uvi Light XTD SCHOTT INSTRUMENTS Enter the calibration point number to be deleted and confirm POR TEAT 0630 3 2 x m x Es u i im n Confirm or do not confirm such deletion The operation may be repeated n 2 times n number of calibration points e Simulation It is possible to enter the values of the blank and the standards with the keyboard They will be printed and displayed as the measures See explanations in this chapter page 81 f Printing For printing the calibration curve press F7 Print g Saving The curve will automatically be saved if the user has already saved the analysis parameters h Calibration anomalies During the calibration curve setting up it is possible that different anomaly messages are displayed See explanation in this chapter page 82 i Recall a stored calibration curve To recall a stored calibration curve see in this chapter page 84 j Measurement Press F8 Next when calibration curve is over Enter the number of samples to be measured Confirm Concentration Mode 1 00 RESULTS Conc gt 4 U 2S Copyright SECOMAM Prepare the multicell holder as indicated Start the reading by pressing F1 Measu
17. tet tcn oue etu E m ubt 135 TOTO SUMMARY TABLE mM nm ace Me d 136 QN iz PING m eM 139 20 1S0 9001 CERTIFICA E indu esi ca boo REED ue RET 141 6 Uvi Light xr SCHOTT INSTRUMENTS 1 REMARKS Uvi Light xr SCHOTT INSTRUMENTS 2 WARRANT Y The new equipment and material sold by SCHOTT INSTRUMENTS is guaranteed against any manufacturing defects for one year unless otherwise stated by SCHOTT INSTRUMENTS with effect From the technical acceptance of the equipment in the factory by the buyer or his designee or failing this For Metropolitan France from the date on the delivery note Forother destinations from the date of factory shipment certified by air waybill consignment note or bill of lading The SCHOTT INSTRUMENTS company guarantee applies exclusively to defectiveness arising from a design fault or from a concealed defect It is strictly limited to the free dispatching of replacement parts except for consumable items or to the repairing of the equipment in our workshops within a deadline of 10 working days shipping delay not included By express agreement the following are strictly excluded from our guarantee All damages notably for staff costs loss of earnings business trouble etc Any breakdown due to an incorrect use of the equipment non adapted mains
18. Uvi Light XTD SCHOTT INSTRUMENTS Absit Abs t x Factor x 60 tm Abs t Abs of the solution at the end of the lag time Abst Abs of the solution at the end of the measuring time Factor Coefficient of molecular absorption im Measuring time Ratio reducing the result to one minute But the results can also be given in comparison with a standard or a calibration range 12 2 1 Programming the parameters For the parameters modification see page 36 Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fl Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure 2ptsKin wavelength Lag time 0 20005 Meas 1 20 100005 Reagent blank Decimal place 0 3 No of standards 0 10 0 10000 0 10000 0 10000 Unit 15 Y axis Absorbance Type of curve Copyright SECOMAM Type of measurement Choose 2ptsKin 2 points kinetic Wavelength Between 190 and 900 nm Lag time from O to 2000 seconds of time The lag time is introduced at the beginning of the reading to take into account the non linearity of a solution The unit performs the reading in the meantime but the delta of absorbance will only be calculated at the end of such time
19. Message An error has occured while backuping the files Retry gt Yes LENT ESC Files copy Ax Message 2 SECOMAM Light Manager Info Current software version vi Message Floppy disk is not a software update or is defective Retry gt Yes CENT ESC Message 3 SECOMAN Uva Light Manager Info Current software version v1 HH B Software update version v2 35 Message Error while creating update directory Retry gt Yes LENT ESC 23 SCHOTT INSTRUMENTS Message 4 SECOMAN Light Manager Info Current software version v1 HHBH B Software update version v2 35 Message Error while copying files from floppy disk Retry gt Ves CENT ESC Files copy Ax Message 5 SECOMAN Uvi Light Manager vi Info Current software version v1 HH Message There is no Floppy disk in drivet Please insert a Floppy disk gt Continue ENT Abort ESC Message 6 SECOMAN Uva Light Manager Info Current software version v1 HH B Software update version v2 35 Message Error while overwriting software files gt Ves CENT ESC Files copy 5 3 4 Set date and time From that screen 24 Light xTD Uvi Light XTD SCHOTT INSTRUMENTS SECOMAM Uvi Light Manager v1 20 Choices F1
20. line diagonally has been minimized and pass by the origin 12 5 PARTICULAR WORKINGS 12 5 1 Simulation It is possible to simulate readings which is very helpful for programs with calibration curves When the user has forgotten to save the calibration can rebuilt it if he knows the concentrations and the absorbances of the standards Press F2 Simulation Enter the absorbances values blank standards samples ranging from 1 000 and 3 000 with the keyboard The display and the printing of the results will be made as a true measurement Example of a calibration curve Enter in the concentration mode Look for the following screen 81 SCHOTT INSTRUMENTS Uvi Light Concentration Mode v1 00 RESULTS Sample Conc Copyright SECOMAM EMIJUN ME Press F2 Simulation to built the calibration curve Concentration Mode v1 00 RESULTS Sample Conc Abs CELL CELL A Blank CELL B Std 1 CELL C Std 2 CELL D Std 3 CELL E Std 4 SC A y NOPQR Space 0 1234567 89 E n nter Copyright RMON fe Enter the different absorbance values of the blank and the standards They will be printed and displayed as true measurements The operator will eventually continue with samples absorbance 12 5 2 Calibration anomalies During the calibration curve setting up it is possible that different anomaly messages are displayed FIX axis FZV axis S Type
21. As regards use with a coefficient a standard or several standards frequent references will be made to the description of the concentration mode Furthermore either of the three modes will be selected by changing one of the parameters in the table Type of measure 12 1 CONCENTRATION MEASUREMENT From the mean menu select Concentration mode Confirm 4 Introduction of datas User s name HOSPITAL Kit number With the help of mouse or virtual keyboard enter User s name 20 characters signs letters and numbers Batch number 8 characters signs letters and numbers Samples origin 30 characters signs letters and numbers Reagent kit number 8 characters signs letters and numbers This information is useful for measurements traceability G L P ISO 9000 and will be printed with the results Press Esc if the user wants to pass this stage 12 1 1 Programmation of parameters See page 29 From main menu Press F2 Parameters and F1 Modification 45 SCHOTT INSTRUMENTS Uvi Light XTD Introduction of datas Concentration Mode v1 00 PARAMETERS of measure Abs wavelength Decimal place 0 3 No of standards 0 10 Up 0 10000 0 10000 Unit X axis Concentration Y axis Absorbance Type of curve Numbering Parameters locke WXYZ Space 01 ight SECOMAM Type of measure Abs 2pt kin Multi WL Calibrated quanti
22. BLANK SAMPLE BLANKS STANDARDS BLANKS AND BLANK OF THE BLANK ARE READ AS AGAINST THE REFERENCE SOLUTION The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Blank Bl Abs 0 052 Time 11 43 am Std bl 1 Abs 0 100 Std bl 2 Abs 0 095 Std bl 3 Abs 0 107 Blank Abs 0 075 Std 1 Conc 10 0G L Abs 0 300 Std 2 Conc 20 0 G L Abs 0 600 Std 3 Conc 30 0G L Abs 0 850 Bias 0 025 Slope 0 025 Correlation 0 995 Sam bl 1 Abs 0 191 Time 11 47 am Sam 1 6 9 Abs 0 359 Sam bl 2 Abs 0 564 Sam 2 Conc 4 6 G L Abs 0 676 Sam bl 3 Abs 0 985 Sam 3 8 3 Abs 1 188 1 4 Abs 1 337 Sam 4 Conc Abs 1 318 Means that the calculation Sam x Sam Bl before to be reported on Y axis is negative Enter the number of samples to be measured Confirm Follow the instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode 12 2 CALIBRATED KINETIC This measuring method also called 2 point kinetic method 2ptskin allows to calculate the absorbance variation of a solution as a function of time As a rule the result is as follows
23. Blank bl Std bl BlanK Std Conc Factor 21 0 Sam bl 1 Sam 1 Conc Sam bl 2 Sam 2 Conc Sam bl 3 Sam 3 Conc 10 7 G L Sam bl 4 Sam 4 Conc Abs 0 010 Abs 0 047 Abs 0 030 10 0 Abs 0 543 Time 11 43 am Time 11 43 am Abs 0 030 Abs 0 438 Abs 0 035 Abs 0 492 Abs 0 072 Abs 0 589 Abs 0 074 Abs 1 009 8 2 G L 9 2 G L Time 11 45 am 19 6 G L If when result is displayed it appears it means that the result of the calculation before multiplication by factor is negative The factor is calculated as follows 67 SCHOTT INSTRUMENTS Uvi Light XTD Std Conc Factor Std Abs Std Bl Blank Bl Blank Values read as against the air The concentration of the sample is SamConc ISamAbs SamBl Blank actor Values read as against the air Enter the number of samples to be measured Confirm Follow the instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode 12 1 5 3 Measurement with several standards a Without reagent blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method IMPORTANT If the operator wants to keep the calibration curve he must first of a
24. Come back to the following screen CE f 2003 11 19 16 15 26 SECOMAM Uvi Light v1 00 SIMULATION Program choices m Absorbance Mode v1 00 04 11 2003 Kinetic mode v1 00 04 11 2003 v1 00 04 11 2003 Spectrum Mode v1 00 04 11 2003 Copyright SECOMAM SECOMAM Uvi Light Manager v0 01 Choices Fl Launch Light software F2 Servicing Tools Time remaining before automatic launch 16 sec Then the user can switch off Uvi Light spectrophotometer 14 Uvi Light XTD SCHOTT INSTRUMENTS 5 3 SERVICING TOOLS Press F2 Servicing Tools SECOMAM Uvi Light Manager 1 00 Choices Launch Uvi Light software F2 Update Uvi Light software Retrieve previous version of Uvi Light software F4 Set date and time FS Set PC keyboard mapping 5 3 1 Update Uvi light software WARNING this update procedure is available only with diskette supplied directly by SCHOTT INSTRUMENTS From this screen 1 SECOMAM Uvi Light Manager 1 00 Choices F1 Launch Light software FZ Update Uvi Light software Retrieve previous version of Uvi Light software 4 Set date and time FS Set PC keyboard mapping Press F2 or valid SECOMAM Uvi Light Manager v1 00 Info mw Current software version v1 00 Message A backup of your files is going to be created
25. Copyright SECOMAM Type of file with the and gt gt keys search for the type of file to be deleted editor program ASCII macro spectra or kinetic file Confirm Number of drive define as above the type of drive A C where the file to be deleted can be found Confirm Select the files to be deleted Confirm The files selected will be deleted When deletion is completed the software returns to the file management menu 17 2 4 Curves transformation in ASCII files This function allows to use the data obtained with Uvi Light in another software For example it is possible to transfer the data of a curve file spectra or kinetic file to a spreadsheet ex Excel to run them Press F4 Curve ASCII The display is 131 SCHOTT INSTRUMENTS Uvi Light XTD Selection Va idat ion Esc Escape files management conversion to ASCII Type of file scanning Separator of decimals Point Name of drive Copyright SECOMAM SUFIAN Type of file with the and keys select the type of file to be converted spectra or kinetic file Confirm Decimal separator point or comma In fact if the values obtained ex in a spectrum wavelength and absorbance have decimals the type of separator must be set forth A number of softwares into which the data will be transferred only accept one form of results The Uvi Light allows to adapt the files in order that they may be compatibl
26. Friday november the 21 2003 Time 02 07 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 BlanK Abs 0 050 Time 02 07 pm Std Conc 10 0G L Abs 0 653 Time 02 07 pm Factor 14 2 Sam Conc 6 7 G L Abs 0 473 Sam Conc 5 8 G L Abs 0 408 Sam Conc 13 9G L Abs 0 975 Sam Conc 12 8 G L Abs 0 902 Sam Conc 10 0G L Abs 0 702 Sam Conc 8 7 G L Abs 0 612 Sam Conc 13 7 G L Abs 0 962 Blanc Abs 0 050 Time 02 08 pm Sam Conc 2 9 G L Abs 0 204 Conc 5 91 G L Abs 0 416 The concentration value of the sample is calculated as follows Std Conc Std Abs Blank The concentration of the sample is computed as follows Std Conc Sam Abs Blank Factor Enter number of samples to be measured Confirm Follow indications displayed to prepare a new multicell holder for another series of measurement Press F1 Measure to read the following series Press ESC to leave the measuring mode Factor 12 1 4 3 Measurement with several standards a Preparing the measurement We will not make any difference between the measurement with a blank or not The displayed values on the curve are read as against the blank or reference solution 53 SCHOTT INSTRUMENTS Uvi Light XTD Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters
27. IMPORTANT In order to benefit from SCHOTT INSTRUMENTS s service application notes SCHOTT INSTRUMENTS information technical assistance etc immediately complete the attached guarantee card and return it to the following address SCHOTT INSTRUMENTS Hattenbergstra 3e 10 D 55122 Mainz GERMANY Phone 49 6131 66 5111 Fax 49 6131 66 5111 E mail support schottinstruments com 11 Uvi Light XTD SCHOTT INSTRUMENTS 4 THEORETICAL REMINDER 4 1 OPTICAL BEAM The polychromatic light emitted by a halogen lamp or deuterium lamp is broken down into monochromatic radiations after reflection on a grating Once the monochromatic light corresponding to the maximum absorption of the solution to be analyzed has been selected it crosses the sample where it will be partially absorbed 4 2 BEERT LAMBERT LAW Let s consider a beam of monochromatic light with an intensity Io which crosses a cell with flat parallel faces containing an absorbing solution Upon crossing the solution the beam with an intensity Io is attenuated in accordance with the Beer Lambert law Where 1 los elC 5 Where E is molar absorption coefficient of the substance isthe length of the cell generally 1 cm C is the concentration of the solution looked for in the cell LIMITS OF THE BEER LAMBERT LAW The light must have a narrow spectrum preferably a monochromatic one The wavelength used must correspond to the maxim
28. Launch Uvi Light software F2 Update Uvi Light software F3 Retrieve previous version of Uvi Light software Set date and time FS Set PC keyboard mapping Press F4 and valid SECOMAM Uvi Light Manager v1 00 Date and time Day DD _ Enter data in following order SECOMAM Uvi Light Manager v1 00 and time Day 00 Month MM Year YYYY Hour Valid data 25 SCHOTT INSTRUMENTS Uvi Light XTD 2003 11 20 SECOMAM Uvi Light Manager v1 00 Choices Fl Launch Light software F2 Servicing Tools Verify at the top right side of the screen inside black frame if date and hour are updating Run software by pressing F1 5 3 5 Set external keyboard mapping Independently of the language used in the software the operator can adapt the external keyboard to the country where he is Example U S A QUERTY keyboard FRANCE AZERTY keyboard And so on Several choices are proposed U S English French German or Italian keyboard From this screen QU 4 16 SECOMAM Uvi Light Manager 1 00 Choices Fi Launch Uvi Light software F2 Update Uvi Light software Retrieve previous version of Uvi Light software F4 Set date and time Press F5 or confirm 26 Uvi Light XTD SCHOTT INSTRUMENTS AU SECOMAM Uvi Light Manager v1 00 PC Keyboard Keyboard map
29. Measuring time from 20 to 10000 seconds Reagent blank YES or NO See meaning of the blank in this chapter page 80 Decimal place from 0 to 3 This parameter defines the number of significant digits after the decimal point Number of standards Factor 73 SCHOTT INSTRUMENTS Uvi Light XTD To work with a factor answer O Choose a value between 10000 1 standard To work with 1 standard answer 1 Choose a value between 0 and 10000 Several standards To work with several standards enter the number of standards from 2 through 10 Choose a value between 0 and 10000 Unit With keyboard enter the abbreviation wanted X axis Concentration axis Choose between linear and logarithmic Y axis Absorbance axis Linear logarithmic 1 Abs 1 Y or Logit See Logit axis meaning at the end of this chapter page 80 Type of curve Linear segmented curve Regre linear regression Regre 0 linear regression going through the origin 0 Spline See explanation on the type of curve at the end of this chapter page 81 Numbering This parameter concerns samples identification Auto Identification of the sample is numerical and increasing Example Sam 1 Sam 2 Unit The user assigns a name to each sample 8 characters and eventually attributes a dilution factor See explanation at the end of this chapter page 88 Table The user prepares a table in which he will enter the number of samples t
30. Save or not the method IMPORTANT If the operator wants to keep the calibration curve he must first of all save the method of analysis F2 Parameters F3 File results 2003 11 27 11 12 55 Concentration Mode v1 00 PARAMETERS of measure Abs wavelength Reagent blank Sample blank Decimal place 0 3 of standards 0 10 Conc 1 0 10000 2 0 10000 20 0 3 0 10000 30 0 4 0 10000 40 0 Unit G L X axis Concentration Lin Y axis Absorbance Lin Type of curve Linear Copyright SECOMAM Press F1 Execute 2003 11 27 11 16 43 Concentration Mode v1 00 RESULTS conc A Blank B Std Std D Std E Std Copyright SECOMAM SESEO Prepare the multicell holder as indicated Start the reading by pressing F1 Measure axis i 2 axis i s ESAE P LEN 405 Fo Del Pointi Print Next E o x m 2 l 0 m 2 o M 2 q 25 20 Concentration b Details on the curves and results The curves are drawn from the standards and the zero concentration point represented by the blank or the reference solution The values displayed on the absorbance axis are the values of the standards or samples read as against the blank or the reference solution The curves will therefore go through z
31. The sample is not stable or too concentrated SOLUTION Change the lamp used Verify that absorbance of the reference solution is not too high the sample is stable sample concentration is not too high there are not bubbles or impurities The spectrophotometer does not do anything any more EXPLANATION The instrument can be carrying out a calculation or a no locatable operation by the user SOLUTION Wait a minute If after one minute the instrument does not do anything any more switch it off and on and re start it again Wavelength abnormalities It is easy to check if the spectrophotometer is well adjusted as a whole in regards to the wavelength Select absorbance mode Check with a sheet of white paper in the cell well the correspondence of the colors with the wavelengths 650 nm red 600 nm orange 5 8 nm yellow 547 nm green 500 nm turquoise blue 450 nm blue 400 nm purple 140 U vi Light SCHOTT INSTRUMENTS 20 ISO 9001 CERTIFICATE E 1 gt J 2 iJ E ig P t4 t e E i d Tel 17 F qt F h gi s X iE T E m 27 I i A Lp aie 1 4 1 E CERTFKATION Dr ENTREPRISES The President of the Certification Committee fre 4 01698 Le President du Comit de Certification This certificate N 2003 CLXV
32. 1 100 The measuring time can be cut into several segments to allow for intermediate slopes in the results Choose from 1 to 100 Segment time 30 13000s It is the time of each segment in seconds Choose from 30 to 13000 s Minimum time for calculating the slope 1 segment x 30s 305 Maximum time for calculating the slope 100 segments x 13000s 1 300 000 s about 361 hours Type of calculation Linear regression It is the most accurate means of calculating the slope for a linear variation When the unit performs a reading it makes a least 300 scannings per second For each second the readings are averaged out The sequence of the absorbances forms a cloud of points between time T1 beginning of the first segment and time T2 end of the last segment The calculation draws a straight line within the points by minimizing the square of the distances between themselves and the straight line The result is obtained with the following formula N total number of points of the kinetic curve nb of segments x segment time 2142434 N N 1 2 Y total of absorbances at points 1 2 3 N Al A2 AN gt XY 2 2 3 AN x 1 4 9 N N N 102 1 6 The slope thus calculated is expressed delta of Abs min 2 Points The unit calculates the difference between the absorbances of the 2 most extreme points end of the lag time and the end of the last seg
33. 10000 conc 3 0 10000 Conc 4 0 10000 Unit 15 Y axis Absorbance Type of curve Copyright SECOMAM Print analysis parameters with F2 Print Save analysis parameters with F3 Save Press Esc Quit to continue Fl1 Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Abs wavelength Reagent blank 5 Sample blank Decimal place 0 3 No of standards 0 10 conc 1 0 10000 conc 2 0 10000 conc 3 0 10000 4 0 10000 Unit X axis Concentration Y axis Absorbance Type of curve Copyright sECOMAM Create a file to save analysis result with F3 File results Press F1 Execute Fo SinuTat fon easure of Zero H64 03716 16 23 31 Concentration Mode 1 PARAMETERS RESULTS measure Abs Conc A 10068 gt 0 1 amp H 1HBHHHB gt amp H 1HBHHB gt axis amp Concentration Y axis CAbsorbance gt Type of curve Copyright SECOMAM AEA ET Insert the Zero Reference cuvette in place Press F1 Measure After measuring remove the Reference cuvette Insert the Blank cuvette in place Press F1 Measure After measuring remove the Blank cuvette 40 Uvi Light XTD SCHOTT INSTRUMENTS E Simistion HH4 63 16 16 31 12 easure of Std Concentration Mode PARAMETERS RESULTS of
34. Central point 113 SCHOTT INSTRUMENTS Uvi Light XTD largeur du segment en secondes 20 1 4 Absorbance Absorbance 1 1 20 Temps secondes He will just have to move the cursor with the direction keys The software will automatically compute the slope on one or several curves within the time range centered around the position of the cursor ELTIECECSUDNEBEREETEDDOTTSUREEDRZDUIRATTA Absorbance 1 1 n S S S 20 30 40 50 Temps en secondes Both times defined by the time range will be displayed together with the name s of the curves and the slope s They are calculated with the last coefficient displayed in the parameter window and reduced to one minute b Extreme points Press F2 Extreme points Set the times with the cursors Confirm Choose the calculation mode regression F1 Regre or 2 points F2 2pts Confirm 114 Uvi Light XTD SCHOTT INSTRUMENTS y 9 0 2 m o 2 1 1 1 aS 20 40 50 Temps en secondes Both times defined by the times range are displayed with the name s of the curves and the slope s They are calculated with the last coefficient displayed in the parameter window and reduced to one minute The command F1 New value allows to select other time intervals For both modes central point or Extreme poin
35. N 0 Example X axis 7 Lin Y axis Lin Type Linear Zero point not used x m 2 y g m 2 o 2 q n 20 25 Concentration Press F4 Y 0 to use the zero point again Changing the X axis Press F1 X axis Choose a linear or logarithmic axis for concentrations Press F1 Lin Linear axis for concentrations Press F2 Log Logarithmic axis for concentrations Example X axis Log Y axis Lin Type Linear Zero point not used 55 SCHOTT INSTRUMENTS Uvi Light XTD X axis axis FP o Del point LY E Absorbance blank 15 20 Concentration Changing the Y axis Press F2 Y axis Choose a linear or logarithmic axis for absorbances or a calibrated axis in 1 Abs or Logit Press F1 Lin Linear axis for absorbances Press F2 Log Logarithmic axis for absorbances Press F3 1 Y reverse of absorbance Press F4 Logit See explanation in this chapter page 80 Example X axis 7 Lin Y axis 1 Y Type Linear Zero point not used X axis i 4a lt gt 0 Fo Del Print S Next x 2 y 9 2 o 2 Changing the type of curve
36. Sam 1 2 87 SCHOTT INSTRUMENTS Uvi Light XTD Entrer number of cell 1 8 aU AB 4 Concentration Mode v1 00 RESULTS Conc Abs Sample CELL Std 10 0 g l Q 173 CELL Reference Factor 57 9 CELL B Std CELL C Sam 1 1 22 1 g l 0 381 CELL D Sam 2 2 35 3 g l 0 610 CELL E Sam 3 Sam 3 42 8 g l 0 739 CELL Sam 4 Sam 4 57 3 g l 0 989 CELL G Sam 5 5 59 0 g l 1 019 CELL Sam 6 Sam 6 69 0 g l 1 192 CELL I Sam 7 Sam 7 87 7 g l 1 514 sc NO Q R UVWX Y Bac 0 123456789 re Enter Copyright SECOMAM 12 5 6 2 Unit identification Select Unit for the Numbering parameter Charge factor It is possible for certain reagents VWR to enter a factor F which is a correction factor corresponding to the reagent batch used The factor ranges between 0 9 and 1 1 and is initialized to 1 The value of the results is multiplied by such factor If the operator does not want to change it he just has to confirm it otherwise he will enter the new value and confirm Dilution factor The dilution factor ranges from 1 to 999 and is initialized at 1 The value of the results is multiplied by such factor When the operator has confirmd the sample name the cursor positions itself on the dilution factor If the operator does not want to change it he will just have to confirm it otherwise he will enter the new value and c
37. Sam Conc Sam Abs Sam Bl Factor Values read as against air Follow the instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode b With a blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fl1 Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure 5 5 Wave length Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Factor 10000 Unit Numbering Parameters locked Press F1 Execute Confirm 62 Copyright 5 U vi Light SCHOTT INSTRUMENTS 2003 12 01 12 12 28 Concentration Mode v1 00 RESULTS Conc CELL CELL A Reference CELL B Blank b1 CELL C Blank Copyright sECOMAM Prepare the multicell holder as indicated Start the reading by pressing F1 Measure Entrer number of cell 1 8 8 2003 12 01 12 14 30 Concentration Mode v1 00 RESULTS Sample conc Abs CELL Blank 1 0 183 CELL A Reference Blank 0 344 CELL B Blank bl CELL Blank QRSTUVWX YZ Back 6 G Enter Copyright SECOMAM Concentration Mode v1 00 RESULTS conc CELL Reference Sam b1 Sam 1 Sam bl Sam 2 Sa
38. XTD SCHOTT INSTRUMENTS Absorbance 1 1 i 1 zo 30 Temps en secondes Confirm Absorbance 20 Temps en secondes To retrieve the initial curve enter F1 Original 14 6 2 3 Manual zoom Press F3 Manual Enter the time scales Example Time mini 10 Confirm Time maxi 20 Confirm Automatic zoom Yes The unit recalculates the absorbance scales automatically No Enter the absorbances scales 14 6 3 Calculation Press F3 Calculation 111 SCHOTT INSTRUMENTS Uvi Light XTD 1 3 Der ivee Uperation o Constante F i Michaelis Esc Quiel g m 2 o 2 1 20 Temps en secondes This function allows to read the absorbance on one or several curves at a given time calculate a slope calculate the derivatives add two curves substract one curve from another multiply a curve by a number add a constant to a curve calculate the Michaelis constant 14 6 3 1 Absorbance Allows to read an absorbance in view of a time on one or several spectra Press F1 Absorbance Absorbance 1 1 20 Temps secondes a With cursor The absorbance of one or several curves can be read simultaneously depending on the position of a cursor moved by the operator Press F1 cursor 112 Uvi Light XTD SCHOTT INSTRUMENTS y o 0 2 0
39. be sent to SCHOTT INSTRUMENTS For equipment with a guaranty card this is only applicable if the card delivered with the equipment is returned to SCHOTT INSTRUMENTS duly completed SOFTWARE GUARANTEE The software is guaranteed by the designer or the distributor of the software under the conditions specified in the literature accompanying the aforementioned software packages Under no circumstances whatsoever will SCHOTT INSTRUMENTS supply any type of guarantee for software packages By express agreement all damages notably for staff costs lost of earnings business trouble etc are strictly excluded from our guarantee The customer is informed that the software cannot be guaranteed exempt from defects or bugs Uvi Light xr SCHOTT INSTRUMENTS 3 INSTALLATION The SCHOTT INSTRUMENTS equipment has been designed manufactured tested and inspected according to the ISO 9001 standards If the unit is not immediately installed it should be stored in a dry and clean area The storage temperature should be between 10 and 35 SCHOTT INSTRUMENTS equipment is carefully inspected before it is packed As soon as you receive your equipment check condition of the packaging and if you notice any problems notify your carrier within 48 hours Then consult the packing list and check that everything is in order Finally if you discover that something is missing or if the goods are damaged immediately notify SCHOTT INSTRUMENTS
40. for security reason gt Continue ENT Abort ESC Inside the upper frame it displays the actual version of the software Press Esc Abort Then the software comes back to the main menu Before to import new version of the software it is important to save the previous version Taper ENT Continue 19 SCHOTT INSTRUMENTS Uvi Light XTD SECOMAM Uvi Light Manager v1 00 Info Current software version v1 00 Message Please insert a Uvi Light update floppy disk gt Continue ENT Abort ESC Insert in floppy drive the new version of the software and press ENT SECOMAM Uvi Light Manager v1 00 Info Current software version Software update version Message Floppy disk files are being copied gt Please wait Files copy SECOMAM Uvi Light Manager v1 00 Info Current software version v1 00 Software update version v7 77 Message Software update process is now finished gt Please press a key Press any key 20 Uvi Light XTD SCHOTT INSTRUMENTS 1 Len 1 2003 11 19 17 16 02 SECOMAM Uvi Light Manager 1 00 Choices Fl Launch Uvi Light software F2 Servicing Tools Run the new software version by pressing F1 5 3 2 Retrieve previous version of Uvi light software From that screen Choices F1 Launch Uvi Light software F2 Update Uvi Light software Set date and time Set P
41. library proposed choose the one to recall confirm and confirm 14 6 1 3 Delete This function allows to delete one or several curves chosen by the user Press F3 Delete Choose the curve s to be deleted confirm and confirm The curve s will be deleted 14 6 1 4 Print Press F4 Print The curve s will be printed horizontally without comments in a 11x21cm format 14 6 1 5 Hardcopy Press F5 Hardcopy The curve s will be printed vertically without comments in a 21x29 7 cm format 14 6 2 Scales This function allows to expand an interesting section of a curve Press S Scales 109 SCHOTT INSTRUMENTS Uvi Light XTD o m 2 o M 2 q 1 20 Temps secondes NOTE Throughout the function the ESC key allows to return to the previous menu 14 6 2 1 Original This command allows to return to the initial curve after a manual expansion or with the cursor Press F1 Original 14 6 2 2 Zoom with cursor Press F2 Cursor o N Absorbance 1 1 1 20 40 Temps secondes Select the left limit of the selection to be expanded by moving the cursor with the and keys Confirm Then select the right limit of the selection to be expanded Frame at the top and at the bottom the section to be expanded by moving the cursor with 7 keys and validating whenever it reaches the correct position Example 110 Uvi Light
42. name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Sam 1 Conc 0 092 G L Abs 0 046 Time 11 43 am Abs 1 0 437 Abs 2 0 460 Sam 2 Conc 0 132 G L Abs 0 066 Abs 1 0 447 Abs 2 0 480 Sam 3 Conc 0 220 G L Abs 0 110 Abs 1 2 0 421 Abs 2 0 476 Sam 4 Conc 0 148 G L Abs 0 074 Abs 1 2 0 442 Abs 2 0 479 12 2 7 The curve The deltas of absorbance defined as follows are on the ordinate axis AAbs mn 5 mn _ Blank IMPORTANT For measures with several standards creation or simulation of calibration curves measure or simulation of absorbance saving or not of methods or calibration curves recall of analyses with stored curves refer to concentration mode Simulation The values entered by simulation are considered deltas of absorbance and multiplied by the factor In such case the absorbance values for the times considered are the last values measured and remain in the display 12 3 MULTI NAVELENGTH 12 3 1 Principle This program allows to make various calculations on a solution measured at several wavelengths ratio Allen s correction and also to know the absorbance value of a solution at different wavelengths up to 10 The formula used is of the following type _ A0 Alx Abs1 A2x Abs2 4 Anx Absn B0 Bl x 1 2 x Abs2 e Bn x Absn R is the result of the calculation 0 1 2
43. ny 20 1 1 i 20 30 Temps en secondes At the top of the screen the time in seconds the name of each curve and the absorbance corresponding to such time below are displayed It is possible to change the position of the cursor by means of the and keys With each new position of the cursor new Time Absorbance couples are displayed b Manual Press F2 manual Enter time in seconds The unit will display the absorbance value for the curve s visualized on the monitor For both modes with cursor and manual the command F4 Hardcopy allows to print the curve s with the values The command F3 Print only print out the values as follows Time 20 0 Sam Abs 0 904 Sam Abs 1 325 Press ESC to return to the previous menu 14 6 3 2 Slope This function allows to read the slope reduced to one minute and multiplied by the coefficient indicated in the parameter window Such calculation can be performed in the regression or 2 poinst mode between 2 times entered by the operator or with reference to a central point around which the operator will have defined a time range on one or several curves Download the curve s on which slope calculations are to be performed Press F2 Slope Absorbance 1 20 Temps secondes a Central point The operator is to define a time range in seconds which will be centered automatically around the measuring point Example 20 Sec Press F1
44. place 0 3 No of standards 0 10 1 0 10000 0 10000 X axis Concentration in Y axis Absorbance Lin Type of curve Linear Numbering 5853348 locked MNO i 96 T Sp 4 1 2 3 456789 Copyright Press ENT to continue the measuring process Press ESC to return to the previous screen 12 5 4 Recalling of a stored analysis 12 5 4 1 Analysis using a factor Load the method 94 Uvi Light XTD SCHOTT INSTRUMENTS Enter the parameters for the traceability Execute Parameters File results Esc Quit CONCENTRATION WITH 4 STANDARDS v1 00 PARAMETERS of measure Abs Wavelength Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Factor 10000 Unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute to start the analysis 12 5 4 2 Analysis using one standard Load the method Enter parameters for the traceability Press F1 execute If the standard value has been saved with the method it displays CONCENTRATION WITH 1 STANDARD v1 00 PARAMETERS RESULTS of measure Abs Sample Conc Wavelength Reagent blank Sample blank Decimal place Q No of standards 0 0 100 Unit Numbering Parameters locked Copyright SECOMAM No new measure Enter F1 Keep Standards The sample measurements will be done using the stand
45. place during the autotest 35 seconds later the display is SECOMAM Uvi Light Manager 0 01 Choices F1 Launch Light software F2 Servicing Tools Time remaining before automatic launch 16 sec 2 choices Run the software by pressing F1 Launch Uvi Light software or let the software run automatically after 20 seconds To have access to the servicing tools by pressing F2 Servicing Tools NOTE If anomalies are detected connection cable disconnected beam completly stopped to prevent doing autotest correctly 4 minutes after launching the application the software will display the following requests Want you to start again the autotest If the user answers NO it displays Warning No measure is possible Pass over the autotest If the user answers YES the software will run in SIMILATION MODE 5 1 LAUNCHING OF THE SOFTWARE Press F1 Launch Uvi Light software Less than 2 minutes later the main menu will be displayed SCHOTT INSTRUMENTS Uvi Light SECOMAM Uvi Light v1 00 SIMULATION Program choices m Absorbance Mode v1 00 04 11 2003 Kinetic mode v1 00 04 11 2003 Concentration Mode v1 00 04 11 2003 Spectrum Mode v1 00 04 11 2003 Copyright SECOMAM If the printer is connected the Light spectrophotometer will print the following autotest report The Uvi Light spectrophotometer is ready to work 5 2 STOP OF UVILIGHT
46. programme parameters Save or not the method Fl1 Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS Abs wavelength 0 3 No of standards 0 10 Concentration 0 10000 Unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute 66 Uvi Light xTD SCHO F2 simulation F3 Parameters TT INSTRUMENTS Concentration Mode v1 00 CELL Reference Blank b1 std m Blank Std sam sam sam sam bl bl 1 2 gt 2 Prepare the multicell holder as indicated Start the reading by pressing F1 Measure Entrer number of cell 1 8 RESULTS Conc Copyright SECOMAM 2003 12 01 16 08 17 Concentration Mode 1 00 Sample RESULTS Conc CELL Reference Blank bl Blank 61 Std bl Blank Std 61 Blank Std sam sam Std Factor bl 1 Sam bl Sam Sam Sam 1 gt 10 0 G L 2170 1 8 2 G L 9 2 G L THE DISPLAYED ABSORBANCES OF THE SAMPLES STANDARD BLANK SAMPLE BLANKS STANDARD BLANK AND BLANK OF THE BLANK ARE READ AS AGAINST THE REFERENCE The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00
47. the bottom of the cuvette is 8 5 mm if there are no bubbles or impurities which block the beam if the date beyond which the reagent is not to be used is not over passed f the blank or reference solution reagent is pure or conform to its use There are some problems during printing EXPLANATION The printer does not receive any data from Uvi Light spectrophotometer SOLUTION Verify that the printer is connected to the main the printer used is a parallel printer the used cable is the correct one SCHOTT INSTRUMENTS reference 0J5706 the cable is correctly connected on the printer and on the spectrophotometer tighten the screws connector the standard of the selected printer corresponds to the used printer The absorbance is too high EXPLANATION Something blocks the beam or there is no beam SOLUTION Verify that the cuvette is setting in the correct position bubbles and impurities does not block the beam selected lamp is working properly 139 SCHOTT INSTRUMENTS Uvi Light XTD The absorbance is negative EXPLANATION The absorbance of solution used to perform the zero is higher than the sample absorbance SOLUTION Verify that the absorbance of solution used to perform the zero is not to high there are no bubbles or impurities inside blank solution Results instabilities EXPLANATION The lamp used still functions but must be close to its end of lifetime
48. the top line of the screen with all the possibilities offered to the operator For example after reading a sample in Spectrum Mode the menu is as follows F1 Measure F2 Parameters F3 Tools Esc Quit To have access to any of the sub routines press the key corresponding to the option selected Example Press F3 Tools The following menu is displayed F1 F2 Scales Calculation F4 Note F5 Windows F6 Abs F7 Trans Esc Quit Press F3 Calculation the following menu is displayed F1 Abs F2 Derivative F3 Maxi Operation F6 Constant F7 Ratio Esc Quit As a rule presentation on the monitor will be as follows A first line with the choice such as F1 Measure F1 I O or F2 Derivative A second line with instructions such as Ready for measure of zero or Authorized measure A large frame with on the right the parameters of the analysis in process and on the left the results or a curve The last line with operations in process such as Storage of the baseline or Saving in process 21 Uvi Light XTD SCHOTT INSTRUMENTS 8 ANALYSIS MODES 8 1 RESIDENT MODES They are the measuring modes included in the software SECOMAM Uvi Light 1 00 SIMULATION Program choices i V 04 11 2060 Concentration Mode 04 11 2003 Spectrum Mode 04 11 2003 Copyright SECOMAM Absorbance continuous measurement of absorbance and transmittance Kinetics follo
49. wavelength 1 nm 190 wavelen chan 320 375 330 100 2400 1800 Absorb Parameters locked No TUVWXYZ ET right SECOMAM Q 5 _ Press F1 Measure to start the measure Enter the number of samples to be measured Confirm After reading the basic line on the cell in A position the spectrophotometer will make the acquisition of the scans one by one starting with the cell on B position It displays one by one every scan progressively with their acquisition The scan will be saved temporarily under the form SP Index The name with the higher index will be the last scan performed by operator 1 F2 Paraneters 3 Tools Esc Quit 0 4 Absorbance 13 5 TOOLS Press F3 Tools This sub routine is used for various processings zoom derivative or calculation on the curves stored in memory 13 5 1 Input output This function allows to load save delete print one or several curves Press F1 I O 93 SCHOTT INSTRUMENTS Uvi Light XTD U 0 m 2 o 2 A 300 Wavelength Cnm 13 5 1 1 Save The curves may be saved one by one or several all together with some comments what is called image NOTE The images are different from curves and it is not possible to superimpose between them or with curves a Curv
50. 0 10000 X axis Concentration Y axis Absorbance Type of curve Numbering Parameters locked sc D H Space 0 1234567 89 Enter file Ex RESULTS Start measurement The results will be saved in ASCII form in the file created by the operator These data be loaded from the DOS and displayed by entering name of ASCII file PRN Example C UVILIGHT Type RESULTS PRN D NUVILIG 1 TYPE RESULTS PRN SECOMAM Spectrophotometer Uvi Light v1 00 Date Thursday december the 11 2003 Time 7 55 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 5426 Method Concentration Mode v1 00 1 2 3 E e 5 Fi D NUVILIG 1 The name of this ASCII file will appear by pressing F3 Tools F1 Files and F1 Directory NOTE If the operator has failed to assign a name to a file for storing the data of a reading in the concentration or kinetic mode the software creates a file by default called CONC PRN or KINET PRN The data formerly will be overlaid by the new ones upon each reading 12 5 6 Samples numbering This parameter is the sample identification There are three ways to proceed Automatic identification One by one identification Identification using a table 12 5 6 1 Automatic identification Select Auto for the Numbering parameter The identification of the sample will be digital and increasing i e
51. 000 Calculation Decimal place 0 3 Factor 10000 ni Speed Substrate unit Micha Numbering Parameters locked Copyright SECOMAM 14 4 SAVING THE ASCII FILES Offer the possibility to keep the results of the analyses in the form of ASCII files Press F3 File results and enter the file name under 8 characters See the explanation page 86 NOTE If the operator has failed to assign a name to a file for storing the data of a reading the software creates a file by default call KINET PRN The data stored formerly will be overlaid by the new ones upon each reading 14 5 MEASUREMENT Press F1 Execute Measure Parameters 3 15 sc Quit Kinetic mode v1 00 PARAMETERS RESULTS Wavelength nm Lag time 0 18000s No of segments 1 100 Segment time 30 13000 Calculation Decimal place 0 3 Factor 10000 Unit Speed Substrate unit Micha Numbering Parameters locked Copyright SECOMAM Prepare the multi cell holder as follows Position A Blank reagent or reference Position B through l Samples Throughout the measuring sequence the multi cell holder will be prepared in the same way 106 Uvi Light XTD SCHOTT INSTRUMENTS Press F1 Measure Enter the number of samples to be measured Confirm The spectrophotometer will make the measurement every 30 seconds of the samples and the reference in A position compensating at the same time
52. 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Table of sample names KITI23A x 20 2 KITI23B x 1 0 3 23 x 4 0 4 KITI23D 3 0 5 2 2 0 6 2 2 0 7 KITI23G x 1 0 8 KITI23H x 3 0 Once the table is ready enter F9 End 89 SCHOTT INSTRUMENTS Uvi Light XTD Measure Simulation Parameters Esc Quit Concentration Mode 1 00 RESULTS Conc CELL erence SACO nNMvVowor Copyright SECOMAM launch the measurement by entering F1 Measure Other lable Remeasure lable sc Quit Concentration Mode v1 00 RESULTS Sample Conc D N 0 315 0 561 0 764 0 871 1 090 1 166 1 971 0 183 Copyright SECOMAM To measure the table again with the same configuration enter F2 Remeasure table To prepare another table enter F1 Other table 12 5 7 Locked parameters Allow to lock the analysis parameters Select Yes for Parameters locked in the parameters window If the operator wants to modify a locked analysis the following message will appear ANALYSIS LOCKED v1 00 PARAMETERS of measure Abs Decimal place No of standards 10 1 Unit Numbering Parameters locked Copyright SECOMAM It is not possible to unlock an analysis You have delete it see page 94 and create a new one
53. 3 2 Retrieve previous version of Uvi light software 2 000020020 000000000000000000 21 9 LI LT LEM 22 S 22 oer date eld mici tam Is NE M M M vated Dd cT c s t Ur M c Les 24 5 3 5 Sel external Keyboard sts ERE LPE 26 CONE IG CRAG IN e c 29 PRINCIPLE FOR MENUS AND COMMAND OF UVI 31 S ANALYSIS MODES aa ie see a a a cc ccs wide we wets nc ee ce wa eed tea eae 33 Gai OID EIN WO DES T 33 92 CUSTOMIZED METHODS ettet dao dte ueber oet 33 5 2 T o Ha e a isa de pda uUa M MM uU AU Is MI ELM MEE 34 9 ALTERING THE PARAMETERS 2 2 24 4 22 4 1 35 SELECTION OFATARAMETER IM Stu Mosca DS EE 36 Su MODIFICATION OFA PARAMETER buie 36 9 2 1 The parameters is entering by the 36 9 2 2 The nature of the parameter is
54. 4 Michaelis Menten On Y axis V On X axis 5 wen V 2 d Eadie Hofstee form Press F5 Eadie Hofstee 119 SCHOTT INSTRUMENTS Uvi Light XTD LA S On Y axis V s Slope Bias Uitesse IS1 1 1 100 150 200 EADIE HOFSTEE Vitesse 4 pMOL L Press F1 Hardcopy It prints the curve and the Michaelis constant value Press ESC ESC Quit It prints the Michaelis constant value and comes back to the previous menu e Simulation The user can rebuilt the curve 5 V 5 and automatically calculate Michaelis constant Press F2 Simulation enter the points number with for every one the substrate concentration and the reaction speed It will display the curve and the Michaelis constant value Proceed the same to print the curve and the constant value f Error messages Problem 8 The substrate concentration values are not increasing and regular Xl X2 X3 Xn Problem 9 The reaction speed values are not increasing and regular YI Y2 Y3 Yn 14 6 4 Note The command F4 Note allows to write comments on the curves These comments are not stored except if the user saves them under image form Enter F4 Note Place the cursor initial position at the top right side of the screen below the strip display by means of the direction keys Confirm its position Enter the text Confirm To print the kinetic curve s with comme
55. 4 Note allows to write comments on the curves The comments are not stored except if the user saves them under Image form See in this chapter Page 102 Enter F4 Note Place the cursor initial position at the top right side of the screen below the strip display by means of arrow keys or mouse Confirm its position Enter the text Confirm For printing the curve s with comments go to F1 I O and enter F5 Hardcopy 13 5 6 Windows The command F5 Windows allows in the graphic mode to return to the parameters The opposite operation is performed by entering F4 Graphic 13 5 7 Absorbance When spectrum is in the transmittance mode on the monitor this command allows to display it in the absorbance mode Press F6 Abs 13 5 8 Transmittance When spectrum is in the absorbance mode on the monitor this command allows to display it in the transmittance mode Press F7 Trans The processings which can be performed on a type of spectrum can also be performed on the other type 7102 7 Uvi Light XTD SCHOTT INSTRUMENTS 1 4 KINETIC MODE NOTE THE KINETIC MODE IS SIMILAR OF THE SPECTRUM MODE IN THE ORGANIZATION OF THE SUB MENUS WE WILL THUS REFER AT THIS PROGRAMS IN THE PRESENTATION OF THE SUB MENUS The kinetic mode is used for measuring the absorbance as a function of time Example determination of enzyme activity evolution of the absorbance of a solution as a function of time On the main menu select Kinetic Mode
56. C keyboard mapping Press F3 or valid SECOMAM Uvi Light Manager 1 00 Info m Current software version Wr or Previous software version v1 00 Message Do you want to retrieve the previous version of the software gt Yes ENT No ESC Press ENT 224 SCHOTT INSTRUMENTS Uvi Light XTD SECOMAM Uvi Light Manager v1 00 Info m Current software version B Previous software version v1 00 Message Restoration of the previous version is being processed gt Please wait Files copy When the previous version is restored the display is SECOMAM Uvi Light Manager v1 00 Info m Current software version v7 77 BH Previous software version v1 00 Message Restoration of the previous version is now finished gt Please press a key Press any key 1 gt 1 2003 11 19 17 16 02 SECOMAM 1 Light Manager 1 00 F2 Servicing Tools Run new software version by pressing F1 5 3 3 Error messages You will find here under different error messages which can appear during software updating In these cases verify if the floppy disk contains the new software version verify if the floppy disk is not damaged switch off and on the spectrophotometer and start again the different operations 22 Uvi Light XTD SCHOTT INSTRUMENTS Message 1 SECOMAN Uvi Light Manager Info Current software version vBH HH
57. Confirm 4 Introduction of datas User s name Batch number Origin 5 HOSPITAL Kit number VWXYZ Back Mea ar a eer ight SECOMAM With the help of virtual keyboard or mouse enter The user s name 20 characters signs letters numbers The batch number 8 characters signs letters numbers The origin 30 characters signs letters numbers Kit number 8 characters signs letters numbers These information are necessary for measurement traceability ISO 9000 G L P and will be printed with the results Press ESC to skip this step 14 1 PROGRAMMING THE PARAMETERS For the parameters modification see page 36 PARAMETERS RESULTS wavelength nm Lag time 0 180005 No of segments 1 100 Segment time 30 13000 Calculation Decimal place 0 3 Factor 10000 Unit Speed Substrate unit Micha Numbering Parameters locked Copyright SECOMAM With Esc key it is possible to return to the previous menu and the main menu 103 SCHOTT INSTRUMENTS Uvi Light XTD Press F2 Parameters then F1 Modification Wavelength Choose from 190 to 900nm Lag time 0 18000s It is a time in seconds that must be introduced at the beginning of the analysis for taking into account the none linearity of a reading The unit performs the scanning in the meantime but the slope calculation will be performed outside the lag time Choose from 0 to 18000s Number of segments
58. E SOLUTION The results will be printed out as follow SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 10 32 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Std Conc 10 0G L Abs 0 174 Time 10 32 am Factor 57 5 24 5 G L Abs 0 427 33 6 G L Abs 0 584 39 4 G L Abs 0 685 Conc 49 8 G L Abs 0 866 57 6 G L Abs 1 002 Conc 81 3 G L Abs 1 414 Conc 99 7 G L Abs 1 560 Conc 10 2G L Abs 0 182 Time 10 33 am 19 2 G L Abs 0 342 Sam Sam Sam Sam Sam Sam Sam Sam 3 The value of the factor is calculated follow 51 SCHOTT INSTRUMENTS Uvi Light XTD Std Conc Factor Std Abs reference The concentration of the sample is computed as follows Std Conc Sam Abs reference F actor Values read as against air Enter number of samples to be measured Confirm Follow indications displayed to prepare a new multicell holder for another series of measurement Press F1 Measure to read the following series Press ESC to leave the measuring mode a With blank Load the method or select Concentration Mode Enter name batch number samples origin and reagent kit number Modify or programme parameters Save or not the metho
59. ED ABSORBANCES OF THE SAMPLES STANDARD SAMPLE BLANC AND BLANK OF THE BLANK ARE READ AS AGAINST THE REFERENCE SOLUTION 65 SCHOTT INSTRUMENTS Uvi Light XTD The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Std bl Abs 0 183 Time 11 43 am Std Conc 10 0G L Abs 0 352 Time 11 43 am Factor 59 1 Sam bl 1 Abs 0 578 Sam 1 Conc 0 1 G L Abs 0 651 Sam bl 2 Abs 0 845 Sam 2 Conc 0 1 G L Abs 1 144 Sam bl 3 Abs 1 282 Sam 3 Conc 0 3 G L Abs 1 520 Sam bl 4 Abs 0 845 Time 11 45 am Sam 4 Conc 0 1 G L Abs 1 144 Sam bl 5 Abs 1 282 Sam 5 Conc 0 3G L Abs 1 520 The value of the factor is calculated as follows Std Conc Std Abs Std Bl Values read as against air Factor The concentration of the sample is Sam Conc Sam Abs Sam Bl JF actor Values read as against air Enter the number of samples to be measured Confirm Follow the instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode b With a blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or
60. ER FA FILTER Tolerance 0 007 Abs Tolerance 0 010 Abs Tolerance 0 010 Abs W L nm Theoretical V Reading value TheoreticalV V Reading value Theoretical V Reading value 137 Uvi Light XTD SCHOTT INSTRUMENTS 1 9 MAINTENANCE The spectrophotometer does not require any specific maintenance Only the halogen lamp or the deuterium lamp must be changed by the user However the spectrophotometer must be kept perfectly clean in particular the cell compartment which must not contain any spilt chemicals the input and output windows of the light beam in the cell compartment must be cleaned regularly with a non fluffy cloth To change the deuterium lamp or halogen lamp see installation guide REF 0M8039 The operator will find here under some probable causes of troubles and some advices to help him to solve these troubles For any no solved problem call SCHOTT INSTRUMENTS or technicians approved by SCHOTT INSTRUMENTS The spectrophotometer does not perform the zero EXPLANATION The photocell does not receive enough energy SOLUTION Verify that the lamp used is still working if not change it if the measure is performed in UV light 190 330 the cuvette is built in quartz QS is written on its side the cuvette is installed in the right direction the optical sides of the cuvette are perfectly clean the distance between the centre of measuring windows when a flow through cell is used and
61. F4New Calibration Concentration Mode v1 00 RESULTS Sample Conc 10 0 g l 20 0 g l 30 0 g l 40 0 g l Copyright SECOMAM EMIJUN ME It is necessary to verify if The concentration values entered in the parameters are monotonous The absorbance values of the standards are monotonous The standards were measured in the correct order The absorbance of the standard at a zero concentration is higher that of all the standards as is the case with a LOGIT absorbance axis 92 Uvi Light XTD SCHOTT INSTRUMENTS The absorbance of the first standard is not nil as is the case with a logarithmic absorbance axis The concentration of the first standard is not nil as the case with a logarithmic concentration axis The operator may Change either of both axis if they were poorly selected Change if need be the concentration of the standards in the parameters window Recalibrate by entering F4 New calibration 12 5 3 Saving the analysis If the user has entered a factor a standard or several standards the analysis saving is the same When parameters are entered it displays Modif ication Printer F3 Save sc Quit PARAMETERS of measure Ab Wavelength Reagent blan Sample blank Decimal place 0 3 No of standards 0 10 Conc 1 0 10000 Conc 0 100 Conc 0 10000 ne 4 0 10000 nit xX axis Concentration Y axis Absorbance Type of curve Li
62. Fse uit Spectrum Mode v1 00 PARAMETERS RESULTS Wavelength 2 nm Wavelength 1 nm Wavelen chan 320 375 Speed 100 2400 15 Parameters locked Copyright Perform baseline zero on the air Stopped the beam with opaque material Launch the measure Absorbance from 190 to 900 nm 2 5 Results 133 SCHOTT INSTRUMENTS Uvi Light XTD 18 6 STRAIGHT LIGHT VERIFICATION AT 340 NM Enter in the absorbance mode Select 340 nm 4 Data introduction Absorbance Mode 1 00 Wavelength RESULTS B Continuous absorbance B p Continuous transmittance 7 sc D H K Space 0123456 89 gt Copyrig Perform the zero on the air Insert SCHOTT INSTRUMENTS standard filter ref 404686 Read the value Results 18 7 VERIFICATION OF MEASURE STABILITY Enter in kinetic mode Enter parameters as follows FI Zero F7 TocIs F3 Paraneters Es c uit Kinetic mode v1 00 PARAMETERS Wavelength nm Lag time 0 18000s o of segments 1 100 RESULTS Segment time 5 13000s Calculation Regre Decimal place 0 3 Factor 10000 1 Unit Speed Substrate unit Micha Numbering Auto Parameters locked No Copyright SECOMAM Perform the zero on the air Launch the measure on the air Make a zoom closer to the curve Read the highest and the lowest a
63. LTS Sample Conc CELL Std Reference Std Std Std Std Std Std Std Std Std Std Std Std Std Std Std gt RSTUVWwWXYZ Back t Enter Copyright SECOMAM Prepare the multicell holder as indicated Start the reading by pressing F Measure Entrer number of cell 1 8 2003 12 01 17 13 12 Concentration Mode 1 00 RESULTS Sample Conc CELL Std bl Reference Std b1 Std 1 Std bl 10 20 30 40 TI B D H I E od e pwne HPOOOOOOOoOOO VWXYZ Back Enter ht SECOMAM THE DISPLAYED ABSORBANCES OF THE SAMPLES STANDARDS SAMPLE BLANKS AND STANDARDS BLANKS ARE READ AS AGAINST THE REFERENCE SOLUTION 69 SCHOTT INSTRUMENTS U vi The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Std bl 1 Abs 0 102 Time 11 43 am Std bl 2 Abs 0 120 Std bl 3 Abs 0 115 Std bl 4 Abs 0 099 Std 1 10 0 G L Abs 0 200 Std 2 Conc 20 0 G L Abs 0 400 Std 3 Conc 30 0 G L Abs 0 600 Std 4 Conc 40 0 G L Abs 0 750 Bias 0 035 Slope 0 017 Correlation 0 994 Sam bl 1 Abs 0 228 Time 11 45 am Sam 1 Conc 0 1 G L Abs 0 400
64. Press F1 Measure 41 SCHOTT INSTRUMENTS Uvi Light XTD Fi Measure F2 Sinulat ion F2 Curve F4 Paraneters Fsc Quit 1 16 16 37 37 eazure nf Sam Concentration Mode vi Hi PARAMETERE RESULTS Type measure 5 fib Conc 16 6 G L 26 6 G L 36 6 G L 46 6 G L standards 75 1 G L 010000 0 1 nian 0 1 A inn A axis Concentration Y axis CAbsorbance gt Type of curve Copyright SECOMAM SRT Insert sample 2 in place Press F1 Measure etc Automatic print out of the values of the Standards and Samples 42 Uvi Light XTD SCHOTT INSTRUMENTS 1 1 ABSORBANCE MODE This mode allows performing a continuous absorbance and transmittance reading Select Absorbance mode with the TAE keys Confirm 11 1 WAVELENGTH CHOICE 4 introduction Absorbance Mode v1 00 RESULTS UVWXYZ Back Enter ight 5 Enter the wavelength and confirm 4 Data introduction Absorbance Mode v1 00 5000 RESULTS Enter SIMULATION 11 2 MEASURE Prepare multicell holder as follow SCHOTT INSTRUMENTS Uvi Light XTD A position Always blanc Reagent or reference solution From B to I Samples To start measurement press F1 Measure F2 Wavelength Absorbance Mode v1 00 500 RESULTS 270 T73 381 610 739 Time 3 02 pm B D E F G H I EFFFFEEFI a At any time the user c
65. Press F3 Type Choose among 4 types of curves Press F1 Linear line segments Press F2 Spline spline curve Press F3 Regression linear regression curve Press F4 Regression 0 linear regression going through the origin See explanation of different types of curve in this chapter page 81 Example Uvi Light xTD X axis 7 Lin Type Spline Absorbance blank 15 20 Concentration Regression curve It is possible to use two different types of curve for the regression mode linear regression linear regression going through the origin Linear regression curve going through the origin Press F3 Type Press F4 Regression 0 Example X axis 7 Lin Y axis Lin Type Regression 0 SCHOTT INSTRUMENTS Zero point used eS EE ELSE Pipe bed point E Pr tink TT fibporbarce blank 5 L cunic Fo irim Regression linear curve Press F3 Type Press F3 Regression Example X axis Lin Y axis Lin Type Regression Zero point used 57 SCHOTT INSTRUMENTS Uvi Light XTD EIE las 0 ope orrelation 4855 m m 2 y 80 2 ps 20 5 20 25 Concentration The sp
66. SE PARAMETERS FOR A CONCENTRATION READING Select the CONCENTRATION analysis mode Press F2 Parameters then F1 Modification Selection Ya idat ion Esc Escape Concentration Mode 1 00 PARAMETERS of measure Abs Wavelength Reagent blank Sample blank Decimal place No of standards Concentration 1 Unit Numbering Parameters locked Copyright SECOMAM The screen displays two windows on the left hand side The first one allows to select the analysis mode measurement of absorbance with reference to a coefficient a standard or a calibration range 2 point kinetic reading multiple wavelength reading The second one is the window linked to the analysis modes In the first window to change the parameter select the measuring mode with lt and keys Then confirm Ex 27 SCHOTT INSTRUMENTS Uvi Light Concentration Mode v1 00 PARAMETERS of measure MultiWl Reagent blank Decimal place No of standards Unit No of M L Wavelength 1 Wavelength 2 GENERAL CALCULATION AQ A1 Abs 1 A2 Abs 2 AnxAbsn Copyright SECOMAM Modify the parameters as stated above NOTE In the Multi WL mode the general calculation formula appears in a window on the right hand side 9 5 PRINTING THE PARAMETERS For printing the parameters when the method of analysis has been loaded press F2 Paramete
67. Sam bl 2 Abs 0 568 Sam 2 Conc 0 1 G L Abs 0 737 Sam bl 3 Abs 0 859 Sam 3 Conc 0 3 G L Abs 1 071 Sam bl 4 Abs 1 208 Sam 4 Conc 0 4 Abs 1 585 Enter the number of samples to be measured Confirm Follow the instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode b With a blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Light xTD IMPORTANT If the operator wants to keep the calibration curve he must first of all save the method of analysis Fl Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS Abs Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Conc 1 0 10000 cone 2 0 10000 Conc 3 0 10000 Unit X axis Concentration Y axis Absorbance Type of curve Numbering Parameters locked Copyright SECOMAM Press F1 Execute Uvi Light xTD CELL Reference Blank 61 Std b1 Std bl Std bl Blank Std Std Std Prepare the multicell holder as indicated Start reading by pressing F1 Measure SCHOTT INSTRUMENTS Concentration Mode v1 00 2003 12 02 07 36 12 RESULTS conc Copyright SECOMAM
68. TRUMENTS _ A0 Alx Abs1 A2x Abs2 0 Blx Abs1 B2x Abs2 Absl Abs Abs2 Abs The calculation will be A0 Al x ADS sonn 2 ADS seo nm 0 Blx 2 0 0 1 155 2 0 757 BO 1 1 0 2 0 _0 1 55 0 757 R 1 0 ADS 86 nm 1 2 ADS 560m Then Cir oteines 1 55 x ADS 0 757 x ADS given in mg mL Another example Allen s correction It allows to make a quantitative measure of a peak in chemical noise The formula is Abs1 Abs3 2 52 Absl Abs3 2 2 Abs Abs2 For three equidistant wavelengths the calculation is A0 Alx Absl A2 x Abs2 Abs3 BO Blx Abs1 B2x Abs2 B3x Abs3 4050 Al 1 42 2 1 BO 2 1 0 B2 0 B3 0 04 1 x Absl 2x Abs2 1 x Abs3 a ee 2 0 Abs1 0x Abs2 Abs3 eff SCHOTT INSTRUMENTS Uvi Light XTD Abs1 Abs3 2 Abs2 Absl Abs3 2 Abs Abs2 12 3 3 Programming the parameters For the parameters modification see page 36 Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fl Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Multiwl Reagent blank z Decima
69. UVI LIGHT XTD User s manual 4 SCHOIT Instruments Uvi Light xr SCHOTT INSTRUMENTS CONTENT ETT E TUI 7 a 9 3 INSTALLA THON CE 11 4 THEORETICAL ska S deu da vate 13 OR TC AL BEAM 13 AZ reat dia d 13 4 DEFINITIONS a a 13 EON M ICI EET 13 72 Bes Waly 1 8 4 Oe B Bus a ae mC n enu aiU A neu a HS oae M 14 A o CoOMDCehllaligl mum sole AMI Lu EI I MILI x M I M ades 14 d OTHO udi ruta e ies M MM LM P M IM M DE I MM cM iu Edda ME 15 43o SCANNING DEREN RETO mmm 15 5 CONNECTION AND 17 S EAUNGHING OF THE SSOFTWAIE etti seo dot 17 52o OP OF A mrt ec esc Un 18 Deo OER VIGING TOOLS m 19 9 3 1 Boni aUos Ko cim 19 5
70. ach of them NOTE The recalibration process consists in shifting the curve in view of the new blank if any and in performing an homothetic transformation which allows to juxtapose the former value of the standard with the new one Delete the previous calibration Yes ENT No ESC NO It keeps in memory the previous calibration curve but it works with the new calibration curve for this sample batch YES The new calibration curve is saved in memory New measure of the whole Enter F2 Redo Std Measure again the blank if any and the standards 12 5 5 Files results Uvi Light offers the possibility to keep the results of the analysis in the form of ASCII files Such files may be retrieved by copying them on floppy disk to run them on other computer systems Excel Execute Parameters File results Esc Quit CONCENTRATION WITH 4 STANDARDS v1 00 PARAMETERS of measure Abs Sample blank Decimal place 0 3 No of standards 0 10 Conc 0 10000 Conc 2 0 100 Conc 3 0 10000 Conc 4 0 10000 Unit A axis Concentration Y axis Absorbance Type of curve Numbering Parameters locked Copyright SECOMAM Press F3 File results It displays 96 Uvi Light XTD SCHOTT INSTRUMENTS aracters CONCENTRATION WITH 4 STANDARDS v1 00 PARAMETERS of measure Sample blank Decimal place 0 3 No of standards 0 10 1 0 100000
71. an change the wavelength by pressing F2 Wavelength THE DISPLAYED BLANC ABSORBANCE IS READ AGAINST THE AIR THE DISPLAYED SAMPLES ABSORBANCES ARE READ AGAINST THE BLANC The results are printed as follow SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Thursday november the 20 2003 Time 3 02 pm Absorbance at 500 nm date Thursday november the 20 2003 Blank W L 500nm Abs 0 299 T 50 3 Time 3 02 pm Sam 1 W L 500 Abs 0 185 T 65 3 Sam 2 W L 500 Abs 0 320 T 47 9 Sam 3 W L 500 Abs 0 575 T 26 6 Sam 4 W L 500 Abs 0 751 T 17 7 Sam 5 W L 500 Abs 0 814 T 15 4 Sam 6 W L 500nm Abs 1 102 T 7 9 Sam 7 W L 500 Abs 1 348 4 5 Sam 8 W L 500 Abs 1 476 T 3 3 Esc key allows to exit out of absorbance mode and come back to the main menu 44 7 Uvi Light XTD SCHOTT INSTRUMENTS 1 2 CONCENTRATION MODE This function allows to use any methods against one or several standards if necessary without standard if a calibration coefficient is entered to obtain concentration results or results processed by a mathematical formula The readings are concentration calibrated kinetics multiple wavelength The concentration programs without standard with standard and with several standards will be studied in detail N B the principle of the other two measuring methods i e calibrated kinetics and multiple wavelength will be defined
72. an eventual unit or reference drift The curves are drawn on the screen following data acquisition They will be saved temporarily under the name KIN index The name with the higher index will be the last scan performed by the operator Absorbance 1 20 Temps secondes At the end of the reading the result is printed out in the following form SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Kinetic Mode v1 00 1 Sam 1 Time 11 43 am 2 Speed 18 G L 3 Slope 0 018 Abs mn 4 time 5sec Abs 0 174 Slope 0 020 Abs mn 5 time 35 Abs 0 184 Slope 0 017 Abs mn 1 2 Time 11 43 am 2 Speed 18 G L 3 Slope 0 018 Abs mn 4 time 5 Abs 0 326 Slope 0 034 Abs mn 5 time 35 Abs 0 343 Slope 0 000 Abs mn Result explanation Line 1 Sample name Line 2 Reaction speed is the total slope per minute multiplied by K factor Line 3 Total slope per minute Line 4 5 Starting time for every segment the corresponding absorbance and intermediate slopes reduced to one minute for visualizing the kinetic evolution To display the results enter F3 Tools then F5 Windows 107 SCHOTT INSTRUMENTS Uvi Light XTD PARAMETERS RESULTS Time Wavelength nm Lag time 0 180005 51 G
73. ard value previously read New measure of the whole Enter F2 Redo 1 Std Measure the standard before measuring the samples Delete the previous calibration Yes ENT No ESC NO It keeps in memory the previous value of the standard but it works with the new standard value for this sample batch YES The new standard value is saved in memory 12 5 4 3 Analysis using several standards Load the method Enter the parameters for the traceability Press F1 Execute If the calibration curve has been saved with the method it will display 85 SCHOTT INSTRUMENTS Uvi Light XTD CONCENTRATION WITH 4 STANDARDS v1 00 PARAMETERS RESULTS of measure Abs Conc Wavelength blank Sample blank Decimal place No of standards 0 1 120 00 0 10000 0 10000 0 10000 X axis Concentration Y axis Absorbance Type of curve Linear Copyright SECOMAM It is possible to retrieve it to work with the curve to recalibrate it or to draw a new one No new measure Enter F1 Keep Standards The curve which was stored is then displayed It is ready for measurement New measure of the whole Enter F2 Redo 1 Std Enter the number of the standard to be re read Confirm Prepare the multicell holder as indicated Press F1 Measure to start the measurement It will consider that all the standards in the range have evolved similary and will recalculate the absorbance of e
74. are a new multicell holder for another series of measurement Press F1 Measure to read the following series Press ESC to leave the measuring mode 12 1 4 2 Measurement with a standard a without blanc Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fl Execute F2 Parameters Fs3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Abs wavelength nm Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Concentration 0 10000 Unit Numbering Parameters locked Press F1 Execute Copyright SECOMAM Uvi Light XTD SCHOTT INSTRUMENTS Concentration Mode v1 00 RESULTS conc CELL Reference Std sam sam sam sam sam sam sam H rOOmmuonour Copyright 5 EMINENS Prepare the multicell holder as indicated Start the reading by pressing F1 Measure Entrer number of cell 1 8 2003 11 24 11 22 04 Concentration Mode 1 00 RESULTS Sample Conc CELL Std 0 Reference Factor 57 9 Std sam sam sam sam sam sam sam sam sam sam sam gt 4 FR RSTUVWXYZ Back 6 Enter JE Copyright SECOMAM SAUFEN THE DISPLAYED ABSORBANCES OF THE STANDARD AND SAMPLES ARE READ AS AGAINST THE REFERENC
75. bsorbance of the solution When the optical path is constant and equal to 1cm therefore constant the concentration of the solution will only depend on the absorbance measured by the instrument and the molar absorption coefficient typical physical constant of the substance to be studied for a given wavelength TWO ALTERNATIVES E is known it is given on the data sheet of the reagent used Then the calculation formula used is F Coefficient or factor When is equal to 1 cm 14 Uvi Light XTD SCHOTT INSTRUMENTS E is not known it can be computed from a standard with a known concentration the absorbance of which will be measured Then it will directly compute the concentration of the different samples to be analyzed _ J talon A talon The concentration of the different samples will be calculated directly 4 3 4 Kinetic It is the continuous measurement of absorbance in relation to time and curve processing see details page 103 4 3 5 Scanning It is the measurement of absorbance or transmission variation in relation to the wavelength and curve processing see chapter page 91 15 Uvi Light XTD SCHOTT INSTRUMENTS 5 CONNECTION AND STARTED To connect the spectrophotometer see installation guide REF 0M8039 Switch on the instrument The blue LED on the unit front starts flashing The user can heard the noise of the filters holder filters which cut straylight when it sets its
76. bsorbance value Do again this operation with wavelength 340 nm Results Estimate this gap and verify if the absorbance value 0 000 is included inside W L 220 nm Tolerance on the entire kinetic 0 005 Abs W L 340 nm Tolerance on the entire kinetic 0 005 Abs 134 Uvi Light XTD SCHOTT INSTRUMENTS 18 8 WAVELENGTH VERIFICATION Enter in the spectrum mode Enter the parameters as follows FT Zero F7 TocIs F3 Paraneters Es c uit Spectrum Mode v1 00 RESULTS PARAMETERS Wavelength 2 Wavelength 1 chan 320 375 PEN 100 2400 1000 5 eters locked Copyright SECOMAM Perform baseline on the air Measure the spectrum of holmium perchlorate solution ref 406817 Make an automatic reading of absorption picks Compare theoretical picks and reading picks Results Verify if the reading wavelengths of the holmium solution picks are included between the maximal and minimal theoretical picks values of this solution _ 2 Uvi Light 2 nm band pass Minimal W L nm Maximal W L nm 288 6 18 9 ABSORBANCE VERIFICATION Enter in absorbance mode Select 340 nm 135 SCHOTT INSTRUMENTS Uvi Light XTD 4 Data introduction Absorbance Mode 1 00 440 r Continuous absorbance E p Continuous transmittance Bac Enter sc V Spac
77. ctor 10000 Unit Speed Substrate unit Micha Number ing Parameters locked n SC A B L H Space 0123456789 87 By means of keys 1 and place the cursor in front of the parameter to be modified Use external keyboard or virtual keyboard with mouse to enter new value Confirm Backspace key allows to delete an error NOTE If the operator does not valid the new value with Enter key and goes to the next parameter with key the parameter will take back its previous value 9 2 2 The nature of the parameter is preset The parameter is chosen among several parameters or values proposed by the Uvi Light 36 Uvi Light SCHOTT INSTRUMENTS Kinetic mode v1 00 PARAMETERS RESULTS Wavelength Lag time 0 18000s No of segments 1 100 4 Segment time 30 13000 30 Calculation Regre Decimal place 0 3 Factor 10000 1000 Unit Speed Q L Substrate unit Micha HMOL L Number ing Unit Parameters locked Copyright SECOMAM In such case choose the parameter or the value by means of the and keys Scroll the parameters Once the selection is completed confirm 9 3 VALIDATION OF ALL THE PARAMETERS It is very important to confirm every parameter line in order to confirm all parameters If the operator press key before to reach last parameter line all parameters modifications will be cancelled 9 4 SPECIFIC CA
78. d Fl Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Abs wavelength nm Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 0 10000 unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute F2 Simulation F3 Parameters Concentration Mode v1 00 RESULTS conc A B D E F G H I Copyright SECOMAM Prepare multicell holder as indicated Start reading by pressing F1 Measure Uvi Light XTD SCHOTT INSTRUMENTS Entrer number of cell 1 8 2003 11 24 11 45 13 Concentration Mode 1 00 RESULTS Sample Conc Abs CELL Blank 0 267 CELL A Blank std 10 0 G L 0 204 CELL B Std Factor 49 1 CELL C Sam 1 CELL D Sam 2 Sam 1 20 4 G L 0 416 CELL E Sam 3 Sam 2 26 1 G L 0 532 CELL Sam 4 Sam 3 34 3 G L 0 699 CELL G Sam 5 Sam 4 40 9 G L 0 833 CELL H Sam 6 Sam 5 54 2 G L 1 104 CELL I Sam 7 Sam 6 64 8 G L 1 320 Sam 7 76 4 G L 1 556 ESC ABCDEFGHIIJIKLMNOPQRSTUVWKXY2Z Back Space 012345678 9 f Enter e 4 6 Copyright SECOMAM THE DISPLAYED ABSORBANCES OF THE STANDARD AND SAMPLES READ AS AGAINST THE BLANK THE DISPLAYED ABSORBANCE OF THE BLANK IS READ AS AGAINST AIR The results will be print out as follows SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date
79. dings spectra absorbance reading for a given wavelength control for an accessory not supplied by SCHOTT INSTRUMENTS The software is made from the editor program 34 Uvi Light XTD SCHOTT INSTRUMENTS 9 ALTERING THE PARAMETERS The procedure for altering the parameters is very simple It is the same whatever the position of the operator in the program and whatever the analysis mode or method used Select a measuring mode Ex KINETIC Confirm With the mouse or virtual keyboard enter User s name 20 characters signs letters and numbers Batch number 8 characters signs letters and numbers Samples origin 30 characters signs letters and numbers Reagent kit number 8 characters signs letters and numbers This information is useful for measurements traceability G L P ISO 9000 and will be printed with the results PARAMETERS RESULTS Wavelength nm Lag time 0 18000s No of segments 1 100 Segment time 30 13000 Calculation Decimal place 0 3 Factor Unit Speed Substrate unit Micha Number ing Parameters locked Copyright SECOMAM The parameters of the analyses can be found in a window on the left hand side of the screen For programming new parameters enter F2 Parameters Modif ication Printer F3 5ave sc Return Kinetic mode 1 00 RESULTS PARAMETERS Wavelength nm Lag time 0 18000s No of
80. e O 123456 89 X Copyright Perform zero on the air Insert F2 filter Set of secondary calibration filters ref 066349 in the cuvette holder Read the value Insert standard filter F3 then standard filter F4 Read the values Do again same operation at 465nm 546 nm 590 nm and 635 nm Results Verify that the reading absorbances of standard filters F2 F3 F4 are included in the tolerance gap expressed as follows Theoretical value of std filter Fx Tolerance lt Reading value of std filter Fx lt Theoretical value of std filter Fx Tolerance Theoretical values of standard filters are given with every standard filters kit um TL Theoretical V 0 007 Abs Theoretical V 0 010 Abs Theoretical V 0 010 Abs Theoretical V 0 007 Abs Theoretical V 0 010 Abs Theoretical V 0 010 Abs 99 Theoretical V 0 007 Abs Theoretical V 0 010 Abs Theoretical V 0 010 Abs 18 10 SUMMARY TABLE BACKGROUND NOISE VERIFICATION ON THE COMPLETE WAVELENGTH RANGE Tolerance 2 5 Abs STRAIGHT LIGHT VERIFICATION AT 340 NM Tolerance 2 5 Abs VERIFICATION OF MEASURE STABILITY W L 220 nm Toler 0 005 Abs W L 340 nm Toler 0 005 Abs WAVELENGTH VERIFICATION 136 Uvi Light XTD SCHOTT INSTRUMENTS Uvi Light Band pass5 Uvi Light Band pass 2 nm ABSORBANCE VERIFICATION FILTERS F2 FILTER F3 FILT
81. e instructions given on the screen to prepare the multicell older for another series Press F1 Measure to read the following series Press ESC to leave concentration mode 12 1 5 2 With a standard a Without a blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method 64 Uvi Light SCHOTT INSTRUMENTS Concentration Mode v1 00 PARAMETERS of measure b wavelength nm Reagent blank Sample blank s Decimal place 0 3 No of standards 0 10 0 10000 unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute F2 Simulation F3 Parameters 2003 12 01 15 23 47 Concentration Mode v1 00 RESULTS Conc CELL Reference Std bl Std Sam bl Sam bl sam sam sam 0 3 020 Copyright 5 Prepare the multicell holder as indicated Start the reading by pressing F1 Measure Entrer number of cell 1 8 2003 12 01 15 26 55 Concentration Mode 1 00 RESULTS Sample Conc CELL Std 1 Reference Std 10 0 G L Std bl Factor 46 6 Std Sam bl Sam bl 1 Sam 11 1 G L Sam bl Sam bl 2 Sam Sam 5 8 G L Sam bl Sam bl 3 Sam Sam 11 7 G L B D E F G H I WWS TUVWXYZ Back 0 _ Enter copyright SECOMAM THE DISPLAY
82. e with programs likely to receive them Select as above the decimal point or the comma Confirm Drive number Precise the type of drive A C in which the file to be converted can be found Confirm Selection i Choice Validate Esc Quit files management curves to convert in ASCII TESTO1 TESTO2 5 03 Copyright SECOMAM Select the file to be converted into ASCII Confirm and confirm When conversion is completed the software returns to the file management menu 132 Uvi Light XTD SCHOTT INSTRUMENTS 1 8 18 1 Background noise verification Straight light at 340 nm verification Measure stability verification Wavelength verification Absorbance verification 18 2 GENERALITY Verification tests are the same for Uvi Light XS Uvi Light XT Dimension Uvi Light PC 18 3 STARTUP Remove cuvette if there is one out of the cuvette holder Switch the spectrophotometer on Wait 45 minutes Warm up Press F1 Configuration Select No for the Option parameter working in monocell mode 18 4 PERIODICITY Verification tests are made once per year regularly or every time the operator wants to be sure that the instrument is working well 18 5 BACKGROUND NOISE VERIFICATION ON THE COMPLETE WAVELENGTH RANGE Enter in spectrum mode Enter parameters as follows FT Zero F7 TecIs F3 Paraneters
83. ectrophotometer gives in the same time with the linear regression curve the three following parameters Bias Absorbance of the intersection point between the regression line and the Y axis Slope Abs Slope of the regression line equal to Correlation The correlation coefficient is a number up to 1 which links the measuring points to the points of regression line The closer it is to 1 the closer the measuring points will be to the points of the theoretical line REMARK The axes of the calibration curve are graduated for absorbances and concentrations whatever the type of axis selected logarithmic 1 Y But the ordinate at origin and the gradient of the regression line are computed according to the type of axis selected d Point deletion It is possible to delete one or several calibration points maximum n 2 n being the number of standards measured which have been poorly measured Upon each deletion the software redraws the calibration curve in the previous configuration axes calculation mode but without the deleted point NOTE The point deleted on the curve will be also deleted from the parameters of the method When the user load an analysis previous stored in memory the deleted calibration points will be only for analysis in process and will not be in the methodology Example Press F4 Del point Point Number between 1 and 4 3 o o o o x m
84. elete conversion hele and listing return to the previous menu Copyright 128 Uvi Light XTD SCHOTT INSTRUMENTS Press F1 Files Directory F2 Save FS Erase F4 Curve DASCIT Ese Guitten files management Directory Listing of files Save Copy of Files Erase Delete files Curve gt ASCII Convert curve in ASCII Copyright SECOMAM GAUETAN 17 2 1 Directory This function is used for displaying the list of files from hard disk or floppy disk Press F1 Directory Selection Va idat ion Esc Escape files management Parameters to list Type of file ECTS Name of drive Copyright SECOMAM Type of file Search for the type of file to be displayed with the Suppr Del key and the and keys There are eight kinds of files refer to the beginning of chapter for the description of files Confirm Choose the drive number as above Drive number Define the type of drive in which the file s to be displayed can be found floppy disk drive C hard disk Selection Validation Esc Escape files management Parameters to list Type of file Name of drive C Copyright SECOMAM 129 SCHOTT INSTRUMENTS Uvi Light XTD When the parameters are defined confirm files management Files MAC PUO Copyright SECOMAM Example List of MACRO files in hard dis
85. ero There are nonetheless two exceptions Uvi Light XTD SCHOTT INSTRUMENTS Inthe event of a regression calculation with non aligned calibration points nthe event of a logarithmic axis for absorbances or concentrations the curve beginning at the first standard NOTE The operator may also decide not to use the zero for calculating the curve if it is not exactly aligned with the calibration points which is the case when standards have fairly high absorbances by reference to the zero and small absorbance differences between themselves The samples are read over a range between the absorbance of the zero standard 10 first standard for a logarithmic axis and the absorbance of the last standard 10 For samples not included in the above range only the absorbances are displayed and the concentration is replaced by c Change of curves parameters When the data acquisition is over the user has possibility to change the parameters He can use or not the zero point for the curve calculation He can change the X axis concentration or the Y axis absorbance He can change the type of calculation He can eventually delete a calibration point Upon each change the curve is redrawn in view of the new parameter See following pages for some examples For the explanations of different parameters LOGIT axis spline curve see at the end of this chapter page 80 Curve calculation without zero point Press F5
86. es saving Maximum curve saving capacity on hard disk 1500 spectra It is possible to increase the memory storage capacity by using the Save routine which allows to export or import curves on a diskette See page 80 Press F2 Save then F1 Curve No curve must be saved return to the previous menu Otherwise choose the curve s to be stored among the curves drawn Confirm Enter the name in 8 digits confirm Return to the previous menu b Images saving Maximum image saving capacity on hard disk 1500 images Press F2 Save then F2 Image Enter the name in 8 digits confirm NOTE It is advised to write as a comment the image file name 13 5 1 2 Load This function is used for loading one or several spectra up to 9 on the screen for various processing zoom or calculation derivatives operations peak read outs NOTE Upon the first entry in the spectra analysis mode and after loading the tools function F3 Tools the display strip shows F1 F2 Calculation Esc Quit If spectra have been saved it is possible to load one of several of them If there is no spectrum in the memory the functions Load and calculation loop back Return to the previous menu with the Esc key a Curves loading Press F1 Load F1 Curve In the spectrum library proposed the spectra saved Example DERIVATIVE DIDYME HOLMIUM and the spectra recently performed SPxx are of different colors The later will be del
87. ess F6 Constant Absorbance o o 1 1 1 60 70 80 Temps en secondes Uvi Light XTD SCHOTT INSTRUMENTS a Multiplication Allows to multiply a curve by a positive or negative number Enter multiplication Choose the curve Enter the multiplying coefficient and confirm The new curve is displayed and stored temporarily under the name KINM index b Addition Allows to add a positive or negative constant to the curve to shift it upwards or downwards for superimposing one or several curves Enter Addition Choose a curve Enter the constant confirm The new curve is displayed and stored temporarily under the name KINC index 14 6 3 7 Km calculation Michaelis constant We have Pl measurements of enzymatic reaction speed obtained by the variation of substrate concentration enzyme concentration is constant We obtain couples of values as follow SLV1 92 V2 3 V3 Sn Vn Michaelis and Menten equation V x18 K s V Reaction speed AAbs mn 5 Substrate concentration K V Maximum reaction speed When 5 00 Michaelis Menten constant inverse ratio to the affinity of enzyme for the substrate Proceed to the various kinetics acquisition for various substrate concentrations Example of 3 samples Press F3 Tools F3 Calculation then F7 Michaelis 1 1 1 1 80 90 100 110 Temps en secondes Press F1 Calcu
88. eted as soon as the spectra mode is closed Choose the spectrum or spectra to be loaded confirm and confirm b Images loading Press F1 Load F2 Image In the image library proposed choose the one to recall confirm and confirm 13 5 1 3 Delete This function allows to delete one or several curves chosen by the user Pres F3 Delete Choose the curve s to be deleted confirm and confirm The curve s will be deleted 94 Uvi Light XTD SCHOTT INSTRUMENTS 13 5 1 4 Print Press F4 Print The curve s will be printed horizontally without comments in a 11x21 cm format 13 5 1 5 Hardcopy Press F5 Hardcopy The curve s will be printed vertically with comments in a 21x29 7 cm format 13 5 2 Scales This function allows to expand an interesting section of a curve Press F2 Scales Absorbance NOTE Throughout the function the Esc key allows to return to the previous menu 13 5 3 Original This command allows to return to the initial curve after a manual or with mouse expansion Press F1 Original 13 5 3 1 Zoom with cursor Press F2 Cursor a LED Se PES EN imi selection Absorbance 1 Select the left limit of the section to be expanded by moving cursors with the and keys Confirm Similarly select the right limit of the section to be expanded Confirm Frame at t
89. explanation at the end of this chapter page 88 46 Uvi Light XTD SCHOTT INSTRUMENTS Parameters locked Allow to locked a method parameters YES or NO See explanation at the end of this chapter page 91 NOTE Three parameters X axis Y axis type of curve can be arbitrarily selected The user can change them when calibration curve is done For each parameter changing the curve will be re drawn by holding account of the new parameter 12 1 2 Impression of parameters Press F2 Printer to print parameters 12 1 3 Method saving Press F3 Save to save method parameters 12 1 4 Measure without sample blank 12 1 4 1 Measure with a factor a without blanc Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fi Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Abs wavelength Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Factor 10000 Unit Numbering Parameters locked Copyright SECOMAM File results See explanation of file results at the end of this chapter page 86 NOTE If the operator has failed to assign name to a file for storing data of a reading the software creates a file by default call CONC PRN The data stored formerly will be overlaid by the new ones u
90. ey F1 Printer Esc Quit F4 Graphic return to the curve F3 Calculation F2 Derivative 1 Order of the derivative 5 00 Differentiation step ESC Quit F1 1 O F5 Hardcopy Esc Quit Press any key to return to the main menu 124 Uvi Light XTD SCHOTT INSTRUMENTS 1 6 UTILIZATION AS A SINGLE CELL HOLDER To install this accessory refer to the Uvi Light installation guide It is possible to use the multicell holder as a single cell holder without to remove it working with a flow through cell To do that proceed as follows From main menu press F1 Config Selection Va idat ion Esc Escape SECOMAM Uvi Light v1 00 SIMULATION Configuration Language Option Printer selection Letter quality Grid 5 Deuterium lamp Stand By White amp blac Virtual keyboard Enabled LCD Contrast 01 Copyright SECOMAM Change Option parameter Choose Confirm all window parameters The spectrophotometer positions the A compartment of the multicell holder right into the beam In that configuration only position A can read the measuring cell The operator alternates the reference solution the blank the standard and the sample as requested by the spectrophotometer The type of holder used is displayed at the bottom of the screen NOTE Upon starting the unit if the No option has been selected the spectrophotometer will check if there is a mult
91. fall attempt at transformation etc or to a lack of maintenance by the user or to poor storage conditions Any breakdown due to the use of parts not supplied by SCHOTT INSTRUMENTS on SCHOTT INSTRUMENTS equipment Any breakdown due to the transporting of the equipment in packaging which is not its original packaging The lamps the cells and generally any item which appears in the accessories section on the price list Our customers are kindly asked to apply for our consent before returning any instrument for repair No return of materials may be accepted whithout the prior written consent of our Sales Management which will precise the terms of such return If the above consent is given articles shall be returned in their original packaging on a prepaid basis to the following address SCHOTT INSTRUMENTS Hattenbergstrafse 10 55122 Mainz Germany We reserve the right to reship all instruments received collect failing such consent Whatever method and conditions of transport are chosen for the shipment of the equipment to be repaired under guarantee in the original packaging the corresponding costs and the insurance costs will be payable by the customer Any damage connected to the return transport of the equipment falls within the framework of the guarantee on the express condition that the customer has sent his complaint within forty eight hours by registered letter with acknowledgement of receipt to the carrier A copy of the letter should
92. flexion must be as small as possible The area between the square of the second derivative curve and the X axis must be as small as possible After 4 or 5 iterations the coefficients converge towards the coefficients specific of the curve searched for Advantages of the spline function This curve does not necessarily run through all the measuring points therefore if a point drifts away from the theoretical curve the algorithm will not cause the curve to go through such point and the calculations will be closer to reality The very definition of the spline function which tends to bring the model of the curve closer to that of a straight line deletes all undulated curves The curve will be closer to the theoretical curves which are regular This type of function must be used in between extreme values No extrapolation is to be made Regression noted Regre To be used mainly when the theoretical curve must be a straight line It is a straight line which goes through the calibration points where the square of distances between such points and the straight line vertically has been minimized Regression passing by the origin noted Regre 0 To be used mainly when the theoretical curve must be a straight line and have to go through the origin It is a straight line calculated by the smallest rectangle mode It is a straight line which goes through the calibration points where the square of distances between such points and the straight
93. he top and the bottom the section to be expanded by moving the cursor with the and keys and validating whenever it reaches the correct position Example 95 SCHOTT INSTRUMENTS Uvi Light XTD pper imi selection 1 1 o U 9 m 2 o M 2 n Confirm Absorbance 310 Wavelength nm To retrieve the initial curve enter F2 Scales then F1 Original 13 5 3 2 Manual zoom Press F3 Manual Enter the wavelength scales Automatic zoom YES The unit recalculates the absorbance scales automatically NO Enter the absorbance scales 13 5 4 Calculation Press F3 Calculation 96 Uvi Light XTD SCHOTT INSTRUMENTS o o Absorbance o This function allows to read the absorbance on one or several spectra at a given wavelength calculate the derivatives determine the peaks or valleys on one or several spectra add two spectra substract one spectrum from another multiply a spectrum by a coefficient add a constant to a spectrum perform a ratio calculation between two absorbances at different wavelengths 13 5 4 1 Absorbance Allows to read an absorbance in view of wavelength on one or several spectra Press F1 Abs i Abs 2 Derivat 3 Max perati ion 6 Constant 7
94. heir name and the dilution factor See explanation at the end of this chapter page 88 Parameters locked Allow to locked a method parameters YES or NO See explanation at the end of this chapter page 73 NOTE Three parameters X axis Y axis type of curve can be arbitrarily selected The user can change them when calibration curve is done For each parameter changing the curve will be re drawn by holding account of the new parameter 12 2 2 Impression of parameters Press F2 Printer to print parameters 12 2 3 Method saving Press F3 Save to save method parameters 12 2 4 Measurement During measurement on the top of the screen will be displayed the count down of the total measuring time The procedure for these measurements is the same as for those performed in the concentration mode The difference will lie in the way of interpreting the readings and the results 12 2 5 The readings The delta of absorbance will be measured by reference to air or by reference to a delta of the blank 12 2 6 The results They are expressed as follows Sam Conc AAbs mn Blank actor If there is a standard the factor will be calculated as follows Std Conc Factor AAbs mn 514 AAbs mn _ Blank Uvi Light XTD SCHOTT INSTRUMENTS The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s
95. hoose between 190 and 900 nm Wavelength 1 Initial wavelength of the spectrum Choose between 190 and 900 nm Wavelength change 320 375 To prevent any switching from the deuterium lamp to the halogen and vice versa on an interesting peak of the spectrum the operator may shift the switching point Choose any value between 320 and 375 nm Speed 100 2400 This is the average speed of wavelengths at small absorbances expressed in nanometers per minute Continuous variation between 100 and 2400 nm mn The step is 1 nm N B 9 2 The smaller the speed the greater the number of elementary measurements and since the background noise is inversely proportional to the square root of the number of measurements it will be even smaller Type of axis The operator may choose between Absorbance and Transmittance for the Y axis Parameters locked Allow to lock the analysis parameters See page 90 At the end of the modification of parameters confirm 13 2 PRINTING THE PARAMETERS Press F2 Printer for printing out the parameters 13 3 SAVING THE METHOD Press F3 Save to save parameters 13 4 MEASUREMENT Prepare the multi cell holder as follows Position A Blank reagent or reference Position B through l Samples Through the measuring sequence the multi cell holder will be prepared in the same way 02 Uvi Light SCHOTT INSTRUMENTS Spectrum Mode v1 00 PARAMETERS RESULTS Wavelength 2 nm 400
96. icell holder installed and will move the A position right into optical beam 125 Uvi Light XTD SCHOTT INSTRUMENTS 1 7 FILES 17 1 FILES DESCRIPTION 17 1 1 Program files They incorporate the measuring programs created by the operator They may be run directly from the main menu These methods can be copied on floppy disk and be filed for ulterior use The parameters of a program can be retrieved from the DOS by entering Name of the program PLN Ex PROT PLN In the main menu programs will appear as follows First left column Name of the method assigned by the operator Second column Name of the file assigned by the operator Example Proteins measurement PROT 17 1 2 Editor files The SPECTRO PASCAL software is a language derived from the PASCAL already developed by SCHOTT INSTRUMENTS It consists of macro instruction commands such as Init window Measure Absorbance Instructions in SPECTRO PASCAL can be found in each of the editor programs To use such programs they must be compiled beforehand This is why there may appear a number of names of editor programs which the operator cannot find in the main menu They have not been compiled These programs can be copied on floppy disk and be filed for ulterior use These programs may be retrieved from the DOS by entering Name of the program TEX Ex MEASURE TEX In the menu of editor programs they will appear under the name assigned to them by the operator or b
97. ich allow to export or import curves on a diskette See explanations page 80 Press F2 Save then F1 Curve No curve must be saved return to the previous menu Otherwise choose the curve s to be stored among the curves drawn Confirm Enter the name in 8 letters confirm Return to the previous menu b Images saving Maximum image saving capacity on hard disk 1500 images Press F2 Save then F2 Image Enter the name in 8 letters confirm NOTE It is advised to write as a comment the image file name 14 6 1 2 Load This function is used for loading one or several curves up to 9 on the screen for various processings zoom or calculations derivatives operations slopes NOTE Upon the first entry in the kinetic analysis mode and after loading the tool function the display strip shows Load Calculation and ESC If kinetic curves have been saved it is possible to load one or several of them If there is no kinetic curve in the memory the functions Load and Calculation loop back Return to the previous menu with the ESC key a Curves loading Press F1 Load then F1 Curve In the kinetic curves library proposed the spectra saved 55 01 5 03 and the kinetic curve recently performed KINxx are of different colors The later will be deleted as soon as kinetic mode is closed Choose the curve s to be loaded confirm and confirm B Images loading Press F1 Load then F2 Image In the images
98. is valid according to the associated contract until April 27 2006 Adjoins Toi la qualit T l 433 08 TH R3 OO oo ASCI QUALITATEM SARI fr CAP VAISE 14 rue Gonge de Loap Fo YON a Pape 1 1 D 141
99. k C 17 2 2 Save This function allows to copy files from one drive to another It is especially useful for saving curves or programs or for loading curves or programs into Uvi light improved capabilities of spectrophotometer spectra libraries software updating Press F2 Save Selection 4 Validation Esc Escape files management Parameters Type of file Name of source drive Name of object drive Copyright SECOMAM Type of file Select the types of file to be copied by means of the lt and keys Confirm Number of source drive Select by means of the and keys the type of drive A C which includes the file to be copied Number of object drive Select as above the type of drive A C into which the selected file will be copied When the parameters have been defined confirm 130 Uvi Light SCHOTT INSTRUMENTS files management Files to copy TESTO1 TESTO2 TESTOS3 Copyright SECOMAM Select the files to be copied Confirm and confirm When the copy is completed the software returns to the file management menu 17 2 3 Erase This function allows to delete files spectra kinetic ASCII editor macro or program files ALWAYS CHECK IF THE FILE TO BE DELETED IS THE RIGHT ONE Enter F3 Erase Selection Ya idat ion Esc Escape files management Parameters to delete Type of file scannin Name of drive
100. l choose the first kinetic curve then confirm and confirm Enter the corresponding substrate concentration Proceed in the same way for the other kinetic curves After the selection it will display Woolf Hanes form the Michaelis and Menten equation curve It will print the value of Km as follows 117 SCHOTT INSTRUMENTS Uvi Light XTD SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method KINETIC Mode v1 00 sam 1 Time 11 43 am 20 uMOL L Speed 494 Slope 0 494 Abs mn Time 5sec Abs 0 546 Slope 0 671 Abs mn Time 15sec Abs 0 658 Slope 0 517 Abs mn Time 25 Abs 0 744 Slope 0 429 Abs mn Time 35sec Abs 0 815 Slope 0 387 Abs mn Michaelis constante Km 5 uMOL L a Woolf Hanes form Press F2 Woolf Hanes IS K 1 0 onYaxis S V On X axis 5 Slope RS Hoo If Hanes Lineweaver Bur Michaelis Menten F5 Eadie S Vitesse 10 15 WOOLF HANES 51 value is displayed the bottom left side of the screen b Linewearver Burk form Press F3 Linewearver Burk 118 Uvi Light XTD SCHOTT INSTRUMENTS 1 K 1 1 V V S V On Y axis tee K p Vn Slope Bias c Michaelis Menten form Press F
101. l place 0 3 of standards 0 10 Concentration 0 10000 Unit No of W L 2 10 wavelength 1 nm GENERAL CALCULATION wavelength 2 nm Coeff comma 0 3 1 51 2 52 5 10000 R 10000 BO B1 AbS1 B2 Abs2 Bn Absn 10000 10000 Copyright SECOMAM The general calculation formula in the multiple wavelength mode is displayed at the bottom left of the screen Type of measure Choose MultiWI Reagent blank YES or NO See meaning of the blank in this chapter page 6080 Decimal place from 0 to 3 This parameter defines the number of significant digits after the decimal point Number of standards Factor To work with a factor answer 0 Choose a value between 10000 1 standard To work with 1 standard answer 1 Choose a value between 0 10000 Several standards To work with several standards enter the number of standards from 2 through 10 Choose a value between 0 and 10000 Unit With keyboard enter the abbreviation wanted X axis Concentration axis Choose between linear and logarithmic Y axis Absorbance axis Linear logarithmic 1 Abs 1 Y or Logit See Logit axis meaning at the end of this chapter page 80 Type of curve Linear segmented curve linear regression Regre 0 linear regression going through the origin 0 Spline See ex
102. ll save the method of analysis Fl Execute F2 Parameters F3 File results Esc Quit Concentration Mode v1 00 PARAMETERS of measure Abs i No of standards 0 10 Conc 1 0 10000 conc 2 0 10000 conc 3 0 10000 Conc 4 0 10000 Unit X axis Concentration Y axis Absorbance Type of curve Numbering Parameters locked Copyright SECOMAM Press F1 Execute F2 Simulation F3 Parameters Concentration Mode v1 00 RESULTS Conc CELL Reference Std 61 Std Std Std Std Std Std Std Copyright SECOMAM Prepare the multicell holder as indicated Start the reading by pressing F1 Measure 68 Uvi Light XTD SCHOTT INSTRUMENTS axis Del point SEE 3E S Next E Absorbance blank 15 20 Concentration DETAILS ON THE CURVE The values displayed on the absorbance axis are the values of the standards or samples calculated as follows Displayed Abs Abs Std or Sam Bl Std or Sam Values read as against the air Refer in this chapter to the page 81 for the processing of the curve How to change the curve axis point deletion Press F8 when the calibration curve is over Enter the number of samples to be measured Confirm Entrer number of cell 1 8 2003 12 01 17 04 28 Concentration Mode v1 00 RESU
103. m bl Sam 3 Sam bl Sam 4 gt Copyright 5 Prepare the multicell holder as indicated Start the reading by pressing F1 Measure 63 SCHOTT INSTRUMENTS Uvi Light XTD Entrer number of cell 1 8 2003 12 01 12 24 04 Concentration Mode 1 RESULTS Sample conc CELL Blank b1 Reference Blank Sam bl Sam bl 1 01 01 bl bl bl bl 0921 RSTUVWXY2Z _ Fo Copyright SECOMAM THE DISPLAYED ABSORBANCES OF THE SAMPLES SAMPLE BLANKS AND BLANK OF THE BLANK ARE READ AS AGAINST THE REFERENCE SOLUTION The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Blank bl Abs 0 185 Time 11 43 am Blank Abs 0 320 Sam bl 1 Abs 0 161 Time 11 45 am Sam 1 Conc Abs 0 355 Sam bl 2 Abs 0 594 Sam 2 Conc Abs 0 776 Sam bl 3 Abs 0 866 Sam 3 Conc Abs 1 102 Sam bl 4 Abs 1 270 Sam 4 Conc Abs 1 546 Sam bl 5 Abs 0 161 Time 11 47 am Sam 5 Conc Abs 0 355 Sam bl 6 Abs 0 594 Sam 6 Conc Abs 0 776 The concentration value of the sample is computed as follows Sam Conc Sam Bl Blank BI Blank F actor Values read as against air Follow th
104. measure Wave length of standards 8 185 0 1 Bin 09 10000 gt 8 1 gt 0 1 gt axis Concentration T axis CAhsorhance gt Type of curve Copyright SECOMAM Insert the Standard 1 cuvette in place Press F1 Measure After measuring remove the Standard 1 cuvette and then Insert the Standard 2 cuvette in place Press F1 Measure After measuring remove the Standard 2 cuvette and then Continue this way till the last of the Standard was read 0 point 85 0 ope orrelation 1 Absorbance blank 5 10 15 20 25 35 Concentration Modify the axis with F1 X axis and F2 Y axis Modify the type of curve calculation with F3 Type Delete the points with F6 Del Point Print the curve with F7 Print Press F8 Next to go to measuring the samples F2 Sinulation 1316 16 34 35 easure of Concentration Mode vi1 Hi PARAMETERS RESULTS af measure Conc 18 8 G L 246 8 G L 30 0 G L 40 0 G L Wave length Cnm 0 3 gt f standards amp H 1H 1 H 1 ninm 1 3 H 1 niin 4 diam axis Concentration Y axis Absorbance Type of curve Copyright SRT Insert Sample 1 place
105. ment AAbs x Coef x 60 104 Uvi Light XTD SCHOTT INSTRUMENTS AAbs Absorbance at the beginning of segment minus absorbance at the end of the segment 2 Sum of absorbances delta of the segments N Number of segments Coef Coefficient of molecular absorption Measuring time 60 2 Ratio reducing the result to one minute The slope thus calculated is expressed in AAbs mn The result is report to one minute Decimal places 0 3 This parameter affects the resolution of the results Choose from O to 3 Factor 10000 The factor can be negative It is used for calculating the concentration The result is obtained as follows Speed AAbs mn x Factor Unit speed reaction Enter the abbreviation of the unit wanted i e n uKatl Unit substrate concentration Enter the abbreviation of the unit wanted i e g l mg l ug l g dl ug dl mMOL I uMOL I Numbering Auto The results will be in the form Sampl 1 Sampl 2 Unit Allows to assign a name to the sample Locked parameters Allow to lock the analysis parameters YES or NO See page 90 14 2 PRINTING THE PARAMETERS Press F2 Printer for printing out the parameters 14 3 SAVING THE METHOD Press F3 Save to save the parameters 105 SCHOTT INSTRUMENTS Uvi Light XTD PARAMETERS RESULTS Wavelength Lag time 0 180005 No of segments 1 100 Segment time 30 13
106. near Copyright SECOMAM Press F3 Save PARAMETERS of measure b 0 10000 X axis Concentration Y axis Absorbance Type of Concentration Mode v1 00 of measure Abs Wavelength nm Reagent blank Sample blank pecina place 0 3 of standards 0 10 1 0 10000 Cane 0 100000 0 100000 0 100000 axis Concentration axis ance Type of curve SCHOTT INSTRUMENTS Uvi Light XTD Enter the full name of the method up to 30 characters This information will appear in the top side of the window in place of CALCULATION MODE Confirm Execute Parameters File results tsc Quit CONCENTRATION WITH 4 STANDARDS v1 00 of measure Ab Wavelength Reagent blan Sample blank Decimal place 0 3 No of standards 0 10 1 0 100000 0 10000 X axis Concentration Y axis Absorbance Type of curve Copyright SECOMAM EMIJUN ME SECOMAM Uvi Light v1 00 SIMULATION Program choices 2 Gent cation Mode 04 11 2003 L CONCENTRATION WITH 4 STANDARDS CONCENTR 10 12 2003 Spectrum Mode i 04 11 2003 Copyright SECOMAM EMIJUN Me If the user does not want to save the analysis he will enter ESC in place of F3 Save CONCENTRATION WITH 4 STANDARDS v1 00 of measure Abs Sample blank Decimal
107. nk Abs 0 020 Time 02 22 pm Conc 293 G L Abs 0 430 Conc 24 0G L Abs 0 358 25 7 G L Abs 0 381 27 3 G L Abs 0 403 Conc 25 2G L Abs 0 375 Conc 23 7 G L Abs 0 355 Sam Conc 33 8 G L Abs 0 492 Sam 22 11 G L Abs 0 332 Blanc Abs 0 020 Time 02 22 pm Sam 9 Conc 27 3 G L Abs 0 403 Sam 10 33 8 G L Abs 0 492 Sam Sam Sam Sam Sam Sam Enter number of samples to be measured Confirm Follow indications displayed to prepare a new multicell holder for another series of measurement Press F1 Measure to read the following series Press ESC to leave the measuring mode 12 1 5 Measurement with a sample blank 60 Uvi Light XTD SCHOTT INSTRUMENTS 12 1 5 1 With a factor a Without blank Load the method or select Concentration Mode Enter user s name batch number samples origin and reagent kit number Modify or programme parameters Save or not the method Fl Execute F2 Parameters rF3 File results Esc Quit 2003 12 01 11 51 47 Concentration Mode v1 00 PARAMETERS of measure wavelength nm Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Factor 10000 Unit Numbering Parameters locked Copyright SECOMAM Press F1 Execute Enter the number of samples to be measured Confirm F2 Simulation F3 Parameters 2003 12 01 11 53 26 Concentration Mode
108. ntain one or several curves with comments or not These files can be copied on floppy disk and be filed for ulterior use These files may be retrieved from the DOS by entering Name of the file IMC IMAGE2 IMC In the menu of image files they appear under the name assigned to them by the operator Ex IMAGE 17 1 7 ASCII files They include in the ASCII format the results of readings if the operator has caused them to be saved They also include in the ASCII format the spectra or kinetic curves which have been converted by sub routine F3 Tools F1 Files and F4 Curve ASCII Such data may be copied on floppy disks and operated by other software Excel These files may be retrieved from the DOS by entering Name of the file PRN Ex RESULTS PRN In the menu of the ASCII file they appear under the name assigned to them by the operator Ex RESULTS 17 1 8 Macro files It includes in an encoded form the sequence of commands which make up the macro instructions These files can be copied on floppy disk and be filed for ulterior use These files may be retrieved from the DOS by entering Name of the file MAC In the menu of the Macrofile they appear under the name of the key used for storing them Ex ALTF1 MAC In the menu of the macro file they appear under the name assigned to them by the operator Ex AItF 1 17 2 FILES SUB ROUTINE From the main menu press F3 Tools Ut ilitarv d
109. ntrast Step from 01 to 63 Explanations of parameters Line 1 Choice of the working language Line 2 Choice of accessories used No accessory connected External peristaltic pump or Multicell holder 29 SCHOTT INSTRUMENTS Uvi Light XTD NOTE If the multicell holder is installed and the corresponding option is not selected Option No the spectrophotometer will detect it and the user will be able to position each cuvette in front of the beam with the keys SHIFT F1 to F10 F1 A Position F9 Position 10 Zero Position WARNING These commands are available only if an external alphanumeric keyboard is connected Line 3 Choice between the following printer standard IBM EPSON or HP PCL It is also possible to work without printer connected So select for no printer connected Line 4 Choice of impression quality Line 5 Possibility of a gradation grid on the curves Line 6 Continuous or standby operation of the deuterium lamp see below Line 7 Allow to select three different contrast of the LCD display Line 8 When no external keyboard is connected to the spectrophotometer it is possible to make a virtual alphanumeric keyboard appears at the bottom of the screen to help the user to program Line 9 Gradual modification of LCD display contrast 01 LCD display black 63 LCD display white Once all parameters have been entered confirm VIRTUAL KEYBOARD Once virtual keyboard has been selected it aut
110. nts press F1 I O and F5 Hardcopy 14 6 5 Windows The command F5 Windows allows in the graphic mode to return to the parameters The opposite operation is performed by entering F4 Graphic 120 Uvi Light XTD SCHOTT INSTRUMENTS 1 5 MACRO COMMANDS WARNING These commands are available only if an external alphanumeric keyboard is connected The software allows for the creation of macro commands by the operator Such macro commands record a combination of keys The software will reproduce automatically such combinations in the order in which the operator has recorded them Macro commands help to save time when a long or repetitive task is to be performed It is for example possible in the spectral analysis program after setting the baseline to measure the spectrum of a sample to search automatically for peaks to draw the derivative and to print values plus curves It is easy to see that if operations must be performed on some twenty samples a macro command reproducing such sequence of operations will be welcome It is also possible to repeat such macro commands a number of times and to set parameters for the interval in between two repetitions 15 1 CREATION OF A MACRO COMMAND For creating a macro command in a user program Enter the analysis mode concerned Go straight to the point in the program where the macro command begins Press keys Ctrl and F1 It will display at the left top of the screen the follo
111. o repeat the same macro command at a specific interval For repeating a macro command on a rated basis Enter the mode of analysis concerned In our example SPECTRUM mode Go to the point where the macro command begins Press the Ctrl and F3 keys It will display at the left top of the screen the following message Hit a macro key In our example Alt F1 It will display at the left top of the screen the following message Repeat Enter the number of times from 2 to 1000 the macro command must be run Confirm It will display at the left top of the screen the following message time between to macros in sec This time corresponds to the interval between two beginnings of macro command Enter a time between 0 and 30 000 seconds Confirm The sequence is run as follows Execution of the macro command Discounting of the time remaining after executing the macro command New launching of the macro command NOTE If the execution time of the macro command is superior to the preset interval the software will repeat the macro command NOTE To escape out the macro command press Esc 15 4 RECALL OF APPLICATION AND LISTING OF A MACRO COMMAND 15 4 1 Recall of application From the main menu enter F3 Tools 122 Uvi Light XTD SCHOTT INSTRUMENTS FI Files FZ Raeros Esc Eit Utilitarv delete conversion hele and listing return to the previous menu Copyright
112. of segments 1 100 2 0 051 Abs mn Segment time 30 13000 j 5 sec Calculation 0 223 Slope 0 068 Abs mn Decimal place 0 3 35 sec Factor 10000 Abs 0 189 Slope 0 026 Abs mn ni Speed Substrate unit Micha Numbering Parameters locked 0 002 Abs mn lt 0 145 Abs mn Copyright SECOMAM IMPORTANT To avoid loosing curve process due power failure or an accidental exit from the kinetic program ANTHELIE Data saves it automatically every minute and stores it under the name LAST index 14 6 TOOLS Press F3 Tools Absorbance 1 1 1 20 Temps en secondes This sub routine is used for various processings zoom derivative or calculation on the curves stored in memory 14 6 1 Input Output This function allows to load save delete and print one or several curves Press F1 I O Absorbance 1 1 1 20 Temps en secondes 108 Uvi Light XTD SCHOTT INSTRUMENTS 14 6 1 1 Save The curves may be saved one by one The curves may also be saved one or several together with some comments so we speak about image NOTE THE IMAGES ARE DIFFERENT FROM CURVES AND IT IS NOT POSSIBLE TO SUPERPOSE BETWEEN THEM OR WITH CURVES a Curves saving Maximum curve saving capacity on hard disk 1500 kinetic curves is possible to increase the memory storage capacity by using the save routine wh
113. omatically appears with each time the user must enter alphanumeric characters The keyboard takes account of capital alphabet letters only numbers signs function keys as Esc Space Back Enter 4 Introduction of datas User s name Batch number Origin Kit number The choice of the different characters of this keyboard is made with the help of the mouse If the external keyboard and the mouse are not connected the choice is made with the help of 1 keys and the character validation including function keys with F10 key SWITCHING OFF THE DEUTERIUM LAMP UV When the Standby mode is chosen by the operator the deuterium lamp switches off after 30 min only if the user stays in the main menu If the user enters in a method of analysis before the deuterium lamp turning off this one will be continually on Upon a solicitation measure between 190 and 330nm in an analysis mode for a punctual reading the deuterium lamp switches on the message Waiting for deuterium light is displayed for 30 secondes and the operator may perform the measure The deuterium lamp will turn off again after 30 min upon returning to the main menu 30 7 Uvi Light XTD SCHOTT INSTRUMENTS 7 PRINCIPLE FOR MENUS AND COMMAND OF UVI LIGHT The overall structure of the program is of the tree type each branch having ramifications and so forth To move through the structure the program proposes at each step a menu on
114. on and are multiplied by the factor 12 4 DEFINITIONS 12 4 1 Meaning of the blank and calculation In a number of environments in particular biological ones there may be spectral interferences with wavelength of the dosage This is the case of colored molecules haemoglobin bilirubin pigments such as carotene medicines such as methylene blue Such interferences which are independent of the parameter to be measured will be eliminated by measuring the sample alone or with reagent which does not cause any coloring It is the sample blank When there is only one reagent and if the color of the reagent may interfere with the measuring wavelength it is deducted from the measurement and it is called a reagent blank When there are two reagents the absorbance of the reagent present in large quantity must be deducted from the blank mixture of two reagents it is the blank of blank NOTE the terminology of the blank is also used for designating the reference solution water a component of the reactional mixture which serves for resetting the measurement Example RI Non triggering reagent R2 Triggering reagent Water xxx Quantit d eau distill e quivalente au produit xxx Bl Blank Abs of the blank of blank Water Sam 2 Blank Abs of the reagent blank x RI R2 T Water Sam Std Bl Abs of the standard blank s RI Std Water R2 Std Abs of the standard s 1 2 Std
115. onfirm Concentration Mode v1 22 RESULTS Sample Conc Std 10 0 4 1 Factor 49 1 ASSAY 1 a 5 b Space 0 123456789 The operator assigns 8characters and if need be dilution factor to sample 12 5 6 3 Identification using a table Select Table for the Numbering parameter Enter or not the charge factor see precedent paragraph The user prepares a table with measuring samples number their name and eventually a dilution factor Press F1 Execute 98 Uvi Light XTD SCHOTT INSTRUMENTS I 1 3 4 5 6 5 8 9 1 Copyright SECOMAM Dimension the table by entering F7 Sample Nb Enter the number of samples from 1 to 120 Place the cursor on the sample to be identified by means of the direction arrows Enter F5 Name and then the 8 characters name Confirm Enter F6 Dilution and then the dilution factor from 1 to 999 The dilution factors are initially 1 Example ete EM IM EM i Hi COCOCOCO T1 20 2U2 X PS Ih COCOCOCoCcoCcoCcC O IDUTIBROWNR 1 Copyright SECOMAM Press F8 Print The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time
116. or downwards for superimosing one or several curves Enter Addition Choose a curve Enter the constant confirm The new curve is displayed and stored temporarily under the name SC index 13 5 4 7 Ratio Without going through the multiple wavelength mode it is possible to perform ratio calculations directly on the spectrum or spectra displayed Press F7 Ratio m 2 m 9 2 Wavelength nm a With cursor Press F1 Cursor Move the first cursor with the direction arrows Confirm The monitor displays the first wavelength W1 Move the second cursor Confirm The monitor displays the second wavelength W2 and the calculation formula R A1 A2 A1 Absorbance at W1 A2 Absorbance at W2 R Ratio The software then gives the value of the ratio for each curve displayed i Move cursor Our t b Manual Press F2 Manual Enter both wavelengths The results will be displayed as above For both modes with cursor and manual Press F3 Printer for printing the results 101 SCHOTT INSTRUMENTS Uvi Light XTD Press F4 Hardcopy for printing the results and the curve s NOTE For any other calculation to be performed on the absorbances of samples at different wavelengths refer to the multiple wavelength mode in page 75 13 5 5 Note The command F
117. ose performed in the concentration mode The difference will lie in the way of interpreting the readings and the results 12 3 7 The readings The absorbances will be measured at different wavelengths as against the blank solution 12 3 8 The results They are the results of calculation ratio sum made on absorbances read at different wavelengths With the results are displayed the absorbance and transmittance values of the samples The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Sam 1 Conc 0 061 Res 0 061 Heure 14h 59 W L 266nm Abs 1 0 240 T 57 5 W L 270nm Abs 1 0 320 T 47 9 W L 274nm Abs 1 0 278 T 53 7 2 Conc 0 110 Res 0 110 W L 266nm Abs 1 0 520 T 30 2 W L 270nm Abs 1 0 602 T 25 0 W L 274nm Abs 1 0 499 T 2 31 7 SCHOTT INSTRUMENTS Uvi Light XTD 12 3 9 The curve The results of calculation are displayed on the Y axis IMPORTANT For measures with several standards creation or simulation of calibration curves measure or simulation of absorbance saving or not of methods or calibration curves recall of analyses with stored curves refer to the paragraph CONCENTRATION Simulation The values entered by simulation are considered as the result of the calculati
118. ping Choice 4 and gt Validate ENT Abort ESC reboot of the machine will be required Chose the type of keyboard with right and left arrow keys Choisir le type de clavier avec les fl ches droite et gauche Confirm with ENT key Cancel the configuration with ESC key 27 Uvi Light XTD SCHOTT INSTRUMENTS 6 CONFIGURATION The whole Uvi Light can be configured when the main menu is displayed Creer File SECOMAM Uvi Light v1 00 SIMULATION Program choices m Absorbance Mode v1 00 04 11 2003 Kinetic mode v1 00 04 11 2003 Concentration Mode v1 00 04 11 2003 Spectrum Mode v1 00 04 11 2003 Copyright SECOMAM Press F1 Configuration Selection lt Validation SECOMAM Uvi Light v1 H2 STIMULATION H64 03716 15 44 15 Conf igyuration Language Option Hulticell Printer selection Letter quality Grid Yes Deuterium lamp mode Always CRT Black White Virtual keyboard Disahled LCD Contrast H1 Copyright SECOMAM See page 29 for changing a parameter The various choices for the different parameters are as follows 1 Language French English 2 Option No Pump Multicell 3 Printer selection No Epson IBM HP B W 4 Letter quality Yes No 5 Grid Yes No 6 Deuterium lamp Always On Stand By 7 White amp black Color1 Color2 8 Virtual keyboard Enable Disable 9 LCD Co
119. planation on the type of curve at the end of this chapter page 81 Number of wavelengths From 2 to 10 Uvi Light XTD SCHOTT INSTRUMENTS Wavelength 190 900 nm Coeff Comma This parameter defines the number of significant digits after the decimal point for coefficients AO A1 BO B1 0 10 Choose a value between 10000 BO B10 Choose a value between 10000 Numbering This parameter concerns samples identification Auto Identification of the sample is numerical and increasing Example Sam 1 Sam 2 Unit The user assigns a name to each sample 8 characters and eventually attributes a dilution factor See explanation at the end of this chapter page 88 Table The user prepares a table in which he will enter the number of samples their name and the dilution factor See explanation at the end of this chapter page 88 Parameters locked Allow to locked a method parameters YES or NO See explanation at the end of this chapter page 90 NOTE Three parameters X axis Y axis type of curve can be arbitrarily selected The user can change them when calibration curve is done For each parameter changing the curve will be re drawn by holding account of the new parameter 12 3 4 Impression of parameters Press F2 Printer to print parameters 12 3 5 Method saving Press F3 Save to save method parameters 12 3 6 Measurement The procedure for these measurements is the same as for th
120. pon each reading Press F1 Execute Enter the number of samples to be measured Confirm F2 simulation F3 Parameters Concentration Mode v1 00 RESULTS conc CELL Reference sam sam sam sam sam sam sam sam gt Copyright SECOMAM Simulation This routine allows to enter by keyboard absorbances of blank standards and samples 47 SCHOTT INSTRUMENTS For explanation see at the end of this chapter page 81 Prepare the multicell holder as indicated above Start measurement by pressing F1 Measure Entrer number of cell 1 8 gt CELL Reference sam sam sam sam sam sam sam Uvi Light XTD 2003 11 21 10 32 41 Concentration Mode v1 00 sample sam sam sam sam sam sam sam sam RESULTS conc MG L MG L MG L MG L MG L MG L MG L MG L 4 U 2 4 RSTUVWXY2Z _ Eoi 42 Copyright SECOMAM Back Enter ABSORBANCES DISPLAYED OF THE SAMPLES ARE READ AS AGAINST REFERENCE The printing will be SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 10 32 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Sam 1 Sam 2 lt Sam 3 4 5 Sam 6
121. re 59 SCHOTT INSTRUMENTS Uvi Light XTD Entrer number of cell 1 8 2003 12 01 11 02 52 Concentration Mode v1 00 RESULTS Sample conc Abs CELL Std 1 10 0 4 1 0 180 CELL A Blank S Ed 2 20 0 09 7 0 380 CELL B Sam T std 3 30 0 4 1 0 580 CELL C Sam 2 Std 4 40 0 g 1 0 780 CELL D Sam 3 Blank 0 270 CELL E Sam 4 Sam 1 9 2 4 1 0 173 CELL F Sam 5 Sam 2 19 9 4 1 0 381 CELL G Sam 6 Sam 3 31 5 4 1 0 610 CELL Sam 7 sam 4 38 1 4 1 0 739 CELL I Sam 8 Sam 5 50 9 g 1 0 989 Sam 6 52 4 9 1 1 019 Sam 7 61 2 0 1 1 192 8 1 514 ESC ABCDEFGHI 3 RS TUVWXYZ Back Space 0123456789 EN UP JAEN Enter Copyright SECOMAM THE DISPLAYED ABSORBANCES OF THE STANDARDS AND THE SAMPLES ARE READ AS AGAINST THE REFERENCE SOLUTION without blank THE DISPLAYED ABSORBANCES OF THE STANDARDS AND THE SAMPLES ARE READ AS AGAINST THE BLANK with a blank THE DISPLAYED ABSORBANCE OF THE BLANK IS READ AS AGAINST AIR The results will be printed out SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 02 21 pm User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Blank Abs 0 020 Time 02 21 pm Std 1 Conc 10 0G L Abs 0 180 Time 02 21 pm Std 2 Conc 20 0G L Abs 0 380 Std 3 Conc 30 0G L Abs 0 580 Std 4 Conc 40 0G L Abs 0 780 Bias 0 008 Slope 0 020 Correlation 1 000 Bla
122. rivative the higher the order the greater the reduction Press F2 Derivative Choose the spectrum to be derived Choose the order of the derivative and the differentiative step The step expressed in nm corresponds to the width of the smoothing window Example Spectrum SP1 Order 1 Step 5 nm Absorbance The derived curve is saved temporarily under the name SD index and may be retrieved for superposition with the initial spectrum Absorbance Wavelength Cnm 13 5 4 3 Maxi This function allows to read automatically the absorbance value of the peaks or valleys of the spectrum spectra loaded on the monitor Press F3 maxi a Peaks Press F1 Peaks Choose the gap Example 0 1A 99 SCHOTT INSTRUMENTS Uvi Light XTD Gap Absorbance value which will define a peak detection threshold Only the peaks the height of which difference between the top and nearest valley is higher than the absorbance value gap will be displayed and printed Fl Printer SCANNING MODE v1 00 PARAMETERS RESULTS Wavelength 2 nm 400 Peaks of 5 wavelength 1 nm 190 325 7 Abs 0 693 wavelen chan 320 375 330 100 2400 1800 Absorb Parameters locked Copyright SECOMAM It is possible to print the results by entering F1 Printer Press ESC twice and enter F4 Graphic to return to the curve b Valleys Pre
123. rs then F2 Printer The parameters are printed as follows SCHOTT INSTRUMENTS UV VIS Spectrophotometer Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Kinetic Mode v1 00 Parameters Wavelength nm Lag time 0 18000s No of segments 1 100 Segment time 30 13000s Calculation Decimal place 0 3 Factor 10000 Unit Speed Substrate unit Micka Numbering Parameters locked 9 6 SAVING THE PARAMETERS For saving parameters proceed as follows Press F2 Parameters F3 Save Give a name 8 characters to the method Confirm Give a title 30 characters to the method Confirm 38 Uvi Light XTD SCHOTT INSTRUMENTS 1 0 HOW USE WITH SINGLE CELL HOLDER PLEASE REMEMBER To use Uvi Light with single cell holder is simpler than with multiple cell holder that is why the description of measuring mode in this manual is given using multiple cell holder However the arborescence of the software is the same in both cases The difference lies in the sequence of measure solution blank standard samples When using multi cell holder user placed the solutions in the cell holder according to the instructions given by the software The solutions will be measured in one series per group of 8 When using single cell holder user placed the solutions in the cell holder one after another successively
124. segments 1 100 4 Segment time 30 13000 30 Calculation Regre Decimal place 0 3 Factor 10000 1000 Unit Speed Q L Substrate unit Micha MMOL L Numbering Auto Parameters locked Copyright SECOMAM Print the parameters by entering F2 Printer For changing one or several parameters enter F1 Modification Press F3 Save to save parameters Press F1 Modification 35 SCHOTT INSTRUMENTS Uvi Light XTD 4 ntroduction of datas Kinetic mode v1 00 PARAMETERS RESULTS Wavelength nm Lag time 0 180005 No of segments 1 100 Segment time 30 13000 Calculation Decimal place 0 3 Factor 10000 ni Speed Substrate unit Micha Number ing Parameters locked sc NOPR Space 0123456789 8 7 Copyrig 9 1 SELECTION OF A PARAMETER cursor blinks on the first line Just move the cursor within the window by means of keys and and place it in front of the parameter to be modified If there are too many parameters for the window they will be displayed by keeping the key down when the cursor is at the bottom of the window 9 2 MODIFICATION OF A PARAMETER There are two alternatives 9 2 1 The parameters is entering by the operator 4 ntroduction of datas Kinetic mode v1 00 PARAMETERS RESULTS Wavelength nm Lag time 0 180005 5 No of segments 1 100 Segment time 30 13000 Calculation Decimal place 0 3 Fa
125. ss F2 Valleys Choose the gap Example 0 2A Gap absorbance value which define a peak detection threshold Only the valleys the depth of which difference in terms of absorbance between the bottom and nearest peak will be printed and displayed It is possible to print the results by entering P Printer Press ESC twice and enter G Graphic to return to the curve is possible to print the results by entering F1 Printer Press ESC twice and enter F4 Graphic to return to the curve 13 5 4 4 Addition Allows to add two curves Enter Operation Choose the first curve confirm Choose the second curve confirm The new curve is displayed and temporarily stored under the name SA index 13 5 4 5 Substraction Allows to substract one curve from another Enter Operation Choose the first curve confirm Choose the second curve confirm The new curve is displayed and temporarily stored under the name SA index 13 5 4 6 Constant Press F6 Constant yo g m 2 0 0 2 q a Multiplication Allows to multiply a curve by a positive or negative number Enter multiplication Choose the curve Enter the multiplying coefficient and confirm The new curve is displayed and stored temporarily under the name SM index 100 Uvi Light XTD SCHOTT INSTRUMENTS b Addition Allows to add positive or negative constant to the curve to shift it upwards
126. t the curve CALCULATION OF DERIVATIVES A window the width of which corresponds to the differentiation step in nanometers is moved over the spectrum the difference between two windows is defined by the measuring step Within each window a mean slope is calculated The value of the slope is entered in the center of the window and point of the derivative curve is thus obtained METHOD OF CALCULATION A method derived from the Stavitzky Golay method is used for obtaining the derivative spectrum a fraction of the curve contained in the calculation window is defined by an equation of the following type u 3 2 From central point of the window coefficients are calculated with the least square method The coefficients help to take into account the distance of the point to the center of the window and to calculate a weighted average along the following formula nl C Y Y 98 Uvi Light XTD SCHOTT INSTRUMENTS Weighting coefficient 1 Absorbance N n Normalization factor This method allows to smooth and derive the curves by simply making a weighted average The signal noise ratio can therefore increase significantly just increase the number of points and therefore expand the calculation window However the spectrum may be distorted owing to the width of the window Actually the value of the derivative decreases when the calculation window is expanded It also depends on the order of de
127. t tic qe Ra E CERE xr OR E XN S re 91 13 2 PRINTING THE PARAMETERS cc etia n e catur itte s Gets d e acie a e dia ei 92 19 9 SAVING THE METHOD N 92 555 ec s Ru e eae a PU E 92 19 9 TOOLS 93 13 3 93 3 6 lol d siat satt tad oan Ses cual uo E zb E 94 TO Dee LOGO TT 94 1351 3 CIO enaa a 94 EON ELE dors ERES 95 VECES 610 6 MERCEDE a ro cM m co E RO CES c EU 95 ro m 95 eo Ey 95 PIL WUT COIS OM mre 95 13 32 MANUAL DUREE 96 CES AICI A E 96 T4 ORA eed Sa cM a M 97 13 59 42 DOT VQUV CS
128. tative methods Choose Abs Wavelength 190 900nm Reagent blank YES NO See blanks meaning at the end of this chapter page 80 Sample blank YES NO See blanks meaning at the end of this chapter page 80 Decimal place From 0 to 3 Precision of the result Standard number Factor To work with factor answer 0 Choose value between 10000 1 Standard To work with 1 standard answer 1 Choose value between 0 and 10000 Several standards To work with factor enter the number of standards from 2 to 10 Choose value between 0 and 10000 Unit With keyboard enter the abbreviation of unit X axis concentration axis Linear or logarithmic Y axis absorbance axis Linear logarithmic 1 Abs 1 Y or Logit See Logit axis meaning at the end of this chapter page 80 Type of curve Linear segmented curve Regre linear regression Regre 0 linear regression going through the origin 0 Spline See explanation on the type of curve at the end of this chapter page 81 Numbering This parameter concerns samples identification Auto Identification of the sample is numerical and increasing Example Sam 1 Sam 2 Unit The user assigns a name to each sample 8 characters and eventually attributes a dilution factor See explanation at the end of this chapter page 88 Table The user prepares a table in which he will enter the number of samples their name and the dilution factor See
129. ticell holder as indicated above Start the reading by pressing F1 Measure Entrer number of cell 1 8 2003 11 21 11 17 43 1 00 RESULTS Sample Conc MG L MG L MG L MG L MG L MG L MG L MG L uU S UJ UJ n2 F2 O UJ Co NUOVI A B D E F G H I RSTUVWXYZ TEQUE NM Copyright SECOMAM THE DISPLAYED ABSORBANCES OF THE SAMPLES ARE READ AS AGAINST THE BLANK THE DISPLAYED ABSORBANCE OF THE BLANK IS READ AS AGAINST AIR 49 SCHOTT INSTRUMENTS The printing will be Uvi Light xTD SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 10 32 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 on Sam 9 Sam 10 0 5 MG L Conc Conc 0 8 MG L 1 4 MG L 1 9 MG L 2 0 MG L 2 8 MG L 3 4 MG L 3 7 MG L 3 0 MG L 2 4 MG L Abs 0 283 Abs 0 185 Abs 0 320 Abs 0 575 Abs 0 751 Abs 0 814 Abs 1 102 Abs 1 348 Abs 1 476 Abs 0 272 Abs 1 204 Abs 0 973 Time Time Time Time 10 32 am 10 32 am 10 33 am 10 33 am The sample concentration will be calculated as follow Sam Conc Sam Abs Blank Factor Values read against the air Enter number of samples to be measured Confirm Follow indications displayed to prep
130. ts the command F2 Print allows to print the result The command F3 Hardcopy prints out the curve s with the results 14 6 3 3 Derivatives Derivate a kinetic curve allows to see the slope evolution versus time CALCULATION OF DERIVATIVES Same calculation as spectrum mode See page 98 Press F3 derivative Choose the kinetic curve to be derived Absorbance T 1 1 1 1 1 1 50 60 80 Temps en secondes Choose the order of the derivative and the differentiation step The step expressed in seconds corresponds to the width of the smoothing window Example Kinetic KIN13 Order 4 Step 75 115 SCHOTT INSTRUMENTS Uvi Light XTD Absorbance 1 1 1 Temps en secondes The derived curve is saved temporarily under the name CD index and may be retrieved for superimposition with the initial spectrum Absorbance 1 1 4 60 Temps en secondes 14 6 3 4 Addition Allows to adds two curves Enter Operation Choose the first curve confirm Choose the second curve confirm The new curve is displayed and temporarily stored under the name KINA index 14 6 3 5 Substraction Allows to substract one curve from another Enter Operation Choose the first curve confirm Choose the second curve confirm The new curve is displayed and temporarily stored under the name KINA index 14 6 3 6 Constant Pr
131. um absorbance of the compound This requirement allows to work with the highest sensitivity There must be no ionization association or solvation of the compound depending on concentration or over time The solution must not be too concentrated for the law only applies below a certain concentration and for a given substance 4 3 D FINITIONS 4 3 1 Transmittance The transmittance T or transmission factor is expressed as a percentage and is equal to 13 SCHOTT INSTRUMENTS Uvi Light a It is generally expressed in 9o Transparent body T 100 Opaque body 0 4 3 2 Absorbance Absorbance A or extinction E is given by the following equation A log E Transparent body body 00 Extinction E or absorbance A are more frequently expressed as the log of the ratio 1 0 1 Absorbance is sometimes called the optical density but such terms are not appropriate It strictly corresponds to the fraction of the internal optical density of a solution due to a number of its components The most common expression is as follows ES 4 3 3 Concentration The concentration of a solution is the quantity of dissolved material per volume unit of solvent It is given by the following equation Pas l Where E is molar absorption coefficient of the substance isthe length of the cell generally 1 cm C is the concentration of the solution looked for in the cell is the a
132. v1 00 RESULTS conc CELL Reference Sam bl Sam 1 Sam bl Sam bl sam sam 1 sam HIO NMOODMDP Copyright SECOMAM EMI ES Prepare the multicell holder as indicated Start the reading by pressing F1 Measure Entrer number of cell 1 8 2003 12 01 11 54 06 Concentration Mode v1 00 RESULTS Samp le Conc CELL Sam bl Reference Sam 0 4 G L Sam bl Sam bl Sam 1 Sam 0 5 G L Sam bl Sam bl Sam Sam 0 5 G L Sam bl Sam bl sam sam 1 1 G L Sam bl sam H D BDmBooooo B D E F G H T RS TUVWAY 2 Back 6 Enter Copyright SECOMAM THE DISPLAYED ABSORBANCES OF THE SAMPLE BLANKS AND SAMPLES ARE READ AS AGAINST THE REFERENCE SOLUTION 61 SCHOTT INSTRUMENTS The results will be printed out Uvi Light xTD SCHOTT INSTRUMENTS UV VIS Spectrophotometer Uvi Light v1 00 Date Friday november the 21 2003 Time 11 43 am User s name SMITH Batch number 123 Origin HOSPITAL Kit number 1256 Method Concentration Mode v1 00 Sam bl 1 Abs 0 186 Time 11 43 am Sam 1 Conc 0 9 Abs 0 377 Sam bl 2 Abs 0 534 Sam 2 Conc 1 7G L Abs 0 776 Sam bl 3 Abs 0 832 Sam 3 Conc 2 6G L Abs 1 002 Sam bl 4 Abs 1 353 Sam 4 Conc 3 6 G L Abs 1 380 Sam bl 5 Abs 0 186 Time 11 44 am Sam 5 Conc 0 9 Abs 0 377 Sam bl 6 Abs 0 534 Sam 6 Conc 1 7G L Abs 0 776 The concentration value of the sample is computed as follows
133. w up measurement of absorbance in relation with time and curve processing Concentration general programs for quantitative readings concentration multiple calibrations linear or not calibrated kinetics multiple wavelength Spectra spectral scanning and curve processing These measuring mode are displayed with their version number and creation date 8 2 CUSTOMIZED METHODS USER S Fi Config Editor fi 3 015 71 4 election SECOMAM Uvi Light v1 00 SIMULATION Program choices Kinetic mode Vl 11 200 L PROTEINES MEASUREMENT 10 12 2003 Concentration Mode L NITRATE MEASUREMENT Spectrum Mode i 04 11 2003 Copyright SECOMAM These are those that the operator has saved under a given name ex PROTEINES MEASUREMENT NITRATE MEASUREMENT to use the same parameters again In the tree structure of main menu every method is saved by the operator as a routine of the mother method used to create it Example PROTEINES MEASUREMENT was created from kinetic mode NITRATE MEASUREMENT was created from concentration mode 33 SCHOTT INSTRUMENTS Uvi Light XTD Short names PROT NO3 which appear on the right side of saved methods are the names of the files contening the methods parameters 8 2 1 Applications created These are original applications created by the operator or SCHOTT INSTRUMENTS from the SPECTRO PASCAL software As a rule they can give rise to several types of rea
134. wing message Beginning of the macro Then run the sequence of commands Ex Draw the spectrum of the sample Print the curve Draw the automatic list of peaks with a difference of more than 0 1 Abs Print the value of the peaks Draw the derivative of order 1 with a differentiation step of 5nm Print the derivative curve Return to the place where the macro command begun Press the Ctrl and F2 keys It will display at the left top of the screen the following message of macro hit a control key Assign a function key to the macro command Which keys can the operator use Keys Alt F1 through Alt F10 For this example we have chosen Alt F1 Once the key has been assigned It will display at the left top of the screen the following message of macro Enter the name of the macro command up to 18 letters and figures Ex SPECTRUM MEAS 15 2 EXECUTION OF A MACRO COMMAND For launching a macro command Enter the mode of analysis concerned In our example SPECTRUM mode 121 SCHOTT INSTRUMENTS Uvi Light XTD Go to the point where the macro command begins Press the function key assigned to the macro command In our example Alt F1 The above mentioned sequence of commands will be run and the unit will return to the point of departure The operator may either launch the macro command again or use the commands directly 15 3 RATED EXECUTION OF A MACRO COMMAND It may be useful t
135. with respect to the measuring order This chapter describes how to measure various solutions using single cell holder when you want to establish a calibration curve in Concentration mode This procedure will be the same for other measuring modes Absorbance Kinetics Spectrum scan For more information on these measuring modes see related chapters for multi cell holder To install the cell holder see installation guide SELECTING SINGLE CELL HOLDER Proceed this way Install the accessory From main menu press F1 F1 Configuration Selection 17 05 Tidatios SECOMAM Light v1 82 SIMULATION 1316 15 51 45 Conf igurat ion Language Option Printer selection Letter quality Grid Deuterium lamp mode Always CRT Black White Uirtual keyboard Disabled LCD Contrast H1 Copyright SRR Modify the parameter OPTION Select No Confirm other parameters in the window 10 1 MEASURING From main menu select a measuring mode example Concentration Mode Program analysis parameter Press F2 Parameters and F1 Modification 39 SCHOTT INSTRUMENTS Uvi Light XTD re save esc quit 2003 12 02 12 04 57 Concentration Mode v1 00 PARAMETERS Abs wavelength nm Reagent blank Sample blank Decimal place 0 3 No of standards 0 10 Conc 1 0 10000 2 0
136. y SCHOTT INSTRUMENTS Ex MEASURE 17 1 3 Spectra files They include in an encoded form the data of the spectra stored These files can be copied on floppy disk and be filed for ulterior use These files may be retrieved from the DOS by entering Name of the file J SPE HOLMIUM SPE In the menu of spectra files they appear under the name assigned to them by the operator Ex HOLMIUM 17 1 4 Kinetic files They include in an encoded form the kinetic data which have been stored These files can be copied on floppy disk and be filed for ulterior use These files may be retrieved from the DOS by entering Name of the file CIN The extension for such files is CIN SAMPLE CIN In the menu of kinetic files they appear under the name assigned to them by the operator Ex SAMPLE 17 1 5 Image files in spectrum mode 127 SCHOTT INSTRUMENTS Uvi Light XTD An image in scan mode is the screen representation where the spectra are displayed It may contain one or several curves with comments or not These files can be copied on floppy disk and be filed for ulterior use These files may be retrieved from the DOS by entering Name of the file IMS Ex IMAGE1 IMS In the menu of image files in spectrum mode they appear under the name assigned to them by the operator Ex IMAGE1 17 1 6 Image files in kinetic mode An image in kinetic mode is the screen representation where the spectra are displayed It may co

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