Quick Guide to the Star Bar

Transcription

1 Saturn View Saturn Writer Quick Guide to the Star Bar Last active Chromatogram in SaturnView Last active method in the Method Editor Click with the right mouse button on the Star Bar to get this menu Sample, Recalc, and Sequence list editor Method Editor 1

2 System Control Manual Control Button Auto Setup Button Temperatures Button Diagnostics Button Acquisition Button Shutdown Button 2

3 Instrument Control The ion trap cannot be turned on when the display is showing red during a filament/multiplier delay segment. The ion trap turns green when it is on. 3

4 Method Editor MS Section includes all Tune and IPM options GC Injector section Tune parameters IPM Parameters Double click on a line to see the details for each compound Compound Table Click on Next or Previous to move through the compounds 4

5 CLICKING THE ENDTIME BUTTON ALLOWS THE END OF THE RUN TO BE EXTENDED Acquisition THE ACQUISITION BUTTON MUST BE PRESSED IN BEFORE THE SATURN MODULE WILL GO READY. Specify default parameters to minimize having to change parameters when making several similar injections Specify default data file naming parameters. Create a new recalc list for each sample, or append to the same list. 5

6 Injecting a Single Sample 1. ACTIVATE SYSTEM CONTROL TO MAKE AN INJECTION 3. CLICK INJECT AND SELECT INJECT SINGLE SAMPLE 2. CLICK ACQUISITION TO PUT SATURN IN THE ACQUISITION MODE. 6. THE STATUS WILL DISPLAY WAITING WHEN IT IS READY FOR AN INJECTION. LEAVE READY FILE BLANK 4. TYPE IN THE NAME OF THE SAMPLE, THIS WILL ALSO BE THE DATAFILE NAME. 4a. CLICK TO CHANGE THE METHOD FILE IF NECESSARY. 4b. SELECT THE DIRECTORY TO STORE THE DATAFILES. MAKE A NEW DIRECTORY IF NECESSARY. 4c. DEFINE THE EXTRA INFO ADDED TO THE DATAFILE NAME 5. CLICK THE INJECT BUTTON 6

7 SaturnView Data Processing Right Click on Plot to add info. To display Select information to add to display. Tick marks showing background correction points. 7

8 Building a Peak Table 1. CLICK THE DATAFILE BUTTON TO OPEN THE LAST ACTIVE FILE OR LAST ACQUIRED DATAFILE. 2. CLICK THE QUANITATION MENU AND SELECT BUILD COMPOUND TABLE. 3. CLICK BUILD METHOD 4. ZOOM THE CHROMATOGRAM IF NECESSARY, AND CLICK ON A PEAK. 5. CLICK ON ADD PEAK. THIS WILL OPEN THE LIBRARY SEARCH WINDOW. RUN A LIBRARY SEARCH AND CLICK OK. ALL OF THE LIBRARY INFO WILL BE ADDED TO THE PEAK. 6. IF A PEAK WAS ADDED BY MISTAKE, HIGHLIGHT THE WHOLE LINE AND CLICK THE DELETE BUTTON. 7. CLICK DONE WHEN ALL PEAKS HAVE BEEN ADDED. THE SOFTWARE WILL PROPMT FOR A METHOD NAME. SELECT A ALREADY BUILT METHOD TO ADD THE PEAK TABLE. ONLY THE PEAK TABLE SECTION WILL BE OVERWRITTEN. 8

9 Editing a Peak Table P1 1. CLICK THE METHOD EDITOR BUTTON AND OPEN A METHOD TO EDIT. THE LAST ACTIVE METHOD CAN BE SELECTED BY CLICKING THE SHORT-CUT BUTTON. 2. DOUBLE CLICK ON A LINE TO EDIT THE COMPOUND. COMPOUND TABLE 3. CLICK THE CALCULATIONS TAB AND MOVE TO THE FIRST COMPOUND IN THE TABLE. 3a. USE THE NEXT / PREVIOUS BUTTONS TO MOVE THROUGH THE COMPOUND LIST. 1a. CLICK AND OPEN COMPOUND TABLE 4. ENTER THE NUMBER OF LEVELS TO BE USED FOR THE CALIBRATION CURVE. 5. ENTER THE AMOUNT OF EACH LEVEL INTO ONE OF THE NUMBERED LEVELS AND THE UNITS INTO THE AMOUNT UNITS BOX. 9

10 Editing a Peak Table P2 6.CLICK THE COMPOUND ATTRIBUTES TAB. 7. MOVE TO THE COMPOUND TO BE USED AS THE INTERNAL STANDARD, AND CLICK THE INTERNAL STANDARD RADIO BUTTON. 8. CLICK THE CLOSE BUTTON. 9. CLICK THE TOP OF THE CALCULATIONS COLUMN TO HIGHLIGHT ALL COMPOUNDS. 10. CLICK THE FILL DOWN BUTTON TO COPY THE SAME LEVELS TO ALL OF THE CONMPOUNDS. 11. DOUBLE CLICK ON THE FIRST INTERNAL STANDARD (IN THE CALCULATIONS COLUMN) THIS WILL OPEN THE CALCULATIONS EDITOR. 12. INTERNAL STANDARDS SHOULD HAVE THE SAME AMOUNT ENTERED FOR EACH LEVEL. ENTER THE AMOUNT INTO THE FIRST LEVEL. 13. CLICK THE COPY AMOUNTS BUTTON TO COPY THIS AMOUNT TO ALL OF THE LEVELS. CLICK CLOSE. 14. REPEAT STEPS 11 THRU 13 FOR EACH INTERNAL STANDARD. 15. CLOSE AND SAVE THE METHOD. 10

11 Processing a Calibration Curve P1 2. FROM QUANTITATION - SELECT PROCESS/REVIEW RECALC LIST 1. OPEN SATURNVIEW 3. CLICK THE BROWSE BUTTON TO CHOOSE THE METHOD FILE CONTAINING THE PEAK TABLE FOR YOUR COMPOUNDS. 4. CLICK THE BROWSE BUTTON TO CHOOSE THE RECALC LIST OF YOUR CALIBRATION DATAFILES, OR CLICK THE EDIT BUTTON TO CREATE A NEW RECALC LIST. 5. CREATE A NEW RECALC LIST AND ADD IN THE DATAFILES FOR EACH LEVEL. 6. INSERT A NEW LINE ON LINE CHANGE THE SAMPLE TYPE TO NEW CALIB BLOCK. 8. SET THE SAMPLE TYPE FOR EACH DATAFILE TO CALIBRATION AND ENTER THE CAL LEVEL. 9. CLOSE THE AUTOMATION EDITOR. NOTE: THE NUMBER IN THE Cal Level COLUMN CORRESPONDS TO THE AMOUNTS IN THE METHOD FILE 11

12 Processing a Calibration Curve P CLICK THE PROCESS BUTTON TO BUILD THE CALIBRATION CURVE IN THE METHOD FILE. 11. TO VIEW EACH POINT INTERACTIVELY HIGHLIGHT THE ROW FOR THE DATA FILE AND CLICK ON RESULTS. 11a. WHEN THE PROCESSING IS DONE, THE CURVES CAN BE VIEWED FOR PRINTING BY CLICKING THE PRINT DATA BUTTON. 12

13 Inspection of Calibration Curves 1. It is essential to adjust all data handling parameters carefully. 2. Sometimes despite best efforts, manual integration is sometimes needed. 13

14 Guidelines For Optimizing Method Integration Parameters Peak Description Pk Width Slope Sens Pk Window should be ~ actual width at 1/2 height should be >= actual span of baseline (Default method values) Area > 1,000; Good to fair pk shape; Little or no pk tailing; Baseline spans single pk; RT's are consistant Area > 1,000; Good to fair pk shape; Little or no pk tailing; Baseline spans fused pk; RT's vary Area ~ ( ); Good pk shape; Base line spans single pk; RT's are consistant 2 to Area ~ ( ); Good pk shape; Base line spans fused pk; RT's vary 2 to Area > 1,000; Fair to poor pk shape; Significant pk tailing Area ~ ( ); Fair to poor pk shape; Some pk tailing 5 to Area ~ ( ); Fair pk shape; Few data points 5 to

15 Integration Parameters Tangent % (0-100; preset = 10) Peaks on the trailing edge of a peak will be integrated via tangent skimming if their heights are less than this % of the parent peak. If not, verticals are dropped to baseline from the valleys. Slope Sensitivity Factor - (1-256; preset = 20) Use Slope Sensitivity, 10 to improve integration of tailing peaks. 15

16 Fine Tune Integration Parameters When adjusting integration parameters - always check final parameters on the lowest level calibration file 16

17 Editing the Compound Table Identification 17

18 Editing the Compound Table Reference Spectrum (be attentive to coelution) After editing click on Redraw Ref Spectrum 18

19 Manual Integration of Target Compounds If data points are Missing, Failed or poorly integrated the Data Handling Method should be revised to achieve proper integration. Alternatively, individual data files can be manually integrated from SaturnView. Highlight the compound of interest and click on view to go to the integration parameters. 19

20 Interactively Moving Peak Events For an event that cannot be moved, such as a Main Apex, the cursor will change to a symbol. When the mouse cursor is placed close to a peak event it will change and a small window will appear with information about the event. For an event that is capable of being moved, such as a Peak Start, the cursor will change to a symbol. You can now click and drag the event to a new position 20

21 Interactively Moving Peak Events continued.. 21

22 Interactively Moving Peak Events the compound results will reflect the change... 22

23 Interactively Moving Peak Events the displayed Compound Report will also be updated Results will be updated here. 23

24 Manual Integration of Target Compounds Reintegrating a data file with a moved event will change the result of the peak affected by the move. Reintegrating a moved peak event of an Internal Standard peak will change the results of all target compounds. These changed results will persist in the data file until it is reintegrated again. When the data file is reintegrated with another data handling method, all moved peak event changes will be lost. 24

25 Manual Integration of Unknown Compounds and TIC s For unknowns and TIC s, manual integration is accomplished from the Chromatogram Integration screen of SaturnView. Highlight the compound of interest and click on view to go to the integration parameters. Note: Time Events are adjustable with non target compounds 25

26 Manual Integration of Unknown Compounds and TIC s Press the Integrate button to reintegrate with the moved event. An information window appears when moving the event. Left click & drag the event to a new position. After the file had been integrated, the results listing will reflect the change. 26

27 Manual Integration of Unknown Compounds and TIC s Results will also be updated here. Integration parameters for unknowns can be adjusted from the Calculations Setup dialog by pressing the Edit Method button. 27

28 Changing Method Integration Parameters Press the Integrate button to process the file with the new parameters. After the file has been integrated, the results will be updated in the report. 28

29 Interactively Adding / Adjusting Time Events Right click here to add a time event. The Time Events window will appear. 29

30 Interactively Adding / Adjusting Time Events continued Select a time event and it will appear in the Time Events window. The beginning time will be where you right clicked the window 2. Click & drag the center bar to move the entire time range. An information window appears when moving or sizing the event. 3. Click & drag and event endpoint to change the time range. 4. Right click the time event and you can choose to delete the event or to edit the event directly in the data file method. 30

31 Adding / Adjusting Time Events Results will be updated on the screen display and also on the Results Report listing 31

32 Most Frequently Adjusted Parameters Tips for missing peaks: Spectral match threshold too high (normal ) Ion Intensity threshold is not set (normal 5%) Reference spectrum is not clean (compound table was created when analytes were coeluting and no editing of the spectrum took place) Too wide identification window Area/height count reject is too high Tips for poorly integrated peaks: Peak width too small Slope sensitivity too high % Tangent too low/too high Identification window too wide/too narrow Check on Help in the software 32

33 What Happens After Manual Integration? Calibration files Change in area count / RRF, RF is recorded in the.sms file and in the ORIGINAL method file. This information is the base of the calibration. Adjustment in compound table parameters are recorded in the original.mth file. Certain parameters can not be changed with manual integration: Analysis runs CAS number of an analyte Designation of IS The improved original.mth file will be used to quantitate the analysis runs. Change in area count/amount is recorded in the.sms file If integration parameters were changed, those can be saved in the original method file or in the copy of the original method file attached to the.sms file. This information will remain with the.sms file until it is reprocessed again. Summary Prepare the compound table with DILIGENCE Verify that identification and integration parameters are set properly for all compound. (do this on the lowest level of calibration) Adjust Peak start and end parameters in the manual integration window Adjust method parameters via the edit method option in manual integration Integrate non-target analytes as in StarWS 33

34 Summary of SaturnView Quantitation 1. Acquire Sample 2. Process Active File 3. Report 34

35 Standard and Custom Reports ICC Report Tuning Report Sample Report Unknowns Report Compound Report Library Search of Unknowns Report 35

36 QUICK START 36