Steps for LCMS-8040 Triple Quadrupole
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1 Steps for LCMS-8040 Triple Quadrupole June 2017 Version 2 A. Data Acquisition 1. Check lights on MS. There should be a total of three lights lit by LED. One of them is blinking. Power Blinking Gas Status 2. Check level of solvents A and B on top of instrument. Be sure there is sufficient solvent volume for your run. Bottle A is aqueous and Bottle B is organic. 3. Check the waste can. 4. If changing solvents: a. Hit Purge button on Autosampler. This is set to purge for 25 minutes. b. Turn black knob on LC-20AD Pump A 90 counterclockwise towards open. Press purge on LC-20AD Pump A. 90 counterclockwise 1
2 c. Turn black knob on LC-20AD Pump B 90 counterclockwise towards open. Press purge on LC-20AD Pump B. d. Turn black knob back to closed on LC-20AD Pump A 90 clockwise when purge is complete (5 minutes). Turn clockwise until you can t turn anymore. The A sticker should be upright. e. Turn black knob back to closed on LC-20AD Pump B 90 clockwise when purge is complete (5 minutes). The B sticker should be upright. 5. Double click on Lab Solutions icon on computer Desktop. 6. Logon as Admin. No password needed. Just hit OK. 7. The Lab solutions windows will open. Click on the Instruments icon 8. Select and Double-click on the LCMS icon 1. Click on Instruments 2. Click on LCMS 2
3 9. Click on Main Tab on far left. Main 10. Click the Data Acquisition icon in the Main assistant toolbar. Data Acquisition 11. Under File, open Method File. Or double-click on your method to load. 12. Click on Download when complete.. This will start Mass Spect. It will not start heating column, if needed, or flow rate. a. Oven Temperature. Want slightly above room temperature. Usually set at 40 C. Hit icon to start warming column. b. Pump ON/Off icon on the top toolbar. Pump Solvent delivery is started. 3
4 B. Create or Edit a Method 1. If you need to create a New Method file, start with MS Tab. 1) Set the MS Instrument Parameters. Click on MS tab. Each individual MS condition is called an event and polarity can be set for each event. The system can be scanned in positive and negative modes simultaneously. This can be accomplished by setting events. Each event has its own parameters MS tab a) Select polarity of the event to set: Positive or Negative. b) Click the button for the data s acquisition mode; MRM(±), Product Ion Scan(±), Precursor Ion Scan (±), or Neutral Loss Scan (±), SIM (±) or scan (±). c) The event is now added to the Event Table. 4
5 ] b.) If choosing SIM, be sure to put in M/Z and change dwell time to 20msec. 5 Add M/Z and change Dwell time to 20msec for SIM.
6 c) Set Same measurement time for each event by right clicking and choosing Set Same Measurement time followed by OK. Set Same Measurement Time d.) Type OK 6
7 2. Set Interface parameters. a. ESI and/or APCI mode. b. Nebulizing Gas flow 2 L/min. (3 L/min for higher flow rates) c. DL temp =250 (300 for higher flow rates) d. Heat Block temp =400C. e. Check off drying gas flow 15 L/min. 3. Set Data Acquisition parameters. A. Enter the LC Stop Time as time from one sample to the next. B. Click Apply to All acquisition time to set end time for each detector to the same time as the LC detector. Apply to All Acquisition Time 7
8 4. Under Pump, keep as binary gradient, even if running isocratic. a. Set Pump Flow rate and initial concentration for gradient systems, Pump B conc (never 100% H2O for C18 RP column). b. Set Pressure Max for your column ( i.e 6000)(Look up on internet). 5. Set Oven Temperature slightly above room temperature. Usually set at 40 C. Hit icon to start warming column 6. Click on the LC Time Prog. tab to enter gradient conditions. A. Set Time, Module, Command, and Value. 8
9 B. Click on Draw Curve to visualize your method. C. Click on the MS Instrument On/Off icon on the top toolbar to start DL heater and heat block temperature control. 7. Click on Download when complete. Download 8. Click on the Pump ON/Off icon on the top toolbar. Pump Solvent delivery is started. 9
10 9. Under Autosampler, change Purge Time to 10 min. Change Purge time to min 10. Under File, choose Save Method File As.. a) Enter the File name and click Save. 10
11 C. Click the Realtime Batch icon 1. A realtime batch table will open. 2. Under File, choose New Batch File. a. Enter Vial #, b. Tray name =1 c. Sample Name d. Sample ID e. Sample Type-unknown or standard f. Method File set up previously g. For Data File write unique name of sample(can t use name already assigned to previous sample) h. Injection volume from 0.1ul to 100µl. 3. Right click on screen and choose Settings or click on Settings icon 4. Go to Shutdown tab under Settings. Enter conditions for automatic shutdown with a cooling time of longer than the running time. Be sure all instruments are checked off to shutdown after run. Shutdown is checked Cool down time longer than run time All instruments checked off 11
12 5. Under File, choose Save Batch method as Select the rows and numbers to be analyzed. 7. Click on Start Realtime Batch icon Select All Rows or Selected Rows Hit Stop button if you need to stop the run. 12
13 D. Data Analysis 1. Click on the Postrun icon 2. Double-click on Postrun.(Browser is used to look at more than one sample at a time) 13
14 3. Drag and drop a file into the window. Or Double-click on file. 4. Average MS Spectrum- a. Click on Average Spectrum on toolbar. A stable spectrum can be displayed by totaling and averaging the spectra within a certain time range. 14
15 5. Subtract MS Spectrum a. Click Subtract Spectrum on MS Data Analysis toolbar. A cleaner-looking spectrum can be displayed by subtracting the background MS spectrum from the averaged spectrum. 15
16 6. Add spectra to Spectrum Process Table a. The spectrum displayed can be entered in the Spectrum Process Table. b. Click on Register to Spectrum Process table in MS Analysis Toolbar. c. Click OK. d. The Spectrum is registered to the Spectrum Processed Table. 16
17 E. Report 1. Save screen as seen a. Under File, choose Print Graph Image, then PDF Output. b. Save on USB drive. F. Shutdown 1. Close Lab Solutions by going under File and choosing Exit. 17
18 2. A shutdown sub-window opens. Be sure everything is checked off. 3. Under File, choose Exit Lab Solutions. 18
19 19
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