QExactive Calibration

Use newest tune file or if problems, go back to POS HESI Tune 01132020_CPC tune file for positive calibration or NEG for negative calibration.
The flow rate should be 3-5ul/min. Calibration tune files have aux heater temp at 0C.
To get a stable spray, increase the sheath gas flow 5-10, Aux gas flow to 0-5 and sweep gas flow rate to 0-2.
Start with positive ion calmix.
The signal with calmix should be >=1E8 with IT of ,<=2ms and TIC variation under 15% for 200 scans. If parameters not met, change ion transfer tube, may need to clean SRIG.
First evaluate using Evaluate tab on left side of Tune page Evaluate Base pos, uncheck Isolation. Then Calibrate the parameters that have failed. Mass calibration needs to be done irrespective of passing or failing. Analyzer accuracy is C trap calibration which should be calibrated before a long run.
Once calibrated with the regular method, calibrate the Custom Calibration, within the Calibration tab. Select only the positive ions while calibrating the +ve mode and vice versa.
After the custom calibration, calibrate only the mass under the regular Calibration once again. This would make sure to calibrate both low and high mass range.
To check if the low and high range is stitched, look under the instrument status →control →Mass calibration data →MCal → Higher order shape → 5pts (74-524) and 6pts( 138-1622). This is found on right side of Tune page.
Repeat procedure steps 5-10 for negative mode with negative ion calmix, TIC variation is higher but should be <=15%. Select only negative ions in custom calibration. Negative should be done once a month if not running negative mode samples.
Run reconditioning runs for both columns, BSA runs, and standby runs. Look for peak intensity and shape, compare to Nov method loaded by default into Freestyle.