Backup current tune file before autotune Running QTOF System Tune in Positive Polarity Begin Running System Tune Backup All Parameters Spraying Calibrant Solution Disable sureMass optimization Setup Dual AJS ESI Source to Achieve Stable Signals for Auto Tune Restore Collision Cell Pressure to its default value 22 psi Reset All Parameters to Their Default Values Baseline Adjustment for 10 GHz Instrument Binary search the linear range of high gain base line Sucessfully found the linear starting point of Amp Offset at 2666.0 DAC Finding High Gain Baseline Using Linear Regression Adjust High Gain Base Line Estimate Hi Gain Amp Offset Found at 2353 Binary search the linear range of low gain base line Sucessfully found the linear starting point of Low Gain Preamp Offset at 1877.9 DAC Finding Low Gain Baseline Using Linear Regression Estimate Low Gain Base Line Optimum Low Gain Amp Offset Found at 1814 Final Optimum Low Gain Pre Amp Offset Found at 1814 DAC Final Optimum High Gain Pre Amp Offset Found at 2353 DAC Waiting for Calibrant to Elute... Monitoring 118.06 m/z Peak for Intensity and Stability Signal Intensity and Stability Satisfied Auto Tune Requirements Performing Coarse TOF Mass Calibration Using 6/10 Calibrant Masses Successfully Perform Mass Calibration with RMS Deviation (ppm): 0.1 Reset Optics 2 and TOF Parameters to Their Default Values Coarse TOF Resolution - Mirror Mid Optimization Change Mirror Mid from -2,270.0 to Optimum Value at -2,265.1 Coarse TOF Resolution - Puller Offset Optimization Change Puller Offset from 40.0 to Optimum Value at 30.8 Performing Coarse TOF Mass Calibration Using 6/10 Calibrant Masses Successfully Perform Mass Calibration with RMS Deviation (ppm): 0.35 Adjusting VCap to reduce signals to reasonable height 100000 Reduce high mass signals by reduce VCap to 1844 V Adjusting Quad AMU to Produce Single Ion Event at MCP of 700 Coarse optimum Quad AMU Found at 9.4 with Peak Height of 4174 Optimum Quad AMU Found at 9.2 with Arrival Frequency of 4.3 % Searching MCP to Produce 20 Counts per Single Ion Counts MCP Frequency Status 30.7 700 4.7 % in Progress 16.5 668 3.9 % in Linear Range 17.7 671 3.9 % in Linear Range 18.4 674 3.6 % in Linear Range 19.0 675 4.4 % in Linear Range 20.6 676 4.0 % in Linear Range 21.2 676 3.9 % in Linear Range 19.7 675 4.2 % in Linear Range 19.4 675 4.3 % in Linear Range Optimum of MCP Found at 675 V Running Transmission Tune for 3200 m/z Range Initializing Parameters for 3200 m/z Range Standard Tune Fragmentor: 165.00 Skimmer: 45.00 Oct 1 DC: 28.50 Lens 1: 27.00 Quad DC: 25.00 PostFilter DC: 25.00 Cell Entrance: 24.00 Hex DC: 23.00 Hex Delta: -7.00 Hex2 DC: 16.00 Hex2 DV: -1.00 Hex3 DC: 14.80 Ion Focus: 10.00 Lens 2: 18.00 Quad AMU: 140.00 Optimizing Optics Rail From Ion Source to Detector Adjusting Optics Rail from Oct 1 DC to TOF for 3200 m/z Range Change Optics Rail from Oct 1 DC to TOF from 0.0 to Optimum Value at 0.1 Optimum of Oct 1 DC Found at 28.6 Adjusting Optics Rail from Lens 1 to TOF for 3200 m/z Range Change Optics Rail from Lens 1 to TOF from 0.0 to Optimum Value at -0.2 Optimum of Lens 1 Found at 26.9 Adjusting Optics Rail from Quad DC to TOF for 3200 m/z Range Change Optics Rail from Quad DC to TOF from 0.0 to Optimum Value at -0.1 Optimum of Quad DC Found at 24.8 Adjusting Optics Rail from PostFilter DC to TOF for 3200 m/z Range Change Optics Rail from PostFilter DC to TOF from 0.0 to Optimum Value at -0.2 Optimum of PostFilter DC Found at 24.6 Adjusting Optics Rail from Cell Entrance to TOF for 3200 m/z Range Change Optics Rail from Cell Entrance to TOF from 0.0 to Optimum Value at -0.3 Optimum of Cell Entrance Found at 23.3 Adjusting Optics Rail from Hex DC to TOF for 3200 m/z Range Change Optics Rail from Hex DC to TOF from 0.0 to Optimum Value at 0.3 Optimum of Hex DC Found at 22.7 Skimmer: 45.00 Oct 1 DC: 28.59 Lens 1: 26.92 Quad DC: 24.79 PostFilter DC: 24.63 Cell Entrance: 23.33 Hex DC: 22.66 Hex Delta: -7.00 Hex2 DC: 15.66 Hex2 DV: -1.00 Hex3 DC: 14.46 Ion Focus: 9.66 Performing Coarse TOF Mass Calibration Using 6/10 Calibrant Masses Successfully Perform Mass Calibration with RMS Deviation (ppm): 0.39 Quad Transmission Optimization Change Quad AMU from 140.0 to Optimum Value at 128.6 Balancing TOF Mirrors and Slits at Puller Offset of 35.0 Mirror Mid Bot Slit Resolution Abundance Score -2,265.6 -46.0 31,519 146,204 128 -2,265.6 -45.0 33,316 226,379 143 -2,265.6 -44.0 34,577 243,817 148 -2,265.6 -44.5 32,843 237,540 145 -2,265.6 -43.5 37,170 214,610 155 -2,265.8 -43.0 40,666 162,751 158 -2,265.8 -42.5 38,936 115,312 146 Local maxima found Bot Slit: -43.0 Refine Puller Ofsset - Puller Offset Optimization Change Puller Offset from 35.0 to Optimum Value at 33.4 Mapping Area 1: Resolution: 39,555 Abundance: 160,858 Score: 158 Balancing TOF Mirrors and Slits at Puller Offset of 45.0 Mirror Mid Bot Slit Resolution Abundance Score -2,265.8 -46.0 22,177 141,482 94 -2,265.3 -45.0 25,307 200,638 111 -2,265.3 -44.0 27,388 191,482 118 -2,265.3 -44.5 26,424 206,379 116 -2,266.3 -43.5 27,937 164,228 118 Local maxima found Bot Slit: -44.0 Refine Puller Ofsset - Puller Offset Optimization Change Puller Offset from 45.0 to Optimum Value at 36.8 Mapping Area 2: Resolution: 35,098 Abundance: 244,304 Score: 153 Best Multidimensional Optimization Solution Found: Bot Slit: -43.00 Mirror Mid: -2,265.84 Puller Offset: 33.43 Refine TOF Resolution - Pusher Offset Optimization Change Pusher Offset from 0.0 to Optimum Value at -39.8 Reoptimizer delta slit with newly found optimum Pusher Offset of -39.8 Balancing TOF Mirrors and Slits at Puller Offset of 33.4 Mirror Mid Bot Slit Resolution Abundance Score -2,265.8 -46.0 31,164 153,047 129 -2,265.4 -45.0 35,755 232,517 152 -2,265.4 -44.0 37,931 240,122 160 -2,265.4 -44.5 35,808 248,296 155 -2,265.4 -43.5 38,605 204,571 161 -2,265.4 -43.0 39,281 159,479 154 Local maxima found Bot Slit: -43.5 Final Multidimensional Optimization Mirror Mid, Bottom Slit, and Puller Offset Optimized Score Improved 2.5 % to 128 Best Solution Found: Bot Slit: -43.64 Mirror Mid: -2,265.35 Puller Offset: 33.32 Refine TOF Resolution - Puller Offset Optimization Change Puller Offset from 33.3 to Optimum Value at 34.5 Refine TOF Resolution - Pusher Offset Optimization Change Pusher Offset from -39.8 to Optimum Value at 30.2 Refine TOF Resolution - Slit Offset Optimization Change Slit Offset from 0.0 to Optimum Value at 3.3 Refine TOF Resolution - Puller Offset Optimization Change Puller Offset from 34.5 to Optimum Value at 32.6 Refine TOF Resolution - Pusher Offset Optimization Change Pusher Offset from 30.2 to Optimum Value at -14.8 Standard TOF Resolution - Lens 3 Optimization Change Lens 3 from -60.0 to Optimum Value at -61.4 Fine TOF Resolution - Extractor DC Optimization Change Extractor DC from -10.0 to Optimum Value at -14.0 Fine TOF Resolution - Ion Focus Optimization Change Ion Focus from 9.7 to Optimum Value at 5.4 Reduce signals before optimization Acc Focus in high resolution mode All Calibrant Peaks Passed Saturation Check - No Abundance Adjustment Needed Do not need to reduce signals Fine TOF Resolution - Acc Focus Optimization Optimum for Acc Focus Found at -2,146.0 Refine TOF Resolution - Puller Offset Optimization Change Puller Offset from 32.6 to Optimum Value at 31.4 Fine TOF Resolution - Mirror Mid Optimization Change Mirror Mid from -2,265.4 to Optimum Value at -2,265.1 Performing Coarse TOF Mass Calibration Using 6/10 Calibrant Masses Successfully Perform Mass Calibration with RMS Deviation (ppm): 0.31 TOF Energy Offset Optimization Change TOF Energy Offset from 0.0 to Optimum Value at -0.2 Enable sureMass optimization Performing Coarse TOF Mass Calibration Using 6/10 Calibrant Masses Successfully Perform Mass Calibration with RMS Deviation (ppm): 0.36 Set Quad State to Initial Calibration Mass Calibrants: 322.0m/z and 2121.9m/z Coarse Quad Calibration for Wide Peak Width Wide - Mass Tolerance = 1. amu Wide - PeakWidth Target = 9 +/- 1 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 -21.70/-52.26 11.48/59.88 0 in Progress 2 01.98/-04.85 00.08/13.95 0 in Progress 3 -01.40/-01.46 01.55/06.87 0 in Progress 4 00.64/01.86 00.38/01.45 4 in Progress 5 00.24/05.74 00.12/-03.43 0 in Progress 6 01.44/02.82 -01.21/-02.25 0 in Progress 7 -00.38/00.10 00.23/-00.18 78 Passed Results for Wide Peak Width: Score of 78 Coarse Quad Calibration for Medium Peak Width Medium - Mass Tolerance = .5 amu Medium - PeakWidth Target = 4 +/- 0.5 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 02.51/01.84 -01.05/-00.81 0 in Progress 2 -00.65/-00.72 00.00/00.04 9 in Progress 3 00.02/00.04 00.10/-00.02 96 Passed Results for Medium Peak Width: Score of 96 Coarse Quad Calibration for Narrow Peak Width Narrow - Mass Tolerance = .2 amu Narrow - PeakWidth Target = 1.3 +/- 0.15 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 01.30/00.78 -00.23/00.20 0 in Progress 2 -00.22/-00.37 00.02/00.30 0 in Progress 3 00.03/-00.02 00.00/00.11 64 Passed Results for Narrow Peak Width: Score of 64 Finish Quad Tune Optimize Lens 2 RF Phase Lens 2 RF Ph Optimization Change Lens 2 RF Ph from 0.0 to Optimum Value at 97.0 Optimize Lens 2 RF as a Function of m/z m/z Lens 2 RF 0118.1 00.0 0322.0 79.5 0922.0 147.8 1522.0 212.8 2121.9 271.2 2721.9 316.1 Optimize Lens 2 DC as a Function of m/z m/z Lens 2 DC 0118.1 00.1 0322.0 20.3 0922.0 15.0 1522.0 07.3 2121.9 03.3 2721.9 03.4 Fine Quad Calibration for Wide Peak Width Wide - Mass Tolerance = 1. amu Wide - PeakWidth Target = 9 +/- 1 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 -00.93/-05.24 01.39/04.61 0 in Progress 2 -00.58/-03.37 00.91/02.90 0 in Progress 3 -00.06/-02.25 00.50/01.94 0 in Progress 4 -00.30/-01.41 00.08/01.24 9 in Progress 5 -00.21/-01.19 00.65/00.90 9 in Progress 6 -00.18/-01.00 -00.26/00.66 31 Passed - Refine 7 -00.12/-00.78 -00.40/00.88 25 Passed - Refine 8 -00.12/-00.65 -00.18/00.49 58 Passed - Refine 9 -00.12/-00.68 00.36/00.68 57 Passed - Refine 10 00.03/-00.41 00.39/00.46 76 Passed - Refine 11 00.06/-00.37 00.11/00.29 84 Passed - Refine 12 -00.18/-00.32 -00.03/00.30 87 Passed - Refine 13 -00.13/-00.21 -00.03/00.34 88 Passed - Refine 14 -00.06/00.04 00.32/00.01 92 Passed - Refine 15 00.14/-00.13 -00.02/-00.15 94 Passed Final Results for Wide Peak Width: Isolation Efficiency: 83, Score: 94 Fine Quad Calibration for Medium Peak Width Medium - Mass Tolerance = .5 amu Medium - PeakWidth Target = 4 +/- 0.5 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 -00.73/-04.39 01.32/03.24 0 in Progress 2 -00.19/-03.46 00.54/03.25 0 in Progress 3 -00.23/-02.40 00.32/02.03 0 in Progress 4 -00.18/-01.50 00.15/01.34 0 in Progress 5 -00.18/-01.08 00.07/01.12 0 in Progress 6 -00.06/-00.70 00.19/00.80 5 in Progress 7 -00.07/-00.38 00.03/00.47 37 Passed - Refine 8 -00.10/-00.43 00.17/00.52 9 in Progress 9 00.00/-00.28 00.11/00.45 48 Passed - Refine 10 -00.06/-00.26 -00.13/00.29 55 Passed - Refine 11 -00.01/-00.26 00.01/00.29 63 Passed - Refine 12 -00.05/-00.22 00.00/00.29 68 Passed - Refine 13 00.01/-00.07 00.01/00.28 77 Passed - Refine 14 -00.05/-00.08 00.05/00.21 88 Passed - Refine 15 -00.01/-00.10 00.08/00.16 92 Passed - Refine 16 00.04/-00.10 00.14/00.12 91 Passed - Refine 17 -00.03/-00.07 -00.01/00.08 97 Passed Final Results for Medium Peak Width: Isolation Efficiency: 74, Score: 97 Fine Quad Calibration for Narrow Peak Width Narrow - Mass Tolerance = .2 amu Narrow - PeakWidth Target = 1.3 +/- 0.15 Iteration Delta m/z Delta PW Score Result (322./2122.) (322./2122.) 1 -00.78/01.94 00.74/-00.38 0 in Progress 2 -00.07/-02.29 00.28/-00.27 0 in Progress 3 -00.09/-03.53 00.05/01.64 0 in Progress 4 -00.19/-03.45 00.13/03.20 0 in Progress 5 -00.32/-02.46 00.23/02.40 0 in Progress 6 -00.24/-01.61 00.20/01.66 0 in Progress 7 -00.16/-01.10 00.09/01.22 0 in Progress 8 -00.12/-00.71 00.08/00.98 0 in Progress 9 -00.07/-00.42 00.12/00.73 0 in Progress 10 -00.02/-00.26 00.07/00.53 0 in Progress 11 -00.04/-00.19 00.08/00.43 0 in Progress 12 -00.02/-00.09 00.07/00.30 6 in Progress 13 00.00/-00.08 00.04/00.29 7 in Progress 14 -00.04/-00.07 00.03/00.23 9 in Progress 15 00.00/-00.03 -00.01/00.23 9 in Progress 16 -00.03/00.00 00.02/00.21 9 in Progress 17 00.02/00.00 00.02/00.18 9 in Progress 18 -00.02/00.00 00.06/00.10 75 Passed - Refine 19 00.02/00.00 00.02/00.09 78 Passed - Refine 20 00.01/00.04 00.01/00.12 59 Passed - Refine Final Results for Narrow Peak Width: Isolation Efficiency: 50, Score: 78 Finish Quad Tune Start Post Tune Calibration(may take a few min)..... Progress 0%...... Progress 3%...... Progress 4%...... Post Tune Calibration failed, abort! Abort called, terminate the calibration Stop Spraying Calibrant Solution System Tune did not complete due to the following reason(s): Post Tune Calibration failed Completed QTOF System Tune in Positive Polarity