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WiSIM DIA Method Set-up Orbitrap Fusion MS MS Instrument Control Software v.1.1 SP1

WiSIM DIA Method Set-up Orbitrap Fusion MS MS Instrument Control Software v.1.1 SP1. WiSIM DIA, how does it work?. Unique to the Orbitrap Fusion MS: Three HR/AM SIM are used to cover precursors ions from 400 to 1000 m/z MS1 ultra high resolution quantification

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WiSIM DIA Method Set-up Orbitrap Fusion MS MS Instrument Control Software v.1.1 SP1

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  1. WiSIM DIA Method Set-upOrbitrap Fusion MSMS Instrument Control Software v.1.1 SP1

  2. WiSIM DIA, how does it work? • Unique to the Orbitrap Fusion MS: • Three HR/AM SIM are used to cover precursors ions from 400 to 1000 m/z MS1 ultra high resolution quantification • In parallel with each HR/AM SIM scan, 17 sequential Ion Trap CID MS/MS with 12 amu isolation windows are acquired to cover the associated 200 amu HR/AM SIM mass range MS2 confirmation tSIM Orbitrap SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu tMS2 Ion Trap 406 806 606 418 618 818 430 630 830 442 642 842 898 598 998

  3. WiSIM DIA, how does it work? OT 240k 1st MS1 SIM 600 – 800 m/z SIM 800 – 1000 m/z SIM 400 – 600 m/z MS2 No. 1 … … … … … … MS2 No.17 17 Windows of 12 amu each Ion Trap OT 240k 2nd MS1 MS2 No. 1 … … … … … … MS2 No.17 17 Windows of 12 amu each Ion Trap 3.6 sec OT 240k 3rd MS1 MS2 No. 1 … … … … … … MS2 No.17 17 Windows of 12 amu each Ion Trap

  4. WiSIM DIA Method Set-up at a glance: 6 experiments: SIM • 400-600, 600-800, 800-1000 m/z • 200 amu ion trap isolation width • Resolution@ 240 000 • AGC: 3e4 • 50 msMax injection • Profile X3 tMS2 • 12 amuquadrupole isolation • 150-1850 m/z mass range • Ion Trap detection • AGC: 5e4 • 35 ms Max Injection • CID 30 • Rapid Scan rate X3

  5. WiSIM DIA Method Set-up: 6 experiments: -Three HR/AM tSIM -Three Ion trap tMS2 CID

  6. Experiment 1: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  7. Experiment 2: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  8. Experiment 2: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  9. Experiment 3: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  10. Experiment 4: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  11. Experiment 4: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  12. Experiment 5: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  13. Experiment 6: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  14. Experiment 6: SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu 806 606 406 818 418 618 430 630 830 442 842 642 898 598 998

  15. WiSIM-DIA advantages • Low detection limits, high selectivity is achieved using 240,000 SIM scan.  • High sensitivity Ion Trap CID MS2 is used for confirmation of the low abundant components. • Unsurpassed speed with 2 detectors working in parallel allows for 3.6 s cycle. • LOD and LOQ down to 10 attomole. • CVs are better than 15 % for 85 % of compounds, better than 20% for 95% of compounds. • Linear dynamic range up to 4 orders of magnitude.

  16. For More details, please see http://planetorbitrap.com/orbitrap-fusion-tips • And: • Application Note 600

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