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WiSIM DIA Method Set-upOrbitrap Fusion MS

MS Instrument Control Software v.1.1 SP1

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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

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

tSIM Orbitrap

tMS2 Ion Trap

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WiSIM DIA, how does it work?

SIM 400 – 600 m/z

MS2 No. 1………………MS2 No.17

1st MS1

SIM 600 – 800 m/z

Ion

Tra

pO

T24

0k

3.6

sec

SIM 800 – 1000 m/z

2nd MS1MS2 No. 1………………MS2 No.17

Ion

Tra

p

3rd MS1MS2 No. 1………………MS2 No.17

Ion

Tra

pO

T24

0kO

T24

0k

17 Windows of 12 amu each

17 Windows of 12 amu each

17 Windows of 12 amu each

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WiSIM DIA Method Set-up at a glance:

SIM• 400-600, 600-800, 800-1000 m/z• 200 amu ion trap isolation width• Resolution@ 240 000• AGC: 3e4• 50 ms Max injection • Profile

tMS2• 12 amu quadrupole isolation• 150-1850 m/z mass range• Ion Trap detection• AGC: 5e4• 35 ms Max Injection • CID 30 • Rapid Scan rate

X3

X3

6 experiments:

406 514418 526430 538442 550454 562466 574478 586490 598502

606 714618 726630 738642 750654 762666 774678 786690 798702

806 914818 926830 938842 950854 962866 974878 986890 998902

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WiSIM DIA Method Set-up:

6 experiments:-Three HR/AM tSIM-Three Ion trap tMS2 CID

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Experiment 1:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 2:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 2:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 3:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 4:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 4:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 5:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 6:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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Experiment 6:

SIM 400-600 amu SIM 600-800 amu SIM 800-1000 amu

406 418 430 442 598 606 618 630 642 898 806 818 830 842 998

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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.

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For More details, please see

Application Note 600

And:

http://planetorbitrap.com/orbitrap-fusion-tips