Here, we are evaluating the performance of Astral compared to the Orbitrap for DIA quantitation. To do this, we have two main experiments:
mzML files are not included on this page due to space considerations and the fact that they are functionally the same as raw files. mzML files can be generated using the ProteoWizard Docker image available on Docker Hub at “ proteowizard/pwiz-skyline-i-agree-to-the-vendor-licenses:3.0.22335-b595b19”.
To re-generate the files in MSConvert, the following command can be used: msconvert *.raw --simAsSpectra --ignoreUnknownInstrumentError --filter "peakPicking true"
Below is a summary of the run types.
File prefix | Sample | Analyzer | Normalized AGC Target | Maximum injection time | Isolation Window | Mass range | Column | Gradient length (min) |
---|---|---|---|---|---|---|---|---|
20230403_OLEP08_EV_1ug_MB_30min_AS_10ms_4Th_ | EV plasma | Astral | 500% | 10 ms | 4 Th | 400-1000 | 110 cm | 30 |
20230403_OLEP08_EV_1ug_MB_30min_AS_3p5ms_2Th_ | EV plasma | Astral | 500% | 3.5 ms | 2 Th | 400-1000 | 110 cm | 30 |
20230403_OLEP08_TP_1ug_MB_30min_AS_10ms_4Th_ | Total plasma | Astral | 500% | 10 ms | 4 Th | 400-1000 | 110 cm | 30 |
20230403_OLEP08_TP_1ug_MB_30min_AS_3p5ms_2Th_ | Total plasma | Astral | 500% | 3.5 ms | 2 Th | 400-1000 | 110 cm | 30 |
20230404_OLEP08_EV_1ug_MB_60min_AS_15ms_4Th_ | EV plasma | Astral | 500% | 15 ms | 4 Th | 400-1000 | 110 cm | 60 |
20230404_OLEP08_EV_1ug_MB_60min_AS_dDIA_15ms_2Th_ | EV plasma | Astral | 500% | 15 ms | 2 Th | 300 Th dDIA | 110 cm | 60 |
20230404_OLEP08_TP_1ug_MB_30min_AS_dDIA_20ms_4Th_ | Total plasma | Astral | 500% | 20 ms | 4 Th | 300 Th dDIA | 110 cm | 30 |
20230404_OLEP08_TP_1ug_MB_60min_AS_15ms_4Th_ | Total plasma | Astral | 500% | 15 ms | 4 Th | 400-1000 | 110 cm | 60 |
20230404_OLEP08_TP_1ug_MB_60min_AS_dDIA_15ms_2Th_ | Total plasma | Astral | 500% | 15 ms | 2 Th | 300 Th dDIA | 110 cm | 60 |
20230405_OLEP08_EV_1ug_MB_30min_AS_dDIA_20ms_4Th_ | EV plasma | Astral | 500% | 20 ms | 4 Th | 300 Th dDIA | 110 cm | 30 |
20230405_OLEP08_EV_1ug_MB_60min_AS_23ms_2Th_ChrLib_ | EV plasma | Astral | 500% | 23 ms | 2 Th | 100 Th GPF | 110 cm | 60 |
20230405_OLEP08_EV_1ug_MB_60min_OT_23ms_2Th_ChrLib_ | EV plasma | Orbitrap | 800% | 23 ms | 2 Th | 100 Th GPF | 110 cm | 60 |
20230406_OLEP08_MMCC_1ug_MB_24min_AS_10ms_4Th_ | MMCC | Astral | 500% | 10 ms | 4 Th | 400-1000 | 110 cm | 24 |
20230406_OLEP08_MMCC_1ug_MB_24min_AS_3p5ms_2Th_ | MMCC | Astral | 500% | 3.5 ms | 2 Th | 400-1000 | 110 cm | 24 |
20230406_OLEP08_MMCC_1ug_MB_24min_AS_dDIA_10ms_2Th_ | MMCC | Astral | 500% | 10 ms | 2 Th | 300 Th dDIA | 110 cm | 24 |
20230406_OLEP08_MMCC_1ug_MB_24min_OT_75Th_23ms_2Th_ | MMCC | Orbitrap | 800% | 23 ms | 2 Th | 500-575 | 110 cm | 24 |
20230410_OLEP08_HeLa_1ug_MB_24min_AS_23ms_2Th_ChrLib_ | 200 ng HeLa | Astral | 500% | 23 ms | 2 Th | 100 Th GPF | 110 cm | 24 |
MMCC: Matrix-matched calibration curve of HeLa into SILAC labeled HeLa. Consists of 9 dilution points injected in triplicate.
EV plasma: 1 ug EV enriched plasma
Total plasma: 1 ug of total plasma
GPF: 6 gas phase fraction runs, each covering 100 Da range
dDIA: dynamic DIA