Masshunter mass profiler 10

For DTIM-MS, Agilent
IM-MS Browser 10.0 was used for single-field calibration, evaluation
of stepped-field ^DTCCS_N2, and manual inspection when required.
Agilent MassHunter Mass Profiler 10.0 was used for feature extraction
(peak picking) of triplicate LC-DTIM-MS measurements following single-field
calibration and direct infusion. PNNL Preprocessor 3.0 (2021.04.21)
was used for demultiplexing and data preprocessing steps.^{47 (link)}For TIM-MS, Bruker TASQ software (version
2021) was used to analyze all data acquired using the timsTOF Pro.
This included automatic recalibration, generation of high-resolution
extracted ion chromatograms (EICs), feature detection (peak picking),
and ^TIMCCS_N2 calculation. Full details of used data processing
can be found in the Supporting Information. Exported data was restructured, analyzed, and visualized using
Microsoft Office (Excel and Powerpoint) and R (4.1.2)⁴⁸ together with RStudio (2021.9.1.372)⁴⁹ (see the Supporting Information).

Single-field calibration for DTIM-MS was applied using Agilent IM-MS Browser 10.0. Single-field calibrated data was demultiplexed and pre-processed using PNNL Preprocessor 3.0 (2021.04.21) [27 ], and Agilent MassHunter Mass Profiler 10.0 was used for peak picking and alignment of triplicate measurements [14 ].
For TWIM-MS, DriftScope V.2.8 included in MassLynx 4.2 software (Waters) was used to determine the ^TWCCS_N2 calibration functions, which were saved into corresponding measurement data files. Individual data files were investigated using DriftScope and MS-DIAL 4.60 [28 (link), 29 ] was used to batch-process TWIM-MS data. To this end, datafiles in raw format were converted to.ibf files using the built-in converter. Settings used for peak picking and alignment are provided in the Electronic Supplementary information. ^TWCCS_N2 values were calculated from arrival times using the Enhanced Duty Cycle (EDC) coefficient to correct arrival times [30 (link)], and a detailed description of the applied calibration approach is presented in the Electronic Supplementary Information.
Finally, Microsoft® Office® (Excel® and PowerPoint®) and R (4.1.2) [31 ] together with RStudio (2021.9.1.372) [32 ] were used for data analysis, visualization and creation of final figures.