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. 2023 Jul 17;14:4263. doi: 10.1038/s41467-023-40046-x

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Extracted ion mobilograms (EIMs) of five GPL standards detected in positive ion mode ([M + H]⁺ for PC, PE, and PS, [M + NH₄]⁺ for PG and PI, upper panel) and negative ion mode ([M–H]⁻ for PG, PI, PS, and PE, [M + HCO₃]⁻ for PC, lower panel). b Negative ion mode MS¹ spectrum of a 10:1 mixture of PC 16:0/18:1 and PC 18:0/16:0, detected as [M + HCO₃]⁻. c Zoomed-in MS² CID spectrum of m/z 822.59 formed in panel b without TIMS separation. Only the m/z region of sn-1 fragment is shown. Inset shows the structure of the sn-1 18:0 fragment (m/z 447) derived from MS² CID of PC 18:0/16:0 ([M + HCO₃]⁻). d EIMs of m/z 822.585 (black trace) formed in panel b with a scan rate of 1 V/ms and 0.06 V/ms during TIMS separation. The +2 Da isotope of PC 16:0/18:1 and the mono-isotope of PC 18:0/16:0 are gaussian deconvoluted and shaded in red and blue, respectively. Resolving power (R) is listed. Mobility-resolved (e) MS¹ spectrum and (f) MS/MS spectrum of m/z 822.59 isolated from 1/K₀ of 1.464–1.482 V•s/cm² in panel d. g Comparison of the purity of PC 18:0/16:0 without and with TIMS separation (N = 3 independent experiments). Bars represent mean values ± standard deviation (SD). (h) Schematic presentation of triFAP PB-MS² CID for generating C = C diagnostic ions. (i) Extracted ion chromatograms (EICs) of unreacted PE 16:0/18:1(n-9) (blue trace) and triFAP-modified PE 16:0/18:1(n-9) (red trace). j EIMs of unreacted PE 16:0/18:1(n-9) (blue trace) and triFAP-modified PE 16:0/18:1 (n-9) (red trace). k MS² CID spectrum of triFAP-derivatized PE 16:0/18:1(n-9) ([^PBM + H] ⁺, m/z 892.6). Source data are provided as a Source Data file.