4.6 Article

A liquid chromatography-mass spectrometry workflow for in-depth quantitation of fatty acid double bond location isomers

期刊

JOURNAL OF LIPID RESEARCH
卷 62, 期 -, 页码 -

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ELSEVIER
DOI: 10.1016/j.jlr.2021.100110

关键词

double bond location isomers; quantitation; tandem mass spectrometry; liquid chromatography; lipidomics; type 2 diabetes; Patern`o-Buchi; charge-tagging; 2-acetylpiridine; Fatty acids

资金

  1. National Key R&D Program of China [2018YFA0800903]
  2. National Natural Science Founda-tion of China [21722506]

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This study developed a workflow for global quantitation of fatty acids, including C=C location isomers, via charge-tagging Patern`o-Buchi (PB) derivatization and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method enabled deep profiling of unsaturated fatty acids in yak milk powder and pooled human plasma, achieving successful quantitation across a wide range of concentrations.
Tracing compositional changes of fatty acids (FAs) is frequently used as a means of moni-toring metabolic alterations in perturbed biological states. Given that more than half of FAs in the mammalian lipidome are unsaturated, quantitation of FAs at a carbon-carbon double bond (C=C) loca-tion level is necessary. The use of 2-acetylpiridine (2-acpy) as the charge-tagging PB reagent led to a limit of identification in the subnanomolar range for mono-and polyunsaturated as well as conjugated FAs. Conjugated free FAs of low abundance such as FA 18:2 (n-7, n-9) and FA 18:2 (n-6, n-8) were quanti-fied at concentrations of 0.61 +/- 0.05 and 0.05 +/- 0.01 mg per 100 g in yak milk powder, respectively. This workflow also enabled deep profiling of eight satu-rated and 37 unsaturated total FAs across a span of four orders of magnitude in concentration, including ten groups of C=C location isomers in pooled human plasma. A pilot survey on total FAs in plasma from patients with type 2 diabetes revealed that the rela-tive compositions of FA 16:1 (n-10) and FA 18:1 (n-10) were significantly elevated compared with that of normal controls. In this work, we have developed a workflow for global quantitation of FAs, including C=C location isomers, via charge-tagging Patern`o-Buchi (PB) derivatization and liquid chromatog-raphy-tandem mass spectrometry (LC-MS/MS).

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