Isotopic exchange during derivatization of platelet activating factor for gas chromatography-mass spectrometry

P. E. Haroldsen, S. J. Gaskell, S. T. Weintraub, R. N. Pinckard

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

One approach to the quantitative analysis of platelet activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycerol-3-phosphocholine; also referred to as AGEPC, alkyl glyceryl ether phosphocholine) is hydrolytic removal of the phosphocholine group and conversion to an electron-capturing derivative for gas chromatography-negative ion mass spectrometry. [2H3]Acetyl-AGEPC has been commonly employed as an internal standard. When 1-hexadecyl-2-[2H3]acetyl glycerol (obtained by enzymatic hydrolysis of [2H3]-C16:0 AGEPC) is treated with pentafluorobenzoyl chloride at 120°C, the resulting 3-pentafluorobenzoate derivative shows extensive loss of the deuterium label. This exchange is evidently acid-catalyzed since derivatization of 1-hexadecyl-2-acetyl glycerol under the same conditions in the presence of a trace of 2HCl results in the incorporation of up to three deuterium atoms. Isotope exchange can be avoided if the reaction is carried out at low temperature in the presence of base. Direct derivatization of [2H3]-C16:0 AGEPC by treatment with pentafluorobenzoyl chloride or heptafluorobutyric anhydride also results in loss of the deuterium label. The use of [13C2]-C16:0 AGEPC as an internal standard is recommended for rigorous quantitative analysis.

Original languageEnglish (US)
Pages (from-to)723-729
Number of pages7
JournalJournal of lipid research
Volume32
Issue number4
StatePublished - 1991
Externally publishedYes

Keywords

  • carbon-13 labeling
  • deuterium exchange
  • electron capture mass spectrometry
  • quantitative analysis

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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