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Development of an LC–MS/MS Method for the Analysis of Free Sphingoid Bases Using 4‐Fluoro‐7‐nitrobenzofurazan (NBD‐F)
Author(s) -
Ishikawa Toshiki,
Imai Hiroyuki,
Maki KawaiYamada
Publication year - 2014
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-013-3871-6
Subject(s) - chemistry , derivatization , chromatography , electrospray ionization , tandem mass spectrometry , mass spectrometry , sphingolipid , reagent , selected reaction monitoring , liquid chromatography–mass spectrometry , high performance liquid chromatography , organic chemistry , biochemistry
The molecular species of sphingoid bases were tagged with the fluorescent amino group reagent, 4‐fluoro‐7‐nitrobenzofurazan (NBD‐F). The NBD‐sphingoid bases were analyzed by a highly selective and sensitive liquid chromatography–electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS) technique capable of reliable detection of several fmol of the derivatives. Lipid extracts from plant samples were derivatized with NBD‐F, and all nine species of free sphingoid bases present in plant sphingolipids were separated and quantified for the first time; a complete baseline resolution was achieved for cis ‐8 and trans ‐8 isomers of sphingoid bases by reversed phase HPLC on a C 18 column. The extraction and derivatization procedures and LC–MS/MS method can facilitate the progress of the studies for seeking the active components of sphingoid bases species in response to biological challenges.