Simultaneous quantitative determination method for sphingolipid metabolites by liquid chromatography/ionspray ionization tandem mass spectrometry

Nariyasu Mano, Yoshiya Oda, Koji Yamada, Naoki Asakawa, Kouichi Katayama

Research output: Contribution to journalArticlepeer-review

121 Citations (Scopus)

Abstract

Sphingolipid metabolites ceramide, sphingomyelin, sphingosine, psycosine, sphingosylphosphorylcholine, and dimethylsphingosine were separated and simultaneously quantitated by liquid chromatography/ionspray ionization tandem mass spectrometry (LC/MS/MS). The use of glassware throughout minimized losses due to adsorption and the pretreatment of this method consisted of simple liquid-liquid extraction procedure with a mixture of chloroform and methanol. After separation on a short C18 silica column eluted in a gradient mode, the metabolites were detected by MS/MS. This assay allows simultaneously quantification of these metabolites over a range of at least 0.1 to 100 ng/106 cells. The LC/MS/MS analyses took 10 to 15 min per sample and we could examine up to 50 samples per day. We also detected endogenous sphingosine 1-phosphate in HL-60 cells. The utility of the method was demonstrated by examining changes in metabolites levels in HL-60 cells after treatment with sphingomyelinase. It was found that sphingomyelinase from Bacillus cereus may have selectivity for acyl chain length.

Original languageEnglish
Pages (from-to)291-300
Number of pages10
JournalAnalytical Biochemistry
Volume244
Issue number2
DOIs
Publication statusPublished - 1997 Jan 15
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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