RRC ID 65067
著者 Hama K, Fujiwara Y, Tabata H, Takahashi H, Yokoyama K.
タイトル Comprehensive Quantitation Using Two Stable Isotopically Labeled Species and Direct Detection of N-Acyl Moiety of Sphingomyelin.
ジャーナル Lipids
Abstract Sphingomyelin (ceramide-phosphocholine, CerPCho) is a common sphingolipid in mammalian cells and is composed of phosphorylcholine and ceramide as polar and hydrophobic components, respectively. In this study, a qualitative liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS/MS) analysis is proposed in which CerPCho structures were assigned based on product ion spectra corresponding to sphingosylphosphorylcholine and N-acyl moieties. From MS/MS/MS analysis of CerPCho, we observed product ion spectra of the N-acyl fatty acids as [RCO2]- ions as well as sphingosylphosphorylcholine. A calibration curve for CerPCho was constructed using two stable isotopically labeled CerPCho species and then used to quantify the CerPCho species in HeLa cells as a proof-of-principle study. The present study proposes an accurate method for quantifying and assigning structures to each CerPCho species in crude biologic samples by LC-ESI-MS/MS/MS analysis.
巻・号 52(9)
ページ 789-799
公開日 2017-9-1
DOI 10.1007/s11745-017-4279-5
PII 10.1007/s11745-017-4279-5
PMID 28770378
PMC PMC5583271
MeSH Acetyltransferases / genetics Acetyltransferases / metabolism Acylation Chromatography, Liquid Fatty Acid Elongases HeLa Cells Humans Spectrometry, Mass, Electrospray Ionization Sphingomyelins / analysis* Sphingomyelins / metabolism Tandem Mass Spectrometry Up-Regulation
IF 1.919
リソース情報
ヒト・動物細胞 HeLa(RCB0007)