Quantification of ceramide levels in mammalian cells by high performance liquid chromatography coupled to tandem mass spectrometry with multiple-reaction-monitoring mode (HPLC-MS/MS-MRM)

摘要

Ceramidesplayanimportantroleinavarietyofcellularfunctionsincludingcelldifferentiationandapoptosis,responsestoDNAdamageandstress,andtranscriptionalevents.Detectionofceramidesinmammaliancellsisrequiredformanybiologicalstudies.Here,wereportavalidatedmethodusingLC-MS/MS-MRMonanAgilent6410triplequadrupolemassspectrometertosimultaneouslyquantifysixceramidesextractedfrommammaliancelllysatesfollowinga5minHPLCgradient.Thismethoddemonstratedoutstandingsensitivity,accuracy,reproducibility,andspeedofanalysis...