An LC/MS/MS method for the simultaneous determination of individual sphingolipid species in B cells

•An LC-ESI–MS/MS method was developed to investigate the profile of sphingolipids.•The extraction method was modified to ensure the high recovery of all sphingolipids.•33 molecular species were simultaneously analyzed in a single chromatographic run.•The distribution of sphingolipids were different in healthy and cancerous cells.•The method is easily adaptable to other biological sample types. Comprehensive profiling of sphingolipids is of great importance for clinical and pharmaceutical studies. An LC/MS/MS method was established for the simultaneous separation and quantification of individual sphingolipid species including ceramides, dihydroceramides, glucosylceramides, sphingosine, sphingosine-1-phosphate, sphinganine and sphinganine-1-phosphate. All target individual sphingolipid species were separated and quantified in a single chromatographic run of