To investigate the biological functions and roles of sphingolipids, sensitive and compound-specific methods are required to measure their levels in biological samples. Liquid chromatography-mass spectrometry (LC-MS) using electrospray ionization (ESI) is suitable for the reliable simultaneous analysis of multiple compounds. In addition, the selected reaction monitoring (SRM) mode of tandem mass spectrometry (MS/MS) is effective to quantify with high sensitivity and selectivity. Therefore, LC-MS/MS came to be utilized for simultaneous analysis of the sphingolipids in vivo. Useful methods for the sphingolipids and related features are also summarized. The following protocol demonstrates information on determination of sphingolipids, especially sphingosine and sphingosine-1- phosphate, by LC-ESI-MS/MS in biological samples such as cell lysates, plasma, serum, orurine.
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