Rapid evaluation of 25 key sphingolipids and phosphosphingolipids in human plasma by LC-MS/MS.
We report on a new, sensitive, and fast LC-MS/MS method for the simultaneous determination of 25 key sphingolipid components in human plasma, including phosphorylated sphinganine and sphingosine, in a single 9-min run. This method enables an effective and high-throughput coverage of the metabolic changes involving the sphingolipidome during physiological or pathological states. The method is based on liquid-liquid extraction followed by reversed-phase LC-MS/MS. Exogenous odd-chain lipids are used as cost-effective but reliable internal standards. The method was fully validated in surrogate matrix and naive human plasma following FDA guidelines. Sample stability and dilution integrity were also tested and verified.
Department of Drug Discovery and Development, Istituto Italiano di Tecnologia, via Morego 30, 16163, Genoa, Italy.
Abdul
A
Basit
Daniele
D
Piomelli
Andrea
A
Armirotti
D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000191:economics / Q000379:methods; D005260:Female; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000191:economics / Q000379:methods; D008297:Male; D010743:Phospholipids / Q000097:blood*; D015203:Reproducibility of Results; D013107:Sphingolipids / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000379:methods*
D010743:Phospholipids; D013107:Sphingolipids