High-throughput phospholipid profiling system based on supercritical fluid extraction–supercritical fluid chromatography/mass spectrometry for dried plasma spot analysis

The dried blood spots (DBS) and dried plasma spots (DPS) analysis is available as a diagnostic tool for genetic diseases and is useful for the screening of biomarkers. In this study, a high-throughput analytical system based on supercritical fluid extraction–supercritical fluid chromatography with tandem mass spectrometry (SFE–SFC/MS/MS) was constructed for phospholipids profiling of DPS. This system is able to simultaneously perform extraction and separation, allowing phospholipids that have common polar head groups to be analyzed. Phospholipids in only 3 μl of plasma can be extracted in 5 min and analyzed within 15 min using this system. A total of 134 phospholipids, including phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, phosphatidylethanolamine and lysophosphatidylethanolamine, were annotated, and 74 phospholipids were analyzed with good repeatability. The SFE–SFC/MS/MS, which is able to perform high-throughput lipid profiling analysis for clinical diagnosis and drug discovery, may also be suitable for the screening of biomarkers.

► We established an SFE–SFC/MS/MS for profiling of phospholipids in DPS.
► Phospholipids in DPS could be analyzed without any pretreatment in 15 min.
► A PC HILIC column was used for trapping and separating phospholipids in this system.
► 134 phospholipids (PC, LPC, SM, PE, LPE) from only 3 μl plasma could be detected.