Optimization of lipid extraction and analytical protocols for UHPLC-ESI-HRMS-based lipidomic analysis of adherent mammalian cancer cells.

摘要: Lipidomics, which reveals comprehensive characterization of molecular lipids, is a rapidly growing technology used in biomedical research. Lipid extraction critical step lipidomic analysis. However, the effectiveness different lipid extract solvent systems from cellular samples still remains unclear. In current study, protocol reverse-phase liquid chromatography mass spectrometry (LC/MS)-based lipidomics was optimized for and detection lipids human pancreatic cancer cell line PANC-1. Four methods were compared, including methanol/methyl-tert-butyl ether (MTBE)/H2O, methanol/chloroform, methanol/MTBE/chloroform, hexane/isopropanol. Data acquired using high-resolution positive negative ion modes respectively. The number total detected identified assessed with aid automated identification software LipidSearch. Results demonstrated that methanol/MTBE/H2O provided better efficiency classes, chosen as system. This validated method enables highly sensitive reproducible analysis variety further applied to an untargeted study on PANC-1 lines. Moreover, this system can be other lines similar chemical physical properties.