Objective: To establish ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of 22 phospholipids in serum. Methods: In September 2022, Using synthetic non endogenous phospholipids as internal standard, phospholipids in serum were extracted by methanol-dichloromethane (2∶1, V/V) protein precipitation method. Chromatographic separation was achieved on an ACQUITY UPLC BEH shield RP18 column, and the mobile phase was methanol/water (5∶95, V/V) containing 10 mM ammonium formate and methanol. Detection was performed in multiple reaction monitoring mode with ion mode switching. And the method was applied by analyzing phospholipids in the serum of coal workers' pneumoconiosis patients. Results: The 22 phospholipids showed good linear relationships in their respective concentration ranges and the correlation coefficients were higher than 0.990. The spiked recoveries of the 22 phospholipids were 81.03%-121.63% at the three spiked levels. The intra-assay were less than 14.52%, and the inter-assay were less than 15.00%. Conclusion: The method with the advantages of simplicity, stability and high sensitivity, and it can be used for the analysis of phospholipids in serum.
目的： 建立血清中22种磷脂的超高效液相色谱-串联质谱（UPLC-MS/MS）检测分析方法。 方法： 于2022年9月，以合成的非内源性磷脂作为内标，用甲醇-二氯甲烷（2∶1，v/v）蛋白沉淀法提取血清中磷脂。用ACQUITY UPLC BEH shield RP18柱分离，流动相为含10 mM甲酸铵的甲醇/水（5∶95，v/v）和甲醇。用多反应监测，在正、负离子模式下分段进行扫描分析。并通过分析煤工尘肺患者血清中的磷脂进行方法应用。 结果： 22种磷脂在各自范围内线性关系良好，相关系数均>0.990，在3个添加水平下方法的加标回收率为81.03%~121.63%，批内精密度≤14.52%，批间精密度≤15.00%。 结论： 该检测方法简单、稳定且灵敏度高，可用于血清中磷脂的分析研究。.
Keywords: Liquid chromatography-tandem mass spectrometry (LC-MS/MS); Phospholipids; Serum.