HPLC-MS/MS法检测人血浆中游离花生四烯酸浓度

目的: 建立测定人血浆中花生四烯酸浓度的液相色谱串联质谱法。 方法: 以奥美拉唑为内标,待测血浆经乙酸乙酯萃取后,采用Agilent SB-C₁₈(2.1 mm×100 mm,3.5 μm)色谱柱,以水-甲醇(10:90,v/v)为流动相,流速0.2 mL·min^-1,柱温25 ℃;采用电喷雾离子源(ESI)负离子模式将样品离子化,多反应监测(MRM)模式下对花生四烯酸(m/z 303.2→259.2)和奥美拉唑(m/z 344.2→191.1)进行测定。 结果: 花生四烯酸在497.88~4997.24 ng·mL^-1范围内线性关系良好(r=0.9989)。低、中、高浓度的平均回收率均大于70%,批内与批间RSD均小于10%。 结论: 本方法简单、快速、灵敏,适用于大样本量血浆中游离花生四烯酸的分析研究。

Objective: To develop an HPLC-MS/MS method for determination of arachidonic acid in human plasma. Methods: Omeprazole was used as internal standard.Plasma was extracted with ethyl acetate and samples were separated on an Agilent SB-C₁₈ column (2.1 mm×100 mm,3.5 μm)with a mobile phase of water-methanol(10:90,v/v).The flow rate was 0.2 mL·min^-1 and the temperature of the column was 25 ℃.The eletrospray ionization(ESI)with negative source of the mass spectrometer and detected in multiple reaction monitor(MRM)mode to detect arachidonic acid(m/z 303.2→259.2)and omeprazole( m/z 344.2→191.1). Results: The calibration curve was linear in the range of 497.88-4997.24 ng·mL^-1(r=0.9989)for arachidonic acid.The recovery of the extraction of arachidonic acid was more than 70%;RSDs of the intra-assay and inter-assay were less than 10%. Conclusion: The method is simple,rapid,sensitive and suitable for determination of arachidonic acid in human plasma.