HPLC-MS/MS法测定人血浆中灯盏乙素的浓度

Determination of scutellarin in human plasma by HPLC-MS/MS

目的:建立测定人血浆中灯盏乙素浓度的HPLC-MS/MS法。方法:采用Platisil ODS色谱柱(250 mm×4.6 mm,5μm),流动相为0.3%甲酸水溶液-甲醇(40∶60),流速为1.0 mL.min-1,以黄芩苷为内标,经乙酸乙酯萃取后,采用电喷雾电离化(ESI)方式和多反应离子监测(MRM)模式进行负离子检测,用于定量分析离子反应分别为m/z 461.1→285.1(灯盏乙素)和m/z 445.1→269.1(黄芩苷)。结果:线性检测范围为0.20～50.00μg.L-1,最低定量浓度0.20μg.L-1,r=0.999 3。低、中、高3个浓度的提取回收率分别为63.5%,69.5%,66.8%,基质效应分别为105.7%,97.5%,96.5%。结论:本法样品处理简单、快速,专属性强,灵敏度高,可用于灯盏乙素在人体内的药代动力学研究。

Objective： To establish an HPLC-MS/MS method for the determination of scutellarin in human plasma. Methods： Scutellarin and an interal standard were separated on a Platisil ODS column （250mm×4.6mm,5μm） ; The mobile phase was consisted of 0.3% formic acid-methanol （40： 60） at a flow rate of 1.0 mL·min^-1. The drug and an internal standard （baicalin） were extracted from plasma with ethyl acetate. A mass spectrometer equipped with electrospray ionization source was used as detector operated in the negative ion mode. A multiple reaction monitoring （MRM） mode using the transition of m/z 461.1→285.1 and m/z 445.1→269.1 was used to quantify scutellarin and interal standard, respectively. Results ： The linear range of scutellarin was 0.20 50.00 μg·L^-1, and the minimum detective concentration was 0.20 μg·L^-1, r = 0. 999 3. The extraction recoveries of the three concentrations were 63.5% , 69.5% and 66.8% , and the matrix effects were 105.7% , 97.5% and 96.5% , respectively. Conclusion： The method is sensitive, rapid, convenient, and is proven to be possible for investigation of scutellarin in human plasma.