LC-MS/MS同时测定大鼠血浆中5种银杏酸浓度及其药动学研究

Simultaneous Determination of Five Ginkgolic Acids in Rat Plasma by LC-MS/MS and the Application in Pharmacokinetics Study

关键词:  银杏酸  液相色谱串联质谱法  血药浓度  药动学

Key words:  ginkgolic acids  LC-MS/MS  plasma drug concentration  pharmacokinetics

摘要:

目的 建立LC-MS/MS同时测定大鼠血浆中5种银杏酸的浓度，并进行药动学研究。方法 大鼠血浆样品用乙酸乙酯萃取后，以水杨酸为内标，选用ZORBAX Eclipse XDB C₁₈（2.1 mm×100 mm，3.5 μm）色谱柱，以乙腈-水（均含0.1%甲酸）为流动相梯度洗脱，梯度流速恒定为0.25 mL·min^-1，柱温为30℃，进样量为10 μL，总分析时间为5.0 min，采用电喷雾离子化源，负离子方式，多反应监测（MRM）扫描方式进行监测。结果 血浆中5种银杏酸浓度线性范围为2.0~1 000 μg·L^-1（r>0.99），定量下限为2.0 μg·L^-1；质控样品准确度为92.8%~108.5%；日内、日间RSD均<15%；提取回收率为73.3%~86.2%。结论 该方法快速、灵敏、准确，专属性强，重复性好，适用于同时测定大鼠血浆中5种银杏酸浓度。

Abstract:

OBJECTIVE To develop an LC-MS/MS method for simultaneous determination of five ginkgolic acids in rat plasma and applied to the pharmacokinetics study. METHODS The plasma samples were processed by ethyl acetate extraction, with salicylic acid as an internal standard. The separation was achieved on ZORBAX Eclipse XDB C₁₈ column (2.1 mm×100 mm, 3.5 μm) and eluted with linear gradient using acetonitrile and water (both of containing 0.1% formic acid) at the flow rate of 0.25 mL·min^-1, the injection volume was 10 μL and the column temperature was 30℃. The total time of the analysis was 5.0 min. Detection of the analytes were achieved using negative ion electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. RESULTS The linear calibration curve of five ginkgolic acids obtained concentration range of 2.0-1 000 mg·L^-1(r>0.99), with the lower limit of quantitation (LLOQ) of 2.0 mg·L^-1. The accuracy of was between 92.8%-108.5%, Intra-day and inter-day relative standard deviations were both below 15%. The extraction recovery of low, middle and high concentrations was 73.3%-86.2%, respectively. CONCLUSION The established method is rapid, sensitive, accurate, specific and reliable, and suitable for simultaneous determination of five ginkgolic acids in rat plasma.