标题：LC-MS/MS法测定大鼠血浆中芹黄春浓度及其在药代动力学研究中的应用

      作者：李海龙 1，2，尹航 2，文琪 2，谭银丰 1，2，陈峰 1，2，张小坡 1，2，赖伟勇 1，2
    (1.海南省药用植物研究与开发重点实验室，海南 海口 571199；
2.海南医学院药学院，海南 海口 571199)

      卷次： 2016年27卷13期

      【摘要】 目的 研究大鼠分别灌胃和静脉给药芹黄春（芹菜素-7-O-β-D吡喃葡萄糖苷）溶液后，血浆中的芹黄春
药物浓度及其药代动力学性质。方法 6只雌性Sprague Dawley大鼠随机分为两组，每组3只，分别经灌胃(10 mg/kg)
和静脉注射(2 mg/kg)给予芹黄春溶液后，按设计的时间点从大鼠眼内眦静脉丛采集血液样品，置于肝素化的离心管
中，低温离心得血浆。采用甲醇沉淀法处理血浆生物样品，利用LC-MS/MS方法对血浆生物样品进行分析。药代动
力学参数由 Kinetica 2000软件进行计算。结果 芹黄春浓度在1~2 000 ng/mL范围内线性关系良好(r=0. 997 4)，定
量下限为1 ng/mL，低(8 ng/mL)、中(100 ng/mL)、高(1 200 ng/mL) 3个浓度的提取回收率均大于90%，日内日间精密度和
稳定性的RSD均小于12.8%，日内日间准确度在91.5%~107%，方法学考察均符合要求。大鼠经静脉注射和灌胃芹黄春
后，Cmax分别为(2 363±96) ng/mL和(13.3±5.8) ng/mL，AUC0~∞分别为(796±201) h· ng/mL和(28.9±9.2) h· ng/mL，大鼠经
静脉注射和后 t1/2为(1.20±0.17) h，芹黄春的口服生物利用度约为0.6%。结论 所建立的LC-MS/MS分析方法方便、快
速，灵敏度高，准确度好，可用于大鼠血浆芹黄春浓度测定及其药代动力学的研究。

      【关键词】 大鼠；芹黄春；液相色谱-串联质谱联用法；药代动力学

      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2016）13—2065—05


Determination of apigetrin in rat plasma by LC-MS/MS and its application in the pharmacokinetic study.
LI
Hai-long 1, 2, YIN Hang 2, WEN Qi 2, TANG Yin-feng 1, 2, CHEN Feng 1, 2, ZHANG Xiao-po 1, 2, LAI Wei-yong 1, 2. 1. Hainan
Provincial Key Laboratory of R&D of Tropical Herbs, Haikou 571199, Hainan, CHINA; 2. School of Pharmacy, Hainan
Medical University, Haikou 571199, Hainan, CHINA

【Abstract】 Objective To quantify apigetrin in rat plasma by LC-MS/MS, and investigate pharmacokinetic
properties of apigetrin after oral or intravenous administration of apigetrin solution to rats. Methods Six female
Sprague Dawley rats were randomly divided into two groups, with three rats in each group, to receive a single intrave-
nous dose (2 mg/kg) or an oral dose of apigetrin solution (10 mg/kg). Serial blood samples were collected from the or-
bital sinus under light ether anesthesia. Each plasma sample was precipitated with the IS-spiked methanol solution.
The mixture was well followed by a vortex shake for 5 min and centrifuged at 13 000 r/min for 10 min. The resulting
supernatant was directly applied for LC-MS/MS analysis. Pharmacokinetic parameters were determined with Kineti-
ca 2000 software. Results Good linearity was found in the range of 1~2 000 ng/mL for apigetrin. The lower limit
determination of apigetrin was 1 ng/mL. The relative extraction recoveries were higher than 90% at three concentrations
(8 ng/mL, 100 ng/mL, 1 200 ng/mL). The RSD values for stability of intraday and interday precision were always less
than 12.8%. Intraday and interday accuracy ranged from 91.5% to 107%. The accuracy, precision, stability, extraction re-
coveries and linearity were found to be within the acceptable criteria. This method was successfully applied to a pharma-
cokinetic study after oral or intravenous administration of apigetrin solution to rats. The Cmax was (2 363±96) ng/mL and
(13.3±5.8) ng/mL respectively. The AUC0~∞ was respectively (796±201) h· ng/mL and (28.9±9.2) h· ng/mL. The plas-
ma t1/2 after apigetrin through intravenous administration was (1.20±0.17) hour and its oral bioavailability was very low
(0.6%). Conclusion The LC-MS/MS method for measurement of apigetrin was developed and validated, which can be
successfully applied in determination of apigetrin in rat plasma and pharmacokinetics study.

      【Key words】 Rat; Apigetrin; LC-MS/MS; Pharmacokinetics
·论 著·
6350.2016.13.001