色谱

色谱 ›› 2014, Vol. 32 ›› Issue (3): 290-293.DOI: 10.3724/SP.J.1123.2013.11027

• 技术与应用 • 上一篇    下一篇

高效液相色谱-串联质谱法测定比格犬血浆中的莫诺苷浓度及其药代动力学

熊山1,2, 李敬来2, 朱秀清2, 王晓英2, 吕圭源1, 张振清2   

  1. 1. 浙江中医药大学, 浙江 杭州 310053;
    2. 军事医学科学院毒物药物研究所, 北京 100850
  • 收稿日期:2013-11-14 修回日期:2014-01-01 出版日期:2014-03-08 发布日期:2014-03-04
  • 通讯作者: 张振清，吕圭源
  • 基金资助:

    科技部“重大新药创制”科技重大专项（2012ZX09301003-001-007）.

Determination of morroniside concentration in beagle plasma and its pharmacokinetics by high performance liquid chromatography-tandem mass spectrometry

XIONG Shan1,2, LI Jinglai2, ZHU Xiuqing2, WANG Xiaoying2, LÜ Guiyuan1, ZHANG Zhenqing2   

  1. 1. Zhejiang Chinese Medical University, Hangzhou 310053, China;
    2. Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China
  • Received:2013-11-14 Revised:2014-01-01 Online:2014-03-08 Published:2014-03-04

PDF

237

被引次数

8

摘要：

运用高效液相色谱-串联质谱联用技术，建立了简单、快速、灵敏的比格犬灌胃莫诺苷后血药浓度的检测方法。血浆样品采用蛋白质沉淀法处理，以芍药苷作为内标，色谱柱为Inertsil ODS-SP色谱柱（50 mm×2.1 mm，5 μm），流动相为水（含1 mmol/L甲酸钠）-乙腈，梯度洗脱，流速0.4 mL/min。采用电喷雾离子源（ESI），正离子多反应监测（MRM）模式。绘制血药浓度-时间曲线，并采用DAS 2.0软件计算药代动力学参数。方法学实验结果表明内源性杂质不干扰莫诺苷和内标的测定，线性范围为2~5000 μg/L（r=0.9966），定量限为2 μg/L。方法精密度、准确度、回收率和基质效应均符合生物样品测定的要求，适合比格犬血浆中莫诺苷浓度的测定，可以应用该方法进行莫诺苷的药代动力学研究。比格犬灌胃莫诺苷3个剂量（5、15、45 mg/kg）后的血药浓度-时间曲线下面积（AUC（0-∞））分别为（1631.20±238.50）、（3984.05±750.38）、（10397.64±3156.34）μg/L·h，与给药剂量之间呈现良好的线性关系。

关键词: 比格犬, 莫诺苷, 血浆, 药代动力学, 液相色谱-串联质谱

Abstract:

A sensitive, simple and specific high performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) method was developed for the determination of morroniside in the plasma of beagles administered via intragastric (ig) doses of morroniside. The method employed paeoniflorin as the internal standard and extracted by simple protein precipitation. The separation was achieved using an Inertsil ODS-SP column (50 mm×2.1 mm, 5 μm) with mobile phases of 1 mmol/L sodium formate aqueous solution and acetonitrile (gradient elution) at a flow rate of 0.4 mL/min. The detection was accomplished by a mass spectrometer using multiple reaction monitoring (MRM) in positive mode. Pharmacokinetic parameters were fitted by software DAS 2.0. The methodological study showed a good linear relationship of 2-5000 μg/L (r=0.9966) with a sensitivity of 2 μg/L as the limit of quantification. The precision, accuracy, mean recoveries and the matrix effects were satisfied with the requirements of biological sample measurement. The method described above was successfully applied to the pharmacokinetic study of morroniside in the beagle plasma samples. The area under the plasma concentration-time curves (AUC(0-∞)) of morroniside after single ig administration doses of 5, 15 and 45 mg/kg were (1631.20±238.50), (3984.05±750.38) and (10397.64±3156.34) μg/L·h. The relationship between dose and AUC showed a good linearity. The pharmacokinetic property of morroniside was proposed to be linear pharmacokinetics.

Key words: beagle, liquid chromatography-tandem mass spectrometry (LC-MS/MS), morroniside, pharmacokinetics, plasma

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