论文总字数：17012字

摘 要

建立了一种快速、灵敏的液相色谱-质谱串联（LC/MS/MS）方法测定大鼠血浆中汉防己甲素的含量, 方法已充分验证，并被成功地应用于SD大鼠单次给药后体内药代动力学研究。通过乙酸乙酯液液萃取处理样品。汉防己甲素和阿霉素（内标）通过BDS HYPERSIL-C₈ 柱分离，流动相为5 mmoL·L^-1 乙酸铵-乙腈（54:46,V/V），含0.3%甲酸。浓度测定是在三重四极质谱的多反应监测模式下进行。离子优化利用ESI离子源（ ）和选择性使用MS / MS分析实现，汉防己甲素和内标的离子对分别是m/z 623.1→381.8和m/z 544.4→397.6。这种定量分析大鼠血浆中汉防己甲素的方法在浓度范围是0.5～400 ng·mL^-1,,具有良好的线性，定量下限为0.5 ng·mL^-1。准确度(RE)是在-0.4%～2.6%之间，日内和日间精密度(RSD)分别在4.6%～7.3%和3.9%～4.9%。分析无内源性化合物的干扰，并且汉防己甲素在整个试验期间是稳定的。该方法已成功应用于汉防己甲素以2.5mg·kg^-1 剂量对大鼠单次尾静脉注射给药的药代动力学研究，结果证实这种方法满足汉防己甲素的药代动力学研究的要求。

关键词：LC/MS/MS 汉防己甲素 药代动力学

Establishment of a liquid chromatographic/mass spectrometry method for quantification of tetrandrine in rat plasma and its application to pharmacokinetic study

Abstract

A rapid and sensitive liquid chromatography-tandem mass spectrometric method (LC/MS/MS) for the determination of tetrandrine in rat plasma has been developed, fully validated and successfully applied to pharmacokinetic study in Sprague–Dawley (SD) rats after a single oral administration.Sample preparation involves a liquid–liquid extraction with ethyl acetate. Tetrandrine and Doxorubicin (internal standard) were well separated by LC with a BDS HYPERSIL-C₈ column using 5 mmoL·L^-1 ammonium acetate-acetonitrile solution （54:46,V/V） containing 0.4% formic acid as mobile phase. Detection was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode. The ionization was optimized using ESI( ) and selectivity was achieved using MS/MS analysis, m/z 623.1→381.8 and m/z 544.4→397.6 for tetrandrine and I.S., respectively. The present method exhibited good linearity over the concentration range of 0.5～400 ng·ml^-1 for tetrandrine in rat plasma with a lower limit of quantification of 0.55 ng·ml^-1. The accuracy ranged from -0.4% to 2.6%,the intra- and inter-day precision were from 4.6% to 7.3% and from 3.9% to 4.9%, respectively.No endogenous compounds were found to interfere with the analysis,and tetrandrine was stable during the whole assay period. The method was successfully applied to a pharmacokinetic study after a tail intravenous injection administration of tetrandrine to SD rats with a single dose of 2.5mg·kg^-1. The results confirm that the assay is suitable for the pharmacokinetic study of tetrandrine.

Keywords:LC/MS/MS;tetrandrine;Pharmacokinetics

目 录

摘 要 Ⅰ

Abstract Ⅱ

第一章 文献综述 1

1.1 汉防己甲素的药理及药效作用 2

1.2汉防己甲素药代动力学研究概况 3

1.3汉防己甲素分析方法研究进展 4

1.4总结 4

第二章 实验 5

2.1 化学药品和试剂 5

2.2 实验动物 5

2.3 LC / MS / MS条件与评价 6

2.4 溶液配制 6

2.5 血浆样品的处理 6

2.6 药代动力学研究 7

2.7 方法验证 7

第三章 结果与讨论 9

3.1 LC / MS / MS方法 9

3.2 验证方法 9

3.2.1 标准曲线和最低定量限 9

3.2.2 特异性和敏感性 10

3.2.3 回收率 11

3.2.4 基质效应 12

3.2.5 准确度（RE）和精密度(RSD) 13

3.2.6 稳定性 14

3.2.7 大鼠药代动力学实验结果 15

3.3 药代动力学研究 15

第四章 结论 17

参考文献 18

致谢 21

第一章 文献综述

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