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首页> 外文期刊>Planta medica: Natural products and medicinal plant research >A Simple and Sensitive LC-MS/MS Method for Determination of Four Major Active Diterpenoids from Andrographis paniculata in Human Plasma and Its Application to a Pilot Study
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A Simple and Sensitive LC-MS/MS Method for Determination of Four Major Active Diterpenoids from Andrographis paniculata in Human Plasma and Its Application to a Pilot Study

机译：一种简单灵敏的LC-MS / MS法测定人体血浆中穿心莲中四种主要活性二萜的含量及其在先导研究中的应用

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摘要

Andrographis paniculata contains four major active diterpenoids, including andrographolide (1), 14-deoxy-11, 12-didehydroandrographolide (2), neoandrographolide (3), and 14-deoxyandrographolide (4), which exhibit differences in types and/or degrees of their pharmacological activity. Previous pharmacokinetic studies in humans reported only the parameters of compound 1 and its analytical method in human plasma. The purpose of this study was to develop a simple, sensitive, and selective liquid chromatography tandem-mass spectrometry technique for the simultaneous determination of all four major active diterpenoids in the A. paniculata product in human plasma. These four diterpenoids in plasma samples were extracted by a simple protein precipitation method with methanol and separated on a Kinetex C18 column using a gradient system with a mobile phase of acetonitrile and water. The liquid chromatography tandem-mass spectrometry was performed in the negative mode, and the multiple reaction monitoring mode was used for the quantitation. The method showed a good linearity over a wide concentration range of 2.50-500 ng/mL for 1 and over the range of 1.00-500 ng/mL for the other diterpenoids with a correlation coefficient R-2 > 0.995. The lower limit of quantification of 1 was found to be 2.50 ng/mL, while those of the other diterpenoids were 1.00 ng/mL. The intraday and interday accuracy (relative error) ranged from 0.03% to 10.03%, and the intraday and interday precisions (relative standard deviation) were in the range of 2.05-9.67%. The extraction recovery (86.54-111.56%) with a relative standard deviation of 2.78-8.61% and the matrix effect (85.15-112.36%) were within the acceptance criteria. Moreover, these four major active diterpenoids were stable in plasma samples at the studied storage conditions with a relative error <=-9.79% and a relative standard deviation <= 9.26%. Hence, this present method was successfully validated and used in the pilot study to determine the pharmacokinetic parameters of all four major active diterpenoids in human plasma after multiple oral doses of the A. paniculata product were administered to a healthy, Thai female volunteer.
机译：穿心莲有四个主要的活性二萜类化合物，包括穿心莲内酯（1），14-脱氧-11、12-二氢穿心莲内酯（2），新穿心莲内酯（3）和14-脱氧穿心莲内酯（4），它们的类型和/或程度存在差异其药理活性。先前在人体中的药代动力学研究仅报告了化合物1在人体血浆中的参数及其分析方法。这项研究的目的是开发一种简单，灵敏，选择性的液相色谱串联质谱技术，用于同时测定人血浆中沙棘曲霉产品中的所有四种主要活性二萜类化合物。血浆样品中的这四种二萜类化合物通过简单的蛋白质沉淀法用甲醇萃取，并使用带有乙腈和水流动相的梯度系统在Kinetex C18色谱柱上分离。液相色谱串联质谱以负模式进行，并使用多反应监测模式进行定量。该方法在2.50-500 ng / mL的宽浓度范围内显示出良好的线性，对于其他二萜在1.00-500 ng / mL的范围内显示出良好的线性，相关系数R-2> 0.995。 1的定量下限为2.50 ng / mL，而其他二萜的定量下限为1.00 ng / mL。盘中和盘中精度（相对误差）的范围为0.03％至10.03％，盘中和盘中精度（相对标准偏差）的范围为2.05-9.67％。相对标准偏差为2.78-8.61％的萃取回收率（86.54-111.56％）和基质效应（85.15-112.36％）均在可接受的标准内。此外，这四种主要活性二萜类化合物在血浆样品中，在研究的储存条件下是稳定的，相对误差<=-9.79％，相对标准偏差<= 9.26％。因此，在对健康的泰国女性志愿者多次口服剂量的拟南芥产品后，成功地验证了本方法并将其用于前期研究以确定人血浆中所有四种主要活性二萜的药代动力学参数。

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