丹参提取物中5种酚酸类成分在大鼠血浆中的LC-MS/MS分析

Methodological Study of Five Kinds of Phenolic Acids from Salvia miltiorrhiza Extract in Rat Plasma by LC-MS/MS

目的：建立LC-MS/MS同时检测大鼠血浆中5种酚酸类成分的分析方法,为丹参提取物血浆样品的测定提供参考。方法：采用LC-MS/MS同时检测丹酚酸A,B,C和紫草酸、迷迭香酸含量,流动相0.1%甲酸水溶液-甲醇和乙腈等体积混合液（含0.1%甲酸）梯度洗脱,流速0.6 m L·min^-1。质谱条件采用多反应监测方式进行负离子监测。定量分析的离子对m/z分别为493.3/295.1,717.5/519.1,491.2/293.2,537.3/295.1,359.1/161.0,5种成分的定量分析可在7 min内完成。结果：丹酚酸A,B和紫草酸线性范围均为0.78-100μg·L-1,丹酚酸C和迷迭香酸线性范围均为0.39-50μg·L-1。日内及日间精密度RSD均〈15%,准确度92%-112%。结论：该法选择性强、灵敏度高、操作简便,适用于丹参总酚酸中5种酚酸类成分在大鼠血浆中的药代动力学研究。

Objective： To establish a method for simultaneously determining plasma concentration of five kinds of phenolic acids from Salvia miltiorrhiza extract in rat plasma by LC-MS/MS. Method： LC-MS/MS was adopted to determine contents of lithospermic acid, rosmarinic acid and salvianolic acid A, B, C with mobile phase of 0. 1% aqueous formic acid-equal volume mixture of methanol and acetonitrile for gradient eluent, flow rate of 0.6 mL·min^-1. Mass spectrum was operated by an electrospray ionization probe in the negative ion mode. Scan was in multiple reaction ion monitoring mode. Ion pairs for quantitative analysis of this five phenolic acids were m/z of 537.3/295.1, 359. 1/161.0, 493.3/295.1, 717.5/519. 1, 491.2/293.2. Determination of this five phenolic acids could be accomplished within 7 min. Result： Linear ranges of salvianolic acid A, B and lithospermic acid were 0. 78-100 μg ·L^-1 , linear ranges of rosmarinic acid and salvianolic acid C were 0. 39-50 μg ·L^-1. Their inter- day and intra-day precisions represented by RSD were within 15%, their accuracies were between 92%-112%. Conclusion： A sensitive, accurate and easy LC-MS/MS method has been successfully developed, which can be applied to pharmacokinetic study of five phenolic acids from S. miltiorrhiza in rat plasma.