LC-MS/MS法同时测定犬血浆中丹酚酸A、丹酚酸B、紫草酸和迷迭香酸及其在丹参提取物的药代动力学研究应用

目的:建立同时检测Beagle犬血浆中丹酚酸A、丹酚酸B、紫草酸和迷迭香酸的分析方法,并应用混合有机溶剂萃取法提取含药血浆中的成分。方法:采用C18反相柱,流动相为甲醇乙腈等比混合液-0.1%甲酸水溶液梯度洗脱,质谱采用多反应监测(MRM)方式进行正离子检测。结果:定量分析的离子对(m/z)为:493.2/295.1(丹酚酸A),717.2/519.1(丹酚酸B),537.2/493.2(紫草酸)和359.2/161.0(迷迭香酸)。测定血浆中这4种成分的线性范围均为0.3125～200μg.L-1,定量下限为0.3125μg.L-1,该方法的精密度、准确度和稳定性均符合要求。结论:该法选择性强、灵敏度高、操作简便,可用于血浆中这4种酚酸类成分的药代动力学研究。

Simultaneous Determination of Salvianolic Acid A, Salvianolic Acid B, Rosmarinic Acid and Lithospermic Acid in Dog Plasma by LC-MS/MS: Application to Pharmacokinetic Study of Salvia Miltiorrhiza Extraction

A sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS) was developed and validated for the simultaneous quantification of Salvianolic Acid A(SAA),Salvianolic Acid B(SAB),Lithospermic Acid(LA) and Rosmarinic Acid(RA) in dog plasma.The compounds were extracted by organic solvent using mixed organic solvent.Chromatographic separation was achieved on a reversed-phase C18 column with the mobile phase of acetonitrile/menthol-water(0.1% acetic acid) and step gradient elution resulted at a flow rate of 0.60 mL·min-1.A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode.Quantification was performed using multiple reaction monitoring(MRM) of the transitions m/z 493.2/295.1(SAA),717.2/519.1(SAB),537.2/493.2(LA) and 359.2/161.0(RA).The method showed excellent linearity over the concentration range 0.3125-200 μg·L-1 of four components.The low limits of quantification were all at 0.3125 μg·L-1.The precision,accuracy and stability of the analytes meet the requirements.This method was sensitive,accurate and rapid.It can be applied in the determination of plasma concentration of four salvianolic acids.