Zhao Liang, Qian Xian, Li Wuhong, Lv Lei, Zhang Hai, Chai Yifeng, Zhang Guoqing
Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, 200438, China; School of Pharmacy, Second Military Medical University, Shanghai, 200433, China.
Biomed Chromatogr. 2014 Dec;28(12):1670-8. doi: 10.1002/bmc.3200. Epub 2014 May 14.
A rapid, simple and sensitive, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of bergenin, chlorogenic acid and four flavonoids in a QingGanSanJie preparation in rat plasma. Puerarin was selected as the internal standard (IS). Plasma samples were precipitated with methanol and separated with a reverse phase Agilent Poroshell 120 EC-C18 column using a gradient mobile phase of methanol-water containing 0.1% formic acid (v/v). A triple quadruple mass spectrometer was used for quantification (limit of detection 0.36-5.55 ng/mL). Intra-day and inter-day precisions were within 15% and the average extraction recoveries ranged from 85 to 115% for each analyte. The method allowed simultaneous quantification for the first time of the pharmacokinetics of bergenin, chlorogenic acid and four flavonoids after intragastric administration of a QingGanSanJie extract in Sprague-Dawley rats. It was found that bergenin and chlorogenic acid had typical extravascular administration concentration-time curves; flavonoids had a bimodal distribution improving bioavailability and extending the pharmacodynamics period.
建立并验证了一种快速、简便、灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于同时测定大鼠血浆中清肝散结制剂中的岩白菜素、绿原酸和四种黄酮类化合物。选择葛根素作为内标（IS）。血浆样品用甲醇沉淀，采用含0.1%甲酸（v/v）的甲醇-水梯度流动相，在反相安捷伦Poroshell 120 EC-C18柱上进行分离。使用三重四极杆质谱仪进行定量分析（检测限为0.36-5.55 ng/mL）。日内和日间精密度均在15%以内，各分析物的平均提取回收率在85%至115%之间。该方法首次实现了对SD大鼠灌胃给予清肝散结提取物后岩白菜素、绿原酸和四种黄酮类化合物药代动力学的同时定量分析。结果发现，岩白菜素和绿原酸具有典型的血管外给药浓度-时间曲线；黄酮类化合物具有双峰分布，可提高生物利用度并延长药效学时间。
