Kang Dian, Shao Yuhao, Yin Xiaoxi, Xiao Jingcheng, Rao Tai, Shen Boyu, Chen Huimin, Zhu Zhangpei, Wang Guangji, Liang Yan
Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Tongjiaxiang 24, Nanjing, 210009, China.
Biomed Chromatogr. 2017 Feb;31(2). doi: 10.1002/bmc.3797. Epub 2016 Aug 16.
Schisandra lignans, mainly including schizandrol A, schizandrol B, schisantherin A, schizandrin A, schizandrin B, etc., are the major active ingredients of Schisandra chinensis. In the present study, a robust liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the simultaneous quantification of schisandra lignans in rat primary hepatocytes. Lovastatin was used as an internal standard, and chromatographic separation was achieved on a Shimadzu C column with a gradient elution at the flow rate of 0.2 mL/min. All of the analytes were detected in multiple reaction monitoring mode with positive electrospray ionization since the sodium adduct ion [M + Na] was observed as the most intensive peak in the MS spectrum. For schizandrol A, schisantherin A and schizandrin A, the dynamic range was within 2-1000 ng/mg protein, and the linear range of schizandrol B and schizandrin B was from 5 to 1000 ng/mg protein. The intra- and inter-day precision was <15% and the accuracy (relative error) ranged from -15 to 15%. No significant variation was observed in the stability tests. The validated method was then successfully applied to the time-dependent uptake study for the Schisandra Lignan Extract in rat primary hepatocytes.
五味子木脂素主要包括五味子醇甲、五味子醇乙、五味子酯甲、五味子甲素、五味子乙素等，是五味子的主要活性成分。在本研究中，建立了一种稳健的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量大鼠原代肝细胞中的五味子木脂素。洛伐他汀用作内标，在岛津C柱上以0.2 mL/min的流速进行梯度洗脱实现色谱分离。所有分析物均在多反应监测模式下采用正电喷雾电离进行检测，因为在质谱图中观察到钠加合离子[M + Na]为最强峰。对于五味子醇甲、五味子酯甲和五味子甲素，动态范围在2-1000 ng/mg蛋白质之间，五味子醇乙和五味子乙素的线性范围为5至1000 ng/mg蛋白质。日内和日间精密度均小于15%，准确度（相对误差）在-15%至15%之间。稳定性试验中未观察到显著变化。经验证的方法随后成功应用于大鼠原代肝细胞中五味子木脂素提取物的时间依赖性摄取研究。
