Bioanalytical assay development and validation for simultaneous quantification of five schisandra lignans in rat primary hepatocytes based on LC‐MS/MS: application to a real‐time uptake study for Schisandra Lignan Extract

• Published in: Biomedical chromatography Vol. 31; no. 2; pp. np - n/a
• Main Authors: Kang, Dian, Shao, Yuhao, Yin, Xiaoxi, Xiao, Jingcheng, Rao, Tai, Shen, Boyu, Chen, Huimin, Zhu, Zhangpei, Wang, Guangji, Liang, Yan
• Format: Journal Article
• Language: English
• Published: England 01-02-2017
• Subjects: Animals; Antineoplastic Agents - analysis; Antineoplastic Agents - pharmacokinetics; Cells, Cultured; Chromatography, High Pressure Liquid - methods; Cyclooctanes - analysis; Cyclooctanes - pharmacokinetics; Dioxoles - analysis; Dioxoles - pharmacokinetics; Hepatocytes - metabolism; LC‐MS/MS; Lignans - analysis; Lignans - pharmacokinetics; Limit of Detection; Male; Polycyclic Compounds - analysis; Polycyclic Compounds - pharmacokinetics; rat primary hepatocytes; Rats; Rats, Sprague-Dawley; Schisandra; Schisandra - chemistry; Schisandra chinensis; schisandra lignans; Tandem Mass Spectrometry - methods; uptake; LC-MS/MS; Schisandra chinensis; uptake; rat primary hepatocytes; schisandra lignans
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.3797

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 C18 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.