Simultaneous determination of nineteen compounds of Dahuang zhechong pill in rat plasma by UHPLC-MS/MS and its application in a pharmacokinetic study

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2020-08, Vol.1151, p.122200, Article 122200
• Main Authors: Wu, Li, Du, Shali, Yang, Furong, Ni, Zihui, Chen, Zhipeng, Liu, Xiao, Wang, Yulan, Zhou, Qigang, Li, Weidong, Qin, Kunming
• Format: Article
• Language: English
• Subjects: Characteristic constituents; Dahuang zhechong pill; Pharmacokinetics; UHPLC-MS/MS
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2020.122200

[Display omitted] •A novel UHPLC-MS/MS method for bioanalysis of Dahuang Zhechong pill(DHZCP).•Nineteen characteristic constituents of DHZCP were simultaneously determined in plasma.•The pharmacokinetic characteristics of multiple components of DHZCP were successfully achieved in rat plasma.•The pharmacokinetic characteristics of DHZCP were studied for the first time. Dahuang zhechong pill (DHZCP) is a famous traditional Chinese medicine prescription, which is widely used in the treatment of liver diseases. However, due to the lack of a dynamic DHZCP profile, the in vivo pharmacokinetics of active ingredients within this medicine remains unknown. In this paper, a rapid, sensitive and reliable UHPLC-MS/MS method was used to determine the content of 19 characteristic constituents of DHZCP in rat plasma, including rhein, emodin, chrysophanol, physcion, aloeemodin, p-methoxyphenylacetic acid, hypoxanthine nucleoside, wogonin, wogonoside, baicalin, norwogonin, naringenin, nutmeg acid, paeoniflorin, verbascoside, rhodiola glucoside, forsythoside A, formononetin, and glycyrrhizic acid. An Agilent Extend-C18 column (2.1 mm × 100 mm, 1.8 μm) was used to separate the 19 characteristic constituents, with a mobile phrase of (A) 0.1% formic acid and (B) acetonitrile. The constituents were detected in negative ion mode with multiple reactions monitoring (MRM). The established UHPLC-MS/MS method had good linearity, with a coefficient of determination (r2) of >0.99. The daytime and intra-day precision were less than 12%, and the accuracy ranged from −9.56% to 7.82%. The stability, extraction recovery, and matrix effect met the requirements. The method was successfully applied to the pharmacokinetic study of these nineteen characteristic constituents after oral administration of DHZCP. UHPLC-MS/MS was used for the first time to study the pharmacokinetics of the characteristic chemical constituents in DHZCP, which provided reference and theoretical guidance for further clarification of its pharmacodynamic basis.