Simultaneous determination of seven coumarins by UPLC–MS/MS: Application to a comparative pharmacokinetic study in normal and arthritic rats after oral administration of Huo Luo Xiao Ling Dan or single-herb extract

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 991; pp. 108 - 117
• Main Authors: Wu, Yun, Wang, Fenrong, Ai, Yu, Ma, Wen, Bian, Qiaoxia, Lee, David Y.-W., Dai, Ronghua
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-06-2015
• Subjects: Animals; Anti-Inflammatory Agents - administration & dosage; Anti-Inflammatory Agents - blood; Anti-Inflammatory Agents - pharmacokinetics; Arthritis; Arthritis - drug therapy; Chromatography, High Pressure Liquid - methods; Coumarins; Coumarins - administration & dosage; Coumarins - blood; Coumarins - pharmacokinetics; Drugs, Chinese Herbal - administration & dosage; Drugs, Chinese Herbal - pharmacokinetics; Huo Luo Xiao Ling Dan; Limit of Detection; Liquid-Liquid Extraction - methods; Male; Pharmacokinetics; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry - methods; UPLC–MS/MS; Coumarins; Huo Luo Xiao Ling Dan; Arthritis; Pharmacokinetics; UPLC–MS/MS
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2015.04.017

•A UPLC–MS/MS method was developed for pharmacokinetic study of Huo Luo Xiao Ling Dan.•Simultaneous determination of seven coumarins.•The lower limits of quantitation (LLOQ) were 0.03–0.25ng/mL for all the analytes.•The validated method was successfully applied to a comparative pharmacokinetic study.•The pharmacokinetic profiles in the normal and arthritic rats were elucidated.
A simple, sensitive and reliable ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method has been developed and validated for simultaneous quantitation of seven coumarins, the bio-active ingredients of Huo Luo Xiao Ling Dan (HLXLD), in rat plasma. The liquid–liquid extraction method with ether–dichloromethane (2:1, v/v) was used to prepare the plasma samples. Analytes and internal standard (IS) of bifendate were separated on a Shim-pack XR-ODS column (75mm×3.0mm, 2.2μm particles) using gradient elution with the mobile phase consisting of methanol and 0.05% formic acid in water at a flow rate of 0.4mL/min. Detection was performed on a triple quadrupole (TQ) tandem mass spectrometry equipped with an electrospray ionization source in the positive ionization and multiple reaction monitoring (MRM) mode. The lower limits of quantitation (LLOQ) were 0.03–0.25ng/mL for all the analytes. Intra- and inter-day precision and accuracy of the seven analytes were well within acceptance criteria (15%). The matrix effect and the mean extraction recoveries of the analytes and IS from rat plasma were all within satisfaction. The validated method has been successfully applied to compare pharmacokinetic profiles of the seven active ingredients in rat plasma between normal and arthritic rats after oral administration of HLXLD, Angelica pubescens extract and Notopterygium incisum extract, respectively. Results showed that there were remarkable differences in pharmacokinetic properties of the analytes among the different groups.