Simultaneous determination of asarinin, β-eudesmol, and wogonin in rats using UPLC-MS/MS and its application to pharmacokinetic studies following administration of standards and Gumiganghwal-tang.

Asarinin, β-eudesmol, and wogonin have common anti-angiogenic activities and have the potential for use in chemotherapy. And, they are multivalent substances that are combined in various herbal medicines. The purpose of this study was to develop a method for simultaneous analysis of asarinin, β-eudesmol, and wogonin, which are representative pharmacological components of Asarum heterotropoides, Atractylodes lancea, and Scutellaria baicalensis, respectively, in rat bio-samples using UPLC-MS/MS. The three components were separated using 5 mM aqueous ammonium acetate containing 0.1% formic acid and acetonitrile as a mobile phase, equipped with a KINETEX core-shell C18 -column. The analysis was quantitated on a triple-quadrupole mass-spectrometer employing electrospray-ionization, and operated in multiple-reaction-monitoring. The chromatograms showed high resolution, sensitivity, and selectivity with no interference with plasma, urine, and feces constituents. The developed analytical method satisfied international guidance criteria and could be successfully applied to the pharmacokinetic studies including oral bioavailability of asarinin, β-eudesmol, and wogonin after oral and intravenous administration and their urinary and fecal excretion ratios in oral administration to rats. Furthermore, the analysis was extended to pharmacokinetic studies following oral administration of Gumiganghwal-tang. This study was the first simultaneous analysis of three constituents in rat plasma, urine, and feces and determination of their pharmacokinetic parameters.