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Bioavailability of wilforlide A in mice and its concentration determination using an HPLC-APCI-MS/MS method.
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 2018 July 16
Wilforlide A (WA), an active compound in Tripterygium wilfordii Hook F (TW) which is a traditional Chinese medicine for treatment of autoimmune diseases, is a quality control marker for TW product. At present, the bioavailability/pharmacokinetics of WA is not known. Such information is not only essential to evaluate the relevance of WA as a quality control maker, but also important for future clinical efficacy studies. Therefore, a high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometric method (HPLC-APCI-MS/MS) was developed and applied to a bioavailability/pharmacokinetic study of WA. WA and celastrol (the internal standard, IS) were extracted by a liquid-liquid extraction method using methyl tert-butyl ether. Multiple reaction monitoring (MRM) scanning in positive ionization mode was used to monitor the transition of m/z 455.1 to 191.3 for WA and 451.3 to 201.2 for IS. This method was validated and applied to a pharmacokinetic study of WA in mice following intravenous administration (IV, 1.2 mg/kg), intraperitoneal injection (IP, 6 mg/kg) and oral administration (PO, 30 mg/kg). The lower limit of quantification (LLOQ) for WA was 10 ng/ml. The intra- and inter-day precision was found to be within 15.4% while the accuracy within 94.1-115.7% for all the quality control and LLOQ samples. The samples were stable under all the usual storage and experimental conditions. The terminal elimination half-lives were 14.7, 9.1 and 22.7 min following IV, IP and PO dosing, while the absolute bioavailability for IP and PO WA were 9.39% and 0.58% respectively. These results indicated that the HPLC-APCI-MS/MS assay was suitable for the pharmacokinetic study of WA. WA was found poorly absorbed when given orally and therefore it may not be a relevant marker for the oral TW products in the market.
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