标题：Determination of brusatol in plasma and tissues by LC–MS method and its application to a pharmacokinetic and distribution study in mice
作者：Guo, Nan ;Zhang, Xiaoran ;Bu, Fanlong ;Wang, Lei ;Cao, Zhanqi ;Geng, Chunmei ;Guo, Ruichen ;Ren, Dongmei ;Wen, Qing
作者机构：[Guo, Nan ;Bu, Fanlong ;Geng, Chunmei ;Guo, Ruichen ] Institute of Clinical Pharmacology, Qilu Hospital of Shandong University, Jinan, China;[Wang, Le 更多
来源：Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
摘要：Objectives The quassinoid brusatol, which can be isolated from Brucea javanica (L.) Merr., becomes popularly studied because of its anti-tumor activity. In order to further investigate brusatol and extend its applications, a sensitive analytical method for determination of brusatol in biological samples is essential. However, few methods had been reported until now. In this study, a highly sensitive and reproducible LC–MS method for simultaneous quantification of brusatol in mouse plasma and tissues was developed and validated. Method Plasma samples and tissue homogenate were extracted with diethyl ether after addition of the internal standard solution(IS). The supernatant was blown to dryness with nitrogen and residual was reconstituted with 100 μl of methanol. The separation was performed on an Intersil ODS-3 column and gradient elution was conducted with the mobile phase of water and methanol (0–5 min 47:53, 5–5.5 min 47:53–10:90, 5.5–9 min 10:90, posttime 4 min 47:53) at a flow rate of 0.8 mL/min. Quantification was performed in the selected ion monitoring (SIM) mode at m/z 543.2 for brusatol and 220.0 for IS (ornidazole). The method was validated by analyzing quality control plasma and tissue homogenate samples, and was applied to analyze samples obtained from mice after injections of brusatol via the tail vein. Results With ornidazole as the internal standard, calibration curve of the method ranged from 10 to 320 ng/ml for plasma and 10–240 ng/ml for tissues. Recovery rate of brusatol from plasma and tissues were between 71.09%–94.91%. Relative standard deviation (RSD) for inter- and intra-day precision was less than 15%, and the accuracy was between 96.1%–111.8%. The pharmacokinetics and distribution study of brusatol in mice after three single doses via the tail vein were carried out based on this method. The concentration of brusatol in plasma decreased rapidly and a more than 10 fold concentration of brusatol was found as compared to that in other tissues. Conclusions This is the first reported LC–MS method for detecting brusatol in tissues and can accurately determine the concentrations of these compounds in plasma and different tissues. Further research on the metabolism of brusatol in vivo is still needed.
© 2017 Elsevier B.V.