Pharmacology Laboratory of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China.
J Mass Spectrom. 2012 Aug;47(8):962-8. doi: 10.1002/jms.3042.
A sensitive rapid analytical method was established and validated to determine the bakkenolide A (BA) in rat plasma. This method was further applied to assess the pharmacokinetics of BA in rats receiving a single dose of BA. Liquid chromatography tandem mass spectrometry in multiple reaction monitoring mode was used in the method, and costundide was used as internal standard. A simple protein precipitation based on methanol was employed. The combination of a simple sample cleanup and short chromatographic running time (2.4 min) increased the throughput of the method substantially. The method was validated over the range of 1-1000 ng/mL with a correlation coefficient > 0.99. The lower limit of quantification was 1 ng/mL for BA in plasma. Intra- and inter-day accuracies for BA were 93-112% and 103-104%, respectively, and the inter-day precision was less than 15%. After a single oral dose of 20 mg/kg of BA, the mean peak plasma concentration (C(max) ) of BA was 234.7 ± 161 ng/mL at 0.25 h. The area under the plasma concentration-time curve (AUC(0-24 h) ) was 535.8 ± 223.7 h·ng/mL, and the elimination half-life (T(1/2) ) was 5.0 ± 0.36 h. In case of intravenous administration of BA at a dosage of 2 mg/kg, the area under the plasma concentration-time curve (AUC(0-24 h) ) was 342 ± 98 h⋅ng/mL, and the elimination half-life (T(1/2) ) was 5.8 ± 0.7 h. Based on the results, the oral bioavailability of BA in rats at 20 mg/kg is 15.7%.
建立并验证了一种灵敏的快速分析方法，用于测定大鼠血浆中的 bakkenolide A（BA）。该方法进一步用于评估单次给予 BA 后 BA 在大鼠体内的药代动力学。该方法采用液相色谱-串联质谱多反应监测模式，以 costundide 为内标。采用甲醇简单的蛋白沉淀法。简单的样品净化和较短的色谱运行时间（2.4 分钟）相结合，大大提高了方法的通量。该方法在 1-1000ng/mL 范围内进行验证，相关系数>0.99。BA 在血浆中的定量下限为 1ng/mL。BA 的日内和日间准确度分别为 93-112%和 103-104%，日内精密度小于 15%。单次口服 20mg/kg BA 后，BA 的平均血浆峰浓度（C(max)）为 0.25 小时时为 234.7±161ng/mL。BA 的药时曲线下面积（AUC(0-24 h)）为 535.8±223.7h·ng/mL，消除半衰期（T(1/2)）为 5.0±0.36h。静脉给予 BA 剂量为 2mg/kg 时，BA 的药时曲线下面积（AUC(0-24 h)）为 342±98h·ng/mL，消除半衰期（T(1/2)）为 5.8±0.7h。根据这些结果，大鼠口服 20mg/kg 的 BA 的生物利用度为 15.7%。
