A simple, sensitive, and rapid LC-MS/MS method for the simultaneous determination of BIC, the M7 and M8 active metabolites in SD rat plasma was developed. After salting-out assisted liquid–liquid extraction (SALLE) with 5 M ammonium acetate solution, the sample was analyzed on a Sciences column (C18 3.0 × 100 mm, 3 μm) using a gradient elution at 40 ℃ within 7 min. The assay displayed excellent linearity in the range of 4–2000 ng/mL for M7 and BIC, and 1–500 ng/mL for M8. The results of this method exhibited that the precision, accuracy, matrix effect, recovery, and stability of BIC, M7 and M8 met all requirements for the quantitation in plasma samples. The pharmacokinetic result showed that the AUC_(0→t) was calculated as 383.1 ± 164.4 (ng/mL·h) for BIC, and 5627 ± 1261 (ng/mL·h) for M7 after oral administration with BIC. Compared with BIC group, the pharmacokinetic parameters in BIC-NPs group were improved. Augmentation in AUC_(0→t) (3.02-fold) and t_1/2 (1.43-fold) for BIC. Meanwhile, double peak phenomenon was observed on the mean plasma concentration–time curves of M7 in BIC and BIC-NPs group. In conclusion, both BIC and BIC-NPs were metabolized to abundant M7 in SD rat, which would provide a basis for researching the treatment mechanism of liver injury.

开发了一种简单、灵敏、快速的 LC-MS/MS 方法，用于同时测定 SD 大鼠血浆中的 BIC、M7 和 M8 活性代谢物。用 5 M 醋酸铵溶液进行盐析辅助液液萃取 (SALLE) 后，在 Sciences 柱 (C18 3.0 × 100 mm, 3 μm) 上使用梯度洗脱在 40 ℃ 7 分钟内进行分析。该测定在 M7 和 BIC 的 4–2000 ng/mL 范围内以及 M8 的 1–500 ng/mL 范围内显示出出色的线性。该方法结果表明，BIC、M7和M8的精密度、准确度、基质效应、回收率和稳定性满足血浆样品定量的所有要求。药代动力学结果显示，口服BIC后，计算出BIC的AUC（0 → _{t）为383.1±164.4（ng/mL·h），M7的AUC（0→t）为5627±1261（ng/mL·h）。}与BIC组相比，BIC-NPs组的药代动力学参数有所改善。_{BIC 的 AUC (0→t)}（3.02 倍）和 t _1/2（1.43 倍）增加。同时，BIC组和BIC-NPs组M7的平均血药浓度-时间曲线上观察到双峰现象。总之，BIC和BIC-NPs在SD大鼠体内均代谢为丰富的M7，这为研究肝损伤的治疗机制提供了基础。