Wu Haiya, Yan Qizhi, Fan Zhehua, Huang Miaoling, He Jiamin, Ma Jianshe, Wang Xianqin
The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
Department of Pharmacy, Shaoxing People's Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, China.
Biomed Chromatogr. 2018 Aug;32(8):e4255. doi: 10.1002/bmc.4255. Epub 2018 May 10.
In this work, a selective and sensitive ultra-performance liquid chromatography tandem mass spectrometry method was established and validated for determination of corypalmine in mouse blood after oral or intravenous administration. A UPLC BEH C column was used to separate corypalmine and berberrubine (internal standard) at 40°C. The mobile phase was composed of acetonitrile and 10 mmol/L ammonium acetate (containing 0.1% formic acid) at a flow rate of 0.4 mL/min, and the total run time was 4.0 min. Electrospray ionization in positive ion mode was applied; target fragment ions m/z 342.2 → 178.0 for corypalmine and m/z 322.1 → 307.0 for berberrubine were identified with multiple reaction monitoring mode. The linear range was 1-1000 ng/mL (r > 0.995) and the lower limit of quantification for corypalmine in plasma was 1.0 ng/mL. The intra- and inter-day precisions were both <14%. The range of accuracy in this method was 97.5-109.0%. Mean recovery was >69.6%, and the matrix effect was 96.8-107.6%. Based on its high sensitivity, specificity and reliability, this method was successfully applied to study the pharmacokinetic parameters of corypalmine in mouse by oral and intravenous administration, and finally, the bioavailability of corypalmine was identified at 4.6%.
本研究建立并验证了一种选择性和灵敏的超高效液相色谱串联质谱法，用于测定小鼠口服或静脉给药后血液中的紫堇明碱。采用UPLC BEH C柱在40℃下分离紫堇明碱和小檗红碱（内标）。流动相由乙腈和10 mmol/L醋酸铵（含0.1%甲酸）组成，流速为0.4 mL/min，总运行时间为4.0 min。采用正离子模式的电喷雾电离；在多反应监测模式下鉴定出紫堇明碱的目标碎片离子m/z 342.2→178.0和小檗红碱的目标碎片离子m/z 322.1→307.0。线性范围为1-1000 ng/mL（r＞0.995），血浆中紫堇明碱的定量下限为1.0 ng/mL。日内和日间精密度均＜14%。该方法的准确度范围为97.5-109.0%。平均回收率＞69.6%，基质效应为96.8-107.6%。基于其高灵敏度、特异性和可靠性，该方法成功应用于研究紫堇明碱在小鼠口服和静脉给药后的药代动力学参数，最终确定紫堇明碱的生物利用度为4.6%。
