Ma Jianshe, Lin Chongliang, Wen Congcong, Xiang Zheng, Yang Xuezhi, Wang Xianqin
Laboratory Animal Centre of Wenzhou Medical University, Wenzhou 325035, China.
The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 15;953-954:143-6. doi: 10.1016/j.jchromb.2014.02.006. Epub 2014 Feb 18.
Bicuculline, a phthalide isoquinoline alkaloid is of current interest as an antagonist of gamma-aminobutyric acid (GABA). A simple and sensitive liquid chromatography mass spectrometry method for determination of bicuculline in rat plasma was developed over the range of 5-500ng/mL. After addition of midazolam as internal standard, protein precipitation with acetonitrile-methanol (9:1, v/v) was used as sample preparation. Chromatographic separation was achieved on a Zorbax SB-C18 (2.1mm×150mm, 5μm) column with acetonitrile -0.1% formic acid in water as mobile phase with gradient elution. Electrospray ionization (ESI) source was applied and operated in positive ion mode; selective ion monitoring (SIM) mode was used for quantification using target fragment ions m/z 368 for bicuculline and m/z 326 for the IS. Linear calibration was obtained with correlation coefficients r>0.99. The CV of the precision measurements was less than 13%. The accuracy of the method ranged from 93.6% to 100.5%. Mean recoveries of bicuculline in plasma were in the range of 80.5-91.8%. The method was successfully applied to the pharmacokinetic study after gavage administration of 15mg/kg bicuculline in rats.
荷包牡丹碱是一种酞基异喹啉生物碱，作为γ-氨基丁酸（GABA）的拮抗剂，目前备受关注。建立了一种简单灵敏的液相色谱-质谱法，用于测定大鼠血浆中5-500ng/mL范围内的荷包牡丹碱。加入咪达唑仑作为内标后，采用乙腈-甲醇（9:1，v/v）进行蛋白沉淀作为样品制备方法。在Zorbax SB-C18（2.1mm×150mm，5μm）色谱柱上进行色谱分离，以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱。采用电喷雾电离（ESI）源，以正离子模式运行；采用选择性离子监测（SIM）模式，以荷包牡丹碱的目标碎片离子m/z 368和内标的m/z 326进行定量。获得线性校准，相关系数r>0.99。精密度测量的变异系数小于13%。该方法的准确度范围为93.6%至100.5%。血浆中荷包牡丹碱的平均回收率在80.5-91.8%范围内。该方法成功应用于大鼠灌胃给予15mg/kg荷包牡丹碱后的药代动力学研究。
