Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
J Pharm Biomed Anal. 2013 Apr 15;77:71-5. doi: 10.1016/j.jpba.2012.12.031. Epub 2013 Jan 5.
A rapid and sensitive bioassay based on liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of eight coumarins in rat plasma. The liquid-liquid extraction method with ethyl acetate was used to prepare the plasma samples after addition of warfarin as an internal standard (IS). Chromatographic separation was performed on an Eclipse plus C18 column (100mm×4.6mm, 1.8μm) using gradient elution when 1mM ammonium acetate aqueous solution - acetonitrile was used as the mobile phase. The lower limit of quantitation (LLOQ) of each coumarin was lower than 2.16ngmL(-1). Intra-day and inter-day precisions were less than 15%. The accuracies were in the range of 88.9-117%. The mean recoveries of coumarins and IS were higher than 84%. The method was successfully applied to a pharmacokinetic study of eight coumarins in rats after oral administration of radix angelicae pubescentis.
建立并验证了一种基于液相色谱-串联质谱（LC-MS/MS）的快速灵敏生物分析方法，用于同时测定大鼠血浆中的 8 种香豆素。在加入华法林作为内标（IS）后，采用乙酸乙酯进行液液萃取法处理血浆样品。采用 Eclipse plus C18 柱（100mm×4.6mm，1.8μm）进行色谱分离，以 1mM 乙酸铵水溶液-乙腈为流动相进行梯度洗脱。每种香豆素的定量下限（LLOQ）均低于 2.16ngmL(-1)。日内和日间精密度均小于 15%。准确度在 88.9-117%范围内。香豆素和 IS 的平均回收率均高于 84%。该方法成功应用于口服透骨草后大鼠体内 8 种香豆素的药代动力学研究。
