LC-MS/MS法检测小鼠静注水溶性紫杉醇偶合物后血浆中的药物浓度

Liquid chromatography-tandem mass spectrometry for determination of paclitaxel in mouse plasma after intravenous administration of poly(L-glutamic acid)-alanine-paclitaxel conjugate

目的建立灵敏、特异的LC-MS/MS方法测定小鼠血浆中紫杉醇浓度,并应用于小鼠静注水溶性多聚(L-谷氨酸)-丙氨酸-紫杉醇偶合物的药物动力学研究。方法用甲基叔丁基醚提取小鼠血浆样品,以多西紫杉醇为内标,在BetaBasicC18色谱柱上以流动相乙腈-水-甲酸(65∶35∶0.1,体积比)分离,流速为0.2ml/min;采用液相色谱-质谱-质谱联用仪,通过电喷雾离子化电离源,以选择反应监测方式进行正离子检测,用于定量分析的离子反应分别为m/z854.2→286.1(紫杉醇)和m/z808.3→527.2(多西紫杉醇)。结果测定紫杉醇的线性范围为0.5～1000ng/ml,相关系数r为0.9995,定量下限为0.5ng/ml,日内日间精密度分别小于11.52%和13.11%,准确度控制在87.05%～100.87%,提取回收率为85.04%～93.63%。结论该法快速、灵敏度高、专一性好,成功应用于小鼠静注10mg/kg水溶性多聚(L-谷氨酸)-丙氨酸-紫杉醇偶合物的药物动力学研究。

Objective To develop a rapid,sensitive and specific liquid chromatography-tandem mass spectrometry（LC-MS/MS）method for the quantification of paclitaxel in mouse plasma and to apply it to pharmacokinetic study of poly（L-glutamic acid）-alanine-paclitaxel conjugate（PG-PTX）in mice.Methods Paclitaxel and docetaxel as internal standards were extracted from mouse plasma using methyl tert-butyl ether and separated on a BetaBasic C18 column using a mobile phase of acetonitrile∶water∶formic acid（65∶35∶0.1,v/v/v）at a flow rate of 0.2 ml/min.Using LC-MS/MS,in the positive electrospray ionization mode,selected reaction monitoring of the precursor-product ion transitions m/z 854.2→286.1 for paclitaxel and 808.3→527.2 for docetaxel was used for quantification.Results The linear calibration curve of paclitaxel was obtained in a concentration range of 0.5-1000 ng/ml with a correlation coefficient of 0.9995.The lower limit of quantification was 0.5 ng/ml based on 100 μl of plasma.The intra-and inter-day precision was less than 11.52% and 13.11%,respectively,with the accuracy ranging from 87.05% to 100.87%.The extraction recovery was between 85.04% and 93.63%.Conclusion The method is rapid,sensitive,specific and has been successfully applied to the pharmacokinetic study of PG-PTX intravenously injected to mice at a dose of 10 mg/kg.