高效液相色谱法测定体液中胺苯吖啶的浓度

DETERMINATION OF m-AMSA BY RP-HPLC IN BIOLOGICAL FLUIDS

本文系统地研究和建立了用反相高效液相色谱法测定生物体液中胺苯吖啶的方法。所建立的方法使用YWG-C_(18)H_(37)(10μm)为固定相,采用甲醇:水:四甲基乙二胺:冰醋酸(50:50:0.5:0.26,水相pH为6.40)的混合溶液为流动相.流速为1.5ml/min,紫外检测波长为254nm,以酰胺咪嗪为内标进行定量测定。本法最低检测限为0.5ng,血清中最低检测浓度为0.004μg/ml。天内、天间的方法平均回收率为101％左右,CV在1.52～5.82％之间。各种指标考核证实该法灵敏、特异、准确、精密,可用于胺苯吖啶临床药理的研究。家兔静脉注射胺苯吖啶100mg/kg,其药代动力学过程符合二室开放式模型,分布相半衰期为0.0340h,消除相半衰期为1.046h。

A rapid and sensitive quantitative analytical method for determination of m-AMSA in biological fluids was developed.Liquid Chromatograph Model KYS(Shanghai,China)packed with a reversed-phase YWG-C_(18)H_(37)(10μm)column and equipped with a 254 nm UV detector was selected. The mobile phase consisted of a mixture of 50ml methanol and 50ml buffered aqueous solution containing 0.5ml N N N' N'-tetramethylenediamine which is adjusted to pH 6.40 with 0.26ml glacial acetic acid was pumped through the column at the rate of 1.5ml/min. Carbamazepine was used as an internal standard. The limit of detection is 0.5ng and the lower limit of determinations is 0.004μg/ml in plasma. The precisions of this method were acceptable with the coefficient of variation ranging from 1.89 to 6.35% at the concentration of 0.0120, 0.1000, 0.5000,and 1.000μg/ml.The accuracies of these levels were 100.3±6.35%, 100.8±4.95%, 101.3±1.89% and 102.6±3.36%, respectively. The pharmacokinetic characteristics of m-AMSA in several rabbits after. 10mg/kg intravenously administered was investigated by this method. The results showed that the data fitted to a two-compartment open model adequately with following parameters: t_(1/2) α=0.0340h, t_(1/2)β=1.048h, k_(10)=1.449h^(-1).