反相高效液相色谱荧光检测法测定血浆中利扎曲普坦浓度

Determination of rizatriptan in human plasma by RP-HPLC with fluorescence detection

目的:建立反相高效液相色谱荧光检测法测定血浆中利扎曲普坦(RZT)浓度。方法:采用 Luna C_8(2)分析柱(250mm×4.6m,5μm),柱温为40℃,流动相为0.02 mol·L^(-1)磷酸二氢钾-乙腈-甲醇(87:10:3),流速1.0mL·min^(-1),样品血浆在碱性条件下用二氯甲烷旋涡混合提取浓集后用蒸馏水溶解进样,用荧光检测器(λ_(ex)290nm,λ_(em)356nm)检测,以佐米曲普坦作内标,按内标法定量。结果:标准曲线在0.25～80μg·L^(-1)范围内有良好线性,最低检测限为0.25μg·L^(-1),RZT 及内标的保留时间分别为7.0min 和8.6min,日内和日间 RSD 均小于9%,方法回收率在92%～111%。结论:本法具有快速简便,灵敏准确等特点,适用于利扎曲普坦血药浓度测定及药代动力学研究。

Objective：To establish an RP - HPLC method with fluorescence detection for the determination of rizatriptan （RZT） in human plasma. Methods：A fluorescence detector and Luna Cs （2） analysis column （250 mm × 4.6 mm ,5μm） were used. The alkalinized plasma sample was extracted with 3.0 mL dichloromethane. After evaporation of the organic layer,the residue was dissolved in distilled water and injected onto the column. The mobile phase of 0. 02 mol·L^1 potassium dihydrogen phosphate- acetonitrile- methanol （87： 10： 3） was pumped at 1.0 mL · min^-1 through the column. The detector at high sensitivity was set at λex 290 nm and λem 356 nm. Results：The retention times for RZT and internal standard （zolmitriptan） were 7.0 min and 8.6 rain, respectively. The standard curve was linear over the concentration range of 0. 25 to 80μg · L^-1. The lowest concentration of detection in plasma was 0. 25 μg · L^-1. The method recoveries were 92% - 111% ; the intra -day RSD and inter - day RSD were less than 9%. Conclusion：This method is found to be simple, rapid, sensitive and accurate for determination of RZT in human plasma.