RP-HPLC法测定人血浆中氯氮平浓度

Concentration Determination of Clozapine in Human Plasma by RP-HPLC

目的:建立测定人血浆中氯氮平浓度的反相高效液相色谱法。方法:以Agilent Herpesil ODS C18柱(250 mm×4 mm,5μm)为色谱柱,流动相为甲醇:水(70:30),每250 ml水中加入0.55 ml三乙胺;流速:1.2 ml.min-1;柱温:40℃;检测波长:254 nm。以乙酸乙酯为提取剂。结果:氯氮平在20～1 000 ng.ml-1浓度范围内,峰面积与其浓度呈良好线性关系;氯氮平的高、中、低浓度相对平均回收率分别为101.55%、98.25%、103.57%;日内、日间RSD均<9%(n=5)。分析方法的检测限10 ng.ml-1;氯氮平的标准曲线回归方程为Y=0.32132509X+1.2727281,r=0.99725(n=8)。结论:该方法灵敏、准确、简单、快捷,可用于临床氯氮平的血浆药物浓度监测和药物动力学研究。

Objective：To estabilish a method for determining the concertration of Clozapine in human plasma by RP - HPLC. Methods ：The drug from plasma was analyzed by a reverse phase HPLC system with a Agilent Herpesil ODS C^8 column （250 mm· 4 mm, 5 μm） , methanol - H20 （0.55 mlTEA ＋ 250 mlH2 O） （70 ： 30） were used as mobile phase; the flow rate was 1.2 ml · min - 1 ; the detection wavelength was at 254 nm; Ethyl acetate was used as extracting solvent. Results： The relation of peak area and concentration showed linear in the range of 20 - 1 000 ng ~ m1-1 for Clozapine, respectively. The average recoveries of Clozapine at high, middle and low concentrations werel01.55 % , 98.25%, 103.57% ,respectively. The intra -day and inter- day variations（RSD） were less than 9% （ n = 5 ）. The minimum detectable concentration was 10.0 ng ·ml - 1. The regression equation of Clozapine was Y = 0. 32132509X ＋ 1. 2727281, r = 0. 99725 （ n = 8 ）, respectively. Conclusions ： The method is sensitive, accurate, simple and quick, which can be used for clinical drug monitoring and pharmacokinetic studies of Clozapine.