高效液相色谱法固相萃取测定全血环孢素A浓度

Determination of cyclosporine A in human whole blood by HPLC solid-phase extration

目的:采用高效液相固相萃取法快速测定全血环孢素A的浓度。方法:全血经低渗溶血,乙腈沉淀蛋白,用SPE固相萃取桂萃取,50％甲醇洗杂质,95％甲醇洗脱,收集洗脱液,挥干,乙腈重溶,上机。本法采用反相C_(18)色谱柱,柱温为72℃,流动相为乙腈-甲醇-水(3:1:1),检测波长210nm。结果:本法在40～1500μg·L^(-1)范围内具有良好的线性关系。全血中CsA平均提取回收率87.8％,RSD小于10％。结论:本方法特异性高,灵敏度好,既适合环孢素A的临床常规监测又适合药动学研究。

OBJECTIVE To develop an reversed-phase liquid chromatographie method, utilizing solid-phase extraction procedure for the determination of cyclosporine A(CsA) in human whole blood. METHODS The drug was extracted from whole blood by solid-phase extraction, then separated isocratically using a C18 column and a mobile phase of acetonitrile-methanol-water(3:1:1). The column temperature was maintained at 72C , the wavelength used for detection was 210 nm. The cyclosporine D was used as internal standard. RESULTS The linear relationship was from 40 fig.L-1 to 1500 ug.L-1. The average recoveries of CsA were 87.8% for whole blood,RSD values of within-day and between-day were all lower than 10% . CONCLUSIONS The method is simple and accurate. It is suitable not only for the study of pharmacokineties of CsA,but also for the routine determination of CsA in clinie.