固相萃取高效液相色谱法测定人血浆中硫酸茚地那韦浓度

Determination of Indinavir Sulfate in Human Plasma by Solid Phase Extraction and HPLC with UV Detection

目的:建立硫酸茚地那韦血药浓度的高效液相色谱分析方法。方法:以对硝基苯胺为内标,血浆样品用固相萃取,色谱柱:Kromasil C_(18)色谱柱(150 mm×4.6mm,5μm),在线过滤器;流动相为0.1%三乙胺水溶液(用20%磷酸调节 pH 为4.3)-乙腈(69.5:30.5);检测波长为210 nm;流速1.0 mL·min^(-1),柱温为25℃。结果:硫酸茚地那韦在0.05～25.60μg·mL^(-1)的范围内有良好的线性关系(r=0.9995),最低检测浓度为0.01μg·mL^(-1)(S/N>3),该方法的相对回收率为97.4%～105.0%(n=5),绝对回收率为93.9%～105.6%(n=5);日内 RSD 为2.6%～3.7%,日间 RSD 为1.4%～4.0%。结论:本方法简便,准确,灵敏,特异性强,重现性好,可用于血浆中硫酸茚地那韦的测定及人体内药代动力学研究。

Objective： To establish HPLC method for determination of indinavir sulfate in human plasma. Methods： The method involves the use of solid - phase extraction followed by HPLC, and p - nitroaniline as the internal standard. The Kromasil Cts （ ODS C18, 150 mm×4.6 mm ,5μm） was used as analytical column with a mobile phase consisted of 0.1% triethylamine （ pH was adjusted to 4.3 with phosphoric acid ） - acetonitrile （69.5： 30.5 ）. The flow rate was 1.0 mL·min^-1 with UV detection at 210 nm. Results： The linear range of calibration curve was within drug plasma concentrations of 0.05 - 25.60μg·mL^-1 （ r = 0.9995 ）, the detection limit was 0.01μg·mL^-1 （ S/N〉 3 ）, the absolute recovery was 93.9% - 105.6% （ n = 5 ）, the method recovery was 97.4% - 105.0% （ n = 5 ）, the within - day RSD and between - day RSD were 2.6% - 3.7% and 1.4% - 4.0% （ n = 5 ）. Conclusions： The method was shown to be sensitive, accurate, simple and fine reproducibility for the determination of indinavir sulfate levels in human plasma. It was suitable for pharmacokinetics study of indinavir sulfate.