柱前衍生高效液相色谱法测定克拉霉素血药浓度

目的采用柱前衍生高效液相色谱法测定克拉霉素血药浓度。方法克拉霉素血浆样品以正己烷-正丁醇混合溶液萃取后,与2,4-二硝基苯肼衍生化后进样。色谱柱为KromasiC18(4.6mm×250mm,5μm),0.05mol/LNH4H2PO4缓冲液(pH=6.5)-乙腈(68:32)为流动相,柱温室温,流速1.0mL/min,检测波长340nm。结果克拉霉素血药浓度在50～3000μg/L范围内,浓度与峰面积比有良好的线性关系(r=0.9998),最低检测浓度为30μg/L。平均方法回收率为(99.2±3.5)%(n=15)。日内RSD≤4.9%,日间RSD≤7.7%。结论本方法简单快速、准确,可用于克拉霉素临床药动学研究。

Determination of Clarithromycin in Human Plasma by HPLC with Pre-column Derivatization

Objective To establish a HPLC method with pre-eolumn derivatization to measure the plasma concentration of elarithromyein. Methods A n-hexane and n-butanol extract of plasma was treated with 2,4-dinitrophenylhydrazine and chromatog raphed on an Kromasil C18 column （4.6mm×250mm,5μm） using an eluent composed of aeetonitrile-0.05 mol/L phosphate buffer （ pH6.5 ） （ 32： 58）. The column temperature was at room temperature and flow rate of eluent was 1.0mL/min. The UV detector was operated at 340 nm. Results The assay procedure was shown to produce linear calibration curves over the range 50 μg/L to 3000μg/L of elarithmmycin in plasma（ r = 0. 9998 ）. The limit of detection was 30 μg/L. Within the range, the recovery rate was （ 99.2 ± 3.5 ） % （ n = 15 ）. The coefficients of variation within a day and between days did not exceed 4.9% ,7.7% ,respectively. Conclusion This method is found to be easy,convenient and precise for clinical pharmacokineties study of clarithmmyein.