HPLC测定人血浆中伊曲康唑及其代谢产物羟基伊曲康唑的浓度

HPLC Determination of Itraconazole and Its Metabolite Hydroxyitraconazole in Human Plasma

目的:采用高效液相色谱-荧光法测定人血浆中伊曲康唑及其代谢产物羟基伊曲康唑的浓度。方法:采用Dikma Dia- monsil C18色谱柱(200 mm×4.6 mm,5μm);柱温为40℃;流动相:0.1%三乙胺缓冲液(用85%磷酸调pH为2.5)-乙腈(30∶ 70);流速:1.2 mL·min-1;荧光检测,激发波长260 nm,发射波长365 nm。以R051012作内标,血样采用甲基叔丁基醚1次提取。结果:伊曲康唑和羟基伊曲康唑的线性范围均为5-500 ng·mL-1。伊曲康唑和羟基伊曲康唑的提取回收率均在80% 以上,日内、日间精密度均小于10%。结论:该方法适用于临床血药浓度监测和药动学研究。

Objective:To establish an HPLC method for the determination of itraconazole and its metabolite hydrox- yitraconazole in human plasma. Methods:Dikma Diamonsil C18 HPLC column (200 mm×4.6 mm,5μm) was used; Column temperature was 40℃; The mobile phase consisted of 0.1% triethylamine buffer(adjusted pH to 2. 5 with 85% phosphoric acid) -acetonitrile(30:70); The flow rate was 1.2 mL·min-1;The detector was fluorescence detector, excitation wavelength and emission wavelength was set at 260 nm and 365 nm respectively. R051012 was used as the internal standard,the plasma samples were extracted by methyl tert - butyl ether. Results: The linear range of itraconazole and its metabolite hydroxyitraconazole were all 5 - 500 ng·mL-1. The extraction recovery of itraconazole and its metabolite hydroxyitraconazole were more than 80% ; Intra-day RSD and inter-day RSD were less than 10%. Conclusion:This method is proved to be sensitive,fast and accurate. Therefore,it can be used for therapeutic drug monitor and pharmacokinetic study.