反相高效液相色谱法测定人血清中伏立康唑的药物浓度

Quantitation of Human Plasma Voriconazole Concentration by RP-HPLC

目的：建立应用反相高效液相色谱法测定人血清中伏立康唑药物浓度的方法。方法：应用Alliance 2695液相色谱仪,X-Terra TM Rp C18柱（250 mm×4.6 mm,5μm）,柱温25℃;流动相为乙腈：冰醋酸-三乙胺缓冲液=42：58（V：V）。流速1.0 m L/min;紫外检测波长255 nm。血清样品用乙腈沉淀蛋白,离心后取上清液进样分析。结果：伏立康唑及内标色谱峰分离良好,无内源性杂质干扰。血清中伏立康唑在0.1~20.0μg/m L范围内线性关系良好,线性方程为：Y=0.2279X-0.03,r=0.9998,平均提取回收率为（100.2±3.0）%,平均方法回收率为（100.72±2.67）%,高、中、低3个浓度的日内精密度RSD为2.58%~6.12%,日间精密度为RSD 4.34%~8.16%。结论：本测定方法准确、重现性好,测定迅速,适用于临床治疗中伏立康唑血药浓度的测定。

Objective To develop a method of reverse phase high pressure liquid chromatography（RP-HPLC） to determine the concentration of voriconazole in human plasma. Methods The sample was analysed with RP-HPLC technique using Waters＇ Alliance 2695 HPLC machine and X-Terra TM Rp C18 column（250mm×4.6mm, 5μm）. Ketoconazole was used as the internal standard. The mobile phase was consisted of Acetonitrile and Acetic acid-triethylamine buffer solution（42：58）. Flow rate was 1ml/min at 25℃. The detecting wavelength was 255 nm. Results The chromatography is good and not interfered by the components of the plasma. The standard curve was linear at the voriconazole concentration range of 0.1 ~ 20.0μg/ml.The regression equation is Y = 0.2279X-0.03,r = 0.9998. The average extraction recovery rate was 100.28 % ± 3.0 %. The average analytical recovery rate was 100.72 % ± 2.67 %. The RSD of the between-day was 2.58 % ~ 6.12 %.and the RSD of the within-day was 4.34 %~8.16 %. Conclusion The method was accurate and stable. It is suitable for the determination of voriconazol concentration in human plasma.