反相高效液相色谱法测定小鼠肺组织中伏立康唑药物浓度

Determination of Voriconazole Concentration in Mice Lung by RP-HPLC

目的建立用反相高效液相色谱法测定小鼠肺组织中伏立康唑浓度的方法。方法用Waters X-Terra Rp C18柱,柱温：25℃,样品室温度：20℃。流动相：乙腈-冰醋酸三乙胺缓冲液（41∶59）,流速1.0 mL/min,检测波长为255 nm。酮康唑作为内标,肺组织匀浆经蛋白沉淀法处理后直接进样测定。结果肺组织匀浆伏立康唑药物浓度标准曲线在0.14~28.0μg/g（湿肺组织）,回归方程：Y=0.1421X＋0.025 7,r=0.999 7。平均提取回收率为102.58%±5.92%,平均方法回收率为102.23%±5.67%,高、中、低3个浓度的日内精密度RSD为1.30%~5.41%,日间精密度RSD为5.42%~5.68%。结论该方法具有良好的准确性和精密性,重现性好,适用于肺组织中伏立康唑药物浓度监测。

Objective To develop a method of reversed phase high performance liquid chromatography （ RP-HPLC ） for the determination of Voriconazole concentration in mice lung. Methods With Ketoconazole as the internal standard, voriconazole was extracted from mice lung samples by protein precipitation with acetonitrile. The X-Terra Rp C18 column （250 mm×4.6 ram, 5 μm） was selected to analyze the samples. The mobile phase consisted of aeetonitrile-glacial acetic acid triethylamine buffer solution （41：59）. Flow rate was 1 mL/min at 25 ℃. The detection wave length was 255 nm. Results The standard curve was linear when voriconazole concentration in the range of 0.14- 28 μg/g （wet lung weight）. The regression equation is 0.142 1×＋0.025 7, r = 0.9997. The average extraction recovery rate was 102.58%±5.92%. The average analytical recovery rate was 102.23%±5.67%. The RSD of the between-day was 1.30%-5.41%. The RSD of the within-day was 5.42%-5.68%. Conclusion The established method was accurate, sensitive and stable. It is suitable for the determination of voriconazol concentration in mice lung.