高效液相色谱法测定血清中奥硝唑浓度

Determination of ornidazole in rabbit serum by HPLC

目的建立测定血清中奥硝唑浓度的高效液相色谱法。方法采用Kromasil C_(18)色谱柱(150 mm×4.6 mm,5μm),以甲醇-水(30:70)为流动相,甲硝唑作内标物。血清样品经乙酸乙酯萃取浓缩后进样,紫外检测波长312nm,按内标法定量。结果奥硝唑线性范围为0.312~40 mg·L^(-1)(r=0.9998),最低检测浓度为0.08 mg·L^(-1),平均提取回收率为96.4%~97.8%,平均方法回收率为99.4%~100.5%,日内、日间精密度RSD小于7.2%。结论本方法简便、快速、灵敏、准确,适用于奥硝唑的药代动力学研究。

OBJECTIVE A high performance liquid chromatography was developed for the determination of ornidazole in rabbit serum using metronidazole as the internal standard.METHODS A variable wavelength UV detector and Kromasil C_(18) column(150mm×4.6 mm,5 m) were used.The serum sample(0.1mL) was extracted with 2 mL of ethyl acetate.After evaporation of the organic layer,the residue was dissolved in mobile phase and injected onto a column.The mobile phase of methanol-water(30:70) was pumped at 1 mL·min^(-1) through the column.The detector at 0.01 AUFS was set at 312 nm.The ornidazole was quantitated by internal standard method.RESULTS The retention time for ornidazole and metronidazole were 7.1 min and 2.8 min,respectively.Standard curve was linear in the range of 0.312~40 mg·L^(-1) with a correlation coefficient of 0.9998.The detection limit in serum was 0.08mg·L^(-1) and the extraction recovery was 96.4%~97.8%,the method recovery was 99.4%~100.5%;intra-day RSD and inter-day RSD were less than 7.2%.CONCLUSION This method is simple,rapaid,sensitive and accurate.It is suitable for the study of pharmacokinetics of ornidazole.