RP-HPLC法测定小鼠血浆中蒿甲醚的浓度

Content analysis of artemether in mice plasma by RP-HPLC

目的建立测定小鼠血浆中蒿甲醚浓度的方法。方法采用RP-HPLC法,色谱柱为Diamonsil C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-水(体积比85∶15);流速为1.0 mL/min;检测波长为210 nm;柱温为室温。利用甲基叔丁基醚对小鼠血浆及组织中蒿甲醚及蒿乙醚进行提取后进行测定。结果蒿甲醚与蒿乙醚的色谱峰分离良好并且出峰时间相近,以蒿乙醚为内标测定血浆中蒿甲醚符合方法学要求,血浆标准曲线在一定范围内线性关系良好(R2=0.9991,n=5),高、中、低3个浓度的回收率均为97%以上,日内及日间RSD均小于3%。结论建立的方法可简单快速测定蒿甲醚在小鼠血浆中的浓度,为内标法测定蒿甲醚在动物血浆中的浓度提供思路。

Objective To establish a method for determining the contents of artemether and arteether in mice plasma by RP-HPLC. Methods Artemether and arteehter in mice plasma were extracted by methyl tert-butyl ether before detection, then analyzed on a Diamonsil C18 column （150 nun × 4.6 mm, 5 μm）. The mobile phase consisted of methanol-water（ 85：15 ）. The flow rate was 1.0 mL/ min, The wavelength was 210 nm and room temperature was selected for determination of artemether. Results The chromatographic peaks of artemether and arteether were totally separated with similar retention times at the selected HPLC condition. There was a good linearity for artemether analysis when arteether was used as the inner standard （r = 0. 9991, n = 6）. The recoveries were all more than 96% during high, middle and low concentration of artemether. RSDs of within-days and between-days were all less than 3%. Conclusion The concentration of artemether in plasma could be analyzed easily and rapidly by the established HPLC method.