HPLC测定CO_2超临界萃取的当归油药效组分

Using HPLC Method to Determine the Active Components Alignment of Oleum Angelicae Sinensis Extracted by Supercritical-CO_2 Fluid

目的:测定CO2超临界萃取的当归油药效组分藁本内酯、正丁烯基苯酞、正丁基苯酞。方法:采用HPLC,ZORBAX-Eclipse XDB-C18色谱柱(4.6 mm×25 cm,5μm),流动相0.05 mol.L-1醋酸铵水溶液(pH 4.0±0.2)(A)-乙腈(B),梯度洗脱(0～10 min,30%B;10～60 min,30～60%B;60～65 min,60%B;65～70 min,60～30%B),检测波长274 nm,流速1.0 mL·min-1,柱温25℃。结果:超临界萃取的当归油药效组分藁本内酯、正丁烯基苯酞、正丁基苯酞分别在0.461 0～20.842 6μg,0.012 6～0.476 8μg,0.022 8～0.825 1μg呈良好的线性关系;平均加样回收率分别为101.27%,99.12%,97.14%。结论:HPLC可用于超临界萃取的当归油药效组分测定,为建立当归油及其制剂与临床疗效对应的质量标准提供了新的思路和方法。

Objective：To simultaneously determine the active components alignment which is Ligustilide-Butylidenephthalide-Butylphthalide in Oleum Angelicae Sinensis extracted by supercritical-CO2 fluid.Method： The HPLC chromatographic analysis was performed on a ZORBAX-Eclipse XDB-C18 column（4.6 mm×25 cm,5 μm） with gradient elution（0-10 min,30% B;10-60 min,30%-60% B;60-65 min,60% B;65-70 min,60%-30% B）of 0.05 mol·L-1ammonium acetate（using acetic acid to adjust pH in the range of 4.0±0.2）（A）and acetonitrile（B）.The detection wavelength was set at 274 nm,the flow rate at 1.0 mL·min-1and the colunm temperature at 25 ℃.Result： The calibration curves were linear with in the range of 0.461 0-20.842 6 μg,0.012 6-0.476 8 μg,0.022 8-0.825 1 μg for ligustilide,butylidenephthalide,butylphthalide,respectively.The average recoveries of the active components alignment were 101.27%,99.12%,97.14%,respectively.Conclusion： This method has been successfully applied to the simultaneous determination of the active components alignment in Oleum Angelicae Sinensis extracted by supercritical-CO2 fluid,which provides new methods and index to build the quality standard of Oleum Angelicae Sinensis and its preparations corresponding to clinical effect.