HPLC双波长法同时测定甘草药渣提取物中甘草苷、异甘草苷、甘草素、异甘草素及甘草酸含量

Simultaneous Determination of Liquiritin, Isoliquiritin, Glycyrrhizin, Isoliquiritigenin and Glycyrrhizic Acid in Licorice Extract by HPLC Dual Wavelength Spectrophotometry

目的建立甘草药渣提取物中甘草苷、异甘草苷、甘草素、异甘草素及甘草酸的测定方法。方法采用HPLC双波长法对甘草药渣提取物中的主要成分甘草苷、异甘草苷、甘草素、异甘草素和甘草酸进行定量分析。色谱柱为Symmetry C18柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.085%磷酸水(B)为流动相,采用梯度洗脱(0~8 min,81%B;8~35 min,81%→50%B;35~60 min,50%B),流速1.0 m L/min,进样量10μL,柱温为室温,双波长检测(λ1=237 nm,λ2=254 nm)。结果甘草苷在0.040 8~0.816μg、异甘草苷在0.052 8~1.056μg、甘草素在0.022 4~0.448μg、异甘草素在0.021 2~0.424μg、甘草酸在0.044 8~0.896μg范围内均呈良好的线性关系,平均回收率分别为98.69%、98.31%、99.10%、98.55%、99.14%,RSD分别为1.39%、1.29%、1.78%、2.14%、1.15%。结论本法准确可靠,专属性强,结果稳定,重复性好,可用于甘草药渣提取物中上述5种成分的测定。

Objective To establish a method for the simultaneous determination of liquiritin,isoliquiritin,glycyrrhizin,isoliquiritigenin and glycyrrhizic acid in licorice extract.Methods Liquiritin,isoliquiritin,glycyrrhizin,isoliquiritigenin and glycyrrhizic acid in licorice extract were determined by HPLC dual wavelength spectrophotometry.Symmetry C18column(4.6mm×250mm,5μm)was used.The mobile phase was acetonitrile(A)-0.085%phosphoric acid water(B),ingradient elution mode(0-8min,81%B;8-35min,81%→50%B;35-60min,50%B)with the flow rate of1.0mL/min.The sample size was10μL,and column temperature was room temperature.Dual wavelength detection,λ1=237nm,λ2=254nm.Results Liquiritin,isoliquiritin,glycyrrhizin,isoliquiritigenin and glycyrrhizic acid were linear in the ranges of0.0408-0.816μg,0.0528-1.056μg,0.0224-0.448μg,0.0212-0.424μg,and0.0448-0.896μg,respectively.The average recovery was98.69%,98.31%,99.10%,98.55%,and99.14%,respectively;RSD was1.39%,1.29%,1.78%,2.14%,and1.15%,respectively.Conclusion The method is accurate,reliable and specific.The results are stable with good repeatability.It can be used for the determine of above5components in licorice extract.