高效液相色谱法测定甘草及其炮制品中5种活性成分的含量

 目的建立甘草及其3种炮制品中芹糖甘草苷、甘草苷、芹糖异甘草苷、异甘草苷和甘草酸的HPLC含量测定方法,并比较炮制前后5种主要活性成分含量的变化。方法采用依利特Hypersil ODS色谱柱,乙腈-3%乙酸梯度洗脱,流速为0.6mL·min^-1,检测波长为254nm,柱温为35℃,对甘草、炙甘草、蜜润甘草、清炒甘草中的5种活性成分进行含量测定。结果芹糖甘草苷、甘草苷、芹糖异甘草苷、异甘草苷和甘草酸分别在0.47～15.04μg(r=0.9996),0.13～4.16μg(r=0.9995),0.11～3.52μg(r=0.9999),0.13～4.16μg(r=0.9998),0.53～16.96μg(r=0.9990)内呈良好的线性关系;平均回收率分别为97.7%,100.8%,99.7%,100.5%和99.6%,RSD分别为0.4%,1.8%,1.5%,2.1%,1.9%。测定甘草及其3种炮制品中5种活性成分的结果表明,除甘草苷含量蜜炙甘草高于蜜润甘草外,5种成分在4种样品中的含量依次为清炒甘草>生甘草>蜜润甘草>蜜炙甘草。结论炮制对甘草5种主要活性成分的含量有明显影响;本方法简便,快捷,可用于甘草及其炮制品的质量控制。

Determination of Five Active Constituents in Licorice and Its Processed Products

OBJECTIVE To establish a HPLC method for the simultaneous quantification of four flavonoids (liquiritin apioside , liquiritin,isoliquiritin apioside, isoliquiritin) and one triterpenoid (glycyrrhizic acid) in Radix Glycyrrhizae (licorice), honey-fried licorice, honey-soaked licorice and fried licorice. METHODS The samples were separated at 35 ℃ on a Hypersil ODS column with gradient elution and the mobile phase consisted of acetonitrile and 3% acetic acid.The flow rate was 0.6 mL·min^-1 and the detection wavelength was set at 254 nm. RESULTS Five active constituents in the licorice were separated and determined simultaneously. The calibration curves of liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin and glycyrrhizic acid were linear over the range of 0.47-15.04 μg (r=0.999 6), 0.13-4.16 μg (r=0.999 5), 0.11-3.52 μg (r=0.999 9), 0.13-4.16 μg (r=0.999 8), 0.53-16.96 μg (r=0.999 0), and the recoveries of the 5 components were 97.7%, 100.8% , 99.7% , 100.5% and 99.6%, respectively. The relative standard deviations(RSD) were 0.4%,1.8%,1.5%,2.1% and 1.9%, respectively. The content of liquiritin in the honey-fried licorice was higher than that in the honey-soaked licorice, the contents of the other constituents were in descending order in the four different samples as follows: dry-fried licorice, licorice, honey-soaked licorice and honey-fried licorice. CONCLUSION The processing process has obvious effects on the contents of the five active constituents of licorice. The method is simple and convenient and it can be used for the quality control of the licorice and honey-fried licorice.