RP-HPLC测定不同产地光果甘草废渣中光甘草定含量

RP-HPLC Determination of Glabridin Content in Different Licorice Residues from Glycyrrhiza glabra L

目的利用反相高效液相色谱法测定由不同产地光果甘草制备的甘草废渣中光甘草定的含量。方法对4种产地的光果甘草用同一种水提法制备甘草废渣;再用不同提取法制备6种总黄酮粗提物;采用SymmetryC18反相色谱柱,流动相为乙腈-水-冰醋酸(55∶44∶1),检测波长为282nm,流速为1.0mL·min-1,柱温为30℃,测定各样品中光甘草定的含量。结果光甘草定在0.09～0.45mg·mL-1内呈良好线性关系(r=0.9997),平均回收率100.2%,RSD为0.89%。6种总黄酮粗提物中光甘草定含量分别为1.31%,1.16%,1.59%,2.93%,0.98%,0.85%;4种甘草废渣中,光甘草定含量分别为0.225%,0.290%,0.211%,0.218%。结论反相高效液相色谱法简便、快速、重复性好,适用于光果甘草中光甘草定的含量测定。4种光果甘草废渣中,新疆和静的光果甘草废渣所含光甘草定含量明显高于其他产地的光果甘草废渣。此外,利用甲醇回流提取法制备光甘草定时,提取率和总黄酮出膏率都高于乙醇回流提取法和甲醇超声提取法。

OBJECTIVE To use the RP-HPLC method for determination of glabridin content in different origines of Glycyrrhiza glabra L （licorice）. METHODS Four samples of licorice residue were prepared from 4 origins of Glycrrhiza glabra L with same water extraction method; six samples of licorice flavonoid crude extracts were prepared from 4 samples of licorice residues with different extraction methods. The Symmetry C18 column was used for the determination of glabridin content; mobile phase was acetonitrile-water-glacial acetic acid （55 ∶ 44 ∶ 1）; detection wavelength was 282 nm, column temperature was 30 ℃. RESULTS There was a good linear relationship when the concentration of glabridin was in the range of 0.09-0.45 mg·mL^-1（r=0.999 7）. The average recovery rate was 100.2% and the RSD was 0.89%. The content of glabridin in the six samples was 1.31%, 1.16%, 1.59%, 2.93%, 0.98% and 0.85%, respectively; and the content of glabridin in four samples of licorice residue were 0.225%, 0.290%, 0.211% and 0.218%, respectively. CONCLUSION The detection by RP-HPLC is a simple and easy method for determination of Glabridin in Glycyrrhiza glabra. The glabridin content in Xinjiang Hejing licorice is higher than those of other sources. Furthermore, the reflux-extraction method by methanol is more effective method for increasing the extract yield and glabridin content than those of the extraction by ethanol or by ultrasonic methanol extraction.