Licorice Liquid Chromatography Fingerprinting And Its Application Research

Developed an HPLC fingerprint and quantitatively determine of glycyrrhizic acid(GA) and liquiritin(L) simultaneously for licorice of various species. Identified major peaks in the fingerprint by DAD and ESI-MS detection. The analytical method developed here can be used for the characterization of large quantities of wild and homegrown licorice plants. UV, DAD and ESI-MS/MS detectors were used and the experimental results were compared with the literature data to identify fingerprint major peaks. The 67% methanol extracts of licorice plants were gradient eluted by a mixture of water (with 0.1% H3PO4 for UV and 1% formic acid for ESI-MS detection, respectively) (solution A) and acetonitrile (solution B) on an ODS column. The stepwise gradient profile of elution was 15%B at a starting flow rate of 0.5 mL/min, followed by 25%B at 0.8 mL/min for 10 minutes, 32%B for 20 minutes and 80%B for 70 minutes. The UV detection wavelengths were 0min 276nm 19min 360nm 24min 248nm . Total of 20 compounds of saponins and flavoids were identified from 17 major peaks in the chromatograms. The corresponding peak number/compound were: saponins 14/licorice-saponin(LS)-A3, 16/LS-G2, 17/glycyrrhizin acid, 18/LS-H2, 19/LS-K2, 21/apioglycyrrhizin(in G. inflata), 22/LS-J2 and araboglycyrrhizin(in G. inflata), and 23/LS-C2; flavones 4/liquiritin apioside, 6/liquiritin, 8/isoliquiritin apioside, 9/licuraside, 10/isoliquiritin and ononin, 11/neoisoliquiritin, 22/uralenol, 25/glycycoumarin, 29/licochalcone A and 30/isoglycyrol.A great number of wild and homegrown licorice plants were analyzed to identify the applicability of the fingerprint method. At first, we analyzed parts of wood, bark, taproot, rootstock and rootlet of cultivated G. Uralensis. Bark brought forth more peaks on the fingerprint after 40 min and higher content of most constituents, GA and L. The content of GA and L in different parts show the same trend: rootlet>taproot>rootstock. In 1~4-year-old cultivated llicorice, the content of GA and L increase continually.Then, three widely used medicinal wild licorice plants from Xingjiang, Glycyrrhiza Uralensis Fisch., Glycyrrhiza Glabra L. and Glycyrrhiza Inflata Bat were analyzed by RF-HPLC fingerprint. Hierarchical Cluster Analysis (HCA) and discriminant examination was carried out on the basis of some characteristic peak areas which chosen by statistic and professional analysis and obtained satisfactory results.More than 80 soilless cultivated licorice samples were analyzed to study the influences of mineral elements on the chemical constituents and content of GA and L.The method provides some useful information about the chemical constituents of licorice and determines the chemical contents prescribed by new China pharmacopoeia. We expect more rules on the quality of licorice from different sources to be uncovered as our study deepens.