Simultaneous Quantitative Determination of Nine Active Chemical Compositions in Traditional Chinese Medicine Glycyrrhiza by RP-HPLC with Full-Time Five-Wavelength Fusion Method

A new, simple, accurate and reliable full-time five-wavelength fusion method for the simultaneous separation and determination of nine active chemical compositions (liquiritin apioside, liquiritin, isoliquiritin apioside, ononin, isoliquiritin, liquiritigenin, calycosin, isoliquiritigenin, Glycyrrhizic acid monoammonium salt) in traditional Chinese medicine Glycyrrhiza was developed using reverse phase high-performance liquid chromatography (RP-HPLC) coupled with a diode-array detector (DAD). The chromatographic separation was performed on an Agilent TC-C18 column with gradient elution using 0.04% methanoic acid (A) and acetonitrile (B) at a flow rate of 1.0 mL.min(-1) and UV detection at 248 nm, 250 nm, 276 nm, 362 nm, 370 nm. The standard curves were linear over the range of 2.1379-12.8272 mu g for liquiritin apioside, 3.9299-23.5794 mu g for liquiritin, 1.0432-6.2592 mu g for isoliquiritin apioside, 0.8764-5.8584 mu g for ononin, 1.0701-6.4205 mu g for isoliquiritin, 1.3685-8.2111 mu g for liquiritigenin, 0.3927-2.3563 mu g for calycosin, 0.2498-1.4986 mu g for isoliquiritigenin, 2.0094-12.0564 mu g for Glycyrrhizic acid monoammonium salt, respectively (r(2) > 0: 9997). The recoveries and relative standard deviation (RSD) varied from 95.09% to 103.54% and 1.09% to 2.36%, respectively. The precision for all the analytes was less than 2.52%. The method indicated good performance in terms of precision, accuracy and linearity. The method enabled the simultaneous determination of nine active chemical compositions for quality control of Glycyrrhiza.