This study focused on developing a set of quality evaluation methods that can reflect the multi-levels and multi-characteristics of traditional Chinese medicine (TCM). Taking licorice (Glycyrrhiza glabra L.) as the method development sample, the feasibility of multi-markers assay by monolinear method (MAML) was explored through the standard curve relationship among active components for the first time. Using glycyrrhizic acid as measurement marker, MAML can simultaneously quantify five components of licorice, including isoliquiritigenin, isoliquiritin apioside, liquiritigenin, liquiritin and liquiritin apioside. Comparing MAML and quantitative analysis of multi-components by single-marker (QAMS) to the external standard method (ESM) respectively, it was found that there was no significant difference in the content of components that were calculated by MAML and ESM (the relative error (RE) was generally less than 2.00%). However, the RE of the component content calculated by QAMS fluctuated greatly, indicating that the MAML was more accurate than QAMS. In addition, UV and THz quantum fingerprints were initiated by the interval erasure method. Taking the systematically quantified fingerprint method as the core, a "Trinity" fingerprint quality evaluation system based on HPLC, UV and THz was developed. The system successfully distinguished the quality differences of licorice samples from 13 producing areas and two ecological models by the comprehensive evaluation results. Simultaneously, the quality information of licorice at different technical levels was revealed. Finally, bivariate correlation analysis was used to examine the linkage between UV/HPLC and antioxidant spectrum efficacy, and the two-dimensional activity spectrum of licorice was provided. It may furnish a more thorough and objective analytical technique for licorice and even other TCMs in chemical fingerprint features, chemical bond vibration characteristics and biological activity information.
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