This study was to evaluate the quality consistency of glycyrrhiza extract and to explore the possible anti-oxidant components in combination with chromatographic fingerprint and bioactivity evaluation. Characteristic fingerprints of glycyrrhiza extract samples from different sources were generated by high performance liquid chromatography with diode array detector (HPLC-DAD) and evaluated using hierarchical clustering and similarity analysis. Compared with the conventional qualitative similarity evaluation method, the averagely linear quantified fingerprint method had an important quantitative similarity parameter supported by quantitative analysis, which was recommended in the fingerprint evaluation. Antioxidant activities of the glycyrrhiza extract samples were determined by DPPH (2, 2-diphenyl-1-picryldrazyl) radical scavenging assays. In addition, the fingerprint-efficacy relationship was investigated by the chemical fingerprints and the anti-oxidant activities utilizing partial least squares model, which was capable of exploring and discovering the bioactive components of glycyrrhiza extracts. Therefore, the present study provided a powerful strategy to evaluate the holistic quality consistency of medicinal plant.