A reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for simultaneous determination of five bioactive components, liquiritin apioside, liquiritin, licuraside, isoliquiritin and glycyrrhizin in licorice with diode-array detection (DAD). The method was performed with a mobile phase consisting of acetonitrile and 0.1% (v/v) phosphoric acid under gradient elution. All calibration curves of the five bioactive components showed excellent linearity (r2 ≥ 0.9997) within the ranges of 91.2 - 1824 μg/mL, 54.4 - 2720 μg/mL, 6.4 - 512 μg/mL, 6.4 - 512 μg/mL and 20.4 - 2040 μg/mL, respectively. The average recoveries ranged from 95.86% to 103.11%. This analytical method was also validated with respect to precision, repeatability and accuracy, and proved to be sensitive and accurate to simultaneously determine the five components in licorice. The developed method was further applied to quantify the contents of five components in forty two batches of either raw or honey-processed licorice samples of Glycyrrhiza uralensis Fisch. and G. glabra L. The results revealed that the contents of five components had descended in toto after the honey process. The content ratio of glycyrrhizin to liquiritin was more than 5.0 in licorice of G. glabra whereas it was less than 3.0 in that of G. uralensis in both raw and honey-processed licorice materials. This content ratio difference may be used to distinguish among licorice materials.