This study systematically evaluated the impact of varying dosages of high-energy electron beam irradiation on both the microbial quantity and physicochemical properties of the herb Glycyrrhizae Radix et Rhizoma. The results showed that irradiation within the range of 4-10 kGy effectively eradicated the total counts of aerobic bacteria, molds, and yeasts present in Glycyrrhizae Radix et Rhizoma. Additionally, it was ascertained that the irradiation did not exert a significant influence on the moisture content, alcohol-soluble extract, and total flavonoid content of the said herb. Post-irradiation, the L* value, which is an indicator of the color transformation of Glycyrrhizae Radix et Rhizoma, gradually decreased, whereas the a* and b* values showed conspicuous augmentation. However, no substantial alterations were detected in the overall color difference. When exposed to irradiation doses ranging from 2 kGy to 10 kGy, the contents of isoliquiritin, liquiritigenin, isoliquiritigenin, and licorice chalcone A in Glycyrrhizae Radix et Rhizoma remained relatively unaltered; and irradiation at 8 kGy and 10 kGy markedly augmented the contents of liquiritin and glycyrrhizic acid. The similarity of the fingerprint spectra of Glycyrrhizae Radix et Rhizoma at different irradiation dosages all exceeded 0.98, and the antioxidant activity was retained at a relatively elevated level. In conclusion, high-energy electron beam irradiation can be utilized for the sterilization and storage of Glycyrrhizae Radix et Rhizoma, but the irradiation dosage should be precisely controlled to prevent any potential quality modifications.