Ssanghwa-tang is a medicinal formula that is widely prescribed in Korea to decrease fatigue after an illness. Fermented herbal medicines might be made more efficacious than conventional herbal medicines by increasing the absorption and bioavailability of the active compounds. In this study, Ssanghwa-tang was fermented to produce bioconversion compositions using Lactobacillus fermentum,and six peaks were decreased, four peaks were increased and one peak newly appeared in the HPLC-DAD chromatogram. The structures of the newly-appearing compound (1)and increased (2-5) compounds were identified as follows using NMR and MS: liquiritigenin (1), nodakenetin (2),cinnamyl alcohol (3), decursinol (4), and benzoic acid (5). The decreased compounds were identified to be paeoniflorin (6), liquiritin (7), nodakenin (8), cinnamaldehyde (9),decursin (10), and decursinol angelate (11) using HPLCDAD analysis with authentic compounds. The high performance liquid chromatography method was used to quantify the eleven constituents in Ssanghwa-tang and fermented Ssanghwa-tang. All calibration curves of the standard compounds exhibited excellent linearity with a R2 >0.9940.