<p>This study aims to identify potential tyrosinase inhibitors and antioxidants in Alpiniae Katsumadai Semen (AKS) using various techniques, including matrix solid-phase dispersion (MSPD), thin-layer chromatography (TLC), bioautography, mass spectrometry (MS), and molecular docking. First, three extracts with varying polarities were prepared using 5% methanol aqueous solution, 60% methanol aqueous solution, and 100% methanol as solvents via MSPD. Subsequently, the tyrosinase inhibition activity and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging capacity of these extracts were evaluated. The results indicated that the 60% methanol extract exhibited superior activity in both assays. Thus, 60% methanol extract was selected for the TLC‒bioautography assay, which successfully identified six components, including three tyrosinase inhibitors (1,7-diphenyl-5-hydroxy-1-heptene-3-one and 1,7-diphenyl-4-hepten-3-one and its isomer) and six antioxidants (alpinetin, 1,7-diphenyl-4-hepten-3-one and its isomer, cardamonin, 1,7-diphenyl-5-hydroxy-1-heptene-3-one, and pinocembrin). The tyrosinase inhibitory activity was validated through molecular docking for three tyrosinase inhibitors, while the antioxidant activity was confirmed through in vitro biological assays for alpinetin, cardamonin, and pinocembrin. Notably, the compounds 1,7-diphenyl-5-hydroxy-1-heptene-3-one and 1,7-diphenyl-4-hepten-3-one and its isomer are reported for the first time in AKS. Furthermore, this study presents a novel approach that allows for the simultaneous screening of tyrosinase inhibitors and ABTS antioxidants on a single TLC plate, thereby advancing methodologies within this research field. In conclusion, this research not only identifies the tyrosinase-inhibiting and antioxidant components in AKS but also provides a scientific basis and theoretical support for its future application in product development.</p>