© 2019 Informa UK Limited, trading as Taylor & Francis Group.In this work, the antioxidant properties of 4′,5,7-trihydroxy-3,6-dimethoxyflavone using several bioanalytical assays is investigated in vitro. These methods are Cu2+-Cu+ reducing, Fe3+-Fe2+reducing, DPPH• scavenging, and ABTS•+ scavenging. The IC50 values of the 4′,5,7-trihydroxy-3,6-dimethoxyflavone for DPPH• and ABTS•+ scavenging assays were 9.36 and 3.55 μg m–1, respectively. Additionally, 4′,5,7-trihydroxy-3,6-dimethoxyflavone compound recorded best inhibitory effects against human carbonic anhydrase (hCA) I and II isoenzymes, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE) enzymes. 4′,5,7-Trihydroxy-3,6-dimethoxyflavone had K i value of 389.56 ± 29 nM against hCA I and 306.90 ± 41 nM toward hCA II, respectively. Also, the K i values of AChE and BChE were found as 1005.37 ± 117 nM and 814.22 ± 87 nM, respectively.