Abstract: The chemical constituents of 50% acetone extract of the dried roots bark of Paeonia suffruticosa Andr. (Moutan Cortex) were isolated and purified using various column chromatographic methods, yielding 16 compounds. According to their physiochemical properties and spectral data, the structures of the isolated compounds were further identified as suffruticoside N (1), 1, 1′-(1R)-2, 2′-dihydroxy-6, 6′-dimethoxy1, 1′-biphenyl-3, 3′-diylbisethanone (2), 5-(1, 2-dihydroxy-1-methylethyl)-4-methylbicyclo3.1.0hexan-2-one (3), 4-(1, 2-dihydroxy-1-methylethyl)-1-cyclohexene-1-carboxylic acid (4), teutenone A (5), dihydroactinidiolide (6), cadina-4, 11-dien-14-oic acid (7), adenosine (8), uridine (9), methyl palmitate (10), 1-(3-hydroxy-4-methoxyphenyl)ethanone (11), 2, 4-dihydroxyacetophenone (12), methyl gallate (13), 2, 3-dihydroxybenzoic acid (14), benzoic acid (15), and pinoresinol (16). Compound 1 was a new biphenyl compound. Compounds 2 and 3 were two new natural products, and compounds 4 and 5 were firstly isolated from Moutan Cortex. Additionally, in preliminary vitro bioassays, all the isolated compounds were tested their inhibitory effects on acetylcholinesterase (AChE). As a result, compound 2 exhibited a moderate AChE with IC₅₀ value of 15.7 ± 2.5 μmol·L^-1.