High-speed counter-current chromatography (HSCCC) with a two-phase solvent system (hexane–ethanol–acetonitrile–water 10:8:1:1, v/v) was applied to examine the leaves of Hortia oreadica, which afforded the known limonoid guyanin ( 1), the alkaloids rutaecarpin ( 2) and dictamnine ( 6), the dihydrocinnamic acid derivatives methyl 5,7-dimethoxy-2,2-dimethyl-2 H-1-benzopyran-6-propanoate ( 3), 5,8-dimethoxy-2,2-dimethyl-2 H-1-benzopyran-6-propanoic acid ( 4), together with the new E-3,4-dimethoxy-α(3-hydroxy-4-carbomethoxyphenyl)cinnamic acid ( 5). The recovery of compounds 1–6 was determined by comparison with LC-atmospheric pressure chemical ionization MS/MS data: 66.2%, 93.1%, 102.5%, 101.2%, 99.0% and 84.9%, respectively. Compound 3 showed IC 50 of 23.6 μM against Plasmodium falciparum and 15.6 μM against Trypanosoma brucei rhodesienses and was not toxic to KB cells (IC 50 > 100 μM).