Isolation of Anti-HIV Components from Canarium album Fruits by High-Speed Counter-Current Chromatography

The fruit of Canarium album, also called Chinese olive, is a popular food and traditional Chinese herb. The ethyl acetate extract of Canarium album fruits was found to exhibit an inhibitory effect on six-helix bundle formation of the human immune deficiency virus (HIV) glycoprotein transmembrane subunit gp41 in our previous study. In this paper, a preparative separation of anti-HIV components from an ethyl acetate extract of Canarium album fruits was performed by high-speed counter-current chromatography, using a solvent system of n-hexane–ethyl acetate–ethanol–0.5% acetic acid aqueous (0.5:9.5:2:8, v/v/v/v). Among the six fractions obtained from a single separation process, three exhibited over 90% purity as determined by HPLC, two compounds were identified as 5-hydroxymethylfurfural and 3,4,5-trihydroxybenzoic acid, and four fractions displayed significant inhibitory effects on HIV-1 IIIB infection. These findings suggest that the high-speed counter-current chromatography is an effective and reliable technique for separating bioactive components from medicinal herbs. Further identification of the active compounds in Canarium album fruits and studying their mechanism of action are warranted.