Isolation of a Biflavone (Agathisflavone) with Antifungal Activity from Anacardium occidentale Leaves (Cajú) by Centrifugal Partition Chromatography (CPC)

Anacardium genus (Anacardiaceae) consists of approximately 20 species, and Anacardium microcarpum, Anacardium humile, and Anacardium occidentale are the most widely studied, because of their medicinal and nutraceutical properties. Their common name is cashew, marañon, or cajú. The leaves and flowers of A. occidentale have been used in traditional medicine to treat skin lesions and diarrhea and as anti-inflammatory agents. Previous results in our research group have shown that the leaf extract of A. occidentale possesses an interesting antifungal activity, inhibiting the proliferation and growth of Candida albicans. To identify a potential active marker for this extract, the purification of one of its major compounds by Centrifugal Partition Chromatography (CPC) was carried out in this work. First, the EtOAc fraction was separated using the CHCl3:MeOH:H2O (4:3.5:2) solvent system. We obtained 243 mg of the EtOAc fraction then it was solubilized in 5 mL of both phases. In a single CPC run, 12.5 mg of the major compound could be isolated from the extract, with a purity of over 90%, which was subsequently identified by LC-PDA-MS/MS as being the biflavone Agathisflavone showed a minimum inhibition concentration (MIC) of 250 μg/mL against C. albicans SC5314 and It did not show cytotoxicity activity in vitro. The CPC analysis described in this work allowed obtaining, in a single run, the major flavonoid of the A. occidentale leaf extract, which is now being evaluated for its antifungal potential against Candida albicans.