Preparative thin-layer chromatographic separation followed by identification of antifungal compound in Cassia laevigata by RP-HPLC and GC-MS.

ABSTRACT

BACKGROUND: Several species of the genus Cassia are known for their antioxidant, antimicrobial and antidiabetic activities, but some of the lesser-known Cassia species, e.g. C. renigera, C. biflora and C. laevigata have not been studied for their biological activities. RESULTS: Methanol extract of C. laevigata was fractionated by preparative thin-layer chromatography. The resulting six different fractions were tested against Fusarium oxysporum and Aspergillus niger for their antifungal activity. Due to higher antifungal activity of fraction 1 of C. laevigata, this was further analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC), resulting in distinct separation of one compound at a retention time of 7.2 min with an absorbance of 252 nm. Further, this compound was analyzed by gas chromatography-mass spectrometry (GC-MS) for its putative structural identification. Mass spectra of this compound resembled the spectra of anthraquinone 1-carboxylic acid by NIST library search. The genomic-level expression of chalcone synthase, a key enzyme involved in the biosynthesis of polyketides, was increased in C. laevigata when compared to other Cassia species. CONCLUSIONS: This study provides an insight into the higher antifungal activity of C. laevigata, including the identification of anthraquinone 1-carboxylic acid, which may be responsible for the antifungal activity.