Abstract
Three polymethoxylated flavones (PMFs) including nobiletin (NOB), tangeretin (TAN) and 5-demethylnobiletin (5-DN) were separated from Ponkan (Citrus reticulata Blanco cv. Ponkan) peel by high-speed countercurrent chromatography (HSCCC) with a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (1:0.8:1:1, v:v:v:v). A total of 6.4 mg of NOB, 20.2 mg of TAN and 2.8 mg of 5-DN with the purity of 97.7, 99.05 and 95.9 %, respectively, were obtained in one-step separation from 70 mg of crude extracts. Their chemical structures were identified by HPLC/ESI–MS and 1H and 13C-NMR. The antifungal activities of the three compounds against Aspergillus niger were evaluated by using a microbroth dilution assay. 5-DN displayed the best inhibitory effect, while NOB showed the lowest potency, with respective minimal inhibitory concentration (MIC) values of 0.1 and 0.8 mg/mL, which indicated that the inhibition against A. niger could be affected by the structural difference in PMFs.
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