Chemical analysis of epicatechin and catechin was conducted by HPLC analysis and a minor modification of a previously published method (Kim et al., 2014 (link)). Briefly, dried samples of F. dibotrys rhizomes were ground into a fine powder using a grinder. Powdered samples (~150 mg) were subjected to extraction via ultra-sonication with 80% (v/v) methanol at room temperature for 60 min. Subsequently, the extracts were centrifuged, and the supernatant was filtered through a 0.45-μm micromembrane for HPLC analysis. A Merck C18 column (250 mm × 4.6 mm × 5 μm) was used as the stationary phase, and a solution of ultrapure water (pH was adjusted to 3.00 ± 0.02 with phosphoric acid) and acetonitrile (90:10) was the mobile phase, and the flow rate was maintained at 1.0 mL·min−1 at 35°C. An injection volume of 20 μL and a wavelength of 280 nm were used for detection. The compounds in the sample were determined using a standard curve. All samples were analyzed in triplicate.
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