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Crude HWHF was chromatographed with flow rate of 1.0 ml/min on a (10 x 250 mm) semi-preparative 5μm C-18 column (Phenomenex, UK). The concentration of the eluting solvent was raised using linear gradients of acetonitrile (0–20% from 0 to 10 min, 20–70% from 10 to 40 min and 70–20% from 40 to 60 min). Compounds were detected by measurement of absorbance at 254-360nm.
