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To prepare the sample for HPLC analysis, about 1 g of extract was mixed with 20 mL of methanol and water (1:1 v/v) and heated on a water bath at 70 °C for 1 h. Centrifugation of sample was conducted at 4000 rpm for 10 min after cooling. Then, 2 mL from sample was filtered with Whattman filter paper into HPLC vials and labelled with proper codes.
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