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Protocol

The Isolation of coumarin derivative from Plants

Ethanol, n-hexane, ethyl acetate (EtOAc) and n-butanol

1. Cut air-dried plant materials (1 kg) into small pieces and macerate the sample in 3 liters 90 % ethanol. 2. Centrifuge the samples soaked for 10 days at room temperature. 3. Concentrate the extract under reduced pressure. 4. Partition the concentrated with n-hexane. 5. Extract the sample with ethyl acetate (EtOAc) and n-butanol. 6. Combine and concentrate the extract (organic fractions). 7. Mix the sample with silica gel (sample/silica gel=1/6) 8. Elute the mixed sample applied to column chromatography using n-hexane and EtOAc and collect an appropriate number of fractions for your study. 9. Analyze the fractions using HPLC, LC-MS/MS and NMR.

The Isolation of coumarin derivative from Plants:

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