How to extract secondary metabolite/ antimicrobial compound from bacterial medium supernatant if do not know the nature of metabolite.
I used ethanol, ethyl acetate, chloroform, hexane for extraction,but my compound is coming in the aqueous phase of all..how to purify further because in aqueous phase, there may be medium component present.
I am not sure how you used ethanol for extraction of aqueous-based supernatant as it is miscible with water. You are correct in that media components are there in the aqueous medium along with hydrophilic metabolites. You can go up higher in polarity by using n-butanol which is not miscible with water and is far higher in polarity profile than ethyl acetate.
Thanks, Dear Solomon Habtemariam for your response. I will think over it.
For the extraction of secondary metabolites you can use ethanol, chloroform, hexane, ethyl acetate but as your compound is water soluble then u can follow the process:
Extract directly with ethanol, concentrate it. Now separate this concentrate with hexane or ethyl acetate and water. In water layer you get your compound as major one because sugar is not extracted in ethanol and fatty acids and sterols get removed by hexane.
Secondly, you can do direct extraction with ethanol. After that you have to extract the ethanolic extract directly with hexane. Now the left residue extract directly with water or ethyl acetate
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