I have tried liquid- liquid extraction method for isolating the active compound for anti inflammation from active microbial broth by different organic solvents ethyl acetate, chloroform, hexane, dichloromethane, methanol and ethanol but none of them is capable for isolating it. These solvents were mixed with broth in 1:1 ratio and vigorously rotate for 2-4hours. What should I do to get the active compound? please help me.
I would also recommend to fractionate your supernatant via HPLC. What kind of molecule is your compound? The exact knowledge of your compound of interest is, of course, very important for choosing proper HPLC conditions. Otherwise, you could loose too much time.
If your potential antiinflammatory bioactives are not getting extracted from the active microbial broth, using different organic solvents mentioned by you, then probably your antiinflammatory compound is proteinaceous/lignan in nature. Have u performed a protein test.
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