There must be a way to ascertain the most active component...Say bioassay guided, in vitro or in vivo, in situ etc...using microorganism (e.g antimicrobial assays), experimental animals (anti-inflammatory or immunomodulatory assays) or cell lines (gene regulation assays). Following crude extraction protocols of choice, GCMS or HPLC may be carried out, TLC spotting; all these give the peaks, Rfs of the 5 components present. Column chromatography may be run, followed by further TLC spotting. Spots with similar Rfs may be pooled together,while the corresponding  fractions may be tested for activity by bioassay guided experiments. Characterization and structural elucidation of the most active component may then be done using IR and NMR techniques.

hi

Mokhtar Benchohra,

of course, compounds need to separate in order to check each of their effectiveness. Run column with Sephadex L20 might helpful.

TLC spotting and bulking fractionz with similar Rf values