Hi Oludare Temitope Osuntokun

First perform qualitative phytochemical testing to find out which type of phytochemicals are present in your extract via chemical tests. Afterwards perform thin layer chromatographic profiling of the extract to see how many compounds might be present in the extract and which one is supposed to be the major one. On the basis of chemical composition and previous findings about the plant, try to figure out the major active compound of your extract or the marker compound. It may be the phytochemical marker or biological marker, not necessary that the activity of the extract will be because of that compound. But it will serve as a identifying parameter for the extract and you can standardize the extract if you have the standard marker with you using HPTLC or HPLC. Now you should try to find out the biological activity of that extract via in-vitro or in-vivo experiments whichever is applicable for particular activity. If activity is evident in the extract, then you can try to isolate the active component via column chromatography or other suitable techniques. Alternatively you can also measure the content of standard compound in the extract and if it has the claimed activity in literature, you can call your extract as bio-active. For characterization of the bioactive fraction or the marker compound, you can use UV, IR, NMR, MS, LCMS, GCMS etc, which will help you elucidate the structure of the marker or standardize the fraction accordingly.

Regards 

Hi Oludare Temitope Osuntokun

First perform qualitative phytochemical testing to find out which type of phytochemicals are present in your extract via chemical tests. Afterwards perform thin layer chromatographic profiling of the extract to see how many compounds might be present in the extract and which one is supposed to be the major one. On the basis of chemical composition and previous findings about the plant, try to figure out the major active compound of your extract or the marker compound. It may be the phytochemical marker or biological marker, not necessary that the activity of the extract will be because of that compound. But it will serve as a identifying parameter for the extract and you can standardize the extract if you have the standard marker with you using HPTLC or HPLC. Now you should try to find out the biological activity of that extract via in-vitro or in-vivo experiments whichever is applicable for particular activity. If activity is evident in the extract, then you can try to isolate the active component via column chromatography or other suitable techniques. Alternatively you can also measure the content of standard compound in the extract and if it has the claimed activity in literature, you can call your extract as bio-active. For characterization of the bioactive fraction or the marker compound, you can use UV, IR, NMR, MS, LCMS, GCMS etc, which will help you elucidate the structure of the marker or standardize the fraction accordingly.

Regards 

Try bio activity guided fractationation to confirm first bio active moiety.

I think that for isolation the method most commonly used is 'traditional' isolation based on extraction, fractionation and column chromatography with simultaneous control of the collected fractions (TLC, HPLC). However, this is a long, time-consuming process, which not always gives the desirable results. On the other hand, a quite quicker alternative is the isolation of target compounds by the use of preparative TLC or HPLC. Using all mentioned methods is also possible, especially if you are going to isolate more than one major compound.

Hope this would help.

Best regards

You can extract by organic solvents (the best methanol) and edification by more method or tool example GC-mass, Hplc, tof-tof mass, .....etc.

Try HPTLC and then for identification you can go for MS by dissolving the bands in right solvents. This is a faster method if you compare with HPLC. Also what kind of bioactivity are you looking for?

It varies from sample to sample. Therefore, you have to develop and standardize an activity-guided method to isolate active compound from your extract using different polarity solvents from non-polar to polar.