I used silicagel column chromtoghraphy for fractionation my chloroform fraction of my plant extraction, and when I analysed some of my fraction with TLC I found some phenolic compound in it. I read some article about polyamide cc but I do not know much about it.
I pack it just like silica gel. I packed it in water and attached it to my flash chromatography system (CombiFlash Rf-200), I ran a gradient starting with water to 100% methanol, another gradient from methanol to methanol to 100% acetone, then acetone to 100% water containing 5% ammonium hydroxide (3 gradients total).
if you use a glass column, this can be done with step gradients as well.
You can use the slurry packing method or the wet packing technique. But prior to packing, please don't forget to soak the polyamides in water. Pre swollen material will not pose problems of air retention and subsequent breaking of the column.
For more details you can refer the following papers:
Wall, M. E., Taylor, H., Ambrosio, L. and Davis, K. (1969), Plant antitumor agents III: A convenient separation of tannins from other plant constituents. J. Pharm. Sci., 58: 839–841. doi: 10.1002/jps.2600580709
M.E. Wall, M.C. Wani, D.M. Brown, F. Fullas, J.B. Olwald, F.F. Josephson, N.M. Thornton, J.M. Pezzuto, C.W.W. Beecher, N.R. Farnsworth, G.A. Cordell, A.D. Kinghorn (1996). Effect of tannins on screening of plant extracts for enzyme inhibitory activity and techniques for their removal. Phytomedicine, Volume 3, Issue 3, Pages 281-285
Regards
Thank you everybody for the suggestions, thank you Dr. alok nahata for these papers, unfortunately I could not access to them, could you please send them to my email address : [email protected]. Thank you so much.
And another question is: is it possible to separate sesquiterpene by this method?
It depends how functionalized they are. If they have many substituents such as hydroxyl groups or phenolic's then potentially yes. If they are very non polar sesquiterpenes stick with silica gel
Please send me informations about flavonoids isolation using polyamide column. if is it necessary make previous treatment of the polyamide ... please send me the method to be used. Thank You.
@Fernanda-
For the polyamide column and flavanoids, I like a gradient starting with water to 100% methanol, followed by another gradient from methanol to 100% acetone, then acetone to 100% water containing 5% ammonium hydroxide (3 gradients total). When I know where my desired compound(s) elute, I can just run the gradient where the compounds elute rather than all 3 gradients.