Hi, I want to isolate compounds from Brassica oleracea pants having n-Buoh fraction. I tried many solvents but could not get the desired results even the TLC plates did not showing the separated spots. from TLC plate its seems that different compounds did not showing proper spot because of uneven separation. Anyone can help me to solve this issue. Thanking in advance for any single effort.
what solvents you have used in TLC?
according to my experience ethanol is more appropriate solvents for extraction of Brassicaceae family than water , petroleum ether or benzene.
It is the same to me. I used extractipn kits or Doyle protocol adptated. Both situations with ethanol and it is ok.
Since you are dealing with a very polar fraction, you expected high molecular weight compounds, I'll suggest you use a reverse TLC plate for separation and also run the column using Sephadex.
I want to know if you did extraction with less polar solvent first, or your extraction was directly with n-BuOH?. If you did the second way you have to extract the polar butanol with less polar solvent in order to separate the less polar components.
By other side, Brassica oleracea has been well studied, may be you can get some previous paper. Good luck.
Dear Zaheer,
As you mentioned that you have selected n-BuoH fraction but it is not clear that whether you have done successive extraction or directly selected n-Buoh fraction but for the case of direct fractionation you may get some components with a nature of polarity nearby 3.9, for such kind of compounds you should try various combination of Toluene with ethyl acetate in TLC system, but i think that you may get some tailing effect which may be because of presence of residual n-Butanol solvent so before initializing the system run , make sure that the spot or band was dried properly.
Good luck
Usually the n-butanol fraction contain polar compounds. To isolate the compounds, I would like to suggest that you use HPLC.
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