i want to separate polar fraction of plant extract. For this i want to know how much amount of plant extract should be loaded in the column. Further, for batter separation what should be the ration of plant extract to resin (Diaion and Sephadex LH-20) should be taken in column chromatography.
Diaion makes both ion exchange resins and polymeric reverse phase (CHP-20).
If using ion exchange (for basic compounds such as alkaloids or acidic compounds such as many antioxidants) the load is limited by the number of ionic sites in the column. Although the links refer to a different material, the concept and calculations will still apply to the Diaion material:
http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN87_RediSep%20Rf_SAX_Column_Applications.pdf
http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN40_Purification_of_Alkaloids_with_RediSep_Rf_SCX_Columns.pdf
For the CHP-20 material, I tend to load 1-10% by weight. I use the higher weight if the sample isn't very clean, and the impurities are likely to be resolved from my active compound(s).
I like to use a 1-5% loading on LH-20.
For Diaion: about 1:20
For sephadex: as you the mechanism of isolation here depends on the molecular size (size exclusion), so the flow rate should be very slow and the column is long as possible
it depends upon your column that which type of column you are using for the 1st time its looking very difficult but actually its very easy t understand that how much extract should put in the column and if you are using polar material make the polar system of solvents
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