Can anyone who used Sephadex LH-20 in separation or purification of compounds share his experience?
How many times can I change the mobile phase in the experiment?
And what about using organic solvents other than methanol?
you can change the mobile phase as often as you want, but you have to take care that its not swelling (hence expanding) in the column you use. So the direction of solvent changes has to be to lower bed volumes.
Approx. bed volumes for different solvents can be looked up here: https://www.gelifesciences.com/gehcls_images/GELS/Related%20Content/Files/1314735988470/litdoc56119097AD_20110830235244.pdf
As you can see you can use almost any inert solvent, also highly non-polar ones.
HI! I want to know when you used Sephadex LH-20??? I need to select an adequate column to separate polyphenols, Do you know where I must obtain that information for take a decision???
Thanks in advance,
Patricia.
Hello!
I don't remember if I used LH-20, but i used it to separate flavonoids. I used the mobile phase MeOH/H2O 9:1 v/v, and later MeOH/H2O 1:1 v/v. the flavonoids enriched were in this last eluate.
@ Patricia Velàsquez: I used methanol to purify flavanoids as shown here:
http://www.isco.com/WebProductFiles/Applications/101/Poster_and_Paper_Reprints/Purification_of_Phenolic_Flavonoids.pdf
Yes flavonoides elute late in the column of sephadex but you can't use sephadex column to separate flavonoides from each other
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