I am working on a plant fraction and its weight is 24 mg.. I tried it on TLC normal phase using CHCl3 methanol system 10;1 and reversed phase using methanol; water 8;2 but there is tailing which makes it difficult to run on HPLC .. so what is the solution
I'm sorry, there isn't enough information to help you properly without making assumptions about your chromatography which may be incorrect.
Are you using silica gel columns in your HPLC? If you are running reverse phase HPLC, the problems may not transfer to that chromatography. What solvent system, including modifiers, are you using for HPLC? How have you purified the material prior to this TLC and HPLC step? The purification method gives us a clue as to the sort of compound you are running.
For your TLC, try adding triethyl amine (TEA), which will make basic compounds elute a little earlier and acids elute later, or a little acid (acetic or formic) which will make acids less polar and bases more polar.
Hi, Do you want to do TLC and or HPLC? In addition of the comment of Jack above, if you use TLC maybe you can the correct system in the Camag TLC data base (for your fraction), maybe someone has ever published. For HPLC I think you can do same thing by searching via Google or Scopus or others. Or do you want to do both TLC and HPLC? Best regards
http://www.camag.com/en/tlc_hptlc/customer_magazine_cbs/ccbs_database.cfm
i want to do HPLc bit before it i should try it on TLC to see what is the best system ..
@Sherif...
Please look at my reply and give some answers. Unless you are running normal phase on both HPLC and TLC, or reverse phase on both TLC and HPLC, you can't see which solvent system is best. The selectivities of the two systems are different, and a method in one (normal phase or reverse phase) won't transfer to the other.
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