Chromatography Forum

I try to differentiate 2 Trihexosylceramides in one sample by TLC.
But the difference is just Gal-alpha-1.4 vs. Gal-alpha-1.3.

Does someone has expierence, which solvent could be best to start?

thanx

Do you have enough substance to R&D on your own?
if yes then start with the CAMAG approach of using 10-12 different solvents on small plates no more then 5 cm run lenght all NP plates

after the first 10-12 trials, you should combine two of the solvents. the one that kept your spots or bands in the best shape and the one that gave you the best separation.

if you feel that you have not achieved sufficient separation then try change your stationnary phase to Di-OH and repeat the process until you get good results. move this in the polarity of the plate until you get to RP-18

if you have no budget problems then go to HPTLC plates from the start. they are a lot better then normal TLC plates and you will get a greater separation power (like going down in particle size for columns).

hope this helps

Thank you very much for your suggestion, I didn´t know the CAMAG prism approache,

Cheers Christian