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it depends greatly on the kind of chromatography.

This answer will deal with the two ways i use acetic acid in silica gel chromatography.

1) most often i use acetic acid when i need to separate/elute carboxylic acids from each other or from other compounds on silica gel. carboxylic acids donate protons pretty readily on silica, and thus become ionized. when ionized they do not move on silica gel. By adding between 0.1% and 10% acetic acid to my solvent mix, i can keep the carboxylic acid protonated so that it will elute off of silica gel like any other organic compound. the acetic acid can then be evaporated away just like any other organic solvent under reduced pressure.

2) for particularly polar organic comppounds, i sometimes use acetic acid as the polar component of my solvent mix, but this is rare-- I've only used it this way 5-7 times in the last 8 years.

in both cases above the exact amount of acetic acid added to my solvent mix (also called the mobile phase) has to be determined experimentally by trial and error.

when i have a relatively non-polar carboxylic acid to isolate, i may start with 85% hexane, 14% Ethyl acetate, 1% acetic acid.

if i have a more polar carboxylic acid to isolate, i might start with 95% dichloromethane, 4% methanol and 15 acetic acid.

I'll try a TLC at that mix, and then visualize the migration of the carboxylic acid by UV if it is UV active, or by staining with bromocresol green if it is not (remember to completely and thoroughly dry the TLC plate before staining -- to evaporate all the acetic acid since it will stain positive with bromocresol green too-- and remember to heat the stained TLC plate (about 20-200 seconds at 200C should work).

trifluoroacetic acid (TFA) also often added at a rate of about 0.1% to mixtures of acetonitrile and water for use in HPLC, BUT when the HPLC is coupled to a mass spectrometer (LCMS) use of trifluoroacetic acid can decrease analyte signal, so 0.1% acetic acid is often substituted.

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13y ago

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