Who has experience with hypercarb coulms? We like to analyze the polar metabolites in cell lysate (by MS). The hypercarb (we just purchsed) shows very nice interaction with quite polar metabolites such as glutamic acid. However, the peakshape is not very nice (tayling). Currently we use water 0.1 % formic acid and acetonitere, other buffer may isuppresd our MS signal, but did not optimize our peakshapes. We don't want to use any kind of ion-pair reagent. Anyone experience with tehese columns?
Hello Christian,
You can give it a try with the following buffers: solvent A: 0.3% formic acid brought to pH 3 with 25% ammonia solution. solvent B: 100% Acetonitrile. We know that this works quite nice for glycans but worth a try for your compounds.
(doi: 10.1007/978-1-4939-2760-9_29.)
Hi Christian,
I used strong base NH3aq and KOH with water+MeOH. MeOH gives better selectivity for closely related compounds. You can also try perfluorobytiric acid and others.
A paper would be useful.
Parameter optimization for the analysis of underivatized protein amino acids by liquid chromatography and ionspray tandem mass spectrometry, K. Petritis, P. Chaimbault, C. Elfakir , M. Dreux, Journal of Chromatography A, 896 (2000) 253–263
All the best,
Janusz
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