Influence of trifluoroacetic acid on retention times of histidine-containing tryptic peptides in reverse phase HPLC.
The tryptic peptides of the aminoethylated alpha- and beta-chain of hemoglobin have been separated on a Partisil-10 ODS-2 column with a linear gradient of acetonitrile containing 0.1% trifluoroacetic acid. The elution profile of the tryptic peptides of the chains obtained with this acetonitrile trifluoroacetate system has been compared with that obtained using phosphoric acid as the ion-pairing reagent. This comparison demonstrated that trifluoroacetate influences the retention times of the histidine-containing tryptic peptides much more than it affects those peptides that do not contain histidine residues. This behavior has been rationalized on the basis of ion-pairing of trifluoroacetate with the histidine residues of the tryptic peptides.
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005463:Fluoroacetates; D006441:Hemoglobin A / Q000032:analysis; D006639:Histidine; D009842:Oligopeptides / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis*; D010710:Phosphates; D014269:Trifluoroacetic Acid; D014357:Trypsin
D005463:Fluoroacetates; D009842:Oligopeptides; D010446:Peptide Fragments; D010710:Phosphates; D006639:Histidine; D006441:Hemoglobin A; D014269:Trifluoroacetic Acid; D014357:Trypsin