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Influence of trifluoroacetic acid on retention times of histidine‐containing tryptic peptides in reverse phase HPLC

1983; Wiley; Volume: 22; Issue: 1 Linguagem: Inglês

10.1111/j.1399-3011.1983.tb02071.x

0367-8377

A. Seetharama Acharya, Alberto Di Donato, Belur N. Manjula, Vincent A. Fischetti, James M. Manning,

Metabolomics and Mass Spectrometry Studies

The tryptic peptides of the aminoethylated α‐and β‐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.