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

A. SEETHARAMA ACHARYA, ALBERTO DI DONATO, BELUR N. MANJULA, VINCENT A. FISCHETTI, JAMES M. MANNING

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18 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)78-82
Number of pages5
JournalInternational Journal of Peptide and Protein Research
Volume22
Issue number1
DOIs
StatePublished - Jul 1983
Externally publishedYes

Keywords

  • hemoglobin
  • ion‐pairing
  • trifluoroacetate

ASJC Scopus subject areas

  • Biochemistry

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