Use of scanning diode array detector with reversed-phase microbore columns for the real-time spectral analysis of aromatic amino acids in peptides and proteins at the subsmicrogram level. Applications to peptide and protein microsequencing

Boris Grego, Edouard C. Nice, Richard J. Simpson

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Abstract

The characteristic zero- and second-order derivative spectra of phenylalanine, tyrosine and trytophan are described and used to identify aromatic residues contained in sub-microgram amounts of polypeptides and proteins during their elution from reversed-phase short microbore columns under gradient conditions. Spectral data were acquired with a commercially available scanning diode array detector. The method allows the non-destructive identification of tryptophan residues in complex polypeptide mixtures such as tryptic maps and enables the selection and isolation of such peptides for amino acid sequence analysis. The sub-microgram level of sensitivity is due to the small peak volumes and cosequent elevated solute concentrations obtained on short (< 10 cm), microbore (2 mm I.D.) reversed-phase columns.

Original languageEnglish
Pages (from-to)359-368
Number of pages10
JournalJournal of Chromatography A
Volume352
Issue numberC
DOIs
Publication statusPublished - 21 Feb 1986
Externally publishedYes

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