Fluorescent techniques for the selective detection of chromatographically separated peptides

✍ Scribed by Timothy D. Schlabach; Timothy C. Wehr

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 663 KB
• Volume: 127
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90167-1

✦ Synopsis

Ruorescence detection has been used for the detection of peptides that contain tyrosine or tryptophan. The strong native fluorescence of these groups makes this method several times more sensitive than detection at 254 nm. Furthermore, when combined in series with a lower wavelength absorbance detector-220 nm, for example-the fluorescence-to-absorbance ratios can be used to distinguish peptides having a single tyrosine residue from those having a tryptophan residue. Reaction detection can also be used to specifically identify peptides with lysine residues. The c-amino group in lysine reacts far better with o-phthalaldehyde than any other peptide group including the primary amino terminus. The reagent is continuously added to the column effluent, generating a chromatogram of the peptides with lysine. Not only is this postcolumn reaction specific, it also detects as little as 10 pmol of some peptides.

' This paper was presented at the International Symposium on HPLC of Proteins and Peptides,