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Rapid assessment of early glycation products by mass spectrometry
Author(s) -
Prabhakaram Malladi,
Smith J. B.,
Ortwerth B. J.
Publication year - 1996
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549600201802
Subject(s) - glycation , mass spectrometry , chemistry , environmental chemistry , chromatography , biochemistry , receptor
In order to detect the early glycation products, we have reacted a model peptide (t‐boc‐lys‐ala‐ala) with L‐threose (a degradation product of ascorbic acid) and analyzed the reaction products by a combination of HPLC and mass spectrometry. Amino group modification, as observed by a fluorescamine assay, indicated complete modification after 3 days of incubation with a 10‐fold excess of threose. As much as 60% of the adducts were acid labile and only 4% of the adducts could be observed by amino acid analysis. However, Fast atom bombardment mass spectrometry (FABMS) of the samples incubated for 6 hr showed relative molecular masses consistent with the formation of adducts corresponding to the addition of one and two molecules of L‐threose to the peptide. Likewise, samples incubated for 12 hr showed peptide adducts with two and three L‐threoses. The number of threose molecules added to the peptide was also confirmed from the FABMS analysis by using [1‐13C]‐threose as the glycating agent.