Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 847, Issue 2, Pages 282-288Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2006.10.015
Keywords
electrospray; FDVA; hydroxylation; Hyl; Hyp; RP-HPLC
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Collagens, the most abundant mammalian proteins, contain a high content of hydroxylated amino acids, such as, 3- and 4-cis-/trans-hydroxyproline (Hyp) and 5-hydroxylysine (Hyl). Whereas the global content of 4-Hyp was studied by amino acid analysis, no technique to determine all five hydroxyamino acids simultaneously in collagens has been reported. Here, we report the separation of all five hydroxyamino acids as well as two Hyp epimers from all other proteinogenic amino acids after derivatization with N-2-(5-fluoro-2,4-dinitrophenyl)-L-valine amide (L-FDVA) by RPC-UV-ESI-MS. The general applicability of this method is shown for three Hyp-containing peptides as well as collagen type I. (c) 2006 Elsevier B.V. All rights reserved.
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