title: Analysis of hydroxyproline isomers and hydroxylysine by reversed-phase HPLC and mass spectrometry.
abstract: 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.
authors.affiliation: Institute of Bioanalytical Chemistry, Center for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Langrock
authors.forename: Natividad
authors.initials: N
authors.lastname: García-Villar
authors.forename: Ralf
authors.initials: R
authors.lastname: Hoffmann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003094:Collagen / Q000737:chemistry; D006901:Hydroxylysine / Q000032:analysis* / Q000737:chemistry; D006909:Hydroxyproline / Q000032:analysis* / Q000737:chemistry; D007536:Isomerism; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D006901:Hydroxylysine; D003094:Collagen; D006909:Hydroxyproline