Print Email Facebook Twitter Pre-column derivatisation method for the measurement of glycosylated hydroxylysines of collagenous proteins Title Pre-column derivatisation method for the measurement of glycosylated hydroxylysines of collagenous proteins Author Bank, R.A. Beekman, B. Tenni, R. Tekoppele, J.M. Gaubius Instituut TNO Publication year 1997 Abstract Measurement of the glycosylated hydroxylysines galactosyl- and glucosylgalactosylhydroxylysine (GH and GGH) in combination with other amino acids has been based on ion-exchange chromatography followed by reaction with ninhydrin. Here, a rapid and sensitive high-performance liquid chromatographic method with fluorimetric detection has been developed and employed to determine the glycosylated hydroxylysine residues in alkaline collagen hydrolysates. After hydrolysis, amino acids were derivatised with 9- fluorenylmethyl chloroformate and separated on a Micropak ODS-80TM reversed- phase column (150 x 4.6 mm). With a multistep gradient system all amino acids were separated in less than 30 min, including the collagen-specific hydroxylysine, hydroxyproline and the glycosylated hydroxylysines. The method was used to evaluate the glycosylation levels of human articular cartilage derived from femoral head, femoral condyle, tibial plateau and ankle. GGH was highest in cartilage from femoral head and ankle. GH showed no differences between the different sources of cartilage. Chemicals/CAS: Collagen, 9007-34-5; Hydroxylysine, 28902-93-4 Subject BiologyBiomedical ResearchCollagenDerivatisation, LCGalactosylhydroxylysineGlucosylgalactosylhydroxyl ysineHydroxylysinescollagenhydroxylysineamino acid analysisanklearticular cartilagecontrolled studyfemur condylefemur headhuman tissueion exchange chromatographypriority journalprotein glycosylationreversed phase high performance liquid chromatographytibiaAnimalsAnkleCartilageCartilage, ArticularCattleChromatography, High Pressure LiquidCollagenFemaleGlycosylationHipHumansHydrolysisHydroxylysineKneeMiddle AgedNosePlacentaPregnancySensitivity and SpecificitySpectrometry, FluorescenceTime Factors To reference this document use: http://resolver.tudelft.nl/uuid:22be2ef4-9346-426a-ac66-b6763451e24d DOI https://doi.org/10.1016/s0378-4347(97)00392-7 TNO identifier 234270 ISSN 1572-6495 Source Journal of Chromatography B: Biomedical Applications, 703 (1-2), 267-272 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.