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Pre-column derivatization and its optimum conditions for quantitative determination of urinary hydroxylysine
K Yoshihara1, N Mochidome, Y Shida
1Tokyo College of Pharmacy, Japan.
Biological & Pharmaceutical Bulletin
|June 1, 1993
Abstract:
A method has been developed for rapid analysis of the urinary hydroxylysine glycosides [glucosyl-galactosyl-hydroxylysine (GGH) and galactosyl-hydroxylysine (GH)] as collagen metabolites. The glycosides were fluorometrically derivatized with 5-dimethylaminonaphthalene-1-sulfonyl chloride, and then subjected to reversed-phase high-performance liquid chromatography using a gradient system. Suitable standard hydroxylysine glycosides which were not diastereoisomers were prepared from human urine. Urinary GGH and GH were analyzed within 1 h, including column washing. This method was applied to the urine samples of 68 healthy persons of different ages.