Pre-column Derivatization and Its Optimum Conditions for Quantitative Determination of Urinary Hydroxylysine Glycosides by High-Performance Liquid Chromatography

Kazuhiro YOSHIHARA; Naritoshi MOCHIDOME; Yasuo SHIDA; Yumiko HAYAKAWA; Minoru NAGATA

doi:10.1248/bpb.16.604

Kazuhiro YOSHIHARA, Naritoshi MOCHIDOME, Yasuo SHIDA, Yumiko HAYAKAWA, Minoru NAGATA

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Keywords: hydroxylysine glycoside, fluorescence derivatization, collagen degradation, urinary excretion level, HPLC, electrospray ionization mass spectrometry

JOURNAL FREE ACCESS

1993 Volume 16 Issue 6 Pages 604-607

DOI https://doi.org/10.1248/bpb.16.604

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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.

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