H-1-NMR PROFILE OF GLYCOSAMINOGLYCANS IN HUMAN URINE

A study was performed on human urinary glycosaminoglycans (GAGs) by one- and two-dimensional 500 MHz proton nuclear magnetic resonance (H-1-NMR) spectroscopy. Urinary GAGs were separated by the precipitation methods with hexadecyl pyridinium and with 85% ethanol, and purified by the beta-elimination reaction in alkaline medium containing sodium borohydride to remove proteins and/or peptides. The spectrum of each purified GAG sample from healthy human urine which included chondroitin sulfates (ChS), dermatan sulfate (DS) and heparan sulfate (HS) showed the characteristic signal(s) corresponding to each GAG chain. The assignments and integrations of the structure specific signals in the spectrum made it possible to perform qualitative and quantitative analyses on human urinary GAG mixtures. Furthermore, analytical results of urinary GAGs in children, adults and the aged indicate that the present method is applicable to investigate the effect of aging on excretion of urinary GAGs.