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Clinical Chemistry 17: 782-788, 1971;
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Clinical Chemistry, Vol 17, 782-788, Copyright © 1971 by the American Association for Clinical Chemistry

Hydroxylysine Glycosides in Human Urine

Marjorie F. Lou 1 and Paul B. Hamilton 1

1 Department of Biochemistry, Alfred I. du Pont Institute, Wilmington, Del. 19899.

Hydroxylysyl-galactosyl-glucose (HGG) and hydroxylysyl-galactose (HG) were isolated from normal human urine and shown to be identical with hydroxylysine glycosides (OHLG's) whose structure had been elucidated in other laboratories. We devised a procedure for separating the OHLG's from many other urinary constituents by preliminary fractionation on a column of Sephadex. The OHLG's in the fractions so obtained were then isolated, uncontaminated with other ninhydrin-positive components, in a single pass through an analytical cation-exchange column equipped for splitting the effluent stream. In addition, a procedure was devised for determining as little as 10-9 mol of the OHLG's in 100 µl of filtrate (corresponding to 83.3 µl, of urine) on standard ion-exchange chromatographic amino acid analyzers. In normal adult urine, 1.5, µmol of each glycoside was present per 100 mg of creatinine; the molar ratio of HHG to HG was about 1.1 to 1.2. Each was present in plasma in about 400- to 500-fold smaller concentration than in urine. After surgical fusion of the spine, excretion of OHLG's increased and the ratio of HGG to HG decreased, changes we interpreted as indicating a more rapid bone-collagen turnover consequent to surgical damage.


Key Words: fractionation with Sephadex • cation-exchange chromatography • bone-collagen turnover • nmol-scale determinations • OH-lysine glycosides in plasma • high-voltage electrophoresis—paper chromatography • sequence studies • effect of surgical damage • ninhydrin-positive compounds • diabetes






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Copyright © 1971 by the American Association for Clinical Chemistry.