Liquid-Chromatographic Determination of Total Hydroxyproline in Urine

In this method for quantifying 4-hydroxyproline in human urine, 50 p.1.. of unne is hydrolyzed, derivatized with phenyl-isothiocyanate (PITC), and then quantified by reversed-phase HPLC wIth ultraviolet detection. The detection limit in urine is 373 pg of hydroxyproline per 50-p.L injection. The total CVs for high- and low-concentration pools are 5.3 % and 3.9%, respectively (10 runs in 10 days). The standard curve of the assay is linear over a range of 0 to 22 nmol per injection. We estimated the normal range for hydroxyproline excretion in men on an unrestricted diet to be 123-308 pffiolI24 h. We also report hydroxyproline concentrations in patients with metastatic bone disease and cirrhosis of the liver. AdditIonal Keyphrases: phenylisothiocyanate- chromato-graphy (reversed phase). reference interval. collage