Liquid-chromatographic determination of total hydroxyproline in urine.
In this method for quantifying 4-hydroxyproline in human urine, 50 microL of urine is hydrolyzed, derivatized with phenylisothiocyanate (PITC), and then quantified by reversed-phase HPLC with ultraviolet detection. The detection limit in urine is 373 pg of hydroxyproline per 50-microL 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 mumol/24 h. We also report hydroxyproline concentrations in patients with metastatic bone disease and cirrhosis of the liver.
Dawson
C D
CD
Division of Environmental Health Laboratory Sciences, Centers for Disease Control, Atlanta, GA 30333.
Jewell
S
S
Driskell
W J
WJ
D002853:Chromatography, Liquid; D006801:Humans; D006909:Hydroxyproline / Q000652:urine*; D017879:Isothiocyanates; D013861:Thiocyanates
D017879:Isothiocyanates; D013861:Thiocyanates; C005441:phenylisothiocyanate; D006909:Hydroxyproline