Urine hydroxyproline: a potential error in quantification and a proposed procedure for its measurement

A modification of the colorimetric method of Bergman and Loxley for the measurement of urinary hydroxyproline using external aqueous hydroxyproline standards instead of individual internal standards is described. We show that this modification leads to an underestimation (average 32%) of hydroxyproline because suppression of colour development occurs in urine samples but not in aqueous standards. Use of an internal standard for each urine test corrects for this suppression. Recovery of hydroxyproline (250 mumol/L) added to 12 patient urine samples averaged 99% and the overall imprecision for the assay was then less than 5%. Modifications to the original hydrolysis and colour development procedures allow linearity to 1500 mumol/L. Details of our procedure are given.