Hydroxyproline is liberated during the intracellular maturation of collagen. Furthermore, degradation of the N-terminal peptides which contain 4-hydroxyproline in type I procollagen is probaby also a source of urinary hydroxyproline.
Therefore, increased collagen synthesis as well as breakdown will result in elevated hydroxyproline levels. In fasting individuals, urinary hydroxproline may therefors be used as a specific index of collagen metabolism.
Our objective was to develop a reliable, optimized and more rapid HPLC method for urinay hydroxyproline analysis. ToTal hydroxyproline in urine is derivatized with 4 chloro 7 nitrobenzofurazan after reaction of the urine with o-phthalaldehyde. The highly fluorescent adducts of hydroxyproline is separated on a μ-Bondapak C_18 reversed phase column using acetonitrile 0.01M sodium phsphate buffer, pH 7.0(10:90, v/v) as mobile phase, followed by fluorometric detection.
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