Analysis of hydroxyproline in urine by high-performance liquid chromatography after dabsyl-chloride derivatization |
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| Authors: | M Ikeda K Sorimachi K Akimoto M Okazaki M Sunagawa A Niwa |
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| Institution: | (1) Laboratory of Medical Sciences, Dokkyo University School of Medicine, 321-02 Mibu, Tochigi, Japan;(2) Department of Microbiology, Dokkyo University School of Medicine, 321-02 Mibu, Tochigi, Japan;(3) Research Center for Cellular and Molecular Biology, Dokkyo University School of Medicine, 321-02 Mibu, Tochigi, Japan;(4) Department of Surgery, Dokkyo University School of Medicine, 321-02 Mibu, Tochigi, Japan |
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| Abstract: | Summary In order to analyse hydroxyproline (HYP) in urine, a high-performance liquid chromatographic method was modified. The primary amino groups were blocked with o-phthalaldehyde, and then the secondary amino groups were derivatized with 4-dimethylaminoazobenzene-4 -sulphonyl chloride. In addition, the dabsylated samples were treated with ethyl acetate to obtain a simple elution profile in high-performance liquid chromatography. The dabsyl-HYP and -proline were eluted at 4.7 min and 8.0 min, respectively. The chromatographic analysis was completed within 10 min, including the time needed for reequilibration of the column. Using the present method, the concentration of HYP in urine was determined to 260 ± 6µmol/l. |
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| Keywords: | Amino acids Hydroxyproline Dabsyl-chloride Urine High-performance liquid chromatography o-Phthalaldehyde |
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