A Quantitative Assay for the Determination of Proline and Hydroxyproline by Capillary Electrophoresis

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Abstract
A quantitative spectrophotometrc assay for the determination of proline and hydroxyproline is described. The assay is based on the derivatization of the imino acids with the fluorogenic chromophore fluorescamine and the separation of the derivatized analytes by capillary electrophoresis. Fluorescamine reacts efficiently with secondary amino acids to form nonfluorescent aminoenone type chromophores which are easily detected at the low UV region. The assay is simple, rapid, and offers a significant improvement in detection sensitivity when compared with standard colorimotric methods for determining these imino acids. Derivatization with fluorescamine also enhances the degree of resolution of the two imino acids, enabling baseline separation of the derivatized analytes.