Sensitive and highly specific quantitative fluorometric...

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G01N 33/70 (2006.01) G01N 33/52 (2006.01)

Patent

CA 2110628

A SENSITIVE AND HIGHLY SPECIFIC QUANTITATIVE FLUOROMETRIC ASSAY FOR CREATININE IN BIOLOGICAL FLUIDS ABSTRACT OF THE DISCLOSURE A new and useful quantitative assay for creatinine has been developed. Creatinine is reacted with 3,5-dinitrobenzoate under alkaline reaction conditions to produce a fluorophore product which has excitation and emission maxima near 410 nm and 475 nm, respectively. Alternatively, 3,5-dinitrobenzoyl chloride or methyl-3,5-dinitrobenzoate may employed as reagents. The chemical reactions proceed in the presence of Group IA bases and/or quaternary alkyl ammonium hydroxides under aqueous, nonaqueous and mixed solvent conditions. Detection limits for creatinine are well below one micromole per liter. Conversely, the reactivity of creatinine and/or base with 3,5-dinitrobenzoate may be employed to quantity 3,5-dinitrobenzoate by fluorescence intensity measurements. The chemical assay is readily adaptable to reagent pad and dry-layered coating technologies employing fluorescence detection/quantitation.

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