title: Fluorimetric determination of alkaline phosphatase activity in human serum by use of a flow-through biosensor.
abstract: A fluorimetric method for the determination of alkaline phosphatase activity based on the use of a flow-through biosensor is reported. The biochemical basis of the method is the hydrolysis of 4-methyl-umbelliferone-phosphate catalyzed by the analyte with fluorimetric monitoring of the 4-methyl-umbelliferone formed (lambda ex = 365 nm, lambda em = 445 nm). The enhancement of sensitivity achieved when the reaction product is retained on the support packed in the flow-cell makes the method suitable for determination of the analytes in serum samples after 1:50 dilution. The linear range is found to be between 0.1 and 20 U l-1, with relative standard deviation less than 2.2%. The use of this biosensor was tested by the determination of the analyte in human serum from healthy and sick individuals with excellent recoveries (94-103%).
authors.affiliation: Department of Analytical Chemistry, Faculty of Sciences, University of Córdoba, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Sánchez-Cabezudo
authors.forename: J M
authors.initials: JM
authors.lastname: Fernández-Romero
authors.forename: M D
authors.initials: MD
authors.lastname: Luque de Castro
mesh_terms: D000469:Alkaline Phosphatase / Q000097:blood*; D015374:Biosensing Techniques; D001709:Biotechnology; D005069:Evaluation Studies as Topic; D006801:Humans; D006923:Hymecromone / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D006923:Hymecromone; D000469:Alkaline Phosphatase