An Automated Fluorometric Alkaline Phosphatase Microassay with 4-Methylumbelliferyl Phosphate as a Substrate

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Abstract
An automated microassay for alkaline phosphatase activity is described. It utilizes 10-3 M 4-methylumbelliferyl phosphate as a substrate and requires 10 μl. of normal serum. Ranges for serum alkaline 4-methylumbelliferyl phosphatase activities of normal adults and children are established. A close correlation between the phenylphosphatase and the 4-methylumbelliferyl phosphatase activities of normal and abnormal sera has been demonstrated. Heat denaturation showed alkaline phosphatase in children’s sera to resemble the alkaline phosphatase in sera of patients with skeletal disorders. The effect of dilution on the heat stability of alkaline phosphatases is discussed briefly.