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Clinical Chemistry 20: 332-336, 1974;
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Clinical Chemistry, Vol 20, 332-336, Copyright © 1974 by the American Association for Clinical Chemistry

Enzymatic Method for Determination of Inorganic Phosphate in Serum and Urine with a Centrifugal Analyzer

Michael A. Pesce 1, Selma H. Bodourian 1, and John F. Nicholson 1

1 Division of Clinical Chemistry, Babies Hospital, The Childrens Medical and Surgical Center of New York at Columbia-Presbyterian Medical Center; and the Department of Pediatrics, College of Physicians and Surgeons of Columbia University, 3975 Broadway, New York, N. Y. 10032.

We describe an enzymatic method for determining inorganic phosphate in serum and urine, with use of the "CentrifiChem." The sample is mixed with a combined enzyme—substrate system consisting of the enzymes phosphorylase a, phosphoglucomutase, and glucose-6-phosphate dehydrogenase, the coenzyme NADP, the substrate glycogen, and adenosine 5'-monophosphate as the activator for phosphorylase. The increase in absorbance at 340 nm as NADPH is formed is linearly proportional to the concentration of inorganic phosphate to 15 mg P/dl. This method circumvents deproteinization and requires only 10 µl of sample. Results obtained with the automated enzymatic method show good correlation with manual and automated molybdenum blue methods.


Key Words: CentrifiChem • comparison of methods • direct spectrophotometry

Submitted on October 1, 1973
Accepted on December 6, 1973






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Copyright © 1974 by the American Association for Clinical Chemistry.