Measurement of alkaline phosphatase levels in body fluids
AssigneeChembro Holdings (Proprietary) Limited
InventorsBarry Clifford Axcell, Cyril Donninger
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Abstract
A method of measuring the alkaline phosphatase level in a body fluid such as serum or plasma comprising hydrolyzing excess monophosphate ester dissolved in a suitable buffer by action of alkaline phosphatase in a predetermined volume of the body fluid to yield an oxidizable substrate, oxidizing the substrate with excess molecular oxygen in the presence of excess oxygenase and measuring the rate of oxygen consumption, preferably by means of an oxygen electrode. The invention also provides a reagent for use in this method which comprises a diethanolamine-HCl buffer having a molarity of 0.1 M to 4.0 M and a pH of 8.6 to 10.8 and containing magnesium ions in a concentration of 100 to 1000 .mu.M and an excess of oxygenase.
Claims15
A method of measuring the alkaline phosphatase level in a body fluid comprising hydrolysing excess monophosphate ester dissolved in a suitable buffer by action of the alkaline phosphatase in a predetermined volume of the body fluid to yield an oxidisable substrate, said ester being a catechol monophosphate ester which may have one or more ring substituents, oxidizing the substrate with excess molecular oxygen in the presence of excess oxygenase and measuring the rate of oxygen comsumption.
A method of measuring the alkaline phosphatase level in a body fluid comprising hydrolysing excess catechol monophosphate dissolved in a diethanolamine-HCl buffer having a molarity of 0.1 M to 4.0 M and a pH of 8.6 to 10.8 and containing magnesium ions in a concentration of 100 .mu.M to 1000 .mu.M by action of the alkaline phosphatase in a predetermined volume of the body fluid to yield catechol, oxidising the catechol and excess molecular oxygen in the presence of excess polyphenol oxidase and measuring the rate of oxygen consumption using an oxygen electrode.
Dependencies are inferred from the claim text.