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Method for analyzing plural oxidizable components in a liquid

  • US 4,725,539 A
  • Filed: 02/28/1984
  • Issued: 02/16/1988
  • Est. Priority Date: 03/08/1983
  • Status: Expired due to Fees
First Claim
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1. A method for analyzing inosinic acid, inosine and hypoxanthine in a liquid containing them, which comprises the steps:

  • (a) adding a volume of the liquid and pH buffer solution at a pH of 10.5 to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,(b) adding alkaline phosphatase enzyme to catalyze the hydrolysis of inosinic acid to inosine,(c) adjusting the pH of said liquid to neutrality and adding xanthine oxidase enzyme to catalyze the oxidation of hypoxanthine to xanthine,(d) recording oxygen sensor output,(e) adding nucleoside phosphorylase enzyme to catalyze the conversion of inosine to hypoxanthine, the xanthine oxidase enzyme present catalyzing the oxidation of the thus formed hypoxanthine to xanthine,(f) recording oxygen sensor output,(g) adding a volume of the liquid identical to that added in step a, the nucleoside phosphorylase and the xanthine oxidase enzymes present catalyzing the conversion of the inosine added to hypoxanthine and the oxidation of the hypoxanthine added and that thus formed to xanthine,(h) recording oxygen sensor output, and(i) calculating the amount of hypoxanthine present from the decrease in oxygen sensor output in step d, calculating the amount of inosinic acid present from the decrease in oxygen sensor output in steps d plus f less the decrease in oxygen sensor output in step h, and calculating the amount of inosine present from the decrease in oxygen sensor output in step h less the decrease in oxygen sensor output in step d.

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