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Clinical Chemistry 18: 943-950, 1972;
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Clinical Chemistry, Vol 18, 943-950, Copyright © 1972 by the American Association for Clinical Chemistry

Application of a New Peroxide Indicator Reaction to the Specific, Automated Determination of Glucose with Glucose Oxidase

Nathan Gochman 1 and Joan M. Schmitz 1

1 Clinical Chemistry Service, Clinical Pathology Department, Clinical Center, National Institutes of Health, Bethesda, Md. 20014.

We describe an automated, colorimetric determination of glucose in biological fluids that combines the specificity of glucose oxidase and of a new peroxide indicator reaction. In the presence of peroxidase, 3-methyl-2-benzothiazolinone hydrazone oxidatively couples with N,N-dimethylaniline to form a stable, intensely colored, water-soluble indamine dye, the concentration of which is proportional to that of the third reactant, hydrogen peroxide. This reaction, used earlier to determine uric acid [Clin. Chem. 17, 1154 (1971)], is substantially less affected by negative interference of reducing substances than are previously described peroxide indicators. Results from use of AutoAnalyzers I and II and this method were compared with those from a manual spectrophotometric hexokinase/glucose-6-phosphate dehydrogenase procedure, and showed good correlation for specimens from patients. The automated methods are suitable for measuring glucose in serum, plasma from fluoride-or iodoacetate-preserved blood, urine (without ion-exchange pretreatment), or cerebrospinal fluid. They eliminate the problem of falsely high results caused by medication or reducing metabolites associated with uremia, in methods in which alkaline ferricyanide or copper—neocuproine is used.


Key Words: glucose in serum, plasma, cerebrospinal fluid, urine • 3-methyl-2-benzothiazolinone hydrazone • N,N-dimethylaniline • AutoAnalyzer • manual hexokinase procedure compared • uremia and drug interference

Submitted on April 14, 1972
Accepted on May 13, 1972




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