Enzymatic determination of glucose
First Claim
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1. An analytical agent for the quantitative enzymatic determination of glucose, consisting essentially of:
- a. glucose dehydrogenase having a dry weight activity of at least 2 U/mg., the NAOH2 oxidase activity of which is less than 0.1% of said glucose dehydrogenase activity;
b. a pyridine coenzyme selected from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide purine dinucleotide, nicotinamide-(6-methylpurine)-dinucleotide and nicotinamide-(2-chloro-6-methylpurine)-dinucleotide in an amount sufficient to provide about a 2.0-5.0 millimole concentration in an analytical test solution;
c. means for maintaining an aqueous test solution of said analytical agent at a pH of about 6.5-8.5; and
d. mutarotase in an amount sufficient to increase the spontaneous mutarotation of α
-glucose to β
-glucose in said test solution by more than 50%.
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Abstract
Glucose is rapidly ad quantitatively determined with an analytical agent containing glucose dehydrogenase having an activity of at least 2 μ/mg and NADH2 oxidase activity less than 0.1%, a pyridine coenzyme, a buffer and mutarotase to increase spontaneous mutarotation of alpha-glucose to beta-glucose. There may be optionally present inhibitors for reduced pyridine coenzyme oxidases and a thermal stabilizing amount of alkali metal chloride.
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Citations
10 Claims
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1. An analytical agent for the quantitative enzymatic determination of glucose, consisting essentially of:
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a. glucose dehydrogenase having a dry weight activity of at least 2 U/mg., the NAOH2 oxidase activity of which is less than 0.1% of said glucose dehydrogenase activity; b. a pyridine coenzyme selected from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide purine dinucleotide, nicotinamide-(6-methylpurine)-dinucleotide and nicotinamide-(2-chloro-6-methylpurine)-dinucleotide in an amount sufficient to provide about a 2.0-5.0 millimole concentration in an analytical test solution; c. means for maintaining an aqueous test solution of said analytical agent at a pH of about 6.5-8.5; and d. mutarotase in an amount sufficient to increase the spontaneous mutarotation of α
-glucose to β
-glucose in said test solution by more than 50%. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification