Method of analysis, reagent composition and use thereof for glucose determination
First Claim
1. A method to quantitatively determine the glucose concentration of a sample of undiluted whole blood, comprising contacting a sample of undiluted whole blood with a reagent in dry form in a microcuvette to obtain hemolyzed undiluted whole blood, the reagent comprising:
- glucose dehydrogenase (GDH);
one or more substances from the group consisting of diaphorase, phenazine methosulphate, phenazine ethosulphate, phenazine phenosulphate and Meldola blue;
one or more substances from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide-purine dinucleotide, nicotinamide-methylpurine dinucleo-tide and nicotinamide-2-chloro-methylpurine dinucleo-tide;
one or more hemolyzing substances from the group consisting of phospholipase, hemolyzing saponins, compounds of hydrophilic mono-, di- or tri-saccharides and aliphatic hydrocarbons having 10-16 carbon atoms; and
a redox indicator dye;
and measuring by transmission spectrophotometry a color change brought about by the reaction of the reagent with glucose in the undiluted whole blood to quantitatively determine the glucose concentration of undiluted whole blood.
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Abstract
A method, a reagent composition and the use thereof for quantitative determination of total glucose in undiluted whole blood are described. A sample of undiluted whole blood is contacted with a reagent composition which is in dry form and comprises:
glucose dehydrogenase (GDH);
one or more substances from the group consisting of diaphorase, phenazine methosulphate, phenazine ethosulphate, phenazine phenosulphate and Meldola blue;
one or more substances from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide-purine dinucleotide, nicotinamide-methylpurine dinucleotide, nicotinamide-2-chloro-methylpurine dinucleotide;
one or more hemolysing substances from the group consisting of phospholipase, hemolysing saponins, and compounds of hydrophilic mono-, di- or tri-saccharides and aliphatic hydrocarbons having 10-16 carbon atoms;
a redox indicator dye; and
optionally mutarotase.
A color change brought about by the reaction of the reagent with glucose in the undiluted whole blood is measured by transmission spectrophotometry.
27 Citations
12 Claims
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1. A method to quantitatively determine the glucose concentration of a sample of undiluted whole blood, comprising contacting a sample of undiluted whole blood with a reagent in dry form in a microcuvette to obtain hemolyzed undiluted whole blood, the reagent comprising:
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glucose dehydrogenase (GDH); one or more substances from the group consisting of diaphorase, phenazine methosulphate, phenazine ethosulphate, phenazine phenosulphate and Meldola blue; one or more substances from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide-purine dinucleotide, nicotinamide-methylpurine dinucleo-tide and nicotinamide-2-chloro-methylpurine dinucleo-tide; one or more hemolyzing substances from the group consisting of phospholipase, hemolyzing saponins, compounds of hydrophilic mono-, di- or tri-saccharides and aliphatic hydrocarbons having 10-16 carbon atoms; and a redox indicator dye; and measuring by transmission spectrophotometry a color change brought about by the reaction of the reagent with glucose in the undiluted whole blood to quantitatively determine the glucose concentration of undiluted whole blood. - View Dependent Claims (2, 7, 9)
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3. A reagent composition to quantitatively determine the total glucose concentration of undiluted whole blood, wherein the reagent composition is in dry form and the reagent composition comprises:
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glucose dehydrogenase (GDH); one or more substances form the group consisting of diaphorase, phenazine methosulphate, phenazine, ethosulphate, phenazine phenosulphate and Meldola blue; one or more substances from the group consisting of NAD, NADP, thio-NAD, thio-NAD, nicotinamide-purine dinucleotide and nicotinamide methylpurine dinucleotide; one or more hemolyzing substances from the group consisting of phospholipase, hemolyzing saponins, saccharides and aliphatic hydrocarbons having 10-16 carbon atoms; and a redox indicator dye. - View Dependent Claims (4, 5, 8, 10, 11)
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6. A reagent composition used to quantitatively determine the glucose concentration of a sample of undiluted whole blood, the reagent composition comprising:
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glucose dehydrogenase (GDH); one or more substances from the group consisting of diaphroase, phenazine methosulphate, phenazine ethosulphate, phenazine phenosulphate and Meldola blue; one or more substances from the group consisting of NAD, NADP, thio-NAD, thio-NADP, nicotinamide-purine dinucleotide, nicotinamide-methylpurine dinucleotide and nicotinamide-2-chloro-methylpurine dinucleotide; one or more hemolyzing substances from the group consisting of phospholipase, hemolyzing saponins, compounds of hydrophilic mono-, di- or tri-saccharides and aliphatic hydrocarbons having 10-16 carbon atoms; and a redox indicator dye; and which composition is in dry form in a microcuvette, for carrying out quantitative determination of total glucose in undiluted whole blood by transmission spectrophotometry to quantitatively determine the glucose concentration of undiluted whole blood. - View Dependent Claims (12)
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Specification