Method and device for detecting glucose

• US 3,964,871 A
• Filed: 12/18/1974
• Issued: 06/22/1976
• Est. Priority Date: 12/18/1974
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Hochstrasser, Harry
• Primary Examiner(s): Serwin, R.E.
• Application Number:
  US05/533,972
• Time in Patent Office: 552 Days
• Field of Search: 23/230 B, 23/253 TP, 195/103.5 R, 195/127, 195/103.5 C, 116/114 AM
• US Class Current: 435/10
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

A disposable indicator is disclosed which is useful for the measurement of substances, e.g., glucose, in biological fluids. The instrument registers the concentration of substance in a given biological fluid with indicia which are directly readable in a convenient notation system (e.g., digital or other symbolic notation). The indicator comprises an improvement over prior art disposable devices which require a comparison of the viewable indication with a color intensity scale to translate the indicia registered to meaningful numerical values or other symbolic notations. The indicator permits the use of chemical reagent concentrations which are at least an order of magnitude greater than those used in prior art, thereby improving stability and drastically lowering the magnitude of interferences. As an example, the use of the indicator of the invention in conjunction with novel reagent compositions in the determination of urinary glucose concentrations is disclosed.

Claims

1. An indicator for the measurement of glucose dissolved in biological fluids, which comprises:

a support member; and

a plurality of indicating reagents for glucose, each of which is located in a separate zone of said member and at least two of which indicate the presence of different concentrations of glucose in solution.

2. The indicator of claim 1 wherein said support member is hydrophobic and said zones are formed of bibulous material.

3. The indicator of claim 1 wherein said reagents are disposed upon said support member in the shape of numerals corresponding to indicia for the level of glucose for which they indicate.

4. The indicator of claim 1 wherein said reagents are disposed upon said member in the order of their sensitivity.

5. The indicator of claim 1 wherein said reagents comprise an enzyme having glucose oxidase activity, peroxidase, an indicator which undergoes a color change in the presence of hydrogen peroxide and peroxidase and a compound which prevents the accumulation of oxidized indicator until such time as said compound has been completely consumed in such reaction.

6. The indicator of claim 1 wherein said support member is mounted on a stiffening member, said stiffening member having apertures therethrough for viewing said separate zones on the underlying support member, said apertures providing, by their shape, appropriate symbols of indicia.

7. The indicator of claim 6 wherein said stiffening member is a strip of polymeric material.

8. The indicator of claim 1 wherein there is also borne on said support member, a reagent which undergoes a color change in the presence of acid, the latter reagent being located in a separate zone of said support member.

9. The indicator of claim 1 wherein said support member is a transparent or translucent sheet.

10. The indicator of claim 1 wherein said indicating reagents comprise glucose oxidase, horseradish peroxidase, a compound which forms a color upon oxidation by hydrogen peroxide, a compound selected from the group consisting of indoxyl sulfate;

Uric acid;

3,4-dihdroxylmandelic acid;

3-methoxy-4-hydroxymandelic acid;

3,4-dihydroxyphenylacetic acid;

3-methoxy-4-hydroxyphenylacetic acid;

3,4-dihydroxyphenylalanine;

3-methoxy-4-hydroxyphenylalanine;

5-hydroxyindole-3-acetic acid;

5-hydroxyindole-2-carboxylic acid;

5-hydroxytryptophane;

creatinine;

2,5-dihydroxyphenylacetic acid;

0-hydroxytyramine (dopamine);

5-hydroxytryptamine; and

2,5-dihydroxybenzoic acid; and

a buffer composition which will maintain the pH of said reagents within the range of from about 4 to about 6 in the presence of a biological fluid.

11. The indicator of claim 10 wherein the compound selected is 2,5-dihydroxybenzoic acid.

12. The indicator of claim 10 wherein the proportion of components of said reagents is:

40 IU or more of glucose oxidase;

60 PU or more of peroxidase;

from 0.05 to 15 mg. of said color forming compound; and

from 0.01 to 5 mg. of the selected compound, all in a final volume of about 100 microliters, 0.01 to 10 microliters of which is applied to each sensitive zone.

13. An assembly for the measurement of glucose dissolved in a biological fluid which comprises;

A. an indicator which comprises;

a support member; and

a plurality of indicating reagents for glucose each of which is disposed in a separate zone of said member and each of which upon exposure to a solution of said glucose will indicate visually if a different minimum concentration of glucose is present in said solution, by a darkening thereof; and

B. a reticule adapted to overlay said indicator and bearing thereon a reference grid corresponding to the order in which said reagents are disposed on said support member.

14. A method of determining the concentration of glucose in a biological solution which comprises:

immersing in said solution an indicator which comprises;

a support member; and

a plurality of indicating reagents for glucose, each of which is located in a separate zone of said member and at least two of which indicate the presence of a different concentration of glucose in solution by registering an indicia in said zone;

removing said indicator from said solution; and

reading the minimum concentration of glucose in said solution directly from the indicia registered.

15. A method according to claim 14 in which the concentration of glucose dissolved in said solution is determined by counting the number of indicia registered while reading.

16. A method according to claim 15 in which the registered indicia are counted in an electromechanical device.

17. A method according to claim 16 in which the zones are arranged in a linear manner and are counted by moving the indicator through the sensing zone of an electromechanical counter operated on the basis of detecting zones by their opacity, reflectance, radioactivity, fluorescence, luminescence, electrical conductivity, or other electromagnetic absorptive or radioactive property.

18. A glucose indicating composition which comprises:

glucose oxidase;

horseradish peroxidase;

a compound which forms a color upon oxidation by hydrogen peroxide;

a compound selected from the group consisting of indoxyl sulfate;

Uric acid;

3,4-dihydroxymandelic acid;

3-methoxy-4-hydroxymandelic acid;

3,4-dihydroxyphenylacetic acid;

3-methoxy-4-hydroxyphenylacetic acid;

3,4-dihydroxyphenylalanine;

3-methoxy-4-hydroxyphenylalanine;

5-hydroxyindole-3-acetic acid;

5-hydroxyindole-2-carboxylic acid;

5-hydroxytryptophane;

creatinine;

2,5-dihydroxyphenylacetic acid;

o-hydroxytyramine (dopamine);

5-hydroxytryptamine; and

2,5-dihydroxybenzoic acid; and

a buffer composition which will maintain the pH of said glucose indicating composition within the range of from about 4 to about 6 in the presence of biological fluids.

19. A composition according to claim 18 wherein said selected compound is 2,5-dihydroxybenzoic acid.

20. A composition according to claim 18 -methoxy-the proportions of components is from 40 IU or more of glucose oxidase;

60 PU or more of peroxidase;

0.05 to 15 mg. of said color forming compound; and

from 0.01 to 5.0 mg. of the compound selected from indoxyl sulfate;

Uric acid;

3,4-dihydroxymandelic acid;

3-methoxy-4-hydroxymandelic acid;

3,4-dihydroxyphenylacetic acid;

3-methoxy-4-hydroxymandelic acid;

3,4-dihydroxyphenylacetic acid;

31methoxy-4-hydroxyphenylacetic acid;

3,4-dihydroxyphenylalanine;

3-methoxy-4-hydroxyphenylalanine;

5-hydroxyindole-3-acetic acid;

5-hydroxyindole-2-carboxylic acid;

5-hydroxytryptophane;

creatinine;

2,5-dihydroxyphenyl-acetic acid;

o-hydroxytyramine (dopamine);

5-hydroxytryptamine; and

2,5-dihydroxybenzoic acid;

per 100 microliters of said buffer.

21. A composition according to claim 20 wherein said selected compound is 2,5-dihydroxybenzoic acid.