Chemical timer for a visual test strip

• US 5,719,034 A
• Filed: 09/03/1996
• Issued: 02/17/1998
• Est. Priority Date: 03/27/1995
• Status: Expired due to Term

• Current Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Original Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Inventors: Kiser, Ernest J., Tomasco, Michael F., Yu, Yeung S., Rice, Edward G.
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US08/706,753
• Time in Patent Office: 532 Days
• Field of Search: 435/4, 435/14, 435/25, 435/28, 435/69.2, 435/184, 422/57, 422/101, 436/904, 436/166, 436/169, 436/170
• US Class Current: 435/14
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

A chemical timer for a direct-reading reagent test strip changes color a predetermined time after a biological fluid is applied to the strip. The strip measures the concentration of an analyte in the fluid. The timer is a dry coating of an indicator, an enzyme-containing reagent that when hydrated can react with glucose to change the color of the indicator, an inhibitor to inhibit the change in color of the indicator, glucose, and optionally, an aldose that does not react with the enzyme in the reagent. Preferably, the reagent and glucose are present in excess in the coating, and the time it takes for the timer color to change can be controlled by the inhibitor concentration. The aldose provides timer stability, probably by interfering with glycosylation by the glucose in the dry state.

Claims

1. A chemical timer for a visual-reading reagent test strip for measuring the concentration of glucose in a biological fluid that is applied to the strip, the timer comprising a dry coating of

a) a colored indicator composition,

b) a reagent that, when hydrated, reacts with glucose to change the color of the indicator,

c) an inhibitor to inhibit the change in color of the indicator, and

d) glucose,

in which the inhibitor and glucose concentrations in the dry coating are selected so that the glucose, over a predetermined time after the biological fluid is applied to the strip, reacts with the reagent to change the color of the indicator.

2. The timer of claim 1 in which the biological fluid is blood.

3. The timer of claim 1 in which the indicator is selected from the group consisting of (a) 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) combined with 3,3-dimethylaminobenzoic acid (DMAB);

(b) MBTH combined with 3,5-dichloro-2-hydroxybenzene-sulfonic acid (DCHBS);

(c) 4-aminoantipyrene (4-AAP) and 5-oxo-1-(p-sulfophenyl)-2-pyrazoline-3-carboxylic acid (OPSP);

(d) 4-AAP and N-(m-tolyl)-diethanolamine (NDA);

(e) 2,2'"'"'-azino-di(3-ethylbenzthiazoline)sulfonic acid (ABTS);

(f) 4-AAP and 4-methoxynaphthol;

(g) pyrogallol red (PGR);

(h) bromopyrogallol red (BPR);

(i) acid green 25 (AG);

or (j) 3-methyl-2-benzothiazolinone hydrazone!N-sulfonyl benzenesulfonate monosodium (MBTHSB), combined with 8-anilino-1-naphthalene sulfonic acid ammonium (ANS).

4. The timer of claim 3 in which the dye is MBTHSB-ANS.

5. The timer of claim 1 in which the reagent comprises an oxidase enzyme.

6. The timer of claim 5 in which the oxidase enzyme is glucose oxidase.

7. The timer of claim 1 in which the inhibitor comprises ascorbic acid.

8. A visual-reading reagent test strip for measuring concentration of glucose in a biological fluid that is applied to a strip, comprising the chemical timer of claim 1.

9. A method for preparing a chemical timer for a visual-reading reagent test strip for measuring the concentration of glucose in a biological fluid that is applied to the strip, comprising the steps of

a) coating onto a porous membrane a solution of a reagent that, when hydrated, reacts with glucose to form a reaction product,

b) drying the coating to form a first layer

c) applying onto the first layer a second layer that contains

(i) glucose,

(ii) an indicator that reacts with the reaction product to cause a color change, and

(iii) an inhibitor to inhibit the change in color of the indicator.

10. The method of claim 9 in which the reagent comprises glucose oxidase.

11. A method for measuring the concentration of glucose in a biological fluid comprising the steps of

(a) applying the fluid to a test strip that comprises

(i) a plurality of result segments that each change color when contacted with fluid containing at least a predetermined amount of glucose, different from the amount of glucose that causes a change in color of the other segments and

(ii) a timer segment that changes color after a time whose duration is substantially independent of the amount of glucose in the fluid and

(b) determining the glucose concentration by observing the result segment that has changed color last when the timer segment changes color.

12. A chemical timer for a visual reading test strip for measuring the concentration of an analyte in a biological fluid that is applied to the strip, the timer comprising a dry coating of

a) a colored indicator composition

b) an enzyme-containing reagent that, when hydrated, reacts with an analyte to change the color of the indicator;

c) an inhibitor to inhibit the change in color of the indicator,

d) glucose, and

e) an aldose that does not react substantially with the enzyme in the reagent.

13. The timer of claim 12 in which the aldose is selected from the group consisting of mannose and galactose.

14. The timer of claim 12 in which the analyte is glucose.

15. The timer of claim 12 in which the biological fluid is blood.

16. The timer of claim 12 in which the indicator is selected from the group consisting of (a) 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) combined with 3,3-dimethylaminobenzoic acid (DMAB);

(b) MBTH combined with 3,5-dichloro-2-hydroxybenzene-sulfonic acid (DCHBS);

(c) 4-aminoantipyrene (4-AAP) and 5-oxo-1-(p-sulfophenyl)-2-pyrazoline-3-carboxylic acid (OPSP);

(d) 4-AAP and N-(m-tolyl)-diethanolamine (NDA);

(e) 2,2'"'"'-azino-di(3-ethylbenzthiazoline) sulfonic acid (ABTS);

(f) 4-AAP and 4-methoxynaphthol;

(g) pyrogallol red (PGR);

(h) bromopyrogallol red (BPR);

(i) acid green 25 (AG);

or (j) 3-methyl-2-benzothiazolinone hydrazone!N-sulfonyl benzenesulfonate monosodium (MBTHSB), combined with 8-anilino-1-naphthalene sulfonate (ANS).

17. The timer of claim 16 in which the indicator is MBTHSB-ANS.

18. The timer of claim 12 in which the enzyme in the reagent comprises an oxidase enzyme.

19. The timer of claim 18 in which the oxidase enzyme is glucose oxidase.

20. The timer of claim 12 in which the inhibitor comprises ascorbic acid.

21. A visual test strip for measuring concentration of an analyte in a biological fluid that is applied to a strip, comprising the chemical timer of claim 12.

22. A method for preparing a chemical timer for a visual reading test strip for measuring the concentration of an analyte in a biological fluid that is applied to the strip, comprising the steps of

a) coating onto a porous membrane a solution of an enzyme-containing reagent that, when hydrated, reacts with an analyte to form a reaction product,

b) drying the coating to form a first layer, and

c) applying onto the first layer a second layer that contains

(i) an indicator that reacts with the reaction product to cause a color change,

(ii) an inhibitor to inhibit the change in color of the indicator,

(iii) glucose, and

(iv) an aldose that does not react substantially with the enzyme in the reagent.

23. The method of claim 22 in which the analyte is glucose and the enzyme comprises glucose oxidase.