Color-forming couplers and their use in the analytical determination of hydrogen peroxide or other analytes

• US 4,672,029 A
• Filed: 12/06/1984
• Issued: 06/09/1987
• Est. Priority Date: 12/06/1984
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Washburn, William N., Cook, Ewell R., Dappen, Glen M.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Deck, Randall E.
• Application Number:
  US06/678,931
• Time in Patent Office: 915 Days
• Field of Search: 435/28, 435/14, 435/25, 435/10, 435/805, 435/810, 546/23, 546/171, 546/172, 546/178, 546/152, 548/413, 548/482, 548/491, 436/66, 436/904
• US Class Current: 435/10
• CPC Class Codes: C07F 9/60 Quinoline or hydrogenated quinoline ring systems
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 1/62 involving uric acid
• CPC Class Codes: C12Q 2326/00 Chromogens for determinations of oxidoreductase enzymes
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Particular heterocyclic compounds are useful as color-forming couplers in analytical compositions, elements and methods. These compounds are represented by the structure: ##STR1## wherein R is a solubilizing group, R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and Z represents the carbon atoms necessary to complete the nucleus of a 5- or 6-membered fused ring. The color-forming coupler is combined with an oxidizable color developing compound, e.g. 4-aminoantipyrine to provide a chromogenic composition. This composition can be used in solution or dry assays of biological fluids to produce a color dye in the presence of hydrogen peroxide or other analytes, e.g. glucose or uric acid, which react to produce hydrogen peroxide.

Claims

1. A chromogenic composition for the determination of hydrogen peroxide in an aqueous liquid, said composition comprising:

(1) a color-forming coupler of the structure;

##STR7## wherein R is a water solubilizing group,

R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring, and

(2) an oxidizable color developing compound which is capable of reacting with said coupler in the presence of both hydrogen peroxide and a substance having peroxidative activity to produce a color dye.

2. The composition of claim 1 wherein said oxidizable color developing compound is 4-aminoantipyrine.

3. The composition of claim 1 wherein R is alkyl, cycloalkyl or aryl, each substituted with one or more sulfo, hydroxy, carboxy, phosphono, quaternary amino, sulfonium or phosphonium groups or equivalent esters or salts.

4. The composition of claim 3 wherein R is alkyl substituted with one or more sulfo, hydroxy, carboxy, phosphono moieties or equivalent esters or salts, R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl of 1 to 3 carbon atoms, alkoxy of 1 to 3 carbon atoms or halo, and Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 6-membered fused ring.

5. The composition of claim 4 wherein R is

6. The composition of claim 1 further comprising an interactive reagent(s) which produces hydrogen peroxide upon interaction with an analyte.

7. The composition of claim 1 further comprising a substance having peroxidative activity.

8. A dry analytical element for the determination of hydrogen peroxide or of an analyte which can react to produce hydrogen peroxide in an aqueous liquid, said element comprising an absorbent carrier material containing the chromogenic composition of claim 1.

9. A dry analytical element for the determination of an analyte in an aqueous liquid, said element comprising a support having thereon one or more zones one of which comprising a porous spreading zone, and containing in any of said zones:

(1) an interactive reagent(s) which produces hydrogen peroxide upon interaction with an analyte,

(2) a color forming coupler of the structure;

##STR8## wherein R is a water solubilizing group,

R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring, and

(3) an oxidizable color developing compound which is capable of reacting with said coupler in the presence of both hydrogen peroxide and a substance having peroxidative activity to produce a color dye.

10. The element of claim 9 further comprising a substance having peroxidative activity.

11. The element of claim 9 wherein R is alkyl substituted with one or more sulfo, hydroxy, carboxy, phosphono moieties or equivalent esters or salts, R1, R2, R3, and R4 are independently hydrogen, alkyl of 1 to 3 carbon atoms, alkoxy of 1 to 3 carbon atoms or halo, and Z represents the carbon atoms necessary to complete the nucleus of a N-substitued 6 membered fused ring.

12. The element of claim 11 wherein R is

13. The element of claim 9 wherein said interactive reagent is uricase.

14. A dry analytical element for the determination of glucose in an aqueous liquid, said element comprising a support having thereon, in order and in fluid contact, a registration layer and a porous spreading/reagent layer, said element containing in any of said layers:

(1) glucose oxidase,

(2) peroxidase,

(3) a color-forming coupler of the structure;

##STR9## wherein R is a water solubilizing group,

R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring, and

(4) an oxidizable color developing compound which is capable of reacting with said coupler in the presence of both hydrogen perioxide and peroxidase to produce a color dye.

15. The element of claim 14 further comprising a radiation-barrier layer between said registration and spreading/reagent layers.

16. The element of claim 14 wherein said oxidizable color developing compound is 4-aminoantipyrine, and said color-forming coupler is ##STR10##

17. A method for determining hydrogen peroxide or an analyte which reacts to produce hydrogen peroxide in an aqueous liquid, said method comprising the steps of:

A. in the presence of a substance having peroxidative activity, physically contacting a sample of a liquid with a colorimetric composition comprising a color-forming coupler of the structure;

##STR11## wherein R is a water solubilizing group,

r₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring, and

an oxidizable color developing compound which is capable of reacting with said coupler in the presence of both hydrogen peroxide and said peroxidative substance

to produce a color dye, and

B. detecting said dye as a result of the presence of hydrogen peroxide or an analyte which reacts to produce hydrogen peroxide.

18. The method of claim 17 wherein said analyte is glucose and said contacting step A occurs in the presence of glucose oxidase.

19. A diagnostic test kit for the determination of hydrogen peroxide, said kit comprising a container means containing:

(1) a color-forming coupler of the structure;

##STR12## wherein R is a water solubilizing group,

R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring, and

(2) an oxidizable color developing compound which is capable of reacting with said coupled in the presence of both hydrogen peroxide and a substance having peroxidative activity to produce a color dye.

20. A color-forming coupling compound of the structure:

##STR13## wherein R is a water solubilizing group having one or more phosphono groups or equivalent esters or salts.

R₁, R₂, R₃ and R₄ are independently hydrogen, alkyl, alkoxy, aryl, aryloxy, a heterocyclic group, halo, amino or R, and

Z represents the carbon atoms necessary to complete the nucleus of a N-substituted 5- or 6-membered fused ring.

21. The compound of claim 20 wherein R is

22. The compound of claim 20 wherein Z represents the carbon atoms necessary to complete the nucleus of a saturated N-substituted 6-membered fused ring.

23. The compound of claim 22 selected from the group consisting of:

##STR14##