Stabilized composition, test device, and method for detecting the presence of a sugar in a test sample

• US 4,318,984 A
• Filed: 11/13/1979
• Issued: 03/09/1982
• Est. Priority Date: 11/13/1979
• Status: Expired due to Term

• Current Assignee: Miles Laboratories, Inc. (Bayer AG)
• Original Assignee: Miles Laboratories, Inc. (Bayer AG)
• Inventors: Tabb, David L., Magers, Thomas A.
• Primary Examiner(s): Warden, Robert J.
• Application Number:
  US06/093,605
• Time in Patent Office: 847 Days
• Field of Search: 435/14, 435/25, 435/28, 23/230 B, 422/56, 252/408
• US Class Current: 435/14
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 2326/14 Ortho-Tolidine, i.e. 3,3'-dimethyl-(1,1'-biphenyl-4,4'-diamine)

Abstract

An improved composition, test device and method for detecting the presence of a sugar in a test sample are disclosed. The composition comprises a sugar oxidase, a peroxidase, a chromogen compound capable of producing a detectable response in the presence of hydrogen peroxide and peroxidase, and an enhancer compound having the structure ##STR1## in which the R substituents, same or different, are hydrogen, lower alkyl, lower alkyloxy, aryl or aryloxy; or in which two of the R substituents bound to a common atom together form a closed ring having 3 to about 6 atoms, said ring being saturated, unsaturated or aromatic. R'"'"' is hydrogen or an alkyl or alkenyl group having 1 to about 20 carbon atoms. The composition can be incorporated with a carrier matrix to form a test device. The method comprises contacting a test solution with the device and observing any detectable response.

Claims

1. In a composition for detecting the presence of a sugar in a test sample, wherein said composition comprises a peroxidase, a sugar oxidase and a chromogen compound having the structure ##STR5## in which R and R*, same or different, are H, lower alkyl, lower alkyloxy, aryl, or aryloxy, or in which the substituents R* together comprise --CH₂ --_n in which n is 1 or 2;

the improvement wherein said composition further comprises an enhancer compound having the structure ##STR6## in which the R substituents, same or different, are as described above;

or in which two of said R substituents bound to a common atom together form a closed ring having 3 to about 6 atoms, said ring being saturated, unsaturated, or aromatic; and

in which R'"'"' is hydrogen or an alkyl or alkenyl group having 1 to about 20 carbon atoms.

2. The improved composition of claim 1 wherein said enhancer compound is R₂ S→

O.

3. The improved composition of claim 1 wherein said enhancer compound is ##STR7##

4. The improved composition of claim 1 wherein said enhancer compound is ##STR8##

5. The improved composition of claim 1 wherein said enhancer compound is R₃ N→

O.

6. The improved composition of claim 1 wherein said enhancer compound is dimethyl sulfone, dimethyl sulfoxide, pentamethylene sulfoxide, formamide, N,N-dimethyl formamide, acetamide, N,N-dimethylacetamide, N-N-di(β

-hydroxyethyl) acetamide, cocoyldiethanolamide, talloyldiethanolamide, tallowyldiethanolamide, trimethylamine oxide, pyridine-1-oxide, 3-hydroxypyridine-1-oxide, 4-methylpyridine-1-oxide, or 2-chloropyridine-1-oxide.

7. The improved composition of any of claims 1-5 or 6 wherein said sugar is glucose and said sugar oxidase is glucose oxidase.

8. The improved composition of any of claims 1-5 or 6 wherein said chromogen compound is o-tolidine.

9. The improved composition of any of claims 1-5 or 6 wherein said chromogen compound is tetramethylbenzidine.

10. The improved composition of any of claims 1-5 or 6 wherein said chromogen compound is 2,7-diaminofluorene.

11. In a test device for detecting the presence of a sugar in a test sample, wherein said device comprises a carrier matrix incorporated with a peroxidase, a sugar oxidase and a chromogen compound having the structure ##STR9## in which R and R*, same or different, are hydrogen, lower alkyl, lower alkyloxy, aryl, or aryloxy, or in which the substituents R* together comprise --CH₂ --_n in which n is 1 or 2;

the improvement wherein said carrier matrix is additionally incorporated with an enhancer compound having the structure ##STR10## in which the R substituents, same or different, are as recited above;

or in which two of said R substituents bound to a common atom together form a closed ring having 3 to about 6 atoms, said ring being saturated, unsaturated, or aromatic, and in which R'"'"' is hydrogen or an alkyl or alkenyl group having from 1 to about 20 carbon atoms.

12. The improved test device of claim 11 wherein said enhancer compound is R₂ S→

O.

13. The improved test device of claim 11 wherein said enhancer compound is ##STR11##

14. The improved test device of claim 11 wherein said enhancer compound is ##STR12##

15. The improved test device of claim 11 wherein said enhancer compound is R₃ N→

O.

16. The improved test device of claim 11 wherein said enhancer compound is dimethyl sulfone, dimethyl sulfoxide, pentamethylene sulfoxide, formamide, N,N-dimethyl formamide, acetamide, N,N-dimethylacetamide, N,N-di(β

-hydroxyethyl) acetamide, cocoyldiethanolamide, talloyldiethanolamide, tallowyldiethanolamide, trimethylamine oxide, pyridine-1-oxide, 3-hydroxypyridine-1-oxide, 4methylpyridine-1-oxide, or 2-chloropyridine-1-oxide.

17. The improved test device of any of claims 11-15 or 16 wherein said sugar is glucose and said sugar oxidase is glucose oxidase.

18. The improved test device of any of claims 11-15 or 16 wherein said chromogen compound is o-tolidine.

19. The improved test device of any of claims 11-15 or 16 wherein said chromogen compound is tetramethylbenzidine.

20. The improved test device of any of claims 11-15 or 16 wherein said chromogen compound is 2,7-diaminofluorene.

21. A method for detecting the presence of a sugar in a test sample, said method comprising contacting said sample with a test device comprising a carrier matrix incorporated with

a peroxidase,

a sugar oxidase,

a chromogen compound having the structure ##STR13## in which the substituents R and R*, same or different, are hydrogen, lower alkyl, lower alkyloxy, aryl, or aryloxy, or in which the R* substituents together comprise --CH₂ --_n in which n is 1 or 2;

and

an enhancer compound having the structure ##STR14## in which the R substituents, same or different, are as recited above, or in which two of said R substituents bound to a common atom together form a closed ring, having 3 to about 6 atoms, said ring being saturated, unsaturated, or aromatic; and

in which R'"'"' is hydrogen or an alkyl or alkenyl group having from 1 to about 20 carbon atoms; and

observing a detectable response produced by said test device.

22. The method of claim 21 wherein said enhancer compound is R₂ S→

O.

23. The method of claim 21 wherein said enhancer compound is ##STR15##

24. The method of claim 21 wherein said enhancer compound is ##STR16##

25. The method of claim 21 wherein said enhancer compound is R₃ N→

0.

26. The method of claim 21 wherein said enhancer compound is dimethyl sulfone, dimethyl sulfoxide, pentamethylene sulfoxide, formamide, N,N-dimethyl formamide, acetamide, N,N-dimethylacetamide, N,N-di(β

-hydroxyethyl) acetamide, cocoyldiethanolamide, talloyldiethanolamide, tallowyldiethanolamide, trimethylamine oxide, pyridine-1-oxide, 3-hydroxypyridine-1-oxide, 4-methylpyridine-1-oxide, or 2-chloropyridine-1-oxide.

27. The method of any of claims 21-25 or 26 wherein said sugar is glucose and said sugar oxidase is glucose oxidase.

28. The method of any of claims 21-25 or 26 wherein said chromogen compound is o-tolidine.

29. The method of any of claims 21-25 or 26 wherein said chromogen compound is tetramethylbenzidine.

30. The method of any of claims 21-25 or 26 wherein said chromogen compound is 2,7-diaminofluorene.