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The invention provides a reagent system for the enzymatic determination of an oxidizable substrate in a fluid sample; the system includes an active amine trap for inactivating high concentrations of aldehyde and ketone oxidation products comprising a combination of at least one primary amine and at least one alpha-effect amine. An example of the oxidizable substrate is alcohol.

Inventors: John L. Palmer, Marsha W. Timmerman, Stephan D. Daubney
Original Assignee: Enzymatics, Inc.
Current U.S. Classification: 435/25; 435/26; 436/131; 436/132
International Classification: C12Q 126

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Citations

Cited PatentFiling dateIssue dateOriginal AssigneeTitle
US3493467Jul 26, 1967Feb 3, 1970REVERSIBLE BIOCHEMICAL REACTION EMPLOYING A TRAPPING AGENT
US4786596Feb 20, 1985Nov 22, 1988Chem-Elec., Inc.Method of preparing a test strip for alcohol testing

Referenced by

Citing PatentFiling dateIssue dateOriginal AssigneeTitle
US5290683Nov 19, 1992Mar 1, 1994Rapid analysis of ethanol in body fluids
US5429931May 24, 1993Jul 4, 1995Eastman Kodak CompanyMultilayer analytical element containing crosslinked binder and method for the determination of ethanol
US5429932May 24, 1993Jul 4, 1995Eastman Kodak CompanyMultilayer analytical element containing niacinamide and method for the determination of ethanol
US5525481May 29, 1992Jun 11, 1996Hoffman-La Roche Inc.Enzymatic Reagents for ethanol assay containing diamino compounds
US5968746Nov 26, 1997Oct 19, 1999Method and apparatus for preserving human saliva for testing
US6291178Aug 30, 1999Sep 18, 2001Method and apparatus for preserving human saliva for testing
US6436716May 30, 2000Aug 20, 2002Integrated Biomedical Technology, Inc.Aldehyde test strip
US6632874May 1, 2001Oct 14, 2003The Coca-Cola CompanyMethod to decrease aldehyde content in polyolefin products
US6686336Dec 22, 2000Feb 3, 2004Federal Government as represented by the Department of Veterans AffairesN-terminal D(-)-penicillamine peptides as aldehyde sequestration agents
US6762275May 27, 2003Jul 13, 2004The Coca-Cola CompanyMethod to decrease the acetaldehyde content of melt-processed polyesters
US7041350Aug 30, 2002May 9, 2006The Coca-Cola CompanyPolyester composition and articles with reduced acetaldehyde content and method using hydrogenation catalyst
US7790467Sep 4, 2007Sep 7, 2010Southwest Sciences IncorporatedDiode laser based ketone and aldehyde detection

Claims

1. A method for trapping an aldehyde or ketone in an assay for alcohol comprising:

reacting an active amine trap comprising a combination of at least one alpha-effect amine and at least one primary amine, wherein each of the amines employed is substantially non-volatile and has a pKa of at least about 7.5, with
a fluid sample comprising an aldehyde or ketone which is an oxidation product of an enzymatic oxidation reaction of an enzyme substrate, wherein the concentration of said enzyme substrate and oxidation product is greater than about 5 mM.

2. The method of claim 1, wherein each of the amines employed has a solubility in water at 25.degree. C. of at least about 100 mM.

3. The method of claim 1, wherein the active amine trap is capable of inactivating a concentration of aldehyde o ketone oxidation product of at least about 20 mM.

4. The method of claim 1, wherein the active amine trap is capable of inactivating a concentration of aldehyde or ketone oxidation product of at least about 50 mM.

5. The method of claim 1, wherein the active amine trap is capable of inactivating a concentration of aldehyde or ketone oxidation product of about 150 mM.

6. The method of claim 1, wherein the alpha-effect amine is a substituted hydrazine or substituted hydroxylamine.

7. The method of claim 1, wherein the primary amine is a naturally-occurring amino acid or a non-naturally occurring amino acid comprising an amino-C.sub.2 -C.sub.10 -carboxylic acid.

8. The method of claim 1, wherein the weight ratio of primary amine to alpha-effect amine is from about 10% w/w to about 60% w/w.

9. The method of claim 6, wherein the weight ratio is from about 20% w/w to about 50% w/w.

10. The method of claim 1, wherein the dehydrogenation or oxidation takes place at a pH of from about 9 to 10.

11. The method of claim 1, wherein the fluid sample comprising the aldehyde or ketone is a biological fluid.

12. The method of claim 11, wherein the biological fluid is blood or saliva.

13. The method of claim 11, wherein the enzyme substrate is an alcohol.

14. The method of claim 13, wherein the alcohol concentration is up to about 150 mM.

15. The method of claim 14, wherein the conversion rate of the alcohol to ketone product is about 20%.

16. The method of claim 14, wherein the conversion rate of the alcohol to ketone product is about 20 to about 100%.

17. A reagent system for trapping an aldehyde or ketone in an assay for alcohol comprising:

an active amine trap comprising a combination of at least one alpha-effect amine and at least one primary amine, wherein each of the amines employed is substantially non-volatile and has a pKa of at least about 7.5; and
a fluid sample comprising an aldehyde or ketone which is an oxidation product of an enzymatic oxidation reaction of an enzyme substrate, wherein the substrate and oxidation product are present in a concentration greater than about 5 mM.

18. The reagent system of claim 17, wherein each of the amines employed has a solubility in water at 25.degree. C. of at least about 100 mM.

19. The reagent system of claim 17, wherein the active amine trap is capable of inactivating a concentration of aldehyde or ketone oxidation product of at least about 20 mM.

20. The reagent system of claim 17, wherein the active amine trap is capable of inactivating a concentration of aldehyde or ketone oxidation product of at least about 50 mM.

21. The reagent system of claim 17, wherein the active amine trap is capable of inactivating a concentration of aldehyde or ketone oxidation product of about 150 mM.

22. The reagent system of claim 17, wherein the alpha-effect amine is a substituted hydrazine or substituted hydroxylamine.

23. The reagent system of claim 17, wherein the primary amine is a naturally-occurring amino acid or a non-naturally occurring amino acid comprising an amino-C.sub.2 -C.sub.10 -carboxylic acid.

24. The reagent system of claim 17, wherein the weight ratio of primary amine to alpha-effect amine is from about 10% w/w to about 60% w/w.

25. The reagent system of claim 17, wherein the weight ratio is from about 20% w/w to about 50% w/w.

26. The reagent system of claim 17 which is buffered to a pH of from about 9 to 10.

27. The reagent system of claim 17, wherein the enzyme substrate is an alcohol.

28. The reagent system of claim 27, wherein the alcohol concentration is up to about 150 mM.

29. A diagnostic kit for the enzymatic determination of an alcohol in a fluid sample, comprising a film of the reagent system of claim 17 disposed on support means for supporting the film.

30. The diagnostic kit of claim 29, wherein the enzyme substrate is an alcohol.

31. The diagnostic kit of claim 27, wherein the alcohol is ethanol.

32. The diagnostic kit of claim 31, wherein the alcohol concentration is up to about 150 mM.

33. The diagnostic kit of claim 29, wherein the fluid sample comprising the aldehyde or ketone is disposed on a dry physical support.