Determination of hydrogen peroxide

• US 4,251,629 A
• Filed: 07/28/1978
• Issued: 02/17/1981
• Est. Priority Date: 07/29/1977
• Status: Expired due to Term

• Current Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Original Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Inventors: Tokuda, Kuniaki, Hanada, Tosiro, Yamanisi, Kazuhiko
• Primary Examiner(s): Wiseman, Thomas G.
• Application Number:
  US05/928,804
• Time in Patent Office: 935 Days
• Field of Search: 252/408 R, 195/99, 195/103.5 R, 195/103.5 C, 195/103.5 U, 23/230 B, 23/230 M, 435/14, 435/12, 435/11, 435/10, 435/20, 435/28
• US Class Current: 435/28
• CPC Class Codes: C09B 53/02 Indamines; Indophenols
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 1/60 involving cholesterol
• CPC Class Codes: C12Q 1/62 involving uric acid
• CPC Class Codes: C12Q 2326/00 Chromogens for determinations of oxidoreductase enzymes
• CPC Class Codes: C12Q 2326/96 4-Amino-antipyrine
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

Colorimetrical determination of hydrogen peroxide in clinical examinations, food additives analyses and general analyses can be carried out accurately, rapidly and economically by using a composition for producing color producing reagents comprising 4-aminoantipyrine, a N-substituted-3-alkylaniline and a hydrogen peroxide activating agent such as peroxidase, said composition forming a coloring material having excellent sensitivity and color stability in contact with hydrogen peroxide.

Claims

1. A composition for producing color producing reagents for determining hydrogen peroxide which comprises

4-aminoantipyrine,

a N-substituted-3-alkylaniline of the formula:

##STR4## wherein R¹ is a member selected from the group consisting of a lower alkyl group having 1 to 5 carbon atoms and a hydroxyalkyl group having 1 to 5 carbon atoms;

R² is a member selected from the group consisting of a lower alkyl group having 1 to 5 carbon atoms and a hydroxyalkyl group having 1 to 5 carbon atoms and R³ is methyl, ethyl or propyl, and an agent for activating hydrogen peroxide.

2. A composition according to claim 1, wherein in the formula (I) R¹ is a lower alkyl group having 1 to 5 carbon atoms and R² is a hydroxyalkyl group having 1 to 5 carbon atoms.

3. A composition according to claim 1, wherein in the formula (I) R¹ and R² are the same or different alkyl groups having 1 to 5 carbon atoms.

4. A composition according to claim 1, wherein in the formula (I) R¹ and R² are the same or different hydroxyalkyl groups having 1 to 5 carbon atoms.

5. A composition according to claim 1, wherein the N-substituted-3-alkylaniline is N-ethyl-N-hydroxyethyl-3-methylaniline.

6. A composition according to claim 1, wherein the N-substituted-3-alkylaniline is N,N-dimethyl-3-methylaniline.

7. A composition according to claim 1, wherein the N-substituted-3-alkylaniline is N,N-bis(β

-hydroxyethyl)-3-ethylaniline.

8. A composition according to claim 1, wherein the agent for activating hydrogen peroxide is peroxidase.

9. A composition according to claim 1, wherein the agent for activating hydrogen peroxide activating agent is a mixture of an iodide and one compound selected from the group consisting of tungstic acid and its salts, molybdic acid and its salts.

10. In a method for determining hydrogen peroxide in an enzymatic reaction system wherein 4-aminoantipyrine is oxidatively condensed with an aniline to produce a color developed material and the optical absorbance of the color developed material is determined, the improvement comprising employing an N-substituted-3-alkylaniline of the formula:

##STR5## wherein R¹ is a member selected from the group consisting of a lower alkyl group having 1 to 5 carbon atoms and a hydroxyalkyl group having 1 to B 5 carbon atoms;

R² is a member selected from the group consisting of a lower alkyl group having 1 to 5 carbon atoms and a hydroxyalkyl group having 1 to 5 carbon atoms and R³ is methyl, ethyl or propyl, in the presence of an agent for activating hydrogen peroxide.