# US 5,702,955 A · Ascorbate resistant detection of hydrogen peroxide

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- Number: US 5,702,955 A
- Document ID: US5702955A
- Status: Granted
- Kind: A
- Application number: 08/446,478
- Filed: 1995-10-06
- Granted: 1997-12-30
- Earliest claimed: 1995-10-06
- Assignees: Bayer Aktiengesellschaft
- Inventors: Michael Pugia
- Web page: https://patentgrep.com/patent/US5702955A

## CPC

- G01N 31/228: {for peroxides}
- Y10T 436/142222: Hetero-O [e.g., ascorbic acid, etc.]
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- Y10T 436/144444: Glucose
- Y10T 436/145555: Hetero-N
- Y10T 436/146666: Bile pigment
- Y10T 436/147777: Plural nitrogen in the same ring [e.g., barbituates, creatinine, etc.]
- Y10T 436/148888: Uric acid
- Y10T 436/20: Oxygen containing
- Y10T 436/200833: Carbonyl, ether, aldehyde or ketone containing
- Y10T 436/203332: Hydroxyl containing
- Y10T 436/206664: Ozone or peroxide
- Y10T 436/25125: Digestion or removing interfering materials
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The present invention is a method for the detection of hydrogen peroxide in biological fluids or an aqueous solution which involves contacting the solution with an oxidation-reduction indicator and a transition metal complex. The transition metal complex is either a creatinine coordinated with iron or a guanidine coordinated with iron.

## Claims

### Claim 1

A method for the detection of hydrogen peroxide in a solution which may contain ascorbate, comprising the steps of:
(a) adding to said solution an oxidation-reduction indicator and a transition metal complex having the structure: ##STR5## wherein X and Y are independently chosen from the group consisting of creatinine, chloride, fluoride, bromide, iodide, sulfate, phosphate, perchlorate, nitrate, oxalate, sulfide, ammonium, gluconate, cyanide, thiocyanate, catechol, tropolone, phenol, pyridine, acetate, citrate, tartaric acid, malonic acid, boric acid, succinic acid, glycerol-2-phosphate, salicylic acid, oxalic acid, and malic acid; and
(b) analyzing the color transition to detect the presence or concentration of hydrogen peroxide.

### Claim 2 (depends on 1)

The method of claim 1 wherein X and Y are chloride.

### Claim 3 (depends on 1)

The method of claim 1 wherein X and Y are acetate.

### Claim 4 (depends on 1)

The method of claim 1 wherein the oxidation-reduction indicator is chosen from the group consisting of benzidine, o-tolidine, 3,3',5,5'-tetraalkylbenzidines having alkyl groups containing from one to six carbon atoms, o-dianisidine, 2,7-diaminofluorene, bis-(N-ethylquinol-2-one)-azine, (N-methylbenzthiazol-2-one)-(1-ethyl-3-phenyl-5-methyltriazol-2-one)-azine , 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), 4-aminoantipyrine, potassium iodine, and combinations thereof.

### Claim 5 (depends on 4)

The method of claim 4 wherein the oxidation-reduction indicator is 3,3',5,5'-tetramethylbenzidine.

### Claim 6 (depends on 1)

The method of claim 1 wherein the solution is a biological fluid or an aqueous solution.

### Claim 7 (depends on 1)

The method of claim 1 wherein the solution also contains ascorbate ion.

### Claim 8 (depends on 1)

The method of claim 1 wherein the concentration of hydrogen peroxide is in the range of from about 0.1 to about 100 mM.

### Claim 9 (depends on 8)

The method of claim 8 wherein the concentration of hydrogen peroxide is in the range of from about 1 to about 50 mM.

### Claim 10 (depends on 1)

The method of claim 1 wherein the oxidation-reduction indicator and the transition metal complex are added to the solution by means of a test strip to which they have been applied.

### Claim 11

A method for the detection of hydrogen peroxide in a solution which may contain ascorbate, comprising the steps of:
(a) adding to said solution an oxidation-reduction indicator and a transition metal complex having the structure: ##STR6## wherein X and Y are independently chosen from the group consisting of creatinine, chloride, fluoride, bromide, iodide, sulfate, phosphate, perchlorate, nitrate, oxalate, sulfide, ammonium, gluconate, cyanide, thiocyanate, catechol, tropolone, phenol, pyridine, acetate, citrate, tartaric acid, malonic acid, boric acid, succinic acid, glycerol-2-phosphate, salicylic acid, oxalic acid, and malic acid, and R.sub.1 and R.sub.2 are independently chosen from the group consisting of C.sub.n H.sub.2n+1, (C.sub.2 H.sub.4 O).sub.n H, C.sub.5 H.sub.4 Z, C.sub.4 H.sub.3 Z, and C.sub.11 H.sub.6 Z, wherein n is an integer from 1 to 20, and Z is O, S, N, or CH; and
(b) analyzing the color transition to detect the presence or concentration of hydrogen peroxide.

### Claim 12 (depends on 11)

The method of claim 11 wherein X and Y are sulfate.

### Claim 13 (depends on 11)

The method of claim 11 wherein X and Y are acetate.

### Claim 14 (depends on 11)

The method of claim 11 wherein n is an integer from 1 to 12.

### Claim 15 (depends on 11)

The method of claim 11 wherein at least one of R.sub.1 and R.sub.2 is substituted with at least one functional group chosen from the group consisting of alcohol, amine, carbonyl, sulfonic acid, carboxylic acid, fluorine, chlorine, bromine, iodine, phosphonic acid, and phosphate.

### Claim 16 (depends on 15)

The method of claim 15 wherein R.sub.1 is H and R.sub.2 is CH.sub.2 CH.sub.2 --SO.sub.3 H.

### Claim 17 (depends on 15)

The method of claim 15 wherein R.sub.1 is H and R.sub.2 is C.sub.6 H.sub.3 (CO.sub.2 H).sub.2.

### Claim 18 (depends on 15)

The method of claim 15 wherein R.sub.1 is H and R.sub.2 is (CH.sub.2).sub.3 CH(NH.sub.2)CO.sub.2 H.

### Claim 19 (depends on 11)

The method of claim 11 wherein the oxidation-reduction indicator is chosen from the group consisting of benzidine, o-tolidine, 3,3',5,5'-tetraalkylbenzidines having alkyl groups containing from one to six carbon atoms, o-dianisidine, 2,7-diaminofluorene, bis-(N-ethylquinol-2-one)-azine, (N-methylbenzthiazol-2-one)-(1-ethyl-3-phenyl-5-methyltriazol-2-one)-azine , 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), 4-aminoantipyrine, potassium iodine, and combinations thereof.

### Claim 20 (depends on 19)

The method of claim 19 wherein the oxidation-reduction indicator is 3,3',5,5'-tetramethylbenzidine.

### Claim 21 (depends on 11)

The method of claim 11 wherein the solution is a biological fluid or an aqueous solution.

### Claim 22 (depends on 11)

The method of claim 11 wherein the solution also contains ascorbate ion.

### Claim 23 (depends on 11)

The method of claim 11 wherein the concentration of hydrogen peroxide is in the range of from about 0.1 to about 100 mM.

### Claim 24 (depends on 23)

The method of claim 23 wherein the concentration of hydrogen peroxide is in the range of from about 1 to about 50 mM.

### Claim 25 (depends on 11)

The method of claim 11 wherein the oxidation-reduction indicator and the transition metal complex are added to the solution by means of a test strip to which they have been applied.