US4900666A
Expired / Lapsed
US
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Colorimetric ethanol analysis method and test device
Assignee: LifeScan Inc
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Filing: 1988-02-18
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Granted: 1990-02-13
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Expires: 2008-02-18
Abstract
Abstract A disposable test strip device for detecting and measuring ethanol in aqueous solutions is disclosed. The test strip includes an inert support pad that contains a stabilized dry form of the enzyme alcohol oxidase, a material having peroxidative activity and an oxygen acceptor that reacts with hydrogen peroxide to give a compound of changed color. The use of this strip to determine ethanol levels colorimetrically is also disclosed. The stabilizer can comprise a solid aliphatic polyhydroxy compound of from 5-8 carbons and 5-7 hydroxyl groups.
Inventors
- 👤 Roger C. Phillips
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 01:46
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26
Claims Analysis
Total Claims: 42
Claim 1
Independent Claim
129 words
A dry room-temperature-stable test device for detecting ethanol in a aqueous fluid, which comprises an inert support pad containing stabilized alcohol oxidase in a dry form, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide in the presence of said substance having peroxidative activity to give a compound of changed color, wherein ethanol in said aqueous fluid reacts with the alcohol oxidase to yield hydrogen peroxide which reacts with the substance having peroxidative activity and the substance which gives the compound of changed color.
Claim 2
Dependent Claim
129 words
A dry room-temperature-stable test device for detecting ethanol in a aqueous fluid, which comprises an inert support pad containing stabilized alcohol oxidase in a dry form, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide in the presence of said substance having peroxidative activity to give a compound of changed color, wherein ethanol in said aqueous fluid reacts with the alcohol oxidase to yield hydrogen peroxide which reacts with the substance having peroxidative activity and the substance which gives the compound of changed color.
Claim 3
Dependent Claim
16 words
The test device of claim 1 wherein said solid aliphatic polyhydroxyl compound is a polyhydric alcohol.
Claim 4
Dependent Claim
23 words
The test device of claim 2, wherein said polyhydric alcohol comprises a member of the group consisting of mannitol, sorbitol, glucitol, and inositol.
Claim 5
Dependent Claim
12 words
The test device of claim 3, wherein said polyhydric alcohol is mannitol.
Claim 6
Dependent Claim
21 words
The test device of claim 1, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with a chelating agent.
Claim 7
Dependent Claim
13 words
The test device of claim 5, wherein said chelating agent comprises a polyaminepolyacid.
Claim 8
Dependent Claim
15 words
The test device of claim 6, wherein the polyaminepolyacid is a salt of ethylenediaminetetraacetic acid.
Claim 9
Dependent Claim
22 words
The test device of claim 5, wherein said polyhydric compound is mannitol and said chelating agent is a salt of ethylenediaminetetraacetic acid.
Claim 10
Dependent Claim
22 words
The test device of claim 1, wherein said stabilized alcohol oxidase additionally comprises a salt that buffers pH to 6 to 8.
Claim 11
Dependent Claim
59 words
The test device of claim 1, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydric alcohol having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a chelating agent, and a salt that buffers pH to 6 to 8.
Claim 12
Dependent Claim
61 words
The test device of claim 1, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydric alcohol having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a chelating agent, a salt that buffers pH to 6 to 8, and a saccharide.
Claim 13
Dependent Claim
20 words
The test device of claim 1, wherein the inert support pad additionally comprises an effective background-neutralizing amount of peroxide scavenger.
Claim 14
Dependent Claim
20 words
The test device of claim 5, wherein the inert support pad additionally comprises an effective background-neutralizing amount of peroxide scavenger.
Claim 15
Dependent Claim
13 words
The test device of claim 13, wherein the peroxide scavenger is ascorbic acid.
Claim 16
Dependent Claim
20 words
The test device of claim 12, wherein the inert support pad additionally comprises an effective color-moderating amount of peroxide scavenger.
Claim 17
Dependent Claim
20 words
The test device of claim 13, wherein the inert support pad additionally comprises an effective color-moderating amount of peroxide scavenger.
Claim 18
Dependent Claim
13 words
The test device of claim 16, wherein the peroxide acavenger is ascorbic acid.
Claim 19
Dependent Claim
17 words
The test device of claim 1, wherein said support pad is overcoated with a whole-cell-impermeable water-permeable material.
Claim 20
Dependent Claim
12 words
The test device of claim 18, wherein said material is ethyl cellulose.
Claim 21
Dependent Claim
18 words
The test device of claim 1, additionally comprising an inert backing upon which said support pad is mounted.
Claim 22
Dependent Claim
53 words
A stabilized dry alcohol oxidase formulation, comprising alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of stabilizer comprising a solid aliphatic polyhydric alcohol having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a chelating agent, and a buffer that buffers pH to 6 to 8.
Claim 23
Dependent Claim
53 words
A stabilized dry alcohol oxidase formulation, comprising alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of stabilizer comprising a solid aliphatic polyhydric alcohol having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a chelating agent, and a buffer that buffers pH to 6 to 8.
Claim 24
Dependent Claim
64 words
The stabilized dry alcohol oxidase formulation of claim 21 comprising alcohol oxidase in intimate admixture with a stabilizer comprising, per 1000 enzyme units of alcohol oxidase, 300 to 2500 mg of a 5 to 8 carbon solid aliphatic alcohol having 5 to 7 hydroxyl groups, 2 to 50 mg of a polyaminepolyacid chelating agent and a salt that buffers pH to 6 to 8.
Claim 25
Dependent Claim
23 words
The stabilized dry alcohol oxidase formulation of claim 22 further comprising from 0.2 to 12 μmoles of peroxide scavenger per 1000 enzyme units.
Claim 26
Dependent Claim
23 words
The stabilized dry alcohol oxidase formulation of claim 22 further comprising from 4 to 80 μmoles of peroxide scavenger per 1000 enzyme units.
Claim 27
Dependent Claim
12 words
The formulation of claim 23, wherein the peroxide scavenger is ascorbic acid.
Claim 28
Dependent Claim
12 words
The formulation of claim 24, wherein the peroxide scavenger is ascorbic acid.
Claim 29
Independent Claim
124 words
A method for determining the concentration of ethanol in an ethanol-containing aqueous sample, which comprises: (a) reacting a dry room temperature stable test device with the ethanol-containing sample, said test device comprising an inert support pad containing stabilized alcohol oxidase, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color whereby a change of color is obtained; and (b) comparing the color to a standard to determine the ethanol concentration in said sample.
Claim 30
Independent Claim
124 words
A method for determining the concentration of ethanol in an ethanol-containing aqueous sample, which comprises: (a) reacting a dry room temperature stable test device with the ethanol-containing sample, said test device comprising an inert support pad containing stabilized alcohol oxidase, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color whereby a change of color is obtained; and (b) comparing the color to a standard to determine the ethanol concentration in said sample.
Claim 31
Dependent Claim
12 words
The method of claim 27, wherein the aqueous sample is human blood.
Claim 32
Independent Claim
131 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting with saliva of said human a dry, room-temperature-stable test device comprising an inert support pad containing stabilized alcohol oxidase, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color whereby a change of color is obtained; and (b) comparing the color to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 33
Independent Claim
131 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting with saliva of said human a dry, room-temperature-stable test device comprising an inert support pad containing stabilized alcohol oxidase, wherein said stabilized alcohol oxidase comprises alcohol oxidase in intimate admixture with an effective alcohol oxidase stabilizing amount of a stabilizer comprising a solid aliphatic polyhydroxyl compound having from 5 to 8 carbon atoms and 5 to 7 hydroxyl groups, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color whereby a change of color is obtained; and (b) comparing the color to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 34
Independent Claim
116 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting saliva of said human with a dry, room-temperature-stable test device comprising an inert support pad containing alcohol oxidase, a solid 5 to 8 carbon aliphatic polyhydric alcohol, a salt that buffers in the range of pH 6-9, a substance having peroxidase activity, and a substance which will react with hydrogen peroxide to give a compound of changed color, whereby a change of color is obtained in the presence of ethanol; and (b) comparing any color so obtained to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 35
Independent Claim
116 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting saliva of said human with a dry, room-temperature-stable test device comprising an inert support pad containing alcohol oxidase, a solid 5 to 8 carbon aliphatic polyhydric alcohol, a salt that buffers in the range of pH 6-9, a substance having peroxidase activity, and a substance which will react with hydrogen peroxide to give a compound of changed color, whereby a change of color is obtained in the presence of ethanol; and (b) comparing any color so obtained to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 36
Dependent Claim
17 words
The method of claim 30, wherein said test device further comprises a polyamine polyacid metal chelating agent.
Claim 37
Dependent Claim
30 words
The method of claim 30, wherein said test device comprises alcohol oxidase in intimate admixture with from 300-2500 mg of said polyhydric alcohol per 1000 enzyme units of alcohol oxidase.
Claim 38
Dependent Claim
28 words
The method of claim 32, wherein said test device further comprises from 2-50 mg of a polyamine polyacid metal chelating agent per 1000 enzyme units of alcohol oxidase.
Claim 39
Dependent Claim
22 words
The method of claim 30, wherein said polyhydric alcohol comprises a member of the group consisting of mannitol, sorbitol, glucitol, and inositol.
Claim 40
Independent Claim
140 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting saliva of said human with a dry, room-temperature-stable test device comprising an inert support pad containing alcohol oxidase in intimate admixture with a stabilizer comprising 300 to 2500 mg of a 5 to 8 carbon solid aliphatic polyhydric alcohol having 5-7 hydroxyl groups per 1000 enzyme units of alcohol oxidase and a salt that buffers in the range of pH 6-8, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color, whereby a change of color is obtained in the presence of ethanol; and (b) comparing the color so obtained to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 41
Independent Claim
140 words
A method for determining the concentration of ethanol in the bloodstream of a human who is suspected of having consumed ethanol, which comprises: (a) reacting saliva of said human with a dry, room-temperature-stable test device comprising an inert support pad containing alcohol oxidase in intimate admixture with a stabilizer comprising 300 to 2500 mg of a 5 to 8 carbon solid aliphatic polyhydric alcohol having 5-7 hydroxyl groups per 1000 enzyme units of alcohol oxidase and a salt that buffers in the range of pH 6-8, a substance having peroxidative activity, and a substance which will react with hydrogen peroxide to give a compound of changed color, whereby a change of color is obtained in the presence of ethanol; and (b) comparing the color so obtained to a standard to determine the ethanol concentration in the bloodstream of said human.
Claim 42
Dependent Claim
33 words
The method of claim 35, wherein said change of color results from the interaction of horseradish peroxidase with 3-methyl-2-benzothiazolenone hydrazone and N,N-dimethylaminobenzoic acid and said support pad further comprises ascorbic acid or cysteine.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1993-02-12 | 1993-08-14 | 1994-02-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-02-12 | 1997-08-14 | 1998-02-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-02-12 | 2001-08-14 | 2002-02-13 | $7700.00 | Window Closed / Lapsed |