US4956276A
Expired / Lapsed
US
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Analytical method of determining a reduced co-enzyme
Assignee: Alfa Wassermann Inc
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Filing: 1986-03-10
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Granted: 1990-09-11
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Expires: 2006-03-10
Abstract
Abstract The sample for analysis containing the co-enzyme to be detemined, inter alia NADH or NADPH, is placed in the presence of a fluorinated aromatic compound. In the presence of oxygen and peroxidase, the fluorinated compound releases fluoride ions at a rate proportional to the quantity of co-enzyme to be determined. The thus-formed F - ions are determined, thus obtaining the desired result.
Inventors
- 👤 Igbal Siddiqi
- 👤 Jean Brochot
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 05:48
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; C12Q1/008; C12Q1/54; C12Q1/58
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
89 words
An analytical method of determining a nicotinamide co-enzyme selected from the group consisting of NADH, NADPH and APADH wherein the co-enzyme is reacted in a reaction medium with a fluorinated-ring aromatic compound selected from the group consisting of 2-, 3- and 4-fluorophenol, tetrafluorophenol, pentafluorophenol and 4-fluoro-aniline in the presence of oxygen and a substance having peroxidase activity, this reaction resulting in the formation of fluoride ions which are determined, the rate at which the ions are formed being quantitatively linked to the original quantity of co-enzyme to be determined.
Claim 2
Dependent Claim
89 words
An analytical method of determining a nicotinamide co-enzyme selected from the group consisting of NADH, NADPH and APADH wherein the co-enzyme is reacted in a reaction medium with a fluorinated-ring aromatic compound selected from the group consisting of 2-, 3- and 4-fluorophenol, tetrafluorophenol, pentafluorophenol and 4-fluoro-aniline in the presence of oxygen and a substance having peroxidase activity, this reaction resulting in the formation of fluoride ions which are determined, the rate at which the ions are formed being quantitatively linked to the original quantity of co-enzyme to be determined.
Claim 3
Dependent Claim
35 words
A method according to claim 1, the reaction medium contains at least one additional constituent capable of quantitatively reacting in a chain reaction involving NAD/NADH coenzyme systems, the method adapted for determining the additional constituent.
Claim 4
Dependent Claim
24 words
A method according to claim 1, wherein the fluoride ions are determined electrometrically by means of an electrode which is specific to said ions.
Claim 5
Dependent Claim
14 words
A method according to claim 3, wherein a buffer at pH 5-7.5 is used.
Claim 6
Dependent Claim
13 words
A method according to claim 4, wherein the buffer is "tris" or acetate.
Claim 7
Dependent Claim
32 words
A method according to claim 4, wherein a cacodylate buffer is used at a molar concentration of 0.05 to 0.5M, the reaction rate depending on the concentration of dimethylarsinate in the buffer.
Claim 8
Dependent Claim
25 words
A method according to claim 4, wherein Mn ++ ions in a concentration of 2×10 -6 -10 -4 M are added to the reaction medium.
Claim 9
Dependent Claim
59 words
A method according to claim 3, wherein it is used for determining glucose in samples of biological fluids, the glucose reacting with ATP in the presence of hexokinase to give glucose-6-phosphate which is oxidized by NAD + in the presence of glucose-6-phosphate dehydrogenase to D-gluconate-6-phosphate with corresponding production of NADH in direct proportion to the glucose to be measured.
Claim 10
Dependent Claim
57 words
A method according to claim 3, wherein it is used for determining urea in biological fluids, which supply ammonia by hydrolysis in the presence of urease, the ammonia supplying L-glutamate by reaction with 2-oxoglutorate in the presence of glutamate dehydrogenase and NADH, the consumption of the latter being in direct proportion to the urea to be measured.
Claim 11
Dependent Claim
15 words
A method according to claim 1 wherein the substance having peroxidase activity is peroxidase enzyme.
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JPH051900A
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-09-10 | 1994-03-12 | 1994-09-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-09-10 | 1998-03-12 | 1998-09-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-09-10 | 2002-03-12 | 2002-09-11 | $7700.00 | Window Closed / Lapsed |