US4629697A
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Test system and procedure for the determination of NAD (P) H
Assignee: Merck Patent GmbH
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Filing: 1983-12-23
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Granted: 1986-12-16
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Expires: 2003-12-23
Abstract
Abstract A test system having an extended range of measurement and an appropriate procedure for the determination of NAD(P)H or of substrates or enzymes which react to form or consume NAD(P)H in fluids is provided. The test system contains, at one and the same time, several substances acting independently of one another as electron acceptors with respect to NAD(P)H and having different electrochemical potentials. Addition of the test system to the sample solution gives rise to different end products which can be analytically differentiated and which are evaluated visually or by other techniques of measurement.
Inventors
- 👤 Berthold Limbach
- 👤 Roland Helger
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 11:50
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/008; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/81; Y10S436/00; Y10S436/904
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
69 words
A test system useful for providing an extended range of measurement for the determination of NADH, NADPH, NAD + or NADP + , or of a substrate or enzyme which reacts with the formation or consumption of NADH or NADPH, comprising effective amounts of at least two redox indicators which, independently of each other, act as electron acceptors with respect to NAD(P)H and which each have different electrochemical potentials.
Claim 2
Dependent Claim
69 words
A test system useful for providing an extended range of measurement for the determination of NADH, NADPH, NAD + or NADP + , or of a substrate or enzyme which reacts with the formation or consumption of NADH or NADPH, comprising effective amounts of at least two redox indicators which, independently of each other, act as electron acceptors with respect to NAD(P)H and which each have different electrochemical potentials.
Claim 3
Dependent Claim
18 words
A test system of claim 1, wherein the redox indicators have a standard potential of -0.3 to +0.5.
Claim 4
Dependent Claim
18 words
A test system of claim 1, wherein the redox indicators have a standard potential of -0.3 to +0.3.
Claim 5
Dependent Claim
20 words
A test system of claim 1 further comprising an effective amount of a buffer which maintains a pH of 4.5-8.0.
Claim 6
Dependent Claim
20 words
A test system of claim 1 further comprising an effective amount of a buffer which maintains a pH of 5.0-7.0.
Claim 7
Dependent Claim
27 words
A test system of claim 5 wherein the buffer is a phosphate buffer, a citrate buffer, 2-N-morpholinoethanesulphonic acid sodium salt, bis(2-hydroxy-ethyl) aminotris(hydroxymethyl)methane, or piperazine-N,N'-bis(2-ethanesulphonic acid) dipotassium salt.
Claim 8
Dependent Claim
16 words
A test system of claim 1 further comprising an effective amount of an electron transfer agent.
Claim 9
Dependent Claim
25 words
A test system of claim 7 wherein the electron transfer agent is, phenazine methosulphate, phenazine ethosulphate, methoxyphenazine methosulphate, Meldola's blue, diaphorase or a mixture thereof.
Claim 10
Dependent Claim
16 words
A test system of claim 1 wherein each redox indicator is dichlorophenol-indophenol or a tetrazolium salt.
Claim 11
Dependent Claim
18 words
A test system of claim 1 wherein each redox indicator is dichlorophenol-indophenol, 3-(4-iodo- phenyl)-2-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide.
Claim 12
Dependent Claim
11 words
A test system of claim 1 further comprising a background dyestuff.
Claim 13
Dependent Claim
13 words
A test system of claim 11 wherein the background dyestuff is titanium yellow.
Claim 14
Dependent Claim
15 words
A test system of claim 1 wherein the redox indicators are dichlorophenol-indophenol and 3-(4-iodophenyl)-2-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride.
Claim 15
Dependent Claim
15 words
A test system of claim 1, wherein said indicators are impregnated within an absorbent material.
Claim 16
Independent Claim
67 words
A method for the determination of NADH, NADPH, NAD + or NADP + in an aqueous sample solution, comprising treating the sample solution with at least two system compatible redox substances which act, independently of one another, as electron acceptors with respect to NADH or NADPH and which have different electrochemical potentials, whereby there are sequentially formed corresponding, different end products, and analytically measuring each end product.
Claim 17
Independent Claim
67 words
A method for the determination of NADH, NADPH, NAD + or NADP + in an aqueous sample solution, comprising treating the sample solution with at least two system compatible redox substances which act, independently of one another, as electron acceptors with respect to NADH or NADPH and which have different electrochemical potentials, whereby there are sequentially formed corresponding, different end products, and analytically measuring each end product.
Claim 18
Dependent Claim
47 words
A method of claim 15 wherein the NADH, NADPH, NAD + or NADP + moiety is derived from the presence of a substrate or enzyme which has reacted to form said moiety, and the measurement of said end products constitutes a determination of the substrate or enzyme.
Claim 19
Dependent Claim
11 words
A method of claim 15 wherein the measurement is conducted photometrically.
Claim 20
Dependent Claim
11 words
A method of claim 15 wherein the measurement is conducted visually.
Claim 21
Dependent Claim
21 words
A method of claim 15 wherein the determination is carried out in a buffered aqueous solution of a pH of 4.5-8.0.
Claim 22
Dependent Claim
73 words
An absorbent material impregnated with a test system useful for providing an extended range of measurement for the determination of NADH, NADPH, NAD + or NADP + , or of a substrate or enzyme which reacts with the formation or consumption of NADH or NADPH, comprising effective amounts of at least two redox indicators which, independently of each other, act as electron acceptors with respect to NAD(P)H and which have different electrochemical potentials.
Claim 23
Dependent Claim
73 words
An absorbent material impregnated with a test system useful for providing an extended range of measurement for the determination of NADH, NADPH, NAD + or NADP + , or of a substrate or enzyme which reacts with the formation or consumption of NADH or NADPH, comprising effective amounts of at least two redox indicators which, independently of each other, act as electron acceptors with respect to NAD(P)H and which have different electrochemical potentials.
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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) | 1989-12-15 | 1990-06-16 | 1990-12-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-12-15 | 1994-06-16 | 1994-12-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-12-15 | 1998-06-16 | 1998-12-16 | $7700.00 | Window Closed / Lapsed |