US4212938A
Expired / Lapsed
US
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Reagent and method for the determination of cholesterol
Assignee: Roche Diagnostics GmbH
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Filing: 1973-05-15
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Granted: 1980-07-15
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Expires: 1993-05-15
Abstract
Abstract Cholesterol is determined by incubating a sample suspected of containing cholesterol in an aqueous medium with cholesterol oxidase and determining either the oxygen consumption, the hydrogen peroxide formed, or cholestenone, as a measure of the initial cholesterol content; reagent compositions comprising cholesterol oxidase and a system for the determination of H 2 O 2 or for the determination of cholestenone are provided.
Inventors
- 👤 Wolfgang Gruber
- 👤 Hans U. Bergmeyer
- 👤 Erich Bernt
- 👤 Alexander Hagen
- 👤 Peter Roeschlau
- 👤 Gunter Lang
- 👤 Klaus Beaucamp
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 11:43
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/60
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
53 words
A test composition for determining total cholesterol in a biological fluid sample comprising a chemical system having cholesterol ester hydrolase activity, a chemical system having cholesterol oxidase activity, and means for determining at least one of the reaction products produced when free cholesterol is contacted with said chemical system having cholesterol oxidase activity.
Claim 2
Dependent Claim
53 words
A test composition for determining total cholesterol in a biological fluid sample comprising a chemical system having cholesterol ester hydrolase activity, a chemical system having cholesterol oxidase activity, and means for determining at least one of the reaction products produced when free cholesterol is contacted with said chemical system having cholesterol oxidase activity.
Claim 3
Dependent Claim
63 words
Method for the determination of cholesterol in a sample comprising incubating the sample in an aqueous medium with cholesterol esterase and cholesterol oxidase, characterized by its ability to convert cholesterol to cholestenone with the formation of hydrogen peroxide, and determining either the oxygen consumption, the H 2 O 2 formed, or cholestenone as a measure of the initial cholesterol content in the sample.
Claim 4
Dependent Claim
63 words
Method for the determination of cholesterol in a sample comprising incubating the sample in an aqueous medium with cholesterol esterase and cholesterol oxidase, characterized by its ability to convert cholesterol to cholestenone with the formation of hydrogen peroxide, and determining either the oxygen consumption, the H 2 O 2 formed, or cholestenone as a measure of the initial cholesterol content in the sample.
Claim 5
Dependent Claim
25 words
Method as claimed in claim 2 wherein the H 2 O 2 formed is determined and the determination is made enzymatically using catalase or peroxidase.
Claim 6
Dependent Claim
24 words
Method as claimed in claim 2 wherein the cholestenone formed is measured, directly at 240 nm or using a ketoreagent such as 2,4-dinitrophenyl hydrazine.
Claim 7
Dependent Claim
20 words
Method as claimed in claim 2 wherein oxygen consumption is determined polarometrically as a measure of the cholesterol initially present.
Claim 8
Dependent Claim
15 words
Test composition as claimed in claim 1 comprising a system for the determination of cholestenone.
Claim 9
Dependent Claim
19 words
Test composition as claimed in claim 1 comprising a system for the determination of H 2 O 2 .
Claim 10
Dependent Claim
17 words
Test composition as claimed in claim 1 comprising a a system for the determination of oxygen consumption.
Claim 11
Dependent Claim
24 words
Test composition as claimed in claim 1 comprising cholesterol oxidase, cholesterol esterase and a system for the determination of H 2 O 2 .
Claim 12
Dependent Claim
30 words
Test composition as claimed in claim 7 wherein said system for the determination of H 2 O 2 comprises a catalase, a β-diketone, a lower aliphatic alcohol and a buffer.
Claim 13
Dependent Claim
20 words
Test composition as claimed in claim 10 wherein the β-diketone is acetyl acetone and the lower aliphatic alcohol is methanol.
Claim 14
Dependent Claim
25 words
Test composition as claimed in claim 7 wherein the system for the determination of H 2 O 2 comprises peroxidase, a chromogen and a buffer.
Claim 15
Dependent Claim
21 words
Test composition as claimed in claim 8 wherein the system for the determination of cholestenone comprises a derivative reacting with keto-groups.
Claim 16
Dependent Claim
13 words
Test composition as claimed in claim 13 wherein said derivative is 2,4-dinitrophenyl hydrazine.
Claim 17
Dependent Claim
54 words
Test composition as claimed in claim 6 comprising: 13 to 150 U of cholesterol oxidase 2×10 4 to 5×10 5 U of catalase 0.5 to 0.2 ml. of acetyl acetone 2 to 10 ml. of methanol in 100 ml. of ammonium ion-containing buffer, pH 5 to 7 0.05 to 0.5 ml of surface--active agent.
Claim 18
Dependent Claim
49 words
Test composition as claimed in claim 12 comprising: 3 to 40 U of cholesterol oxidase 2×10 2 to 1×10 4 U of commercially available peroxidase 50 to 200 mg. of 2,2'-aminobenzthiazoline sulfonic acid 0.05 to 0.5 ml. of surface-active agent in 100 ml. of buffer, pH 6 to 8.
Claim 19
Dependent Claim
33 words
Test composition as claimed in claim 13 comprising: 0.1 to 1 U of cholesterol oxidase 1to 5 ml. of a 1 mM solution of 2,4-dinitrophenyl hydrazine and 0.005-0.1 ml. of a surface agent.
Claim 20
Dependent Claim
13 words
Test composition as claimed in claim 1 additionally containing a surface active agent.
Claim 21
Dependent Claim
13 words
Method as claimed in claim 2 wherein said sample is a body fluid.
Claim 22
Dependent Claim
57 words
Method for the determination of cholesterol in a biological fluid sample which comprises contacting said sample with a chemical system having cholesterol ester hydrolase activity, a chemical system having cholesterol oxidase activity, and means for determining at least one of the reactants or one of the reaction products produced as a measure of the initial cholesterol content.
Claim 23
Dependent Claim
57 words
Method for the determination of cholesterol in a biological fluid sample which comprises contacting said sample with a chemical system having cholesterol ester hydrolase activity, a chemical system having cholesterol oxidase activity, and means for determining at least one of the reactants or one of the reaction products produced as a measure of the initial cholesterol content.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-07-15 | 1984-01-14 | 1984-07-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-07-15 | 1988-01-14 | 1988-07-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-07-15 | 1992-01-14 | 1992-07-15 | $7700.00 | Window Closed / Lapsed |