Patent US4368261A successfully refreshed from Google Patents.
US4368261A
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Method and reagent for the determination of triglycerides
Assignee: Roche Diagnostics GmbH
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Filing: 1980-12-05
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Granted: 1983-01-11
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Expires: 2000-12-05
Abstract
Abstract A method for the determination of triglycerides by ester cleavage utilizing lipase and optionally esterase with the formation of fatty acids and glycerol, phosphorylation of the glycerol with adnosine triphosphate in the presence of glycerol kinase with the formation of glycerol-1-phosphate and adenosine diphosphate and determination of one of the latter two products. The glycerol kinase used is from Bacillus stearothermophilis and acts in combination with at least one activator selected from the group consisting of detergents, phenol derivatives and aniline derivative.
Inventors
- 👤 Sigmar Klose
- 👤 Albert Roder
- 👤 Walter Schneider
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 06:27
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/61; C12Q1/34; C12Q1/44; C12Q1/48; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/91; G01N2333/912; G01N2333/91205; G01N2333/9121; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/8215; Y10S435/822; Y10S435/832
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
79 words
In a method for the determination of triglycerides which comprises combining sample, lipase, glycerol kinase, adenosine triphosphate, and at least one lipase activator selected from the group consisting of polyethylene oxide ester surfactants, polyethylene oxide ether surfactants, bile acid surfactants, alkyl sulphate surfactants, phenol derivatives, and analine derivatives, under conditions suitable to form glycerol-1-phosphate and adenosine diphosphate and determining the presence or concentration of either glycerol-1-phosphate or adenosine diphosphate, the improvement comprising using a glycerol kinase from Bacillus stereothermophilis.
Claim 2
Dependent Claim
79 words
In a method for the determination of triglycerides which comprises combining sample, lipase, glycerol kinase, adenosine triphosphate, and at least one lipase activator selected from the group consisting of polyethylene oxide ester surfactants, polyethylene oxide ether surfactants, bile acid surfactants, alkyl sulphate surfactants, phenol derivatives, and analine derivatives, under conditions suitable to form glycerol-1-phosphate and adenosine diphosphate and determining the presence or concentration of either glycerol-1-phosphate or adenosine diphosphate, the improvement comprising using a glycerol kinase from Bacillus stereothermophilis.
Claim 3
Dependent Claim
14 words
Method as claimed in claim 1 wherein said activator is a polyethylene oxide ester.
Claim 4
Dependent Claim
14 words
Method as claimed in claim 1 wherein said activator is a polyethylene oxide ether.
Claim 5
Dependent Claim
13 words
Method as claimed in claim 1 wherein said activator is a bile acid.
Claim 6
Dependent Claim
13 words
Method as claimed in claim 1 wherein said activator is an alkyl sulphate.
Claim 7
Dependent Claim
27 words
Method as claimed in claim 1 wherein said activator is a phenol substituted with a substituent selected from the group consisting of carboxyl, sulphonic, halogen and nitro.
Claim 8
Dependent Claim
27 words
Method as claimed in claim 1 wherein said activator is an aniline substituted with a substituent selected from the group consisting of carboxyl, sulphonic, halogen and nitro.
Claim 9
Dependent Claim
15 words
Method as claimed in claim 1 wherein said Bacillus stearothermophilis is Bacillus stearothermophilis NCIB 8924.
Claim 10
Dependent Claim
55 words
Method as claimed in claim 1 wherein said adenosine diphosphate is phosphorylated with phosphoenol pyruvate in the presence of pyruvate kinase with the formation of adenosine triphosphate and pyruvate, and the latter is reduced with nicotinamide-adenine dinucleotide (reduce form) in the presense of lactate dehydrogenase to give nicotinamide-adenine dinucleotide (oxidized form) which is then determined.
Claim 11
Dependent Claim
51 words
Method as claimed in claim 1 wherein glycerol-1- phosphate is further reacted with oxygen in the presence of glycerol-1-phosphate oxidase to give dihydroxyacetone phosphate and hydrogen peroxide which is reacted with 4-aminoantipyrine and a phenol or aniline derivative in the presence of peroxidase to form a colored material which is determined.
Claim 12
Dependent Claim
70 words
Method as claimed in claim 1 wherein adenosine diphosphate is phosphorylated with phosphoenol pyruvate in the presence of pyruvate kinase to form adenosine triphosphate and pyruvate, the so formed pyruvate is reacted with oxygen and pyruvate oxidase to form acetyl phosphate, carbon dioxide and hydrogen peroxide and the latter is reacted with a phenol or aniline derivatives in the presence of peroxidase to give a colored material which is determined.
Claim 13
Dependent Claim
57 words
Reagent for the determination of triglycerides comprising lipase, adenosine triphosphate, glycerokinase, a reagent system for determining glycerol-l-phosphate or adenosine diphosphate and a glycerokinase from Bacillus stearothermophilis, with at least one lipase activator selected from the group consisting of polyethylene oxide ester surfactants, polyethylene oxide ether surfactants, bile acid surfactants, alkyl sulphate surfactants, phenol derivatives and aniline derivatives.
Claim 14
Dependent Claim
57 words
Reagent for the determination of triglycerides comprising lipase, adenosine triphosphate, glycerokinase, a reagent system for determining glycerol-l-phosphate or adenosine diphosphate and a glycerokinase from Bacillus stearothermophilis, with at least one lipase activator selected from the group consisting of polyethylene oxide ester surfactants, polyethylene oxide ether surfactants, bile acid surfactants, alkyl sulphate surfactants, phenol derivatives and aniline derivatives.
Claim 15
Dependent Claim
14 words
Reagent as claimed in claim 12 wherein said activator is a polyethylene oxide ester.
Claim 16
Dependent Claim
14 words
Reagent as claimed in claim 12 wherein said activator is a polyethylene oxide ether.
Claim 17
Dependent Claim
13 words
Reagent as claimed in claim 12 wherein said activator is a bile acid.
Claim 18
Dependent Claim
13 words
Reagent as claimed in claim 12 wherein said activator is an alkyl sulphate.
Claim 19
Dependent Claim
27 words
Reagent as claimed in claim 12 wherein said activator is a phenol substituted with a substituent selected from the group consisting of carboxyl, sulphonic, hydrogen and nitro.
Claim 20
Dependent Claim
27 words
Reagent as claimed in claim 12 wherein said activator is an aniline substituted with a substituent selected from the group consisting of carboxyl, sulphonic, halogen and nitro.
Claim 21
Dependent Claim
14 words
Reagent as claimed in claim 12 wherein said activator is a sodium dodecyl phosphate.
Claim 22
Dependent Claim
24 words
Reagent as claimed in claim 12 wherein said reagent system for determining adenosine diphosphate comprises phosphoenol pyruvate, pyruvate kinase, nictotinamide-adenine dinucleotide and lactate dehydrogenase.
Claim 23
Dependent Claim
11 words
Reagent as claimed in claim 12 wherein said activator is dichlorophenol.
Claim 24
Dependent Claim
19 words
Reagent as claimed in claim 12 wherein said activator is sodium cholate and an alkyl aryl polyethylene oxide ester.
Claim 25
Dependent Claim
17 words
Reagent as claimed in claim 12 wherein said activator is 2,4-dichlorophenol, isotridecylpolyethylene oxide ether and sodium cholate.
Claim 26
Dependent Claim
18 words
Reagent as claimed in claim 12 wherein said system for determining glycerol-1-phosphate comprises glycerol phosphate oxidase, peroxidase and-4-aminoantipyrine.
Claim 27
Dependent Claim
83 words
Reagent as claimed in claim 12 comprising 60 to 200 U lipase, 0.4 to 10 U esterase, 0.5 to 10 U glycerokinase from Bacillus stearothermophilis, 0.5 to 10 U pyruvate kinase, 2 to 10 U lactate dehydrogenase, 0.25 to 0.7 μMol phosphoenol pyruvate, 0.25 to 0.7 μMol ATP, 0.15 to 0.3 μMol NADH, 0.3 to 0.5 μMol sodium dodecyl sulphate, 4to 100 μMol MgSO 4 and 20 to 100 μMol phosphate buffer (pH 6.8 to 7.5) referred to 1 ml. of reagent solution.
Claim 28
Dependent Claim
96 words
Reagent as claimed in claim 12 comprising 0.6 to 10 U esterase, 60 to 300 U lipase, 0.5 to 10 U glycerokinase from Bacillus stearothermophilis, 0.5 to 10 U pyruvate kinase, 2 to 10 U lactate dehydrogenase, 0.15 to 3 μMol NADH, 0.25 to 0.7 μMol phosphoenol pyruvate, 0.25 to 0.7 μMol ATP, 1.5 to 4 μMol 2,3-dichlorophenol, 1.5 to 5.0 mg. sodium cholate, 0.8 to 2 μl. octylphenylpolyethylene oxide ester, 150 to 300 μMol MgSO 4 and 40 to 100 μMol tris/powdered tartaric acid (pH 7.0 to 7.6) referred to 1 ml. of reagent solution.
Claim 29
Dependent Claim
32 words
Reagent as claimed in claim 12 comprising 0.6 to 10 U esterase, 60 to 300 U lipase, 0.5 to 10 U glycerokinase from Bacillus stearothermophilis, 2 to 10 U glycerol phosphate oxidase,
Claim 30
Dependent Claim
44 words
3 to 1 μMol ATP, 3 to 10 μMol 2,4-dichlorophenol, 1.5 to 5.0 mg. sodium cholate, 1.5 to 10 μl. isotridecylpolyethylene oxide ether, 150 to 300 μMol magnesium sulphate, 40 to 100 Mol buffer (pH 7.5 8.5) referred to 1 ml. of reagent solution.
Claim 31
Dependent Claim
44 words
3 to 1 μMol ATP, 3 to 10 μMol 2,4-dichlorophenol, 1.5 to 5.0 mg. sodium cholate, 1.5 to 10 μl. isotridecylpolyethylene oxide ether, 150 to 300 μMol magnesium sulphate, 40 to 100 Mol buffer (pH 7.5 8.5) referred to 1 ml. of reagent solution.
Claim 32
Dependent Claim
14 words
Method of claim 1 wherein, along with lipase, esterase is used for the determination.
References Cited (Backward Citations)
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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) | 1986-01-10 | 1986-07-12 | 1987-01-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-01-10 | 1990-07-12 | 1991-01-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-01-10 | 1994-07-12 | 1995-01-11 | $7700.00 | Window Closed / Lapsed |