US3862009A
Expired / Lapsed
US
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Determination of triglycerides
Assignee: Roche Diagnostics GmbH
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Filing: 1973-05-30
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Granted: 1975-01-21
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Expires: 1993-05-30
Abstract
Abstract Triglycerides are determined by enzymatic saponification with lipase and measurement of the liberated glycerol by carrying out the saponification in the presence of carboxylesterase and an alkali metal or alkaline earth metal alkyl sulfate; reagents are provided for carrying out the process and comprise a buffer and, as a saponification agent, lipase, carboxylesterase and the alkali metal or alkaline earth metal alkyl sulfate, and a glycerol detection system.
Inventors
- 👤 August Wilhelm Wahlefeld
- 👤 Hans Mollering
- 👤 Wolfgang Gruber
- 👤 Erich Bernt
- 👤 Peter Roeschlau
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 19:04
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/48; C12Q1/34; C12Q1/44; C12Q1/61
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
58 words
Reagent for the determination of triglycerides by enzymatic saponification which reagent comprises a saponification agent, a buffer, and a system for the detection of glycerol, wherein the saponification agent comprises a lipase obtained from Rhizopus arrhizus, carboxylesterase, and an alkali metal or alkaline earth metal alkyl sulfate, wherein the alkyl group contains from 10 to 15 carbon atoms.
Claim 2
Dependent Claim
58 words
Reagent for the determination of triglycerides by enzymatic saponification which reagent comprises a saponification agent, a buffer, and a system for the detection of glycerol, wherein the saponification agent comprises a lipase obtained from Rhizopus arrhizus, carboxylesterase, and an alkali metal or alkaline earth metal alkyl sulfate, wherein the alkyl group contains from 10 to 15 carbon atoms.
Claim 3
Dependent Claim
15 words
Reagent as claimed in claim 1 wherein the said carboxylesterase is a pig liver esterase.
Claim 4
Dependent Claim
36 words
Reagent as claimed in claim 1 wherein the said system for the detection of glycerol comprises nicotinamideadenine-dinucleotide in reduced form, adenosine triphosphate, phosphoenol pyruvate, lactate dehydrogenase, pyruvate kinase, glycerokinase, cobalt and/or magnesium ions and a buffer.
Claim 5
Dependent Claim
19 words
Reagent as claimed in claim 1 wherein the buffer is a 0.1M triethanolamine buffer of pH 7 to 8.5.
Claim 6
Dependent Claim
15 words
Reagent as claimed in claim 1 wherein the alkyl sulfate is an alkali metal sulfate.
Claim 7
Dependent Claim
17 words
Reagent as claimed in claim 1 wherein the alkyl sulfate is an alkaline earth metal alkyl sulfate.
Claim 8
Dependent Claim
13 words
Reagent as claimed in claim 1 wherein the reagent additionally contains serum albumin.
Claim 9
Dependent Claim
15 words
Reagent as claimed in claim 1 additionally comprising 0.5 to 1.0 mM of cobalt ions.
Claim 10
Dependent Claim
88 words
Reagent as claimed in claim 1 comprising: 0.1 to 10.0 mg./ml. lipase from Rhizopus arrhizus 0.5 to 20.0 mg./ml. carboxylesterase 0.01 to 0.2 mg./ml. sodium dodecyl sulfate 1 to 20 mM nicotinamide-adenine-dinucleotide in reduced form 10 to 100 mM adenosine triphosphate 2 to 20 mM phosphoenol pyruvate 0.5 To 5 mg./ml. lactate dehydrogenase 0.2 to 5 mg./ml. phosphoenol kinase 0.05 to 10 mg. glycerokinase 0.1 to 2.0 mg./ml. serum albumin 3 to 30 mM magnesium ions and 0.03 to 0.3 M buffer solution of pH 6 to 9.
Claim 11
Dependent Claim
14 words
Process as claimed in claim 10 wherein the carboxylesterase is a pig liver esterase.
Claim 12
Dependent Claim
19 words
Method as claimed in claim 10 wherein the saponification is carried out in the additional presence of serum albumin.
Claim 13
Dependent Claim
19 words
Method as claimed in claim 10 wherein the saponification is carried out at a pH from 6 to 9.
Claim 14
Dependent Claim
19 words
Method as claimed in claim 10 wherein the saponification is carried out at a pH between 7 and 8.5.
Claim 15
Dependent Claim
25 words
Method as claimed in claim 10 wherein the said alkyl sulfate is used in an amount of from 0.01 to 1.0 mg./ml. of reagent basis.
Claim 16
Dependent Claim
23 words
Method as claimed in claim 12 wherein the serum albumin is used in an amount of 0.1 to 2.0 mgs./ml. of reagent solution.
Claim 17
Dependent Claim
112 words
Method as claimed in claim 10 wherein the determination is carried out with a combined reagent comprising the saponification agent, the system for the detection of glycerol, and a buffer comprising: 0.1 to 10.0 mg./ml. lipase from Rhizopus arrhizus 0.5 to 20.0 mg./ml. carboxylesterase 0.01 to 0.2 mg./ml. sodium dodecyl sulfate 1 to 20 mM nicotinamide-adenine-dinucleotide in reduced form 10 to 100 mM adenosine triphosphate 2 to 20 mM phosphoenol pyruvate 0.5 to 5 mg./ml. lactate dehydrogenase 0.2 to 5 mg./ml. phosphoenol kinase 0.05 to 10 mg. glycerokinase 0.1 to 2.0 mg./ml. serum albumin 3 to 30 mM magnesium ions and 0.03 to 0.3 M buffer solution of pH 6 to 9.
References Cited (Backward Citations)
US3759793A
Cited
US3703591A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-01-20 | 1978-07-22 | 1979-01-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-01-20 | 1982-07-22 | 1983-01-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-01-20 | 1986-07-22 | 1987-01-21 | $7700.00 | Window Closed / Lapsed |