US4066508A
Expired / Lapsed
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Process and reagent for determining triglycerides
Assignee: Roche Diagnostics GmbH
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Filing: 1976-07-21
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Granted: 1978-01-03
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Expires: 1996-07-21
Abstract
Abstract Triglycerides are determined by enzymatic saponification with lipase in a buffer in the presence of a carboxylesterase and an alkali metal or alkaline earth metal (10-15 carbon atom) alkyl sulfate, in the additional presence of a compound of the general formula RO(CH 2 CH 2 ) n H, in which R is alkyl or alkenyl containing 14 to 20 carbon atoms and n is a number of from 7 to 20; this determination prevents turbidity and does not disadvantageously influence enzyme activity.
Inventors
- 👤 Elli Rauscher
- 👤 Erich Bernt
- 👤 Wolfgang Gruber
- 👤 Helmut Determann
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 00:35
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/48; C12Q1/34; C12Q1/44; C12Q1/61; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/911; Y10S435/939
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
85 words
Process for the determination of triglycerides comprising enzymatically saponifying the triglycerides with lipase in the buffer in the presence of a carboxylesterase and of an alkali metal of alkaline earth metal alkyl sulfate, the alkyl radical of which contains 10 to 15 carbon atoms, in the additional presence of a compound of the formula RO(CH 2 CH 2 O) n H, in which R is alkyl or alkenyl of from 14 to 20 carbon atoms and n is an integer of from 7 to 20.
Claim 2
Dependent Claim
85 words
Process for the determination of triglycerides comprising enzymatically saponifying the triglycerides with lipase in the buffer in the presence of a carboxylesterase and of an alkali metal of alkaline earth metal alkyl sulfate, the alkyl radical of which contains 10 to 15 carbon atoms, in the additional presence of a compound of the formula RO(CH 2 CH 2 O) n H, in which R is alkyl or alkenyl of from 14 to 20 carbon atoms and n is an integer of from 7 to 20.
Claim 3
Dependent Claim
15 words
Process as claimed in claim 1 wherein n is an integer from 9 to 15.
Claim 4
Dependent Claim
10 words
Process as claimed in claim 1 wherein n is 12.
Claim 5
Dependent Claim
35 words
Process as claimed in claim 1 wherein the compound of the formula RO(CH 2 CH 2 O) n H is added in an amount of from 0.005 to 0.2 wt. percent per volume of buffer.
Claim 6
Dependent Claim
35 words
Process as claimed in claim 1 wherein the compound of the formula RO(CH 2 CH 2 O) n H is added in an amount of from 0.01 to 0.03 wt. percent per volume of buffer.
Claim 7
Dependent Claim
13 words
Process as claimed in claim 1 wherein there is additionally present serum albumin.
Claim 8
Dependent Claim
14 words
Process as claimed in claim 1 wherein said lipase is obtained from Rhizopus arrhizus.
Claim 9
Dependent Claim
13 words
Process as claimed in claim 1 wherein said buffer is a phosphate buffer.
Claim 10
Dependent Claim
20 words
Process as claimed in claim 8 wherein a 0.02 to 0.1M phosphate buffer of pH 6.5 to 8.0 is used.
Claim 11
Dependent Claim
75 words
Reagent for the determination of triglycerides, comprising lipase, carboxylesterase, an alkali metal or alkaline earth metal alkyl sulfate of from 10 to 15 carbon atoms in the alkyl radical, a buffer, a system for the detection of glycerol, and a compound of the general formula RO(CH 2 CH 2 O) n H wherein R is alkyl or alkenyl of from 14 to 20 carbon atoms and n is an integer of from 7 to 20.
Claim 12
Dependent Claim
75 words
Reagent for the determination of triglycerides, comprising lipase, carboxylesterase, an alkali metal or alkaline earth metal alkyl sulfate of from 10 to 15 carbon atoms in the alkyl radical, a buffer, a system for the detection of glycerol, and a compound of the general formula RO(CH 2 CH 2 O) n H wherein R is alkyl or alkenyl of from 14 to 20 carbon atoms and n is an integer of from 7 to 20.
Claim 13
Dependent Claim
33 words
Reagent as claimed in claim 10 wherein the system for the detection of glycerol comprises nicotinamideadenine-dinucleotide in reduced form, adenosine triphosphate, phosphoenol pyruvate, lactate dehydrogenase, pyruvate kinase, glycerokinase, magnesium ions and a buffer.
Claim 14
Dependent Claim
10 words
Reagent as claimed in claim 10 additionally comprising serum albumin.
Claim 15
Dependent Claim
14 words
Reagent according to claim 10 wherein the lipase used is lipase from Rhizopus arrhizus.
Claim 16
Dependent Claim
104 words
Reagent as claimed in claim 10 comprising: 0.1 to 10.0 mg./ml. lipase from Rhizopus arrihizus, 0.01 to 20.0 mg./ml. carobxylesteresterase from micro-organisms, 0.01 to 0.2 mg./ml. sodium dodecyl sulfate, 0.015 to 0.3 mg./ml. cetyl stearyl polyglycol ether containing 12 mole ethylene oxide, 1to b 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 l 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 0.012 to 0.3 M buffer solution of pH 6 to 9.
Claim 17
Dependent Claim
22 words
Reagent according to claim 11 wherein the buffer used is a 0.02 - 0.1 M phosphate buffer of pH 6.5 to 8.0.
Claim 18
Dependent Claim
15 words
Reagent according to claim 15 wherein the buffer is a 0.05 M sodium phosphate buffer.
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) | 1981-01-02 | 1981-07-04 | 1982-01-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-01-02 | 1985-07-04 | 1986-01-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-01-02 | 1989-07-04 | 1990-01-03 | $7700.00 | Window Closed / Lapsed |