Patent US5392775A successfully refreshed from Google Patents.
US4259440A
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Hydrolysis and assay of triglycerides
Assignee: Bayer Corp
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Filing: 1979-05-21
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Granted: 1981-03-31
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Expires: 1999-05-21
Abstract
Abstract A method and composition for the hydrolysis and assay of triglycerides are disclosed. The method includes the steps of adding lipase and cholesterol esterase to a triglyceride in combination with a glycerol assay system and determining the amount of triglycerides present based on the amount of glycerol produced. The composition includes a mixture of lipase, cholesterol esterase and a glycerol assay system.
Inventors
- 👤 Surendra K. Gupta
- 👤 Panna R. Chaudhari
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 22:43
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/61; C12Q1/34; C12Q1/44; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/81
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
55 words
A method for determining the amount of triglycerides present in an aqueous fluid which comprises contacting the fluid with a mixture of a lipase and cholesterol esterase for a time sufficient to hydrolyze the triglyceride to glycerol and free fatty acids and determining the amount of triglycerides present based on the amount of glycerol produced.
Claim 2
Independent Claim
55 words
A method for determining the amount of triglycerides present in an aqueous fluid which comprises contacting the fluid with a mixture of a lipase and cholesterol esterase for a time sufficient to hydrolyze the triglyceride to glycerol and free fatty acids and determining the amount of triglycerides present based on the amount of glycerol produced.
Claim 3
Dependent Claim
26 words
A method as claimed in claim 1 wherein the lipase is produced by a microorganism selected from the group consisting of Rhizopus delemar, and Chromobacterium viscosum.
Claim 4
Dependent Claim
22 words
A method as claimed in claim 1 wherein the cholesterol esterase is obtained from the microorganism Pseudomonas aeruginosa or from beef pancreas.
Claim 5
Dependent Claim
43 words
A method as claimed in claim 1 wherein the lipase is produced from a microorganism selected from the group consisting of Rhizopus delemar, Rhizopus arrhizus and Chromobacterium viscosum and the cholesterol esterase is obtained from the microorganism Pseudomonas aeruginosa or from beef pancreas.
Claim 6
Dependent Claim
16 words
A method as claimed in claim 1 wherein the amount of glycerol present is determined enzymatically.
Claim 7
Dependent Claim
30 words
A method as claimed in claim 1 wherein the hydrolysis is carried out at a pH of from 6 to 9 and a temperature of from 20° to 40° C.
Claim 8
Dependent Claim
20 words
A method as claimed in claim 1 wherein a buffer is added to the mixture of lipase and cholesterol esterase.
Claim 9
Dependent Claim
20 words
A method as claimed in claim 1 wherein a surfactant is added to the mixture of lipase and cholesterol esterase.
Claim 10
Dependent Claim
14 words
A method as claimed in claim 1 wherein the fluid is a body fluid.
Claim 11
Dependent Claim
15 words
A method as claimed in claim 9 wherein the body fluid is serum or plasma.
Claim 12
Dependent Claim
22 words
A method as claimed in claim 1 wherein there is present from 0.01 to 5.0 U cholesterol esterase per 10 U lipase.
Claim 13
Dependent Claim
22 words
A method as claimed in claim 1 where there is present from 0.35 to 1.8 U cholesterol esterase per 10 U lipase.
Claim 14
Dependent Claim
43 words
A method as claimed in claim 1 wherein the amount of glycerol present is determined by contacting the liberated glycerol with a glycerol assay system comprising: adenosine triphosphate; phosphoenolpyruvate; pyruvate kinase; lactate dehydrogenase; reduced nicotinamide adenine dinucleotide; glycerol kinase and a magnesium salt.
Claim 15
Dependent Claim
40 words
A method as claimed in claim 1 wherein the amount of glycerol present is determined by contacting the liberated glycerol with a glycerol assay system comprising: adenosine triphosphate; glycerol kinase; α-glycerolphosphate dehydrogenase; oxidized nicotinamide adenine dinucleotide; diaphorase and a dye.
Claim 16
Dependent Claim
22 words
A composition for the determination of triglycerides by enzymatic hydrolysis comprising a mixture of lipase, cholesterol esterase and a glycerol assay system.
Claim 17
Dependent Claim
22 words
A composition for the determination of triglycerides by enzymatic hydrolysis comprising a mixture of lipase, cholesterol esterase and a glycerol assay system.
Claim 18
Dependent Claim
13 words
A composition as claimed in claim 15 wherein there is present a buffer.
Claim 19
Dependent Claim
13 words
A composition as claimed in claim 15 wherein there is present a surfactant.
Claim 20
Dependent Claim
29 words
A composition as claimed in claim 15 wherein the assay system comprises: adenosine triphosphate; phosphoenolpyruvate; pyruvate kinase; lactate dehydrogenase; reduced nicotinamide adenine dinucleotide; glycerol kinase and a magnesium salt.
Claim 21
Dependent Claim
27 words
A composition as claimed in claim 15 wherein the assay system comprises: adenosine triphosphate; glycerol kinase; α-glycerol phosphate dehydrogenase; oxidized nicotinamide adenine dinucleotide; diaphorase and a dye.
Claim 22
Dependent Claim
25 words
A test device for the determination of triglycerides in a fluid which comprises a carrier incorporated with lipase, cholesterol esterase and a glycerol assay system.
Claim 23
Dependent Claim
25 words
A test device for the determination of triglycerides in a fluid which comprises a carrier incorporated with lipase, cholesterol esterase and a glycerol assay system.
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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) | 1984-03-30 | 1984-09-29 | 1985-03-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-03-30 | 1988-09-29 | 1989-03-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-03-30 | 1992-09-29 | 1993-03-31 | $7700.00 | Window Closed / Lapsed |