US4056442A
Expired / Lapsed
US
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Lipase composition for glycerol ester determination
Assignee: Dow Chemical Co
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Filing: 1976-06-01
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Granted: 1977-11-01
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Expires: 1996-06-01
Abstract
Abstract A synergistic composition of microbial lipases from Rhizopus arrhizirs and Candida cylindracea is used for the hydrolysis of glycerol esters such as triglycerides in the determination of glycerol esters present in an aqueous medium such as body fluid.
Inventors
- 👤 Charles Y. Huang
- 👤 Alejo V. Roy
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 20:15
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/16; C12N9/18; C12N9/20; C12Q; C12Q1/00; C12Q1/34; C12Q1/44; C12Q1/48; C12Q1/61; C12Q2326/00; C12Q2326/90; C12Q2326/92; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/902; G01N2333/904; G01N2333/91; G01N2333/912; G01N2333/91205; G01N2333/9121; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/911; Y10S435/921; Y10S435/939
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
44 words
A composition useful for the hydrolysis of a glycerol ester in an aqueous medium comprising a mixture of from 15 to 95 units of Rhizopus arrhizus lipase and from 5 to 85 units of Candida cylindracea lipase per 100 units of total lipase present.
Claim 2
Dependent Claim
44 words
A composition useful for the hydrolysis of a glycerol ester in an aqueous medium comprising a mixture of from 15 to 95 units of Rhizopus arrhizus lipase and from 5 to 85 units of Candida cylindracea lipase per 100 units of total lipase present.
Claim 3
Dependent Claim
37 words
The composition of claim 1 wherein the composition contains from about 35 to 80 units of Rhizopus arrhizus lipase and from about 20 to 65 units of Candida cylindracea lipase per 100 units of total lipase present.
Claim 4
Dependent Claim
18 words
The composition of claim 2 wherein the ratio of Candida cylindracea lipase to Rhizopus arrhizus lipase is 0.9.
Claim 5
Dependent Claim
84 words
A reagent composition useful for the determination of triglyceride concentration in serum by measuring the light absorbance of the serum following hydrolysis of the triglycerides to glycerol and fatty acids which comprises an enzyme mixture having from 15 to 95 units of Rhizopus arrhizus lipase and from 5 to 85 units of Candida cylindracea lipase per 100 units of total lipase and chemical means for measurably changing the spectroscopic absorbance of the composition in proportion to the amount of glycerol released in the serum.
Claim 6
Dependent Claim
84 words
A reagent composition useful for the determination of triglyceride concentration in serum by measuring the light absorbance of the serum following hydrolysis of the triglycerides to glycerol and fatty acids which comprises an enzyme mixture having from 15 to 95 units of Rhizopus arrhizus lipase and from 5 to 85 units of Candida cylindracea lipase per 100 units of total lipase and chemical means for measurably changing the spectroscopic absorbance of the composition in proportion to the amount of glycerol released in the serum.
Claim 7
Dependent Claim
14 words
The reagent composition of claim 4 which contains adenosine-5-triphosphate, glycerokinase, phosphoenol pyruvate, and nicotinamide-adenine-dinucleotide.
Claim 8
Dependent Claim
18 words
The reagent composition of claim 4 which contains adenosine-5'-triphosphate, nicotinamide-adenine-dinucleotide, a tetrazolium salt, glycerokinase, α-glycerophosphate dehydrogenase, and diaphorase.
Claim 9
Dependent Claim
6 words
The lyophilized composition of claim 6.
Claim 10
Independent Claim
89 words
A method for hydrolyzing a glycerol ester and for determining the amount of glycerol ester present in an aqueous medium which comprises incubating the glycerol ester at a pH of 5.5-9.0 with a lipase mixture having the proportions of 15 to 95 units of a lipase from Rhizopus arrhizus and 5 to 85 units of a lipase from Candida cylindracea per 100 units of total lipase present for a time sufficient to hydrolyze the glycerol ester and determining the amount of glycerol or fatty acids released by the hydrolysis.
Claim 11
Independent Claim
89 words
A method for hydrolyzing a glycerol ester and for determining the amount of glycerol ester present in an aqueous medium which comprises incubating the glycerol ester at a pH of 5.5-9.0 with a lipase mixture having the proportions of 15 to 95 units of a lipase from Rhizopus arrhizus and 5 to 85 units of a lipase from Candida cylindracea per 100 units of total lipase present for a time sufficient to hydrolyze the glycerol ester and determining the amount of glycerol or fatty acids released by the hydrolysis.
Claim 12
Dependent Claim
13 words
The method of claim 8 wherein the aqueous medium is a body fluid.
Claim 13
Dependent Claim
11 words
The method of claim 9 wherein the body fluid is serum.
Claim 14
Dependent Claim
16 words
The method of claim 10 further comprising determining the amount of glycerol released by the hydrolysis.
Claim 15
Dependent Claim
55 words
The method of claim 11 wherein the gycerol is determined by reacting the glycerol in the serum with adenosine-5-triphosphate in the presence of glycerokinase to form glycerol-1-phosphate, reacting the glycerol-1-phosphate with nicotinamide-adenine-dinucleotide in the presence of α-glycerophosphate dehydrogenase to form reduced nicotinamide-adenine-dinucleotide, reducing a tetrazolium salt with the reduced nicotinamide-adenine-dinucleotide, and measuring the color produced.
Claim 16
Dependent Claim
12 words
The method of claim 12 wherein the tetrazolium salt is 2-p-indophenyl-3-p-nitrophenyl-5-phenyltetrazolium chloride.
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) | 1980-10-31 | 1981-05-02 | 1981-11-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1984-10-31 | 1985-05-02 | 1985-11-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1988-10-31 | 1989-05-02 | 1989-11-01 | $7700.00 | Window Closed / Lapsed |