EP0091026A2
Expired / Lapsed
EP
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Improved time-stable liquid cholesterol assay compositions
Assignee: Individual
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Filing: 1983-03-24
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Granted: —
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Expires: 2003-03-24
Abstract
Abstract There is provided a stable cholesterol assay composition which comprises an aqueous solution of at least one bile acid or salt thereof being present in an amount of up to about 5mM; a nonionic surfactant present in an amount of from about 0.15 to about 1.5 percent volume by volume; a buffer in a concentration of from 0 to about 65 mM; and cholesterol oxidase in a concentration of at least about 0.2 KIU/I. Solution pH is from about 5.5 to about 8.5. Addition of cholesterol esterase, phenol, peroxidase and 4-aminoantipyrine provides a total cholesterol chromogen system.
Inventors
- 👤 Ivan Endre Modrovich
- 👤 Karen Rae Caris
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 03:13
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/60
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
108 words
A stable cholesterol assay composition which comprises an aqueous solution of: a) at least one acidic compound selected from the group consisting of a bile acid and a salt of a bile acid, the total of said acidic compound being present in an amount of up to about 5mM; b) a nonionic surfactant present in a concentration of from about 0.15 to about 1.5 percent volume by volume; c) a buffer in a concentration of from 0 to about 65 mM; and d) cholesterol oxidase in a concentration of at least about 0.02 KIU/l, said cholesterol assay solution having a pH of from about 5.5 to about 8.5.
Claim 2
Dependent Claim
108 words
A stable cholesterol assay composition which comprises an aqueous solution of: a) at least one acidic compound selected from the group consisting of a bile acid and a salt of a bile acid, the total of said acidic compound being present in an amount of up to about 5mM; b) a nonionic surfactant present in a concentration of from about 0.15 to about 1.5 percent volume by volume; c) a buffer in a concentration of from 0 to about 65 mM; and d) cholesterol oxidase in a concentration of at least about 0.02 KIU/l, said cholesterol assay solution having a pH of from about 5.5 to about 8.5.
Claim 3
Dependent Claim
25 words
A stable cholesterol assay composition as claimed in claim 1 which includes microbial cholesterol esterase present in a concentration of at least about 0.07 KIU/1.
Claim 4
Dependent Claim
22 words
A stable cholesterol assay composition as claimed in claim 1 or 2 which includes a chromogen system for determination of hydrogen peroxide.
Claim 5
Dependent Claim
51 words
A stable cholesterol assay system as claimed in claim 3 in which the chromogen system comprises phenol in a concentration of from about 8 to about 35 mM, and peroxidase and 4-aminoantipyrine in a concentration sufficient to provide a colormetric quantitative determination of the hydrogen peroxide formed from oxidation of cholesterol.
Claim 6
Dependent Claim
59 words
A stable cholesterol assay system as claimed in claim 1 or 2 which includes a chromogen system for determining hydrogen peroxide and which comprises phenol in a concentration of from about 8 to about 35 mM, 4-aminoantipyrine in a concentration of from about 0.25 to about 0.35 mM, and peroxidase in a concentration of at least about 30 KIU/1.
Claim 7
Dependent Claim
18 words
A stable cholesterol assay composition as claimed in any one of the previous claims which includes a bacteriocide.
Claim 8
Dependent Claim
23 words
A stable cholesterol assay composition as claimed in claim 6 which includes 2,4 dichlorophenol in a concentration of up to about 1 mM.
Claim 9
Dependent Claim
24 words
A stable cholesterol assay composition as claimed in any one of the previous claims in which the nonionic surfactant is polyethylene glycol p-isooctylphenyl ether.
Claim 10
Dependent Claim
22 words
A stable cholesterol assay composition as claimed in any one of the previous claims in which the buffer is potassium dihydrogen phosphate.
Claim 11
Dependent Claim
26 words
A stable cholesterol assay composition as claimed in any one of the previous claims in which the acidic compound is a metal salt of cholic acid.
Claim 12
Dependent Claim
141 words
A stable total cholesterol chromogen assay composition comprising an aqueous solution having a pH of from about 6.5 to about 8.5 and comprising: a) phenol in a concentration of from about 8 to about 35 mM; b) a metal salt of cholic acid present in a concentration of up to about 5 mM; c) a nonionic surfactant present in a concentration of from about 0.2 to about 1.5 percent volume by volume; d) a buffer present in a concentration of from 0 to about 65 mM; e) 4-aminoantipyrine; f) microbial cholesterol esterase present in a concentration of at least about 0.07 KIU/1; g) cholesterol oxidase present in a concentration of at least about 0.02 KIU/I; and h) peroxidase, the amount of peroxidase and 4-aminoantipyrine being sufficient to enable quantitative determination of the amount of hydrogen peroxide formed from oxidation of cholesterol.
Claim 13
Dependent Claim
141 words
A stable total cholesterol chromogen assay composition comprising an aqueous solution having a pH of from about 6.5 to about 8.5 and comprising: a) phenol in a concentration of from about 8 to about 35 mM; b) a metal salt of cholic acid present in a concentration of up to about 5 mM; c) a nonionic surfactant present in a concentration of from about 0.2 to about 1.5 percent volume by volume; d) a buffer present in a concentration of from 0 to about 65 mM; e) 4-aminoantipyrine; f) microbial cholesterol esterase present in a concentration of at least about 0.07 KIU/1; g) cholesterol oxidase present in a concentration of at least about 0.02 KIU/I; and h) peroxidase, the amount of peroxidase and 4-aminoantipyrine being sufficient to enable quantitative determination of the amount of hydrogen peroxide formed from oxidation of cholesterol.
Claim 14
Dependent Claim
29 words
A stable total cholesterol chromogen assay composition as claimed in claim 11 in which the buffer is potassium dihydrogen phosphate and pH is from about 6 to about 7.5.
Claim 15
Dependent Claim
41 words
A stable total cholesterol chromogen assay composition as claimed in claim 11 or 12 in which the nonionic surfactant is present in a concentration of from about 0.2 to about 0.4 percent volume by volume and is polyethylene glycol p-isooctylphenyl ether.
Claim 16
Dependent Claim
43 words
A stable total cholesterol chromogen assay composition as claimed in any one of the previous claims in which peroxidase is present in a concentration of at least about 30 KIU/1 and in which 4-aminoantipyrine is present in a concentration of about 0.3 mM.
Claim 17
Dependent Claim
147 words
A stable total cholesterol chromogen assay composition comprising an aqueous solution of: a) phenol in a concentration of about 17 mM; b) 2,4 dichlorophenol present in a concentration of about 0.5 mM; c) a metal salt of cholic acid present in a concentration of up to about 5 mM; d) polyethylene glycol p-isooctylphenyl ether present in a concentration of from about 0.2 to about 0.6 percent volume by volume; e) KH 2 P0 4 present in a concentration of about 12.5 mM; f) peroxidase present in a concentration of about 30 KIU/1; g) cholesterol oxidase present in a concentration of at least about 0.05 KIU/l; h) microbial cholesterol esterase present in a concentration of at least about 0.1 KIU/l; and i) 4-aminoantipyrene present in a concentration of about 0.3 mM, said stable total cholesterol chromogen assay composition having a pH of from about 6.0 to about 7.5.
Claim 18
Dependent Claim
147 words
A stable total cholesterol chromogen assay composition comprising an aqueous solution of: a) phenol in a concentration of about 17 mM; b) 2,4 dichlorophenol present in a concentration of about 0.5 mM; c) a metal salt of cholic acid present in a concentration of up to about 5 mM; d) polyethylene glycol p-isooctylphenyl ether present in a concentration of from about 0.2 to about 0.6 percent volume by volume; e) KH 2 P0 4 present in a concentration of about 12.5 mM; f) peroxidase present in a concentration of about 30 KIU/1; g) cholesterol oxidase present in a concentration of at least about 0.05 KIU/l; h) microbial cholesterol esterase present in a concentration of at least about 0.1 KIU/l; and i) 4-aminoantipyrene present in a concentration of about 0.3 mM, said stable total cholesterol chromogen assay composition having a pH of from about 6.0 to about 7.5.
Claim 19
Independent Claim
176 words
A method for preparing a stable cholesterol assay solution which comprises forming a base solution and an enzyme solution, then combining the base solution and the enzyme solution to form a net solution, and in which: a) the base solution is formed by dissolving in water: i) at least one acidic compound selected from the group consisting of a bile acid and a salt of a bile acid to provide the total of said acidic compound in the net solution in an amount of up to about 5 mM; ii) a nonionic surfactant determined to provide an amount in the net solution a nonionic surfactant concentration of from about 0.15 to about 1.5 percent volume by volume; b) the enzyme solution being formed by dissolving in water containing a portion of the total nonionic surfactant, cholesterol oxidase to provide in the net solution cholesterol oxidase in a concentration of at least about 0.02 KIU/l, the base solution being adjusted if required to provide a net solution having a pH of from about 5.5 to about 8.5.
Claim 20
Independent Claim
176 words
A method for preparing a stable cholesterol assay solution which comprises forming a base solution and an enzyme solution, then combining the base solution and the enzyme solution to form a net solution, and in which: a) the base solution is formed by dissolving in water: i) at least one acidic compound selected from the group consisting of a bile acid and a salt of a bile acid to provide the total of said acidic compound in the net solution in an amount of up to about 5 mM; ii) a nonionic surfactant determined to provide an amount in the net solution a nonionic surfactant concentration of from about 0.15 to about 1.5 percent volume by volume; b) the enzyme solution being formed by dissolving in water containing a portion of the total nonionic surfactant, cholesterol oxidase to provide in the net solution cholesterol oxidase in a concentration of at least about 0.02 KIU/l, the base solution being adjusted if required to provide a net solution having a pH of from about 5.5 to about 8.5.
Claim 21
Dependent Claim
39 words
A method as claimed in claim 16 in which there is added to the enzyme solution microbial cholesterol esterase in an amount to provide in the net solution cholesterol esterase in a concentration of at least about 0.07 KIU/1.
Claim 22
Dependent Claim
82 words
A method as claimed in claim 16 or 17 in which there is added to the base solution phenol in an amount sufficient to provide in the net solution phenol in a concentration of from about 8 to about 15 mM, and 4-aminoantipyrine, and in which there is added to the enzyme solution peroxidase, the amount of peroxidase and 4-aminoantipyrine being sufficient in the net solution to enable quantitative colormetric determination of the amount of hydrogen peroxide formed from oxidation of cholesterol.
References Cited (Backward Citations)
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