Integral element for the analysis of cholesterol
First Claim
1. An integral element for analysis of total cholesterol in a liquid, the element I comprising a spreading layer in fluid contact with a reagent layer and II containinga. a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease;
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the cholesterol ester hydrolyzing composition and cholesterol oxidase being disposed within the element such that, in liquid applied to the element, cholesterol esters are saponified and cholesterol is depomposed producing a detectable change related to the total cholesterol content of the liquid.
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Abstract
An integral analytical element for analysis of liquids for their cholesterol content is disclosed. The element is of the type which comprises at least two superposed layers including a spreading layer and a reagent layer in fluid contact and, optionally, a support. Cholesterol oxidase and a composition for the hydrolysis of cholesterol esters comprising lipase having cholesterol esterase activity and protease are included in the element such that cholesterol esters contained in a sample applied to the spreading layer are saponified to free cholesterol and free cholesterol is decomposed in the presence of cholesterol oxidase to produce a detectable change related to the total cholesterol content of the sample.
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Citations
69 Claims
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1. An integral element for analysis of total cholesterol in a liquid, the element I comprising a spreading layer in fluid contact with a reagent layer and II containing
a. a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease; - and
b. cholesterol oxidase; the cholesterol ester hydrolyzing composition and cholesterol oxidase being disposed within the element such that, in liquid applied to the element, cholesterol esters are saponified and cholesterol is depomposed producing a detectable change related to the total cholesterol content of the liquid.
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2. An integral element for analysis of total cholesterol in a liquid, the element I comprising a spreading layer in fluid contact with a reagent layer and II containing
a. a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease; - and
b. cholesterol oxidase; the cholesterol ester hydrolyzing composition and cholesterol oxidase being disposed within the element such that, in liquid applied to the element, cholesterol esters are saponified and cholesterol is decomposed producing a detectable change related to the total cholesterol content of the liquid, the spreading layer being capable of spreading within itself a substance selected from the group consisting of cholesterol, cholesterol esters or decomposition products of cholesterol to provide a uniform concentration of such substance at the surface of the spreading layer facing the reagent layer, and the reagent layer being of substantially uniform permeability to such substance or a reaction product of such substance. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact:
1. a spreading layer containing a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease; and - View Dependent Claims (18, 19, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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17. a reagent layer interposed between the spreading layer and the support and containing cholesterol oxidase and an indicator composition that can interact with a decomposition product of cholesterol to produce in the element a detectable change related to the total cholesterol content of the liquid.
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20. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact:
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1. a spreading layer containing a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease; and 2. a reagent layer interposed between the spreading layer and the support and containing cholesterol oxidase and an indicator composition that can interact with a decomposition product of cholesterol to produce in the element a detectable change related to the total cholesterol content of the liquid the spreading layer being capable of spreading within itself a substance selected from the group consisting of cholesterol, cholesterol esters or decomposition products of cholesterol to provide a uniform concentration of such substance at the surface of the spreading layer facing the reagent layer, and the reagent layer being of substantially uniform permeability to such substance or a reaction product of such substance.
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34. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact;
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1. a spreading layer buffered to a pH in the range of between about 5.5 and 9.0 and containing (a) from about 90,000 to about 270,000 U/m2 of a microbial lipase having cholesterol esterase activity and (b) from about 36,000 to about 105,000 U/m2 of protease; and 2. a reagent layer containing (a) from about 100 to about 5000 U/m2 of cholesterol oxidase derived from a microorganism selected from the group consisting of NRRL 5767 and NRRL 5768 and (b) an indicator composition comprising a substance having peroxidative activity and a composition that reacts to produce a color change in the presence of hydrogen peroxide and the substance having peroxidative activity, the reagent layer being interposed between the spreading layer and the support and buffered at a pH in the range between about 5.5 and 8.5. - View Dependent Claims (36)
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35. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact;
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1. a spreading layer containing (a) a cholesterol ester hydrolyzing composition comprising a lipase having cholesterol esterase activity and protease and (b) cholesterol oxidase; and 2. a reagent layer interposed between the spreading layer and the support and containing an indicator composition that can interact with a decomposition product of cholesterol to produce in the element a detectable change related to the total cholesterol content of the liquid. - View Dependent Claims (37, 39, 41, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 63, 64, 66, 67, 68, 69)
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38. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact;
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1. a spreading layer containing (a) a cholesterol ester hydrolyzing composition comprising a lipase having cholesterol esterase activity and protease and (b) cholesterol oxidase; and 2. a reagent layer interposed between the spreading layer and the support and containing an indicator composition that can interact with a decomposition product of cholesterol to produce in the element a detectable change related to the total cholesterol content of the liquid the spreading layer being capable of spreading within itself a substance selected from the group consisting of cholesterol, cholesterol esters or decomposition products of cholesterol to provide a uniform concentration of such substance at the surface of the spreading layer facing the reagent layer, and the reagent layer being of substantially uniform permeability to such substance or a reaction product of such substance. - View Dependent Claims (40, 42)
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60. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact:
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1. a spreading layer buffered to a pH of from about 5.5 and 8.5 and containing (a) from about 90,000 to about 270,000 U/m2 of a microbial lipase having cholesterol esterase activity, (b) from about 36,000 to about 105,000 U/m2 of protease and (c) from about 100 to about 5000 U/m2 of cholesterol oxidase derived from a microorganism selected from the group consisting of NRRL 5767 and NRRL 5768; and 2. a. reagent layer interposed between the support and the spreading layer containing an indicator composition comprising a substance having peroxidative activity and a composition to produce a color change in the presence of hydrogen peroxide and the substance having peroxidative activity.
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61. An integral element for analysis of total cholesterol in a liquid, the element comprising a support upon which are superposed in fluid contact:
1. a spreading layer containing a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease;
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62. a reagent layer interposed between the spreading layer and the support and containing cholesterol oxidase and an indicator composition that can interact with a decomposition product of cholesterol to produce in the element a detectable change quantitatively related to the total cholesterol content of the liquid;
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3. interposed between the spreading layer and the reagent layer a barrier layer which inhibits passage of protease to the reagent layer.
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65. An integral element for analysis of total cholesterol in a liquid, the element comprising in fluid contact:
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A. a spreading layer comprising blushed cellulose acetate; and B. a reagent layer comprising a hydrophilic colloid the element containing 1. a cholesterol ester hydrolyzing composition comprising lipase having cholesterol esterase activity and protease; and 2. cholesterol oxidase the cholesterol ester hydrolyzing composition and cholesterol oxidase being disposed within the element such that, in liquid applied to the element, cholesterol esters are saponified and cholesterol is decomposed producing a detectable change related to the total cholesterol content of the liquid.
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Specification