Multilayer analytical elements for use in the assay of...

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150/15.1, 150/15

G01N 33/52 (2006.01)

Patent

CA 1056282

Abstract of the Disclosure An analytical element for use in the quantitative assay of complex fluids for cholesterol content comprising a support, a reagent layer comprising a substance having cholesterol oxi- dase activity dispersed in a suitable binder coated on one side of the support, and a porous medium comprised of one or more layers which serve to spread the sample in a manner providing a substantially constant volume of sample per unit area to the reagent layer in spite of variation in the volume of sample applied to the element coated over the reagent layer. According to a preferred embodiment, the reagent layer also contains an indicator system which reacts with one or more products of the cholesterol oxidase induced decomposition of cholesterol to produce a spectrophotometrically quantizable energy absorption shift. According to a further preferred embodiment, the indicator system comprises a substance having peroxidative activity and a composition which undergoes an energy absorption shift, pre- ferably a color change, in the presence of hydrogen peroxide and said substance having peroxidative activity. Another preferred embodiment provides for the inclusion of a cholesterol ester hydrolyzing system comprising a lipase having cholesterol esterase activity and a protease in either the porous medium or the reagent layer to hydrolyze cholesterol esters which may be present in the complex solution under analysis. In a highly preferred embodiment, the enzymatic cho- lesterol ester hydrolyzing system and the cholesterol oxidase are contained in the porous medium which is separated from the reagent layer containing the indicator system by a hydrophilic barrier layer.

220407

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