Method, reagent and kit for determination of cholesterol in remnant-like particles (rlp)
AssigneeKyowa Medex Co., Ltd.
InventorsKazuhito Miyauchi, Mayumi Fujinaka
Life of this patent
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Abstract
The present invention provides a method, a reagent and a kit for simple and sensitive determination of cholesterol in remnant-like particles without separation of components of a sample. A method for quantitatively determining remnant-like particle cholesterol in a sample, which comprises: in an aqueous medium containing the sample and in the presence of a combination of specific surfactants and a phospholipid-hydrolyzing enzyme, allowing remnant-like particle cholesterol in the sample to react with cholesterol esterase and cholesterol oxidase or cholesterol dehydrogenase (in the presence of oxidized coenzyme); and determining the formed hydrogen peroxide or reduced coenzyme.
Claims24
A method for quantitatively determining remnant-like particle cholesterol in a sample, which comprises:
- in an aqueous medium containing the sample and in the presence of (a)(i) a combination of one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxybutylene copolymer, a polyoxyethylene styrenated-phenyl ether and a polyoxyethylene-polyoxypropylene long-chain branched alkyl ether [surfactant (d1)] and one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxypropylene alkylaryl ether, a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d2)] or (ii) a combination of a polyoxyethylene-polyoxypropylene copolymer [surfactant (d3)] and one or two kinds of surfactants selected from the group consisting of a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d4)] and (b) a phospholipid-hydrolyzing enzyme, allowing (c) cholesterol esterase and cholesterol oxidase or (d) in the presence of oxidized coenzyme, cholesterol esterase and cholesterol dehydrogenase to act on remnant-like particle cholesterol in the sample to form hydrogen peroxide or reduced coenzyme;
- and determining the formed hydrogen peroxide or reduced coenzyme.
A reagent for the quantitative determination of remnant-like particle cholesterol in a sample comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase, cholesterol oxidase, and (i) a combination of one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxybutylene copolymer, a polyoxyethylene styrenated-phenyl ether and a polyoxyethylene-polyoxypropylene long-chain branched alkyl ether [surfactant (d1)] and one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxypropylene alkylaryl ether, a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d2)] or (ii) a combination of polyoxyethylene-polyoxypropylene copolymer [surfactant (d3)] and one or two kinds of surfactants selected from the group consisting of a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d4)].
A reagent for the quantitative determination of remnant-like particle cholesterol in a sample comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase, cholesterol dehydrogenase, oxidized coenzyme, and (i) a combination of one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxybutylene copolymer, a polyoxyethylene styrenated-phenyl ether and a polyoxyethylene-polyoxypropylene long-chain branched alkyl ether [surfactant (d1)] and one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxypropylene alkylaryl ether, a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d2)] or (ii) a combination of polyoxyethylene-polyoxypropylene copolymer [surfactant (d3)] and one or two kinds of surfactants selected from the group consisting of a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d4)].
A kit for the quantitative determination of remnant-like particle cholesterol in a sample which comprises a first reagent comprising a combination of one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxybutylene copolymer, a polyoxyethylene styrenated-phenyl ether and a polyoxyethylene-polyoxypropylene long-chain branched alkyl ether [surfactant (d1)] and a second reagent comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase and cholesterol oxidase, said kit further comprising one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxypropylene alkylaryl ether, a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d2)] in at least one of the first reagent and the second reagent.
A kit for the quantitative determination of remnant-like particle cholesterol in a sample which comprises a first reagent comprising polyoxyethylene-polyoxypropylene copolymer [surfactant (d3)] and a second reagent comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase and cholesterol oxidase, said kit further comprising one or two kinds of surfactants selected from the group consisting of a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d4)] in at least one of the first reagent and the second reagent.
A kit for the quantitative determination of remnant-like particle cholesterol in a sample which comprises a first reagent comprising a combination of one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxybutylene copolymer, a polyoxyethylene styrenated-phenyl ether and a polyoxyethylene-polyoxypropylene long-chain branched alkyl ether [surfactant (d1)] and a second reagent comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase and cholesterol dehydrogenase, said kit further comprising one or more kinds of surfactants selected from the group consisting of a polyoxyethylene-polyoxypropylene alkylaryl ether, a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d2)] in at least one of the first reagent and the second reagent, and further comprising oxidized coenzyme in at least one of the first reagent and the second reagent.
A kit for the quantitative determination of remnant-like particle cholesterol in a sample which comprises a first reagent comprising polyoxyethylene-polyoxypropylene copolymer [surfactant (d3)] and a second reagent comprising a phospholipid-hydrolyzing enzyme, cholesterol esterase and cholesterol dehydrogenase, said kit further comprising one or two kinds of surfactants selected from the group consisting of a polyoxyethylene alkylaryl ether, a polyoxyethylene-polyoxypropylene alkyl ether and a polyoxyethylene-polyoxypropylene short-chain branched alkyl ether [surfactant (d4)] in at least one of the first reagent and the second reagent, and further comprising oxidized coenzyme in at least one of the first reagent and the second reagent.
Dependencies are inferred from the claim text.