Denka Seiken Co., Ltd.

Denka Company Limited

Hokkaido University

Yasuki Itoh

Noriyuki Satoh

Maiko Higuchi

Noriyuki Sato

Hitoshi Chiba

Miki Fujimura

Yuhko Hirao

Tsutomi Hirano

Tsutomu Hirano

Makoto Ikaida

Tsutomu First Dept Of Internal Medicine Hirano

A method that enables quantification of cholesterol in high-density lipoprotein 3 (HDL3) in a test sample without requiring a laborious operation is disclosed. The method for quantifying cholesterol in HDL3 comprises: Step 1 wherein phospholipase and/or sphingomyelinase is/are allowed to act on a test sample to transfer cholesterol to the outside of the reaction system; and Step 2 wherein cholesterol remaining in the reaction system is quantified. The method enables specific quantification of HDL3 cholesterol in a test sample using an automatic analyzer without requirement of a laborious operation such as ultracentrifugation or pretreatment. Further, quantification of the HDL2 cholesterol level can also be carried out by subtracting the HDL3 cholesterol level from the total HDL cholesterol level obtained by a conventional method for quantifying the total HDL cholesterol in a test sample.

Method for quantifying the amount of cholesterol in high-density lipoprotein 3

Disclosed are a method and a reagent for quantifying HDL3 in a test sample without requiring laborious operations. The method for quantifying cholesterol in high-density lipoprotein 3 comprises reacting a test sample with one or more surfactants which react specifically with high-density lipoprotein 3, and quantifying cholesterol. When one surfactant is used, the surfactant is one selected from the group consisting of polyoxyethylene polycyclic phenyl ethers having an HLB of 12.5 to 15. When two or more surfactants are used, at least one of the surfactants is at least one selected from the group consisting of polyoxyethylene polycyclic phenyl ethers, and the two or more surfactants are combined so as to provide the total HLB of 12.5 to 15 of the combined surfactants.

Method and reagent for quantifying cholesterol in high density lipoprotein 3

Disclosed is a method for quantifying HDL2 cholesterol in a test sample without requiring laborious operations. The method for quantifying cholesterol comprises allowing phospholipase to act on HDL to quantify cholesterol. Also disclosed is a method comprising: a first step of transferring cholesterols other than high-density lipoproteins in a test sample to the outside of the reaction system; and a second step of quantifying high-density lipoprotein 2 cholesterol among the high-density lipoproteins remaining in the reaction system; wherein, by performing the second step by the above method, high-density lipoprotein 2 cholesterol in the test sample can be quantified.

Method for quantifying cholesterol in high-density lipoprotein 2, and reagent kit for the method

Disclosed is the provision of a method for quantifying HDL3 in a test sample without requiring a laborious operation. The method for quantifying cholesterol in high-density lipoprotein 3 comprises allowing a surfactant(s) which specifically react(s) with a high-density lipoprotein 3 to react with a test sample and quantifying cholesterol, and the surfactant(s) is(are) at least one selected from the group consisting of polyoxyethylene polycyclic phenyl ether and polyoxyethylene styrenated phenyl ether.

Method for quantifying cholesterol in high density lipoprotein 3

Provided is a method for separately or simultaneously quantifying whole HDL-C and cholesterol in HDL subfractions: ApoE-Containing HDL-C and ApoE-deficient HDL-C. A method for enzymatically and separately quantifying cholesterol in the ApoE-deficient HDL comprising: adding a surfactant composed of a polyoxyethylene benzyl phenyl ether derivative to a test sample in a final concentration of 0.05 to 0.10%, reacting the test sample with cholesterol esterase and cholesterol oxidase, and quantifying hydrogen peroxide generated. A method for enzymatically and separately quantifying cholesterol in ApoE-Containing HDL comprising: adding a surfactant composed of a polyoxyethylene benzyl phenyl ether derivative so as to obtain a final concentration of 0.15 to 0.75%, reacting the test sample with cholesterol esterase and cholesterol oxidase, and quantifying hydrogen peroxide generated.

Method for quantifying subfraction of cholesterol (&#x2212;C) in high-density lipoprotein (HDL)

A method that enables quantification of cholesterol in high-density lipoprotein 3 (HDL3) in a test sample without requiring a laborious operation is disclosed. The method for quantifying cholesterol in HDL3 comprises reacting, with a test sample, a surfactant that specifically reacts with high-density lipoprotein 3, and quantifying cholesterol. The method enables specific quantification of HDL3 cholesterol in a test sample using an automatic analyzer without requirement of a laborious operation such as ultracentrifugation or pretreatment. Further, quantification of the HDL2 cholesterol level can also be carried out by subtracting the HDL3 cholesterol level from the total HDL cholesterol level obtained by a conventional method for quantifying the total HDL cholesterol in a test sample.

This invention provides a method for separately or simultaneously quantifying cholesterol in a total amount of HDL-C, cholesterol in an HDL subfraction of apoE-containing HDL-C, and cholesterol in an HDL subfraction of apoE-deficient HDL-C. The method comprises enzymatically and separately quantifying cholesterol in apoE-containing HDL and cholesterol in apoE-deficient HDL by adding a surfactant selected from the group consisting of a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of 0.7 to less than 1.3, a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of less than 0.7, and a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of 1.3 or more to a test sample, allowing cholesterol esterase and cholesterol oxidase to react therewith, and quantifying the hydrogen peroxide generated.

Method for assaying cholesterol in ApoE-containing HDL

A method that enables quantification of cholesterol in high-density lipoprotein 3 (HDL3) in a test sample without requiring a laborious operation is disclosed. The method for quantifying cholesterol in HDL3 having: Step 1 wherein a surfactant that reacts with lipoproteins other than high-density lipoprotein 3 is reacted with a test sample to transfer cholesterol to the outside of the reaction system; and Step 2 in which cholesterol remaining in the reaction system is quantified. The method enables specific quantification of HDL3 cholesterol in a test sample using an automatic analyzer without requirement of a laborious operation such as ultracentrifugation or pretreatment. Further, quantification of the HDL2 cholesterol level can also be carried out by subtracting the HDL3 cholesterol level from the total HDL cholesterol level obtained by a conventional method for quantifying the total HDL cholesterol in a test sample.

A method for fractional measurement of small, dense LDL, which is adaptable for an autoanalyzer, and a reagent for measurement, are provided, making it possible to conduct rapid and convenient analysis with good sensitivity without pretreatment of a specimen. The method for quantitatively determining small, dense LDL cholesterol in a sample comprises the steps of:

Method and kit for quantitatively determining small, dense LDL cholesterol

A rapid and convenient method capable of performing fractional measurement of small, dense LDLs without pretreatment of a specimen, which is adaptable for an autoanalyzer, is provided. A method for quantitatively determining small, dense LDL cholesterol is provided, which comprises adding enzymes for cholesterol measurement to a test sample in the presence of a polyoxyethylene-polyoxypropylene copolymer or a derivative thereof, causing the polyoxyethylene-polyoxypropylene copolymer or the derivative thereof to selectively act on small, dense LDLs among lipoproteins, and then measuring the amount of cholesterol generated.

Method and kit for quantitative determination for small, dense particle low density lipoproteins

An object of the present invention is to provide an excellent method for detection and diagnosis of familial combined hyperlipidemia. The present invention relates to a method for detection of familial combined hyperlipidemia by measuring the concentration of small, dense LDL cholesterol in a sample collected from a subject.

Method for detection of familial combined hyperlipidemia

A reagent for fractional measurement of small, dense LDLs without pretreatment of a specimen, which is adaptable to a rapid and convenient autoanalyzer, is provided. A method for quantitatively determining cholesterol in small, dense LDLs in a sample is further provided. The method includes first step of eliminating lipoproteins other than small, dense LDLs in a sample in the presence of cholesterol esterase and 0.05 g/L to 1.0 g/L of a surfactant that acts on the lipoproteins other than small, dense LDLs, and a second step of quantitatively determining cholesterol in small dense LDLs that remain after the first step.

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Niigata, Japan

Yasuki Itoh