Method for quantification of cholesterol in small dense low-density lipoprotein

A technology of cholesterol and cholesteryl ester, applied in the field of quantification of cholesterol in small and dense low-density lipoprotein, can solve problems such as time required

Active Publication Date: 2009-08-19
KYOWA MEDEX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is easier to operate than the ultracentrifugation method, b

Method used

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  • Method for quantification of cholesterol in small dense low-density lipoprotein
  • Method for quantification of cholesterol in small dense low-density lipoprotein
  • Method for quantification of cholesterol in small dense low-density lipoprotein

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0090] Buffer, such as MOPS (pH7.0)

[0091] Cholesterol esterases such as LPL3

[0092] Cholesterol oxidase, such as CHO-PEL

[0093] Reagents for the determination of hydrogen peroxide, such as peroxidase, N-ethyl-N-(3-methylphenyl)-N'-succinylethylenediamide (EMSE) and 4-aminoantipyrine (4- AA)

[0094] Surfactants to study

example 2

[0096] Buffer, such as MOPS (pH7.0)

[0097] Cholesterol esterases such as LPL3

[0098] Cholesterol dehydrogenases such as CHDH5

[0099] Oxidized coenzymes such as NAD

[0100] [A reagent for the determination of reduced coenzymes, such as 2-(4-iodophenyl)-3-(2,4-dinitrophenyl)-5-(2,4-disulfophenyl)-2H - tetrazole monosodium salt (WST-3)]

[0101] Surfactants to study

example 3

[0103] Buffer, such as MOPS (pH7.0)

[0104] Cholesterol esterases such as LPL3

[0105] Cholesterol dehydrogenases such as CHDH5

[0106] Oxidized coenzymes such as NAD

[0107] Reduced coenzyme oxidases such as NADH oxidase

[0108] Hydrogen peroxide assay reagents such as peroxidase, EMSE and 4-AA

[0109] Surfactants to study

[0110] In addition, each of the above-mentioned reagents may contain an aqueous medium, a stabilizer, a preservative, an interfering substance eliminating agent, a reaction accelerator, and the like, which will be described later, if necessary.

[0111] 3) Judgment method of surfactant A

[0112] Add each lipoprotein fraction as a sample to the reaction cell (2 μL), then add the reagent (0.15 mL) described in the above 2) to start the reaction, heat at 37°C for 5 minutes, and measure the absorbance of the reaction solution to form a straight line Increased time, for example, the change in absorbance of the reaction solution after 5 minutes of ...

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PUM

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Abstract

A method for determination of sdLDL-C, which comprises the steps of: (i) in a reaction solution containing a surfactant which preferentially inhibits the reaction of sdLDL-C with enzymes for cholesterol measurement such as cholesterol ester hydrolase, allowing the enzymes for cholesterol measurement to act on a sample to eliminate HDL-C, VLDL-C, CM-C and LgLDL-C in the sample; (ii) adding a reagent which causes the reaction of sdLDL-C remaining in the reaction solution after the above step (i) to form hydrogen peroxide or reduced coenzyme, and measuring the formed hydrogen peroxide or reduced coenzyme; and (iii) determining the sdLDL-C concentration in the sample from the measurement value in (ii) and a previously prepared calibration curve.

Description

technical field [0001] The present invention relates to a method for quantifying cholesterol (hereinafter abbreviated as sdLDL-C) in small dense low-density lipoprotein (hereinafter abbreviated as sdLDL) in a sample and a kit for quantification. Background technique [0002] Lipoproteins present in the blood are broadly classified into high-density lipoprotein (abbreviated as HDL hereinafter), low-density lipoprotein (abbreviated as LDL hereinafter), very low-density lipoprotein (abbreviated as VLDL hereinafter) and chyle Microparticles (hereinafter abbreviated as CM) 4 types. The lipid composition constituting each lipoprotein is also different, and the role in the living body varies greatly depending on the type of apolipoprotein constituting each lipoprotein. In addition, as a lipoprotein produced in the process of metabolism from VLDL to LDL, there is an intermediate density lipoprotein (hereinafter abbreviated as IDL) having a density intermediate between VLDL and LDL,...

Claims

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Application Information

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IPC IPC(8): C12Q1/60C12Q1/26C12Q1/32C12Q1/44G01N33/50G01N33/92
CPCG01N33/92C12Q1/60C12Q1/26C12Q1/32
Inventor 荒武知子片山有基三岛慎刚
Owner KYOWA MEDEX CO LTD
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