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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

AI Technical Summary

Problems solved by technology

This method is easier to operate than the ultracentrifugation method, but it takes time to measure because fractional sedimentation is required

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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