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Method of measuring lipid in specific lipoprotein

A lipoprotein, specific technology, applied in the direction of biological testing, material inspection products, etc., to achieve the effect of easy operation

Inactive Publication Date: 2009-02-25
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since VLDL cholesterol is estimated from neutral fat, it cannot be used by people with a neutral fat concentration exceeding 400mg / dl and patients with type III hyperlipidemia, and there is also a problem of negative effects when neutral fat is temporarily increased due to meals.

Method used

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  • Method of measuring lipid in specific lipoprotein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 (measurement of HDL cholesterol)

[0050] According to the method of the present invention, HDL cholesterol was measured with a Hitachi 7170 automatic analyzer, and its measured value was compared with that of a commercially available HDL cholesterol measuring reagent. 15 cases of human serum were used as samples. The reagent of the present invention is to add 240 μL of reagents containing 0.01% 4-aminoantipyrine and 100 mM PIPES buffer (pH 6.5) to 2.4 μL of serum, and after incubating at 37° C. for 5 minutes, add the surface active agent containing the present invention 1%, 1 unit / mL cholesterol oxidase (Oriental yeast), 1 unit / mL cholesterol esterase (Asahi Kasei), 1 unit / mL peroxidase, 0.04% N,N-disulfobutyl m-toluidine , 80 μL of the reagent of 100 mMPIPES buffer (pH 6.5), and measure the change in absorbance at 37° C. at a subwavelength of 700 nm / main wavelength of 600 nm. Using Cholestest N HDL (manufactured by Daiichi Chemical Co., Ltd.) as a comme...

Embodiment 2

[0054] Embodiment 2 (measurement of HDL cholesterol)

[0055] According to the method of the present invention, HDL cholesterol was measured with a Hitachi 7170 automatic analyzer, and its measured value was compared with that of a commercially available HDL cholesterol measuring reagent. 15 cases of human serum were used as samples. The reagent of the present invention is to add 240 μ L of reagents containing 0.01% 4-aminoantipyrine, 45 μ M digitonin (Tokyo Chemical), 100 mM PIPES buffer (pH6.5) in 2.4 μ L of serum, and incubate at 37° C. for 5 Minutes later, add surfactant containing 1% of the present invention, 1 unit / ml cholesterol oxidase (Oriental yeast), 1 unit / mL cholesterol esterase (Asahi Kasei), 1 unit / mL peroxidase, 0.04% by weight N , N-disulfobutyl m-toluidine, 100 mM PIPES buffer (pH 6.5) reagent 80 μL, at 37 ° C at a sub-wavelength of 700 nm / main wavelength of 600 nm to measure the amount of change in absorbance. Using Cholestest NHDL (manufactured by Daiichi...

Embodiment 3

[0059] Embodiment 3 (measurement of HDL cholesterol)

[0060] According to the method of the present invention, HDL cholesterol was measured with a Hitachi 7170 automatic analyzer, and its measured value was compared with that of a commercially available HDL cholesterol measuring reagent. 15 cases of human serum were used as samples. The reagent of the present invention is to add 240 μL of reagent containing 0.01% 4-aminoantipyrine, 0.04% sodium phosphotungstate (Kishida), 0.2% magnesium chloride, and 100 mM PIPES buffer (pH6.5) to 2.4 μL of serum, After incubating at 37°C for 5 minutes, add 1% surfactant containing the present invention, 1 unit / mL cholesterol oxidase (Oriental yeast), 1 unit / mL cholesterol esterase (Asahi Kasei), 1 unit / mL peroxidase , 0.04% by weight N,N-disulfobutyl-m-toluidine, 100 mM PIPES buffer (pH 6.5) reagent 80 μL, at 37 ° C at a sub-wavelength of 700 nm / main wavelength of 600 nm to measure the amount of change in absorbance. Using Cholestest N HDL...

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Abstract

The invention relates to a method for measuring a lipid in a specific lipoprotein in characterized by using a polycyclic polyoxyalkylene derivative at least in a step of determining the specificity of the measurement of a target lipid, wherein the target lipid is cholesterol in LDL, and the polycyclic polyoxyalkylene derivative is preferably a nonionic surfactant which acts preferentially on lipoproteins other than LDL and has an HLB of 15.1 to 18.

Description

[0001] This application is the PCT international application number PCT / JP2003 / 015080, the international application date is November 26, 2003, and the national application number is 200380109195.3 after the PCT international application entered the Chinese phase. The title is "Lipid determination method in specific lipoproteins" divisional application of the Chinese patent application. technical field [0002] The present invention relates to a method and a reagent for efficiently separating and quantifying lipids present in a specific site using a small amount of sample and through simple operations. Background technique [0003] Cholesterol, neutral fat, and phospholipids combine with apoproteins in plasma to form lipoproteins. Lipoproteins can be classified into chylomicrons, very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) according to different physical properties. LDL is sometimes subdivided into intermediate-dens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/92
Inventor 山本祥子山本光章中西一夫斋藤和典
Owner SEKISUI MEDICAL CO LTD