Method for quantitating triglycerides in lipoproteins

a technology of lipids and triglycerides, applied in the field of lipid quantitation methods, to achieve the effect of convenient quantitation of triglycerides

Inactive Publication Date: 2005-02-24
KYOWA MEDEX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0005] The purpose of the present invention is to provide a method for conveniently quantitating triglycerides (TGs) contained in various lipoproteins.

Problems solved by technology

This method, however, is very troublesome because it requires a great deal of time and effort for ultracentrifugation.

Method used

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  • Method for quantitating triglycerides in lipoproteins
  • Method for quantitating triglycerides in lipoproteins
  • Method for quantitating triglycerides in lipoproteins

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0058]

Measurement of TG in LDLReagent 1 (pH 6.25)Buffer [Piperazine-1,4-bis(2-50mMethanesulfonic acid)(PIPES)]TOOS (Dojindo Laboratories)0.3g / LATP 2Na salt (Wako Pure Chemical2.5g / LIndustries, Ltd.)Ascorbic acid oxidase3kU / L(Asahi Kasei Corporation)GK (Toyobo Co., Ltd.)1kU / LGPO (Asahi Kasei Corporation)8kU / LPeroxidase (Toyobo Co., Ltd.)20kU / LPEG modified LPL1.5kU / L(Toyobo Co., Ltd.)LPL III (Amano)60kU / LNonion NS-230 (NOF Coporation)0.1%Magnesium sulfate heptahydrate0.5g / L(Wako Pure ChemicalIndustries, Ltd.)Reagent 2 (pH 6.25)Buffer (PIPES)50mMEmulgen 7090.6%Triton DF-160.3%4-Aminoantipyrine0.5g / L(Wako Pure ChemicalIndustries, Ltd.)

[0059] The specificity was confirmed by tracing the time course under the condition of the following parameters using a Hitachi Auto-Analyzer 7170. As a sample, HDL, LDL and VLDL fractions fractionated from human serum by ultracentrifugation were used. FIG. 1 shows the time course measured with a reagent for measuring total TGs (Kyowa Medex Co., Ltd.) for ...

example 2

[0069]

Measurement of TG in HDLReagent 1 (pH 6.25)Buffer (PIPES)50mMTOOS (Dojindo Laboratories)0.3g / LATP 2Na salt (Wako Pure2.5g / LChemical Industries, Ltd.)Ascorbic acid oxidase3kU / L(Asahi Kasei Corporation)GK (Toyobo Co., Ltd.)1kU / LGPO (Asahi Kasei Corporation)8kU / LCatalase (Sigma)300kU / LSodium dextransulfate0.2g / LMagnesium sulfate heptahydrate0.5g / L(Wako Pure ChemicalIndustries, Ltd.)Reagent 2 (pH 6.25)Buffer (PIPES)50mMEmulgen B-6620g / L(Kao Corporation)Calcium chloride dihydrate0.1g / L4-Aminoantipyrine0.5g / L(Wako Pure ChemicalIndustries, Ltd.)Sodium azide0.5g / LLPL (Asahi Kasei Corporation)1000kU / LPeroxidase (Toyobo Co., Ltd.)20kU / L

[0070] The specificity was confirmed by tracing the time course under the condition of the following parameters using a Hitachi Auto-Analyzer 7170. As a sample, the same HDL, LDL and VLDL fractions fractionated from human serum by ultracentrifugation as Example 1 were used. As shown in FIG. 3, in treatment with the reagent 1 in the first reaction, free gl...

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Abstract

The present invention relates to a method for conveniently quantitating TGs contained in various lipoproteins. A method for quantitating trigyceride (TG) in a particular lipoprotein which comprises eliminating free glycerol from a sample containing free glycerol and TG in the particular lipoprotein, then allowing the resulting sample to react with lipoprotein lipase and an enzyme system which generates hydrogen peroxide from free glycerol, and quantitating the generated hydrogen peroxide, is provided.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for quantitating triglycerides (TG) in lipids, which are significant in a field of clinical laboratory test, particularly, as a risk factor of arteriosclerosis. BACKGROUND ART [0002] At present, in a field of clinical laboratory test, cholesterol in high density lipoprotein (HDL) is frequently determined as a risk factor, i.e., negative factor, of arteriosclerosis, while cholesterol in low density lipoprotein (LDL) is also determined as a positive factor. On the other hand, it has been elucidated by many epidemiological searches that hyperlipemia is a primary factor in development of an arteriosclerotic disease which is accompanied by an ischemic heart disease as a major symptom. In addition, among apoproteins E of very low density lipoproteins (VLDL), E1 is incorporated in the receptor, but E2 relating to III type hyperlipemia is not [Arteriosclerosis, 25 (11 / 12), 415-420 (1998); Journal of Clinical and Experimental M...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/44C12Q1/61
CPCC12Q1/44C12Q1/61G01N2333/918G01N2333/908G01N2333/904
Inventor MIYAUCHI, KAZUHITOTAKADA, SHIZUYOMURAKAMI, TOMOMIMIIKE, AKIRA
Owner KYOWA MEDEX CO LTD
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