Cyclophorase determination method for triglyceride in serum

A triglyceride and measurement method technology, which is applied in the field of determination including enzymes, can solve the problems of endogenous glycerol interference, etc., and achieve the effect of increasing reagent cost and high accuracy

Inactive Publication Date: 2014-07-09
TIANJIN BAODI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0049] In order to solve the problem of endogenous glycerol interference in the determination method of TG in serum in the prior art, the present invention provid

Method used

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  • Cyclophorase determination method for triglyceride in serum
  • Cyclophorase determination method for triglyceride in serum
  • Cyclophorase determination method for triglyceride in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Composition of reagents:

[0061] a. Reagent I:

[0062] Reagent I Each liter of Tris-HCl buffer contains Tris200mmol, 2,4-dichlorophenol 2.0mmol, MgSO 4 12.0mmol, sodium cholate 4.0mmol, adenosine triphosphate 2.0mmol, glycerol kinase 130U, glycerol phosphate oxidase 1600U, peroxidase 1200U, 4-aminoantipyrine 2.0mmol, glycerol-3-phosphate dehydrogenase 1600U, NADH1 .0mmol, Proclin300 preservative 200μl.

[0063] b. Reagent II:

[0064] Reagent II contains Tris200mmol, lipoprotein lipase 1950U, Triton X-1000.12g, and Proclin300 preservative 200μl per liter of Tris-HCl buffer.

[0065] The pH value of the Tris-HCl buffer in the above reagent I and reagent II is 7.6±0.2.

[0066] c. Standard solution: 2.0mmol / L trioleate aqueous solution.

[0067] Among them, MgSO 4 It is an activator of glycerol kinase, Triton X-100 is a surfactant, Proclin-300 is a liquid high-efficiency preservative, and sodium cholate is a bacteriostatic agent.

Embodiment 2

[0069] Measurement procedure

[0070] On the Japanese OLYMPUS AU2700 fully automated biochemical analyzer, the instrument automatically adds 1.5 μl of sample to 200 μl of reagent I and mixes it, incubates at 37°C for 3 minutes, adds 50 μl of reagent II and mixes it, and incubates at 37°C for 5 minutes. Detect at a wavelength of 500nm, and the instrument automatically calculates the TG result, see Table 1 for details.

[0071] Table 1 Automatic biochemical analyzer test conditions of the present invention

[0072]

[0073] response OD TG Calculated value = OD 2 -OD 1 ×[(SV+R 1 V 1 ) / (SV+R 1 V 1 +R 2 V 2 )]

[0074] Triglyceride concentration = F × OD TG

[0075] where OD TG is the absorbance produced by triglycerides. OD 1 is the absorbance measured after adding reagent I to the sample, OD 2 is the absorbance measured after adding reagent II to the sample, SV is the volume of serum, R 1 V 1 is the volume of reagent I, R 2 V 2 is the volume of reagent II. ...

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Abstract

The invention discloses a cyclophorase determination method for triglyceride in serum, and belongs to a method for measuring materials by measuring color change generated by reactions. A technical solution lies in that a reagent I contains effective components of glycerol kinase, 3-phosphoglycerol dehydrogenase, phosphoglycerol oxidase, peroxidase, adenosine triphosphate, NADH, 4-aminoantipyrine and 2,4-dichlorophen; and a reagent II only contains an effective component of lipoprotein lipase. Free glycerol in the serum is subjected to circular reactions with adenosine triphosphate, NADH, 4-aminoantipyrine and 2,4-dichlorophen in the reagent I under catalysis of glycerol kinase, 3-phosphoglycerol dehydrogenase, phosphoglycerol oxidase and peroxidase to generate quinoneimine; and after the reagent II is added, triglyceride is hydrolyzed to generate glycerol, and then the circular reaction is carried out. The content of triglyceride is calculated by an instrument based on quinoneimine generated by reactions in the reagent II, by using quinoneimine generated by reactions in the reagent I as a blank.

Description

technical field [0001] The invention belongs to a method for measuring enzymes; or a method for testing materials by using visible light to produce a color change as a result of a test reaction, in particular to a method for measuring circulating enzymes using a biochemical analyzer to detect triglycerides in serum . Background technique [0002] The determination methods of triglyceride (TG) in serum can be generally divided into three categories: chemical method, enzymatic method and chromatographic method. The present invention introduces a cyclic enzymatic method to measure triglyceride in serum. Triglyceride (TG) generates glycerol and fatty acid under the catalysis of lipoprotein lipase (LPL), and glycerol and ATP generate glycerol under the catalysis of glycerol kinase (GK). -3-Phosphate (G-3P), which is brought into the circulation reaction, is oxidized to dihydroxyacetone phosphate (DHAP) by glycerol phosphate oxidase (GPO), and DHAP is oxidized by glycerol-3 phosp...

Claims

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Application Information

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IPC IPC(8): G01N33/92G01N21/31G01N30/06
CPCG01N33/92G01N21/31G01N30/02G01N2405/00
Inventor 李立和郑宝永王彦平李铮刘果艳
Owner TIANJIN BAODI HOSPITAL
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