Method for testing triglyceride in serum by using glycerol dehydrogenase

A technology of glycerol dehydrogenase and triglyceride, which is applied in the field of determination of glycerol dehydrogenase, can solve the problems of endogenous glycerol interference, etc., and achieve the effect of economy, convenience, low reagent cost and high accuracy

Inactive Publication Date: 2014-02-26
TIANJIN BAODI HOSPITAL
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0010] In order to solve the problem of interference of endogenous glycerol in the determination method of TG in serum in the prior art, the present invention ...

Method used

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  • Method for testing triglyceride in serum by using glycerol dehydrogenase
  • Method for testing triglyceride in serum by using glycerol dehydrogenase
  • Method for testing triglyceride in serum by using glycerol dehydrogenase

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Embodiment 1

[0020] Composition of reagents:

[0021] a. Reagent I: Dissolve potassium carbonate 200mmol, sodium bicarbonate 200mmol per liter of carbonate buffer, dissolve NAD + Solution 10.0mmol, glycerol dehydrogenase 45000U, Proclin-300 preservative 200μl.

[0022] b. Reagent II: Dissolve potassium carbonate 200mmol, sodium bicarbonate 200mmol, lipoprotein lipase 1950U, Triton X-100 0.12g, Proclin-300 preservative 200μl per liter of carbonate buffer.

[0023] c. Standard solution: 2.0mmol / L trioleate aqueous solution.

[0024] Among them, K 2 CO 3 middle k + It is an activator of glycerol dehydrogenase, Triton X-100 is a surfactant, and Proclin-300 is a liquid high-efficiency preservative.

[0025] Measurement procedure

[0026] On the Japanese OLYMPUS AU2700 fully automated biochemical analyzer, the instrument automatically adds 2 μl of sample to 150 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 ...

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Abstract

The invention discloses a method for testing triglyceride in serum by using glycerol dehydrogenase, belonging to a method for testing a material through color change caused by a test reaction result by utilizing visible light. The technical scheme is as follows: a reagent I simultaneously contains glycerol dehydrogenase and NAD (Nicotinamide Adenine Dinucleotide)+, and a reagent II contains the effective component of lipoprotein lipase. The method comprises the steps of firstly, subjecting the serum and the reagent I to warm bath at the temperature of 37 DEG C for 3-5min; subjecting free glycerol in the serum and the reagent I to reaction to generate NADH (Nicotinamide Adenine Dinucleotide Hydrogen); adding the reagent II, and then, carrying out warm bath at the temperature of 37 DEG C for 4-7min; hydrolyzing the triglyceride to generate glycerol, and subjecting the glycerol to reaction to generate NADH; detecting on the position with the wavelength of 340nm by using an instrument; with the NADH generated by the reaction of the reagent I as a blank, calculating the content of the triglyceride according to the NADH generated by the reaction of the reagent II. When used for detecting, the method is not affected by endogenous glycerol; the method for testing the triglyceride is same as other enzyme methods in use and range, few in tool enzyme, little in hybrid enzyme interference, low in reagent cost, economic, convenient, feasible and higher in accuracy.

Description

technical field [0001] The present invention belongs to a kind of determination method that comprises enzyme; Or utilize visible light, produce the method for testing material through the result of test reaction color change, especially relate to a kind of glycerol dehydrogenase that uses biochemical analyzer to detect triglyceride in serum test methods. 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. Early methods of determination were estimated as the difference between total lipid and cholesterol and phospholipids. More accurate is the dichloromethane-silicic acid-chromotropic acid method (Van Handel-Caslson method). This method can extract completely, can remove the interference of phospholipids and glycerin, and has high color sensitivity and stable color development of chromotropic acid. It is still a disease in the United...

Claims

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Application Information

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IPC IPC(8): C12Q1/44C12Q1/32
Inventor 李立和张超白会仓丁弘郭秋红
Owner TIANJIN BAODI HOSPITAL
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