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Method for detecting peroxidase (LOP) in milk product

A technology for peroxidase and dairy products, which is applied in the direction of testing dairy products, biochemical equipment and methods, and testing food, etc. Simple, easy-to-use effects

Inactive Publication Date: 2008-07-02
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

For example, use aniline or pyrogallol to measure the activity of peroxidase, but these methods need to add more reagents, the operation process is cumbersome, the error is large, and it is not easy to standardize; many people use the guaiacol method, but the reagents are expensive and difficult. control response

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  • Method for detecting peroxidase (LOP) in milk product
  • Method for detecting peroxidase (LOP) in milk product
  • Method for detecting peroxidase (LOP) in milk product

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Embodiment Construction

[0024] The method for detecting lactoperoxidase uses ABTS as a substrate, and under the catalysis of lactoperoxidase, a green oxidized ABTS substance is generated, and the colorimetric method is used for continuous detection under visible light, and the change of absorbance value rate to characterize enzyme activity.

[0025] Through the study of the method itself, find out the influence of the highest absorption wavelength of the reaction product, pH, enzyme concentration, substrate concentration, temperature and other factors on the reaction, and determine the best reaction conditions. And apply this method to the detection of lactoperoxidase in milk and other dairy products

[0026] A method for detecting peroxidase (LPO) in dairy products, comprising the following steps:

[0027] 1) Take two cuvettes, each of which is a control tube and a test tube, and add 2.2mL of phosphate buffer solution to adjust the pH to 5.5;

[0028] 2) Add 0.7mL of 2mM ABTS solution;

[0029] 3...

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Abstract

The invention provides a method for detecting lactoperoxidase (LPO) in milk products with the advantages of high sensitivity, low error, simplicity and rapidness. The method comprises the following steps of: (1) taking two cuvettes used as a contrast tube and a test tube respectively, and adding 2.2mL phosphate buffer solution (pH 5-6) respectively; (2) adding 0.5-4mM ABTS solution 0.7mL respectively; (3) adding 1-30mM H2O2 solution 0.1mL respectively; (4) adding 0.2mM dipotassium hydrogen phosphate solution 0.05mL to the contrast tube, and adding 0.05mL sample to the test tube; and (5) mixing, zeroing a blank tube, continuously detecting the absorbance at 416nm in the first 120s at room temperature with spectrophotometer using dynamic curve mode, outputting and printing to obtain one curve approximate to straight line can be obtained for each detection, calculating the slope y of the curve, and calculating enzyme activity x according to the equation x=256.4y+0.0256. The invention uses ABTS reagent, and has the advantages of low cost and easiness for generalization.

Description

technical field [0001] The invention relates to the field of staged food detection, specifically a method for detecting peroxidase (LPO) in dairy products Background technique [0002] Milk is rich in protein, fat, carbohydrates, vitamins, enzymes, minerals and other nutrients necessary for human growth, development and metabolism, and is also conducive to human digestion and absorption, so it is called the closest to perfect food. With the improvement of modern life, people's demand for milk is also increasing. Before drinking milk, it is very important to properly sterilize the milk. [0003] Lactoperoxidase is a type of peroxidase. As one of the endogenous enzymes in milk, LPO exists in the whey of milk. LPO has relatively high thermal stability, and its enzyme activity will be lost after heat treatment at 80°C. If the dairy product is not properly sterilized, the activity of lactoperoxidase will be significantly increased. Lactoperoxidase, thiocyanate and hydrogen pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/28G01N33/04
Inventor 杨国宇韩立强
Owner HENAN AGRICULTURAL UNIVERSITY
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