Method for analyzing heterocyclic amine formation mechanism by using dry and hot soy protein isolate as model

A technology of soybean protein isolate and analysis method, which is applied in the direction of material separation, material analysis, material stimulation analysis, etc., and can solve the problems of heterocyclic amine formation mechanism limitation, complex composition, incompatible protein decomposition and supplementary amino acid content, etc.

Pending Publication Date: 2021-03-26
HENAN UNIVERSITY OF TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still a lack of understanding of the factors that affect the formation of HAAs on the structural modification of plant proteins and chemical reactions at high temperatures, and there is no systematic study on the changes in the formation of heterocyclic amines during protein heat treatment.
In the past, there were mainly two ways to study the formation mechanism of heterocyclic amines. One was to prepare solutions such as amino acid-reducing sugar-creatine (anhydride) or dry heat in proportio

Method used

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  • Method for analyzing heterocyclic amine formation mechanism by using dry and hot soy protein isolate as model
  • Method for analyzing heterocyclic amine formation mechanism by using dry and hot soy protein isolate as model
  • Method for analyzing heterocyclic amine formation mechanism by using dry and hot soy protein isolate as model

Examples

Experimental program
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Effect test

Embodiment 1

[0047] In order to determine the best conditions for simultaneous analysis of protein physicochemical properties and heterocyclic amine production content, the dry heat treatment conditions of soybean protein isolate were designed. Weigh 5g of DuPont 603 soybean protein isolate, place it in a glass petri dish with a diameter of 10cm, and place it in a blast drying oven with a constant temperature for 30min, heat it at 90-230°C for 10min and 60min respectively, and then culture it Take out the dish, put it in a sealed bag after it cools down naturally, and store it in a -20°C refrigerator for later use.

[0048] Because of the obvious color change of dry heat soybean protein isolate (soybean protein isolate after the above dry heat treatment), the L, a, and b values ​​of soybean protein isolate after dry heat were determined.

[0049]L value represents lightness (black and white), a represents red-green, b represents yellow-blue, among them, L: if it is a positive value, it mea...

Embodiment 2

[0063] The dry-heated soybean protein isolate obtained in Example 1 was acid-hydrolyzed to obtain heterocyclic amines, and the heterocyclic amines were purified and concentrated in combination with solid-phase extraction; The content of each heterocyclic amine, the experimental results are as follows image 3 shown, where image 3 The vertical axis of all represents the heterocyclic amine content, and the unit is ng / g.

[0064] Accurately weigh 1 g of each dry-heated soybean protein isolate sample that has been heat-treated at 90-230°C in Example 1, mix them with 10mL6mol / L hydrochloric acid in a 10mL hydrolysis tube, and hydrolyze in a blast drying oven at 110°C for 24 hours. Water was added to the solution to nearly 100mL, and the hydrolyzed solution was adjusted to 100mL after suction filtration with an organic filter membrane.

[0065] Take 10mL of the above diluted hydrolyzate for solid phase extraction. The solid-phase extraction column was pre-activated with 10 mL of...

Embodiment 3

[0073] Analysis of changes in dry heat soy protein isolate using SDS-PAGE:

[0074] A gel rapid preparation kit from Biosharp was used to prepare a 4% stacking gel and a 15% separating gel for SDS-PAGE gel. The soybean protein isolate after the heat treatment in Example 1 was dissolved in PBS, and the supernatant was collected by centrifugation for gel electrophoresis experiment.

[0075] Fluorescein-5-thiosemicarbazide (FTC) can bind to carbonyl compounds in proteins, so FTC can be used to label carbonyl proteins to visualize the degree of protein oxidation and oxidation sites.

[0076] The specific operation is: mix 0.4 mL of the protein (0.5 mg / mL) in Example 1 with 0.1 mL of 5 mmol / L FTC solution, and incubate at 37° C. for 150 min in the dark, stirring gently every 30 min. Proteins were then precipitated with 20% pre-cooled trichloroacetic acid. The mixture was then incubated at 4°C for 10 min in the dark and centrifuged at 12000 rpm for 10 min at 4°C. The precipitate ...

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Abstract

The invention relates to a method for analyzing a heterocyclic amine formation mechanism by using dry and hot isolated soy protein as a model. The method comprises the following steps: carrying out dry heat treatment on the isolated soy protein; determining the content of heterocyclic amine; analyzing protein changes; analyzing a secondary structure and a tertiary structure of the protein; analyzing the oxidation degree of the protein; analyzing the degradation degree of the protein; analyzing the change of the content of formaldehyde and acetaldehyde in the protein pyrolysis process; and evaluating the oxidation resistance change of a protein system. The method can be used for simultaneously detecting the correlation between the content of heterocyclic amine after dry-heat soybean proteinisolate and the physicochemical properties of the protein after heat treatment, is complete in experimental method, has important academic significance, has important significance for reasonably adding the soybean protein isolate in the food industry, and has guiding significance in properly treating the food quality safety problem faced by the development of the food taking the soy isolate protein as a main raw material.

Description

technical field [0001] The invention relates to a method for analyzing the formation mechanism of heterocyclic amines by using dry-heated soybean protein isolate as a model, and belongs to the technical field of food processing. Background technique [0002] Heat transfer is an important purpose of food thermal processing, which can achieve sterilization, ripening, and provide unique flavor and other effects. It is generally believed that the protein content per 100 grams of solid food is not less than 12 grams, which is a high-protein food, and high-protein foods will produce proteins including heterocyclic amines due to changes in protein oxidation, degradation, and aggregation at high temperatures during thermal processing. (HAAs) including many hazards. Toxicology experiments have shown that the toxicity of heterocyclic amines is far higher than that of typical carcinogens such as aflatoxin B1, polycyclic aromatic hydrocarbons, and nitrite. Currently, more than 30 hete...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/33G01N21/25G01N30/02G01N30/06G01N30/86
CPCG01N21/6428G01N21/33G01N21/25G01N30/02G01N30/06G01N30/8675G01N2030/062G01N2021/6432
Inventor 席俊陈阳付杨陈慧彬汪学德马宇翔
Owner HENAN UNIVERSITY OF TECHNOLOGY
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