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Method for evaluating sensitization of natural bee pollen and enzymic wall-broken bee pollen

A technology of natural bee pollen and evaluation method, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problem of lack of an enzymatic method for evaluating the allergy of bee pollen with broken walls, and achieves the effect of high accuracy

Active Publication Date: 2021-09-03
BEE RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the lack of evaluation methods for the allergenicity of bee pollen by enzymatic method in the prior art, and proposes an effective evaluation method

Method used

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  • Method for evaluating sensitization of natural bee pollen and enzymic wall-broken bee pollen
  • Method for evaluating sensitization of natural bee pollen and enzymic wall-broken bee pollen
  • Method for evaluating sensitization of natural bee pollen and enzymic wall-broken bee pollen

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preparation example Construction

[0059] Preparation of test samples

[0060] The samples applicable to the evaluation method of the present invention are natural rapeseed bee pollen freeze-dried powder without broken wall treatment, and two kinds of broken rapeseed bee pollen freeze-dried powder treated by enzymatic hydrolysis, and the samples can be obtained through commercial purchase , or homemade.

[0061] The samples of the present invention can be prepared by the following method.

[0062] Sample 1: Broken rape bee pollen freeze-dried powder treated by enzymolysis with cellulase, pectinase and papain;

[0063] Its preparation method, concrete steps are as follows:

[0064] 1) Accurately weigh 5.0g of ground rapeseed bee pollen powder (100-300 mesh), add 10mL of distilled water, vortex and mix well, seal and pack in a food-grade ziplock bag, and then pasteurize at 63°C , 30 min to obtain bee pollen solution;

[0065] 2) After the sterilization is completed, transfer the bee pollen solution to a ste...

Embodiment 1

[0072] This embodiment provides a method for evaluating the allergenicity of rapeseed bee pollen, the specific steps are as follows:

[0073] 1) Protein extraction to obtain precipitated protein:

[0074] Accurately weigh 100.0 mg of samples 1 to 3 respectively, and add 1 mL of protein extractant;

[0075] Ultrasonic crushing, the condition is ultrasonic 3s, interval 3s, repeat 30 times, continue to shake at room temperature for 30-60min after ultrasonication;

[0076] Then centrifuge at 4°C and 13000g for 15min, absorb 400μL of supernatant, add 2mL of acetone and place it on ice for 60min to precipitate protein, remove supernatant, and obtain precipitated protein;

[0077] Among them, the protein extractant consists of 25mmol / L Tris hydrochloride at pH 7.4, 150mmol / L sodium chloride, 1mmol / L ethylenediaminetetraacetic acid, 1% ethylphenyl polyethylene glycol and 5% glycerin composition.

[0078] 2) Proteolytic digestion to obtain peptides:

[0079] 2.1) Reconstitution of ...

Embodiment 2

[0109] In order to improve the accuracy of sensitization evaluation, the present invention performs dot immunoblotting detection on samples 1-3. The specific detection steps are as follows:

[0110] 1. Test group:

[0111] Natural rapeseed bee pollen group (BP), namely sample 3, prepare 3 parallel samples;

[0112] Broken rape bee pollen (EBP-A) group treated with cellulase and pectinase enzymolysis, i.e. sample 2, prepared 3 parallel samples;

[0113]Broken rape bee pollen (EBP-B) group treated by cellulase, pectinase and papain enzymolysis, i.e. sample 1, prepared 3 parallel samples;

[0114] 2. Test steps:

[0115] (1) Use the vegetable protein extraction kit to extract protein from samples 1 to 3 of the same quality according to the operating instructions;

[0116] Then use the bicinchoninic acid (BCA) protein quantification kit to measure the protein concentration of the protein extract of each group of samples according to the operating instructions;

[0117] (2) Pr...

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Abstract

The invention relates to the technical field of food, in particular to a method for evaluating sensitization of natural bee pollen and wall-broken bee pollen based on Cystatin and Profibrin indexes. According to the invention, Cystatin and Profilin are used as sensitization evaluation indexes of natural bee pollen or wall-broken bee pollen for the first time, and a specific evaluation method is provided according to the evaluation indexes. Meanwhile, the invention further optimizes the application of the dot immunoblotting method in determining the sensitization of the natural bee pollen and the wall-broken bee pollen, and the dot immunoblotting method is used as a mutual verification means. According to the evaluation method, the sensitization of the natural bee pollen and the wall-broken bee pollen can be rapidly and accurately evaluated, and an evaluation basis is provided for standardized production and quality control of the bee pollen.

Description

technical field [0001] The invention relates to the field of food technology, in particular to a method for evaluating the allergenicity of natural bee pollen and enzymatically broken bee pollen based on cystatin and prefibrin. Background technique [0002] Based on reducing the allergenicity of bee pollen, releasing the nutritional components of bee pollen more fully, and improving the nutritional value of bee pollen, the prior art usually processes bee pollen, such as wall breaking, fermentation, enzymatic hydrolysis, etc. . For example, CN102948656A discloses a method for pollen wall breaking, desensitization, and water-soluble component extraction, which improves the wall breaking rate of pollen through enzymatic treatment, and at the same time solves the problem of pollen allergy to a certain extent. [0003] However, during the actual use of this type of broken bee pollen, consumers are allergic to enzymatically broken bee pollen from time to time; fast and accurate ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 李强强陶宇逍尹舒婷吴黎明薛晓锋王妙
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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