Method for evaluating sensitization of natural bee pollen and fermented bee pollen

A natural bee pollen and bee pollen technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the lack of allergenicity of bee pollen, the inability to accurately evaluate the degree of bee pollen allergy, and restrict the standardized production and quality of bee pollen fermented products Control and other issues to achieve high accuracy

Active Publication Date: 2021-07-27
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

However, due to the lack of scientific methods for evaluating the allergenicity of bee pollen in the existing technology, the allergenicity evaluation methods of conventional food cannot accurately evaluate the allergenicity of bee pollen, thus restricting the standardized production and quality control of bee pollen fermented products

Method used

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  • Method for evaluating sensitization of natural bee pollen and fermented bee pollen
  • Method for evaluating sensitization of natural bee pollen and fermented bee pollen
  • Method for evaluating sensitization of natural bee pollen and fermented bee pollen

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preparation example Construction

[0111] Preparation of test samples

[0112] The sample applicable to the evaluation method of the present invention is untreated natural bee pollen freeze-dried powder or fermented bee pollen freeze-dried powder, and the sample can be purchased from the market or self-made.

[0113] The samples of the present invention can be prepared by the following method.

[0114] Sample 1: Freeze-dried powder of fermented bee pollen prepared by yeast;

[0115] Its preparation method, concrete steps are as follows:

[0116] 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.

[0117] Note: The rapeseed bee pollen is commercially available untreated conventional rapeseed bee pollen.

[0118] 2) After the sterilization is completed, transfer the bee pollen solution to a sterile centrifuge tube in a ste...

Embodiment 1

[0127] The present embodiment provides a method for evaluating the allergenicity of rape bee pollen freeze-dried powder, comprising the following steps:

[0128] 1. Protein extraction and enzyme digestion

[0129] 1.1. Protein extraction

[0130] Protein extraction was performed on the bee pollen freeze-dried powder obtained from samples 1-4 by using a protein extraction kit;

[0131] 1.2. Enzyme digestion

[0132] Use mass spectrometry-grade trypsin, according to protein mass: trypsin mass = 30:1, and incubate at 37 °C for 12 h for enzymatic digestion to obtain protein extracts.

[0133] 2. Detection of proteomics technology

[0134] The conditions for proteomic detection of protein extracts are:

[0135] An EASY-nLC 1000 nanoliter liquid chromatograph was used in series with an LTQ-Orbitrap Elite combined mass spectrometer (Thermo Fisher Scientific, USA).

[0136] The enrichment column (100 μm × 2 cm, 5 μm) and the analytical column (75 μm × 15 cm, 3 μm, 100 Å) were rev...

Embodiment 2

[0150] In order to improve the accuracy of allergenicity evaluation, the present invention further detects the levels of oligopeptides and amino acids in the bee pollen freeze-dried powder of samples 1-4.

[0151] The specific detection steps are as follows:

[0152] 1. Preprocessing

[0153] Accurately weigh 300 mg of unfermented bee pollen freeze-dried powder, yeast-fermented bee pollen freeze-dried powder, lactic acid bacteria-fermented bee pollen freeze-dried powder and mixed bacteria-fermented bee pollen freeze-dried powder of equal mass respectively, add 1.5 mL of methanol, and vortex. Mix well, sonicate in ice-water bath for 5 min, centrifuge at 13000g for 15 min, take an equal amount of supernatant, and filter it with a 0.22 μm nylon filter for testing;

[0154] 2. Mass spectrometry detection

[0155] UPLC-Q-TOF / MS was used for detection with ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF / MS), and the param...

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Abstract

The invention relates to the technical field of food, and particularly relates to a method for evaluating sensitization of natural bee pollen and fermented bee pollen based on oleosin and glutaredoxin characteristic peptide indexes. According to the invention, the oleosin B2 and the glutaredoxin are used as evaluation indexes for evaluating the sensitization of the untreated natural bee pollen or the fermented bee pollen for the first time, and a specific evaluation process is provided according to the evaluation indexes for evaluating the sensitization of the untreated natural bee pollen or the fermented bee pollen. Meanwhile, on the basis of a specific protein evaluation method, the invention further provides a supplementary means for detecting the contents of oligopeptide and amino acid in the methanol extract of the bee pollen, and further verifies the evaluation result of the sensitization of the bee pollen. The evaluation method provided by the invention has the advantages of high accuracy, convenience and rapidness, and provides an evaluation basis for standardized production and quality control of bee pollen.

Description

technical field [0001] The invention relates to the technical field of food, in particular to a method for evaluating the allergenicity of natural bee pollen and fermented bee pollen based on oleosin and glutaredoxin characteristic peptide indexes. Background technique [0002] Bee pollen has been used in food, health products, pharmaceutical industries, and animal husbandry due to its rich nutritional value, high edible and medicinal effects. However, with the expansion of the scope of application of bee pollen, the safety problems brought about by the consumption of bee pollen have gradually attracted people's attention. [0003] Bee pollen comes from plant pollen. Although some allergenic substances are changed or degraded after the treatment of worker bees, the allergenicity is improved compared with that of plant-derived pollen. However, data show that there are still some susceptible people who will produce bee pollen after eating bee pollen. Clinical allergy symptoms...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 李强强吴黎明陶宇逍尹舒婷薛晓锋赵柳微
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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