A method for evaluating the allergenicity of natural bee pollen and fermented bee pollen

A natural bee pollen, bee pollen technology, applied in the direction of instruments, measuring devices, 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: 2021-11-19
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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  • A method for evaluating the allergenicity of natural bee pollen and fermented bee pollen
  • A method for evaluating the allergenicity of natural bee pollen and fermented bee pollen
  • A method for evaluating the allergenicity 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] This embodiment provides a method for evaluating the allergenicity of rapeseed bee pollen freeze-dried powder, comprising the following steps:

[0128] 1. Protein extraction and digestion

[0129] 1.1. Protein extraction

[0130] Utilize protein extraction kit to carry out protein extraction to the lyophilized powder of bee pollen obtained in sample 1-4;

[0131] 1.2. Enzyme digestion

[0132] Mass spectrometry-grade trypsin was selected, according to protein quantity: trypsin quantity = 30:1, and incubated at 37°C for 12 h for enzymatic digestion to obtain protein extracts.

[0133] 2. Proteomics technology detection

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

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

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

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 freeze-dried powder of bee pollen described in samples 1-4.

[0151] The specific detection steps are as follows:

[0152] 1. Pretreatment

[0153] Accurately weigh 300 mg each 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 respectively, add 1.5mL methanol, vortex Mix well, sonicate in an ice-water bath for 5 minutes, centrifuge at 13,000 g for 15 minutes, take an equal amount of supernatant, and filter it with a 0.22 μm nylon filter membrane before testing;

[0154] 2. Mass spectrometry detection

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

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Abstract

The invention relates to the field of food technology, 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. This invention proposes for the first time that oleosin (Oleosin B2) and glutaredoxin (Glutaredoxin) are used as the evaluation index of untreated natural bee pollen or fermented bee pollen allergenicity evaluation, and provides a specific evaluation process accordingly . At the same time, on the basis of the specific protein evaluation method, the present invention further proposes to further verify the evaluation results of the allergenicity of bee pollen by detecting the content of oligopeptides and amino acids in the methanol extract of bee pollen as a supplementary means. The evaluation method of the invention has the advantages of high accuracy, convenience and quickness, and provides evaluation basis for standardized production and quality control of 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 fermented bee pollen based on oleosin and glutaredoxin characteristic peptide indicators. Background technique [0002] Because of its rich nutritional value, high edible and medicinal effects, bee pollen has been used in food, health care products, pharmaceutical industries, and animal husbandry. However, with the expansion of the scope of application of bee pollen, the safety problems brought about by eating bee pollen have gradually attracted people's attention. [0003] Bee pollen is derived from plant pollen. Although some allergenic substances are changed or degraded after treatment by worker bees, the allergenicity is improved compared with its plant source pollen; but the data show that there are still some susceptible people who will develop Clinical allergy symptoms. [0004] Fermentation has been recog...

Claims

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Application Information

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Patent Type & Authority Patents(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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