Method for determining apis cerana Fabricius MRJP1 protein content in honey by utilizing liquid chromatography tandem mass spectrum

A technology of liquid chromatography and tandem mass spectrometry, which is applied in the field of food testing, can solve the problems of lowering the price of middle and medium honey and reducing the enthusiasm of beekeepers for production.

Inactive Publication Date: 2019-12-20
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

Due to the flood of adulterated honey in the market, maliciously lowering the price of honey, not only seriously hit the enthusiasm of beekeepers, but also led to consumers' confidence in the consumption of honey.

Method used

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  • Method for determining apis cerana Fabricius MRJP1 protein content in honey by utilizing liquid chromatography tandem mass spectrum
  • Method for determining apis cerana Fabricius MRJP1 protein content in honey by utilizing liquid chromatography tandem mass spectrum
  • Method for determining apis cerana Fabricius MRJP1 protein content in honey by utilizing liquid chromatography tandem mass spectrum

Examples

Experimental program
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Effect test

Embodiment 1

[0100] Example 1 Method for Determining the Protein Content of Apis MRJP1 in Honey Using Liquid Chromatography Tandem Mass Spectrometry

[0101] 1. Sample source

[0102] Buy real honey samples and syrups from markets or beekeepers.

[0103] 2. Experimental steps

[0104] (1) Solution preparation

[0105] 5M Urea solution: Weigh 4.5g of Urea, and dilute to 15mL with ultrapure water;

[0106] 100mM DTT: Weigh 308.5mg of DTT, 40mM NH 4 HCO 3 Dilute the solution to 20mL, dispense 1 mL into each tube, and store in a -20°C refrigerator until use.

[0107] 40mM NH 4 HCO 3 Solution: weigh 0.316g NH 4 HCO 3 , dilute to 100 mL with ultrapure water, and store in a 4°C refrigerator until use.

[0108] 100mM IAA solution: Weigh 0.39g of IAA, add 40mM NH 4 HCO 3 The solution was fixed to 20mL and stored in a -20°C refrigerator until use.

[0109] Activation solution: 500 μL ACN, 3 μL TFA, dilute to 1 mL with ultrapure water, store at 4°C.

[0110] Equilibrium solution: 1 μL T...

Embodiment 2

[0169] Quantitative detection of embodiment 2 honey adulteration

[0170] 1. Sample source

[0171] Buy real honey samples and syrups from markets or beekeepers.

[0172] 2. Experimental steps

[0173] (1) Solution preparation

[0174] With embodiment 1.

[0175] (2) Blending of honey: mix syrup into honey in different proportions.

[0176] Mix syrup into honey according to the ratio of 5%, 10%, 20%, 50%, 80%.

[0177] (3) Pretreatment of honey samples

[0178] ① Accurately weigh 10g of the honey to be tested into a 50mL centrifuge tube, add 10mL of deionized water, and vortex until the honey is fully dissolved. Centrifuge at 12000rpm, 4°C for 10min. Collect the supernatant in a new 2mL centrifuge tube.

[0179] ② Pipette 200 μL of protein solution, add 800 μL 40mM NH 4 HCO 3 mix. Add 100 μL of 30 mM DTT solution to the above mixed solution, react at room temperature for 60 min, and then add 500 μL of 100 mM IAA solution and react in dark at room temperature for 60 ...

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Abstract

The invention provides a method for determining apis cerana Fabricius MRJP1 protein content (major royal jelly protein 1) in honey by utilizing liquid chromatography tandem mass spectrum. The method comprises the following steps: selecting characteristic peptide fragment, manufacturing a standard curve, pretreating the sample, performing liquid chromatographic separation and tandem mass spectrum detection quantitation and like steps. The detection method of apis cerana Fabricius MRJP1 content in honey has high accuracy, precision and sensitivity, and strong in stability, and suitable for the accurate quantitation of the apis cerana Fabricius MRJP1. The method further can be used for the truth evaluation of the honey, thereby assisting to identify whether the honey is adulterated, and is ofgreat significance for maintaining the healthy development of the honey consumption industry and the rights and interest of the honey consumer.

Description

technical field [0001] The invention relates to the field of food testing, in particular to a method for measuring the protein content of the honeybee MRJP1 in honey by liquid chromatography tandem mass spectrometry. Background technique [0002] Medium honey, also known as soil honey, is a kind of honey brewed by the native bees in my country, Apis chinensis, which collect honey plants. At present, there are about 3 million bee colonies in my country, which are mainly distributed in the mountainous areas of southern provinces and cities in my country. Italian honey bees mainly adopt the method of chasing flowers and chasing honey to produce honey from a single flower; while Chinese bees mainly raise and produce honey from a variety of flowers in a fixed place. The output of Chinese honey is far lower than that of Italian honey, which leads to a higher market price of Chinese honey, which is about 5-10 times that of Italian honey. In order to pursue high profits, unscrupul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/36G01N30/72G01N30/86G01N33/68
CPCG01N30/02G01N30/06G01N30/34G01N30/36G01N30/7266G01N30/8679G01N33/6818G01N33/6848G01N2030/027
Inventor 杨术鹏李熠丛晓蕾周金慧张金震金玥
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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