Method for quantifying Apis cerana cerana honey MRJP2 through liquid chromatography-tandem mass spectrometry

A technology of tandem mass spectrometry and liquid chromatography, applied in the field of honey MRJP2 in the quantification of liquid chromatography tandem mass spectrometry, can solve the problems of disturbing the normal production and sales order of honey, and achieve good accuracy and precision, strong specificity and high sensitivity Effect

Inactive Publication Date: 2020-02-28
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

In order to pursue high profits, unscrupulous companies often use Italian honey to pretend to be Chinese honey, or blend Italian honey with Chinese honey for adulteration, or even mix it with cheap fructose syrup, which seriously disrupts the normal production and sales of honey.

Method used

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  • Method for quantifying Apis cerana cerana honey MRJP2 through liquid chromatography-tandem mass spectrometry
  • Method for quantifying Apis cerana cerana honey MRJP2 through liquid chromatography-tandem mass spectrometry
  • Method for quantifying Apis cerana cerana honey MRJP2 through liquid chromatography-tandem mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Method for Determining the Content of Bee MRJP2 Using Liquid Chromatography Tandem Mass Spectrometry

[0059] 1. Sample source

[0060] A total of 20 samples of real medium honey were purchased from the market or beekeepers.

[0061] 2. Experimental procedure

[0062] (1) Solution preparation

[0063] 5M Urea solution: Weigh 4.5 g of Urea, and dilute to 15 mL with ultrapure water.

[0064] 100 mM DTT: Weigh 308.5 mg DTT, 40 mM NH 4 HCO 3 Dilute the solution to 20 mL, dispense 1 mL into each tube, and store in a -20°C refrigerator until use.

[0065] 40 mM NH 4 HCO 3 Solution: weigh 0.316 g NH 4 HCO 3 , dilute to 100 mL with ultrapure water, and store in a 4°C refrigerator until use.

[0066] 100 mM IAA solution: Weigh 0.39 g of IAA, add 40 mM NH 4 HCO 3 The solution was fixed to 20 mL, and stored in a -20°C refrigerator until use.

[0067] Activation solution: 500 μL ACN, 3 μL TFA, dilute to 1 mL with ultrapure water, store at 4°C.

[0068] Equil...

Embodiment 2

[0095] Example 2 Quantitative detection of the authenticity of Chinese honey samples by MRJP2

[0096] 1. Sample source

[0097] Buy real samples of honey from the market

[0098] 2. Experimental procedure

[0099] (3) Pretreatment of honey samples

[0100] ① Accurately weigh 10 g of the honey to be tested into a 50 mL centrifuge tube, add 10 mL of deionized water, and vortex until the honey is fully dissolved. Centrifuge at 12000 rpm for 10 min at 4°C. Collect the supernatant in a new 2 mL centrifuge tube.

[0101] ② Pipette 200 μL of the protein solution and add 4 μL of 100 μg / mL internal standard peptide (IVNNDFNFNDVNFR*), and mix with 800 μL of 40 mM 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 min.

[0102] ③Add 30 μL of trypsin solution to each sample, and digest at 37°C overnight. When the digestion react...

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Abstract

The invention provides a method for quantifying Apis cerana cerana honey MRJP2 through liquid chromatography-tandem mass spectrometry. The characteristic peptide fragment of royal jelly major protein2 (MRJP2) is screened out for Apis cerana cerana honey, a quantitative method of liquid chromatography tandem mass spectrometry is developed, and the method comprises the steps of screening of the characteristic peptide fragment, making of a standard curve, performing sample pretreatment, performing liquid chromatography separation and tandem mass spectrometry detection, and carrying out softwarequantification of the peptide fragment and the like. The quantitative method for the Apis cerana cerana honey MRJP2 provided by the invention has the advantages of high accuracy, precision and sensitivity and strong stability, and is suitable for accurately quantifying the Apis cerana cerana honey MRJP2. The method can be used for assisting Apis cerana cerana honey authenticity evaluation. In addition, the quantitative method has important practical significance in protecting rights and interests of honey consumers and maintaining healthy development of the bee product industry.

Description

technical field [0001] The invention relates to the field of food detection, in particular to a method for quantifying honey MRJP2 in liquid chromatography tandem mass spectrometry. Background technique [0002] Medium honey, also known as soil honey, is the honey brewed by collecting nectar source plants from our country's native bees, Apis chinensis. 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, unscrupulous companies often use Italian hone...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/8679G01N2030/047
Inventor 杨术鹏李熠周金慧丛晓蕾张金震金钥杨宇辉黄京平袁媛赵文
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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