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Method for quantifying Apis mellifera honey MRJP3 by liquid chromatography-tandem mass spectrometry

A technology of tandem mass spectrometry and liquid chromatography, which is applied in the field of liquid chromatography tandem mass spectrometry to quantify Italian honey MRJP3, and achieves the effects of good accuracy and precision, high sensitivity, and the meaning of safeguarding rights and interests.

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

There is no relevant report on how to accurately quantify the content of MRJP3 in honey

Method used

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  • Method for quantifying Apis mellifera honey MRJP3 by liquid chromatography-tandem mass spectrometry
  • Method for quantifying Apis mellifera honey MRJP3 by liquid chromatography-tandem mass spectrometry
  • Method for quantifying Apis mellifera honey MRJP3 by liquid chromatography-tandem mass spectrometry

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1 Liquid chromatography tandem mass spectrometry method for determining the protein content of Italian bee MRJP3 in honey

[0057] 1. Sample source

[0058] A total of 20 samples of real Italian honey were purchased from the market or beekeepers.

[0059] 2. Experimental procedure

[0060] (1) Solution preparation

[0061] 5M Urea solution: Weigh 4.5 g of Urea, and dilute to 15 mL with ultrapure water.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] Activation solution: 500 μL ACN, 3 μL TFA, dilute to 1 mL with ultrapure water, store at ...

Embodiment 2

[0090] Example 2 Authenticity detection of Italian honey samples by quantifying Italian bee MRJP3

[0091] 1. Sample source

[0092] Buy real samples of honey from the market

[0093]2. Experimental procedure

[0094] (3) Pretreatment of honey samples

[0095] 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.

[0096] ② Pipette 200 μL of the protein solution and add 4 μL of 100 μg / mL internal standard peptide (LTVAGESFTVK*), 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.

[0097] 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 mellifera honey MRJP3 through liquid chromatography-tandem mass spectrometry. Aiming at the Apis mellifera honey, characteristic peptide fragmentsof royal jelly major protein 3 (MRJP3) in the Apis mellifera honey are screened out, a quantitative method of liquid chromatography tandem mass spectrometry is developed, and the method comprises thesteps of screening the characteristic peptide fragments, making a standard curve, carrying out sample pretreatment, carrying out liquid chromatography separation, carrying out tandem mass spectrometrydetection, carrying out software quantification on the peptide fragments and the like. The method for quantifying the MRJP3 in the Apis mellifera honey, provided by the invention, has the advantagesof higher accuracy, precision and sensitivity and strong stability, and is suitable for accurately quantifying the MRJP3 in the Apis mellifera honey. The method can be used for assisting authenticityevaluation of Apis mellifera honey. In addition, the quantitative method has important practical significance in protecting rights and interests of honey consumers and maintaining healthy developmentof the bee product industry.

Description

technical field [0001] The invention relates to the field of food testing, in particular to a method for quantifying Italian honey MRJP3 by liquid chromatography tandem mass spectrometry. Background technique [0002] Honey is a kind of sweet food that is formed after bees collect nectar or honeydew from nectar source plants and their secretions, and brew for a period of time. As a traditional natural food, honey has health care functions such as beautifying, beautifying and calming the nerves, and is deeply loved by the people. The production of honey has high labor intensity, long cycle, and the output is easily affected by factors such as nectar source and weather, which leads to high production cost of honey and limited total output, resulting in a shortage of honey on the market. In order to pursue huge profits, some unscrupulous companies manufacture adulterated honey by mixing sucrose, invert sugar, fructose, glucose, fructose syrup, etc. in the production, processin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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