Method for measuring content of apis mellifera MRJP1 protein in honey by using liquid chromatography-tandem mass spectrometry
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A technology of liquid chromatography and tandem mass spectrometry, applied in the field of food testing
Inactive Publication Date: 2019-12-20
BEE RES INST CHINESE ACAD OF AGRI SCI
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However, there is no relevant report on how to accurately quantify the content of MRJP1 in honey
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Embodiment 1
[0101] Example 1 Method for determining the protein content of Italian bee MRJP1 in honey by liquid chromatography tandem mass spectrometry
[0102] 1. Sample source
[0103] A total of 80 common honey samples were purchased from the market or beekeepers, such as acacia honey, wattle honey, rapeseed honey, jujube honey, linden honey, etc.
[0104] 2. Experimental steps
[0105] (1) Solution preparation
[0106] 5M Urea solution: Weigh 4.5g of Urea, and dilute to 15mL with ultrapure water;
[0108] The 100mM DTT 100mL system configuration is as follows: Weigh 48.06g of urea, 4% sulfur CHAPS, 20mM Tris base, 15.22g, CHAPS 4.00g, Tris base 0.24g and DTT 0.46g, dissolve with ultrapure water, mix and make up to 100mL, store in -20°C refrigerator until use.
[0109] 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. ...
Embodiment 2
[0179] Quantitative detection of embodiment 2 honey adulteration
[0180] 1. Sample source
[0181] Buy real honey samples and syrups from markets or beekeepers.
[0182] 2. Experimental steps
[0183] (1) Solution preparation
[0184] With embodiment 1.
[0185] (2) Blending of honey: mix syrup into honey in different proportions.
[0186] Mix syrup into honey according to the ratio of 5%, 10%, 20%, 50%, 80%.
[0187] (3) Pretreatment of honey samples
[0188] 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.
[0189] 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 measuring the content of apis mellifera MRJP1 protein (major royal jelly protein 1) in honey by using liquid chromatography-tandem mass spectrometry. The method comprises the steps of selecting characteristic peptides, preparing a standard curve, pre-treating a sample, performing liquid chromatography separation, and detecting and quantifying tandem mass spectrometry. The method for detecting the content of apis mellifera MRJP1 in honey provided by the invention has the advantages of strong specificity, high sensitivity, good accuracy and precision, and is suitable for accurate quantification of apis mellifera MRJP1 in honey. Since the content of MRJP1 in honey is relatively constant, the method can be used for evaluating the authenticity of honey basedon the content and assisting in identifying whether honey is adulterated or not. In addition, apis cerana honey does not contain apis mellifera MRJP1 and thus can be used for identifying whether apismellifera is mixed in the apis cerana honey. The method is of great significance for maintaining the healthy development of the honey consumption industry and the rights and interests of honey consumers.
Description
technical field [0001] The invention relates to the field of food testing, in particular to a method for measuring the protein content of Italian bee MRJP1 in honey 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, unscrupulous companies manufacture adulterated honey by mixing sucrose, invert sugar, fructose, glucose, fructose syrup, etc. in th...
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