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Method of detecting casein in dairy products by using tetraphenylethylene derivatives

A technology of tetraphenylene and derivatives, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc., can solve the problems of poor specificity, low sensitivity, easy interference of milk protein determination methods, etc., to prevent the effect of interference

Inactive Publication Date: 2014-03-12
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] Therefore, the purpose of the present invention is to provide a method for detecting casein in milk products using tetraphenylethylene derivatives, which can effectively Determination of casein content in dairy products with high sensitivity and specificity

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  • Method of detecting casein in dairy products by using tetraphenylethylene derivatives
  • Method of detecting casein in dairy products by using tetraphenylethylene derivatives
  • Method of detecting casein in dairy products by using tetraphenylethylene derivatives

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Embodiment 1

[0055] Preparation of standard curve: Mix 100 μl of casein solution with concentrations of 5, 2.5, 1.25, 0.625, 0.3125, and 0 mg / ml with 100 μl of 1 μM BSPOTPE molecules, excite at 370 nm, emit at 450 nm to measure fluorescence, and make a standard for fluorescence and concentration changes curve. The result is shown in Figure 1.

Embodiment 2

[0057] The test sample is whole milk powder. Take 5 mg of whole milk powder 1, whole milk powder 2, whole milk powder 3 and whole milk powder 4 respectively, and dissolve them in 1 ml of water respectively to make a 5 mg / ml milk solution, and put the solution in a centrifuge at 2000rpm Centrifuge for 10 minutes to skim off the fat on the surface of the solution. The pH of the lower layer solution was adjusted to 4.7 with 2M acetic acid, at this time the casein would precipitate, and it was centrifuged in a centrifuge at 5000rpm for 10 minutes to take out the precipitate. The precipitate was washed with ethanol and ether at a ratio of 1:1. Dissolve in 1 ml of water after washing. Mix 100 μl of casein solution with 100 μl of 1 μM BSPOTPE molecules, excite at 370 nm, and emit at 450 nm to measure fluorescence. The fluorescence value was compared with the standard curve to read out the casein content. The results are shown in Table 1 below.

[0058] Table 1 Comparison of the ...

Embodiment 3

[0061] The test sample is whole milk, which is Yili pure milk produced in China. Dilute 100 μl of milk in 1 ml of water, centrifuge the solution in a centrifuge at 2000 rpm for 10 minutes to skim off the fat on the surface of the solution. The pH of the lower layer solution was adjusted to 4.7 with 2M acetic acid, at this time the casein would precipitate, and it was centrifuged in a centrifuge at 5000rpm for 10 minutes to take out the precipitate. The precipitate was washed with ethanol and ether at a ratio of 1:1. Dissolve in 1 ml of water after washing. Mix 100 μl of casein solution with 100 μl of 1 μM BSPOTPE molecules, excite at 370 nm, and emit at 450 nm to measure fluorescence. The fluorescence value was compared with the standard curve to read out the casein content. It can be obtained that the casein content per 100ml of whole milk is 2.0 grams. The label of the packaging bag is that the protein content of 100ml of milk is 3.0g.

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Abstract

The invention provides a method of detecting casein in dairy products by using tetraphenylethylene derivatives, and particularly provides an application of the tetraphenylethylene derivatives in preparing products used for detecting the casein in the dairy products. The structural formula of the tetraphenylethylene derivatives is shown as the structural formula I. The method includes preparing a standard curve; precipitating the casein in the dairy products; dissolving the precipitated casein; mixing the dissolved casein with a solution of the tetraphenylethylene derivatives and measuring the fluorescence value; and comparing the measured fluorescence value with the standard curve to obtain the content of the casein. According to the method, the tetraphenylethylene derivatives capable of aggregation and luminescence are adopted. When encountering the casein in the dairy products, the tetraphenylethylene derivatives can enter into the protein structure to form an aggregation state and emit fluorescence to avoid interference, and therefore the content of the casein in the dairy products, such as milk and milk powder can be effectively measured. Formula (I).

Description

technical field [0001] The invention relates to the fields of bioengineering and food safety. It specifically relates to a detection method for casein, in particular to a detection method for detecting casein in dairy products by using tetraphenylethylene derivatives. Background technique [0002] At present, the principle of the quantitative detection method of protein in milk is relatively simple, and no new detection method has appeared for many years. Basically, there are only several standard methods such as Kjeldahl method, biuret method, and Coomassie brilliant blue detection. The protein content is calculated by measuring the nitrogen content in the protein sample content. The quantification of biuret protein is quantified through the complex reaction of the special peptide bond structure contained in the protein and the copper ion to change color. The Coomassie Brilliant Blue method is quantified by the specific binding of Coomassie Brilliant Blue molecules to pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 蒋兴宇王卓刘颖昳张伟张德清
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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