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SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) detection method of true protein of dairy products and identification atlas database of method

A detection method and technology of true protein, which is applied in the field of establishing a dairy protein SDS-PAGE identification map library, can solve the problems of adulteration, cumbersome detection of multiple adulterants, and inability to accurately reflect the protein content, source and composition of dairy products, etc. To achieve the effect of good application prospects

Inactive Publication Date: 2014-03-05
郑秋月 +3
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prominent problem of dairy products in my country is adulteration. Unscrupulous businessmen are driven by cost and profit, and take advantage of the lack or limitations of dairy protein detection technology and standards. In addition to mixing chemical raw material "melamine" into dairy products to obtain huge profits, Incorporation of non-dairy protein ingredients that are not easily detected by ordinary methods in food
[0003] The current conventional protein detection methods cannot accurately reflect the content, source and composition of dairy protein. For new adulteration methods such as mixing cheap hydrolyzed protein into milk or mixing other animal milk into milk, traditional detection methods are difficult to detect. out
Most of the current research on dairy adulteration testing is qualitative and quantitative for a single hypothetical substance, and the detection of multiple adulterants is cumbersome, especially for unknown substances that may appear in dairy products.

Method used

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  • SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) detection method of true protein of dairy products and identification atlas database of method
  • SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) detection method of true protein of dairy products and identification atlas database of method
  • SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) detection method of true protein of dairy products and identification atlas database of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, whole milk powder, whey powder and the relative content of each protein component of beverage containing milk

[0045] The common whole milk powder, whey powder and milk beverages in the market were selected as test objects, and the protein composition in 20 kinds of whole milk powder was analyzed ( figure 1 ). According to the extrapolation of the standard analysis amount, the content ratios of αs1-CN, αs2-CN and β-CN in whole milk powder were determined to be (34.4±1.64)%, (8.35±0.25)% and ( 38.50±2.32)%, the content ratio of α-lactalbumin and β-lactoglobulin is (9.57±0.54)%, (1.33±0.37)%;

[0046] Each protein composition in the tested whey powder (WPI) ( figure 1 Middle 2), the relative contents of bovine serum albumin (BSA), αs2-CN, αs1-CN, β-CN, β-LG and α-LA in WPI were (3.542±0.105)%, (18.532±1.203 )%, (6.303±2.4)%, (22.513±2.77)%, (33.848±0.32)% and (12.446±0.158)%.

[0047] The relative contents of αs2-CN, αs1-CN, β-CN, β-LG and α-LA in the...

Embodiment 2

[0048] Embodiment 2, the absolute content of each protein component of milk-containing beverage B

[0049] In order to accurately determine the absolute content of α-LA and β-LG in the milk-containing beverage, the present invention uses SDS-PAGE electrophoresis combined with gel imaging analysis to establish a standard curve. The α-LA and β-LG standards were dissolved in buffer at a concentration of 10 mg / mL, and the loading volumes were 5, 10, 15, and 20 μL, respectively, and contained 50, 100, 150, and 200 μg of protein, respectively, and electrophoresis was carried out. The results are as follows: figure 2 Place. The electrophoresis film was placed in a UVP gel imager for analysis, and the density of each protein spot was obtained, which was correlated with the relative content and absolute content of α-LA and β-LG. The corresponding standard curve was obtained by plotting the absolute content of the standard protein against the protein spot density.

[0050] According ...

Embodiment 3

[0051] Example 3, the SDS-PAGE electrophoresis spectrum library of dairy brand proteins

[0052] In this example, commercially available milk powder and liquid milk were widely collected, numbered from 1 to 22 (only some examples of results). The SDS-PAGE method of the present invention is used to detect the components and contents of the true protein, and the SDS-PAGE electrophoresis pattern library of the dairy brand protein is established. as attached Figure 3-5 shown.

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Abstract

The invention discloses an SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) detection method of true protein of a dairy product and an identification atlas database of the method. The SDS-PAGE detection method comprises the steps: after pretreating a sample to be detected, detecting by using discontinuous polyacrylamide gel electrophoresis, shooting by using a UVP gel imager and analyzing by combining with Labwork software, associating the protein spot density of a lane with the concentration of true protein in a sample, and performing contrast with a standard curve so as to calculate a protein content. According to the SDS-PAGE detection method of the true protein of the dairy product, provided by the invention, the constitution and content of the true protein in the dairy product can be rapidly, sensitively and conveniently detected, and the sample obtained by treatment in the process can be directly applied to measurement of the content of the total protein in the dairy product. The purpose of obtaining a target result by one-time measurement is actually realized. Meanwhile, by adopting the method, an SDS-PAGE identification atlas database of the true protein of the dairy product is established. The SDS-PAGE detection method can be used for comparing SDS-PAGE of the dairy product to be detected so as to be directly judged and evaluated, and has a very good application prospect.

Description

technical field [0001] The invention uses polyacrylamide gel electrophoresis (SDS-PAGE) detection technology to establish an accurate detection method for the true protein components and contents in dairy products, and simultaneously establishes a dairy protein SDS-PAGE identification map library. Background technique [0002] The quality of dairy products is a major issue related to the national economy and people's livelihood. In 2008, the "melamine poisoned milk powder incident" occurred in our country, which seriously threatened people's health and life. China's dairy industry suffered the most profound industry damage and the most serious crisis of trust. Some local and national famous brands were also on the "blacklist". In 2011, "leather hydrolyzed protein" milk powder made a comeback, and the news of "leather milk" made people talk about milk. At present, the prominent problem of dairy products in my country is adulteration. Unscrupulous businessmen are driven by co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 郑秋月曹际娟徐君怡徐杨
Owner 郑秋月
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