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Method for detecting content of beta-lactoglobulin in dairy product

A detection method and a lactoglobulin technology are applied in the detection field of beta-lactoglobulin content, which can solve the problems of inaccurate determination of beta-lactoglobulin, long analysis time, poor accuracy, etc. Good and accurate results

Inactive Publication Date: 2012-07-04
上海德诺产品检测有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the high cost, poor accuracy, long analysis time and inaccurate determination when the content of β-lactoglobulin in the sample is low. Defects, providing a method for the detection of β-lactoglobulin content in dairy products

Method used

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  • Method for detecting content of beta-lactoglobulin in dairy product
  • Method for detecting content of beta-lactoglobulin in dairy product
  • Method for detecting content of beta-lactoglobulin in dairy product

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1 Determination of β-lactoglobulin in infant milk powder

[0041] ①Centrifugal degreasing: Accurately weigh 10g of infant milk powder (commercially available) sample, dissolve it completely in 100mL of water, and then centrifuge at 4000r / min for 10min at 4°C to remove the floating fat on the upper layer to obtain a degreased sample;

[0042] ②Purification: Take 5mL of 0.10mol / L acetate buffer solution with a pH value of 4.6 and 1mL of degreased sample in a 10mL centrifuge tube, mix well with a vortex shaker for 2min, and centrifuge at 8000r / min for 10min at 4°C , take the supernatant;

[0043] ③Filtration: The supernatant is filtered through a 0.45μm filter membrane to obtain the supernatant;

[0044] ④ Take 10 μL of the upper machine solution into the high-performance liquid chromatograph.

[0045] The content of β-lactoglobulin in infant milk powder was measured to be 15.8mg / g.

Embodiment 2

[0046] Example 2 Determination of the content of β-lactoglobulin in milk beverage

[0047] ①Centrifugal degreasing: take 500ml of milk drink sample (commercially available), centrifuge at 4°C at a speed of 4000r / min for 10min, remove the floating fat on the upper layer to obtain a degreasing sample;

[0048]②Purification: Take 5mL of 0.10mol / L acetate buffer solution with a pH value of 4.6 and 1mL of degreased sample in a 10mL centrifuge tube, mix well with a vortex shaker for 2min, and centrifuge at 8000r / min for 10min at 4°C. Take the supernatant;

[0049] ③Filtration: The supernatant is filtered through a 0.45μm filter membrane to obtain the supernatant;

[0050] ④ Take 10 μL of the upper machine solution into the high-performance liquid chromatograph.

[0051] The content of β-lactoglobulin in the milk drink was measured to be 2.2 mg / ml.

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Abstract

The invention provides a method for detecting the content of beta-lactoglobulin in a dairy product. The method comprises the following steps of: extracting beta-lactoglobulin from the dairy product; and measuring the content of the beta-lactoglobulin by adopting a high performance liquid chromatography method. According to the high performance liquid chromatography method, a C18 chromatographic column is adopted, gradient elution is performed, and the detection wavelength is 210-230 nanometers; an elution solution consists of a mobile phase A and a mobile phase B; the mobile phase A is an ultrapure aqueous solution of trifluoroacetic acid with a volume fraction of 0.05 to 0.15, and the mobile phase B is a mixed solution of acetonitrile, ultrapure water and trifluoroacetic acid; the volume ratio of the mobile phase A to the mobile phase B is 80: 20 to 82: 18; and in the mobile phase B, the volume ratio of the acetonitrile to the ultrapure water is 320: 100 to 480: 100, and the volume of the ultrapure water to trifluoroacetic acid is 100: 0.4 to 100: 0.6. The method is low in cost, short in target peak apperance time, high in separation degree and accuracy, and is favorable for detection of the content of beta-lactoglobulin in the dairy product; the peak shape is acute and symmetric; and the influence from environment change is low.

Description

technical field [0001] The invention relates to a method for detecting the content of β-lactoglobulin in dairy products. Background technique [0002] β-lactoglobulin (β-lactoglobulin) is a kind of protein in fresh milk, accounting for about 7-12% of fresh milk protein. It has been found in some cell and animal experiments that the hydrolyzed substances or molecular modifications of β-lactoglobulin have physiological activities such as lowering cholesterol and anti-oxidation. [0003] At present, there are many methods for determining the content of β-lactoglobulin in dairy products, such as ELISA, electrophoresis and capillary electrophoresis. [0004] However, the above methods have certain limitations in the determination of β-lactoglobulin. For example, ELISA has high selectivity to reagents, and there are many types of proteins in milk, so it is difficult to detect effectively and accurately; There are great limitations; capillary electrophoresis has poor reproducibil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 孟瑾郑小平韩奕奕何亚斌楼飞韩惠雯
Owner 上海德诺产品检测有限公司
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