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High-performance affinity chromatography for determining lactoferrin in dairy product

A technology of lactoferrin and high-performance liquid chromatography, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high cost of lactoferrin content, unsatisfactory separation, complicated pretreatment, etc., and achieve specificity High, low cost, simple pre-treatment effect

Inactive Publication Date: 2013-01-02
青岛谱尼测试有限公司
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  • Abstract
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Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the defects of high cost, complicated pre-treatment, poor accuracy or insufficient separation degree in determining the content of lactoferrin in dairy products in the prior art, and the affinity chromatography column Used in high performance liquid chromatography system to provide a simple, fast, accurate and low-cost method for the determination of lactoferrin in dairy products

Method used

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  • High-performance affinity chromatography for determining lactoferrin in dairy product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The mensuration of lactoferrin content in the formula milk powder of embodiment 1

[0021] 1. Sample extraction: Weigh 5.0g of the sample into a 50mL colorimetric tube, add 15mL of ultrapure water, vortex and mix, and then ultrasonically extract for 30min. After the extraction is completed, adjust the pH value to 4.6 with acetic acid, let stand for 10 minutes to precipitate casein, add ultrapure water to make the volume of the extract solution 25mL, vortex and mix, transfer 10mL extract solution to a 50mL centrifuge tube, and set the temperature at 8000r / min After centrifugation for 5 minutes, the supernatant was passed through a 0.45 μm water filter membrane, and the filtrate was tested.

[0022] 2. Determination by high performance liquid chromatography system: measure the sample and standard working solution according to the conditions of liquid chromatography, correct the concentration of the sample solution with the standard curve, and quantify with the external st...

Embodiment 2

[0034] The mensuration of lactoferrin content in embodiment 2 milk and human milk

[0035] 1. Sample extraction: with embodiment 1.

[0036] 2. Determination conditions of high performance liquid chromatography system: same as example 1.

[0037] 3. Linear relationship: with embodiment 1.

[0038] 4. Actual sample measurement, precision and accuracy:

[0039] The lactoferrin content in a certain milk sample was measured in parallel for 6 times, and the precision was calculated. The content of lactoferrin in this milk sample was 14.06mg / 100g, and the relative standard deviation was 1.5%.

[0040] Two human milk samples were measured in parallel, and the calculated result was that the content of lactoferrin in A human milk was 351.4mg / 100g, and the content of lactoferrin in B human milk was 100.6mg / 100g.

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Abstract

The invention discloses a high-performance affinity chromatography method for determining lactoferrin in a dairy product. The method comprises the steps of: (1) extracting the dairy product with water, adjusting pH value by organic acid to precipitate casein, and centrifuging to obtain a test solution; and (2) separating and detecting the test solution by a high-performance liquid chromatography system, by using an affinity chromatography column as a chromatographic column, and a diode array detector as a detector. A mobile phase of the high-performance liquid chromatography system comprises A:0.01mol / L of a sodium phosphate buffer solution with pH of 7.0, and B:0.01mol L of a sodium phosphate buffer solution with pH of 7.0 and containing 1mol / L sodium chloride; and gradient elution is employed. A correlation coefficient of a regression equation of the method is higher than 0.999, and a minimum detectable amount is 2mg / 100g. The invention adopts the affinity chromatography method to solve the problems of complex sample matrix and difficulty in accurate quantifying, and has technical characteristics of simple pretreatment, good separation effect, strong specificity, high accuracy, and little change caused by environmental influence.

Description

technical field [0001] The invention relates to a method for measuring lactoferrin content, in particular to a high-efficiency affinity chromatography for measuring lactoferrin in dairy products. Background technique [0002] Lactoferrin (Lactoferrin, Lf) is an iron-binding multifunctional glycoprotein with a molecular weight of 70-80kD. It is also called lactoferrin. Because of its red crystal, it is also commonly called "globin". Lf widely exists in milk, saliva, tears and other exocrine fluids or plasma neutrophils. Lactoferrin is a multifunctional protein with broad-spectrum antibacterial and antiviral infection effects, and can regulate the balance of iron in the body; regulate the production of bone marrow cells and promote cell growth; regulate the body's immune function and enhance the body's disease resistance; inhibit The role of human tumor cells; it can work synergistically with a variety of antibiotics and antifungal agents to treat diseases more effectively. ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 宋薇
Owner 青岛谱尼测试有限公司
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