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Determination method for lactoferrin content in dairy products

A method for measuring lactoferrin, which is applied to measuring devices, instruments, scientific instruments, etc., can solve problems such as large errors, poor quantitative repeatability, and instrument damage, and achieve the effects of reducing testing costs, improving accuracy, and avoiding damage

Active Publication Date: 2012-07-18
上海德诺产品检测有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to overcome the poor quantitative repeatability of the enzyme-linked immunosorbent assay method, the large error of the ultraviolet spectrophotometry method, the gel chromatography is only suitable for high-purity samples and the use of a large amount of NaCl on the instrument in the determination of high-performance liquid chromatography. Defects with large damage, providing a method for the determination of lactoferrin content in dairy products

Method used

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  • Determination method for lactoferrin content in dairy products
  • Determination method for lactoferrin content in dairy products
  • Determination method for lactoferrin content in dairy products

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0046] The content determination of lactoferrin in embodiment 1 fresh milk

[0047] (1) Accurately weigh 10g of fresh milk sample, dissolve it completely in 30mL of water, make it up to 50ml, and then centrifuge at 4°C for 10min at a centrifugal speed of 8000r / min to remove fat to obtain a skim emulsion;

[0048] (2) Over-affinity chromatography column: the affinity chromatography column is preactivated by using a sodium phosphate buffer solution with a pH of 6.8, and 10 ml of skim emulsion is passed through the affinity chromatography column, and the pH of 5 mL is 6.8 sodium phosphate Wash the column with buffer, then elute with 5mL of eluent ①, collect the eluate, and filter it with a 0.45μm pore size aqueous phase filter membrane to obtain the sample solution; wherein, the eluent ① contains 1.0mol / L sodium chloride in sodium phosphate buffer, pH 6.8;

[0049] (3) Take 20 μL of the sample solution and put it into the high performance liquid chromatograph for gradient elutio...

Embodiment 2

[0057] The content determination of lactoferrin in embodiment 2 baby milk powder

[0058] (1) Accurately weigh 5g of infant milk powder sample, dissolve it completely in 40mL of 30°C water, set the volume to 50ml, and then centrifuge at 0°C for 20min at a centrifugal speed of 5000r / min to remove fat to obtain a skim emulsion;

[0059] (2) Over-affinity chromatography column: the affinity chromatography column is preactivated by using a sodium phosphate buffer solution with a pH of 7.4, and 10 ml of skim emulsion is passed through the affinity chromatography column, and the pH of 5 mL is 7.4 sodium phosphate Wash the column with buffer, then elute with 5mL of eluent ①, collect the eluate, and filter it with a 0.45μm pore size aqueous phase filter membrane to obtain the sample solution; wherein, the eluent ① contains 1.0mol / L sodium chloride in sodium phosphate buffer, pH 7.4;

[0060] (3) Take 20 μL of the sample solution and put it into the high performance liquid chromatogra...

Embodiment 3

[0068] The content determination of lactoferrin in embodiment 3 bigger infant milk powder

[0069] (1) Accurately weigh 5g of a larger infant milk powder sample, dissolve it completely in 40mL of 30°C water, set the volume to 50ml, and then centrifuge at 4°C for 15min at a centrifugal speed of 6000r / min to remove fat to obtain a skim emulsion;

[0070] (2) Over-affinity chromatography column: the affinity chromatography column is preactivated by using a sodium phosphate buffer solution with a pH of 7.0, and 10 ml of skim emulsion is passed through the affinity chromatography column, and the pH is 7.0 with 5 mL of sodium phosphate Wash the column with buffer, then elute with 5mL of eluent ①, collect the eluate, and filter it with a 0.45μm pore size aqueous phase filter membrane to obtain the sample solution; wherein, the eluent ① contains 1.0mol / L sodium chloride in sodium phosphate buffer, pH 7.0;

[0071] (3) Take 20 μL of the sample solution and put it into the high-perform...

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Abstract

The invention discloses a determination method for lactoferrin content in dairy products, which includes the steps: (1) degreasing mixed liquid of dairy products and water in centrifugal mode, removing fat to obtain separated milk; (2) passing an affinity column: the separated milk passes through the pre-activated affinity column, washing the column through sodium phosphate buffer solution with pH ranging from 6.8 to 7.4, eluting with eluent (1), collecting the eluent to obtain sample introduction solution; the eluent (1) is sodium phosphate buffer solution containing 1.0 mol / L sodium chloride, and the pH of the eluent (1) is 6.8-7.4; (3) detecting the sample introduction solution through high performance liquid chromatography: eluent (2) of the high performance liquid chromatography is formed by a moving phase A and a moving phase B, the moving phase A is acetonitrile, the moving phase B is 0.1wt% trifluoroacetic acid solution, gradient elution is conducted, and a chromatographic column is a C18 chromatographic column. The determination method has the advantages of specificity and repeatability and capability of removing other disturbance objects and environment change effects and the like, reduces detecting cost, is high in accuracy, and simplifies a lactoferrin detecting method.

Description

technical field [0001] The invention relates to a method for measuring lactoferrin content in dairy products. Background technique [0002] Lactoferrin (Lactoferrin LF), also known as "lactotransferrin", is an iron-binding glycoprotein with many physiological activities. It widely exists in milk, saliva, tears and other exocrine or plasma neutrophils. [0003] In terms of nutritional content, whey protein accounts for about 80% of the total protein in breast milk. The amino acid composition of this type of protein is comprehensive and sufficient with an appropriate ratio, and it also contains active ingredients such as α-lactalbumin, lactoferrin, immunoglobulin, and serum albumin. Among them, lactoferrin is an active ingredient with high nutritional value in whey protein. Lactoferrin was first discovered in breast milk by scientists Montreuij and Johansonb in 1960. Studies by the American Dairy Association and some scientists have shown that lactoferrin is a multifunctio...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 孟瑾郑小平韩惠雯王霞马颖清何亚斌
Owner 上海德诺产品检测有限公司
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