Method for detecting basic protein content of milk

A detection method, milk basic protein technology, applied in the field of analysis, can solve problems such as inability to measure milk basic protein, undiscovered detection methods and detection standards, etc.

Inactive Publication Date: 2013-01-09
INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since milk contains a large amount of protein, none of the above methods can determine milk basic protein
Retrieval of domestic and international materials did not find relevant testing methods and testing standards

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Determination of milk basic protein by micro Kjeldahl

[0049] 1) Precipitation and separation of casein and fat in milk added with milk basic protein

[0050] Accurately weigh 100.00g of milk added with milk basic protein, adjust the pH to 4.6 with 1mol / L HCl, let it stand for 30min, centrifuge (10000r / 20min) to remove casein, obtain supernatant, and use 0.5mol / L Adjust the pH to 6.5 with NaOH, centrifuge (10000r / 15min), and use the supernatant for later use, and select a 100mL centrifuge tube.

[0051] 2) Adsorption of milk basic protein

[0052] Take 20ml~25mL CM sephrose FF of cation exchange resin, put it into a 16x 400mm column, and remove the air bubbles left in the resin by decompression method, so that the material of the chromatographic column is evenly distributed. Then equilibrate with four times the column volume of 0.01mol / L phosphate buffer (pH 6.5) to stabilize the baseline of nucleic acid and protein detection (the model of nucleic acid and...

Embodiment 2

[0059] Embodiment 2: liquid chromatography carries out lactoferrin determination

[0060] 1) Precipitation and separation of casein and fat in milk added with milk basic protein

[0061] Accurately weigh 25.00g of milk added with milk basic protein, adjust the pH to 4.6 with 1mol / L trichloroacetic acid, let it stand for 30min, centrifuge (10000r; 20min) to remove casein, obtain supernatant, and use 0.5mol / L NaOH to adjust the pH to 6.5, centrifuge (10000r; 15min), and the supernatant is set aside.

[0062] 2) Adsorption of lactoferrin

[0063] Take 20ml~25mL CM sephrose FF of cation exchange resin, put it into a 16x400mm column, and remove the air bubbles left in the resin by decompression method, so that the material of the chromatographic column is evenly distributed. Then equilibrate with four column volumes of 0.01mol / L phosphate buffer (pH 6.5) to stabilize the baseline of nucleic acid and protein detection.

[0064] Take 15-20mL supernatant, load it on a cation excha...

Embodiment 3

[0081] Embodiment 3: liquid chromatography carries out milk basic protein determination

[0082] 1) Precipitation and separation of casein and fat in milk added with milk basic protein

[0083] Accurately weigh 100.00g of milk added with milk basic protein, adjust the pH to 4.6 with 1mol / L HCl, let it stand for 30min, centrifuge (10000r; 20min) to remove the casein, obtain the supernatant, and use 0.5mol / L NaOH for the supernatant Adjust the pH to 6.5, centrifuge (10000r; 15min), and use the supernatant for later use.

[0084] 2) Adsorption of milk basic protein

[0085] Take 0.5g of CM sephadex C50 cation exchange resin, and soak it in phosphate buffer solution (pH 6.5, 0.01mol / L) for 48h to fully swell the resin. Put it into a 16x400mm column, and remove the air bubbles remaining in the resin by a decompression method, so that the material of the chromatographic column is evenly distributed. Then equilibrate with four column volumes of 0.01mol / L phosphate buffer (pH 6.5) ...

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Abstract

The invention relates to a method for detecting the basic protein content of milk, and provides a method for determining basic protein substances in the milk, in particular for the determination of milk basic proteins. The method comprises the following steps of: 1) removing caseins and fats in an acid regulation centrifugation way; 2) adsorbing the caseins by using cationic resin; 3) collecting eluent; and 4) determining the proteins by utilizing a micro Kjeldahl method. The invention also provides a method for collecting the milk basic proteins by utilizing chromatography equipment instead of manual collection.

Description

technical field [0001] The invention relates to a method for detecting the content of basic protein in milk, in particular to a method for detecting the content of basic protein in milk, and belongs to the field of analysis technology. Background technique [0002] Basic protein refers to a type of protein with an isoelectric point greater than 7. Milk basic protein (milk basic protein) was approved as a new resource food in my country in 2009. Milk basic protein is a product made from fresh cow's milk through defatting, filtration, concentration, removal of acidic proteins such as casein, cationic chromatography, and freeze-drying (Ministry of Health Announcement No. 18, 2009). The U.S. FDA describes the production process of bovine milk basic protein fraction (BMBPF) as follows: pass pasteurized skimmed milk through a cation exchange resin to remove acidic milk protein and lactose to obtain milk basic protein, and then Sodium chloride is used to elute the milk basic prot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N30/02G01N27/447G01N33/68
Inventor 陈伟李志伟钱文涛季国志高增丽李洪亮于景华高连胜
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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