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Milk alpha-casein content checking method

A detection method and technology of casein, which are applied in measurement devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of long separation time of capillary column, high price of coated capillary column, and harsh use environment, etc. The effect of good separation effect, low environmental requirements and low price

Active Publication Date: 2012-05-02
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

[0008] Both of the above two methods use a longer coated capillary column, but the separation time of the longer capillary column is also longer, the price of the coated capillary column is relatively high, easy to damage, and has harsh requirements on the use environment

Method used

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  • Milk alpha-casein content checking method
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  • Milk alpha-casein content checking method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 detects α-casein content in milk (using coated quartz capillary column)

[0042] 1. Preparation of sample buffer: add 40mmol / L 3-morpholinopropanesulfonic acid, 60mmol / L disodium edetate, and 7mol / L urea to 160mmol / L trimethylaminomethane buffer solution. Add 5 ul / ml of β-mercaptoethanol, 0.05% methyl hydroxyethyl cellulose, and use 0.1 M sodium hydroxide solution to adjust the pH value to 8.5.

[0043] 2. Preparation of electrophoresis buffer: 0.32mol / L citric acid, 20mmol / L trisodium citrate, 7mol / L urea, 0.05% hydroxypropyl methylcellulose.

[0044] 3. High-pressure capillary electrophoresis instrument (P / ACE MDQ capillary electrophoresis instrument of Beckman Coulter Company in the United States), detection parameters: the detection temperature is controlled at 30°C, and a coated quartz capillary column with a diameter of 50 μm and a length of 20 mm is used, and a diode array Detector (DAD detector), pressure sampling, sampling pressure is 0.5psi, samp...

Embodiment 2

[0048] Example 2 Detection of α-casein content in milk (using uncoated quartz capillary column)

[0049] 1. Preparation of sample buffer: add 40mmol / L 3-morpholinopropanesulfonic acid, 60mmol / L disodium edetate, and 7mol / L urea to 160mmol / L trimethylaminomethane buffer solution. Add 5ul / ml of β-mercaptoethanol, 0.05% of methyl hydroxyethyl cellulose. The pH was adjusted to 8.5 using 0.1 M sodium hydroxide.

[0050] 2. Preparation of electrophoresis buffer: 0.32mol / L citric acid, 20mmol / L trisodium citrate, 7mol / L urea, 0.05% hydroxypropyl methylcellulose.

[0051] 3. Capillary electrophoresis instrument (P / ACE MDQ capillary electrophoresis instrument of Beckman Coulter Company in the United States), detection parameters: the detection temperature is controlled at 30°C, and an uncoated quartz capillary column with a diameter of 75 μm and a length of 600 mm is used for DAD detection Device, pressure sampling, the injection pressure is 0.5psi, the injection time is 5s, and the ...

Embodiment 3

[0055] Example 3 A detection method for α-casein content in pasteurized milk

[0056] Use the same sample buffer, electrophoresis buffer, and high-pressure capillary electrophoresis instrument as in Example 1, and use the same instrument detection parameters as in Example 1. The standard curve obtained in Example 1 was adopted.

[0057] The sample processing and detection steps are as follows: add the pasteurized milk sample into a 15ml centrifuge tube, and centrifuge at 4500r / min for 10min. Accurately extract 100ul of the supernatant of the pasteurized milk sample after centrifugation and put it into a freezing tube (or other container). Extract 300ul of sample buffer into cryovials (or other containers), and use a vortex mixer to mix. Accurately extract 40ul processed samples and add them into sample tubes, place them in an ultrasonic cleaner for 3 minutes, and then measure them with a capillary electrophoresis instrument.

[0058] Quantitative reading of the sample: afte...

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Abstract

The invention relates to a detection method for the content of Alpha-casein in emulsion, comprising the following steps: an emulsion sample which needs detection and sample buffer solution are done with mixed processing and the processed sample is done with detection by a capillary electrophoresis apparatus; the preparation method of the sample buffer solution comprises the following steps: trimethyl aminomethane buffer is added with 3-morpholinopropane sulphonic acid, ethylene diaminetetraacetic acid disodium and urea; when the sample is processed, Beta-mercaptoethanol and methyl hydroxyethyl cellulose are added; wherein, the processing steps of the emulsion sample comprises: centrifuge processing of the emulsion sample, taking the supernatant liquid of the processed sample to be mixed with the sample buffer solution and then washed by ultrasonic wave; the method of the invention has good separating effect and fast speed and can use an uncoated capillary column; the related coefficient of an equation of linear regression of the method of the invention is more than 0.98, the lowest detection line is 0.15ng, the lowest detection concentration is 0.005mg / ml and the precision is up to more than 90 percent.

Description

technical field [0001] The present invention relates to a method for detecting α-casein content in milk, especially a method for detecting α-casein content in milk by capillary electrophoresis, especially about samples in the process of detecting α-casein content in milk by capillary electrophoresis Improvements to the processing steps. Background technique [0002] Casein products are one of the main products of the dairy industry. Due to their excellent functional properties and high nutritional value, they have been widely used in the food industry and other industries. Casein has high nutritional value and can be added to other foods to strengthen protein. Casein can form a colloid in water, has a certain viscosity and good water holding properties, and is used in ice cream production. Sodium caseinate is added as a stabilizer, which has the functions of enhancing fat emulsification, moisturizing and improving viscosity. There are obvious hydrophobic regions and hydro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 孙国庆康小红胡新宇刘卫星
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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