Milk beta-casein content checking method

A detection method, casein technology, applied in the direction of measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of high price of coated capillary columns, long separation time of capillary columns, harsh use environment requirements, etc., to achieve centrifugal Good separation effect, increase in separation absorbance value, and the effect of low environmental requirements

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 beta-casein content checking method
  • Milk beta-casein content checking method
  • Milk beta-casein content checking method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Detection of β-casein content in milk (using coated quartz capillary column)

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

[0043] 2. Configuration 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 (Beckman Coulter, P / ACE MDQ), detection parameters: the detection temperature is controlled at 30°C, 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 injection, the injection pressure is 0.5psi, the injection time is 5s...

Embodiment 2

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

[0049] 1. Configuration of sample buffer: add 40mmol / L 3-morpholinopropanesulfonic acid, 60mmol / L disodium ethylenediaminetetraacetic acid, 7mol / L urea to 160mmol / L trimethylaminomethane buffer solution, when processing samples 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. Configuration of electrophoresis buffer: 0.32mol / L citric acid, 20mmol / L trisodium citrate, 7mol / L urea, 0.05% hydroxypropyl methylcellulose.

[0051] 3. Capillary electrophoresis instrument (Beckman Coulter, P / ACE MDQ), detection parameters: the detection temperature is controlled at 30°C, using an uncoated quartz capillary column with a diameter of 75 μm and a length of 600 mm, a DAD detector, and pressure sample, the injection pressure is 0.5psi, the injection time is 5s, and the working voltage is 25kV.

[0052...

Embodiment 3

[0055] Example 3 A detection method for β-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 used.

[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: after t...

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Abstract

The invention relates to a method which uses capillary electrophoresis to detect the content of beta-casein protein in milk; and the method includes the following steps: mixing a milk sample to be detected and sample buffer solution; and detecting the sample through the capillary electrophoresis after hydrid processing. The preparation steps of the sample buffer solution is as follows: adding 3-morpholine substituting propane sulfonic acid, disodium EDTA and urea into the trimethyl tris buffer solution; and adding beta-mercaptoethanol and methyl hydroxyethylcellulose when the sample is processed. The steps of processing the milk sample comprise centrifugally processing of the milk sample, mixing the supernatant of the sample after being processed centrifugally and the sample buffer solution, and cleaning the mixture with ultrasonic waves. The method has the advantages of excellent effects of separation and high speed; the capillary column without coating can be applied in the method. The correlation coefficient of linear regression equation of the method is more than 0.98; the minimum detectable concentration is 0.005mg / ml; and the accuracy is above 90 percent.

Description

technical field [0001] The present invention relates to a method for detecting the content of β-casein in milk, especially a method for detecting the content of β-casein in milk by capillary electrophoresis, especially about samples in the process of detecting the content of β-casein 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 hydropho...

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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