Liquid-phase detection method for casein phosphopeptides in milk

A technology of casein phosphopeptide and phosphopeptide solution, which is applied in the field of liquid phase detection of casein phosphopeptide (CPP), which can solve the problem of inability to meet the needs of convenient, fast, and high-sensitivity detection methods, and the inability to detect trace amounts of casein phosphopeptide in milk and other issues to achieve the effect of removing magazine interference, improving sensitivity and ensuring accuracy

Active Publication Date: 2014-04-30
GREENCREAM BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on the fact that the current method cannot detect the content of trace casein phosphopeptide (CPP) in milk, and can no longer meet the needs of a convenient, fast, and high-sensitivity detection method, a high-performance liquid chromatography method was established for the determination of casein phosphopeptide (CPP) in milk. CPP) content method is imminent, which also shows certain advantages of high performance liquid chromatography

Method used

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  • Liquid-phase detection method for casein phosphopeptides in milk
  • Liquid-phase detection method for casein phosphopeptides in milk
  • Liquid-phase detection method for casein phosphopeptides in milk

Examples

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Embodiment 1

[0040] Embodiment 1: The method for determining the content of casein phosphopeptide (CPP) added in milk provided by the present invention comprises the following steps:

[0041] (1) Put the milk to be tested in a beaker, add a casein phosphopeptide (CPP) standard solution with a concentration of 5 mg / mL, mix well, and add the standard solution to form a 100ppm casein phosphopeptide dispersion solution;

[0042] (2) Deproteinization. The method is to take 5g of pre-prepared casein phosphopeptide dispersion solution in a 50mL plastic centrifuge tube, add 5g of water, mix well, then add 5g of 5‰ trichloroacetic acid, vortex and ultrasonic for 5min, then centrifuge at 4800rpm for 10min, centrifuge Finally, put the centrifuge tube into a water bath at 70°C for 10 minutes, and then centrifuge at 4800 rpm for 10 minutes. Take the supernatant and filter it with filter paper for later use;

[0043] (3) Decalcify through 001×7(732) strongly acidic styrene-based cation exchange resin ...

Embodiment 2

[0048] Embodiment 2: The method for determining the content of casein phosphopeptide (CPP) added in milk provided by the present invention comprises the following steps:

[0049] (1) Put the milk to be tested in a beaker, add a casein phosphopeptide (CPP) standard solution with a concentration of 5 mg / mL, mix well, and add the standard solution to form a 10ppm casein phosphopeptide dispersion solution;

[0050] (2) Deproteinization. The method is to take 10g of the pre-prepared casein phosphopeptide dispersion solution sample in a 50mL plastic centrifuge tube, add 10g of water, mix well, then add 10g of 1‰ trichloroacetic acid, vortex, mix and ultrasonic for 10min, and then centrifuge at 4000rpm for 15min. After centrifugation, put the centrifuge tube into a water bath at 75°C for 15 minutes, and then centrifuge at 4000 rpm for 15 minutes. Take the supernatant and filter it with filter paper for later use;

[0051] (3) Decalcification by Mitsubishi WK60L cation exchange resi...

Embodiment 3

[0056] Embodiment 3: The method for determining the content of casein phosphopeptide (CPP) added in milk provided by the present invention comprises the following steps:

[0057] (1) Put the milk to be tested in a beaker, add casein phosphopeptide (CPP) standard solution with a concentration of 5 mg / mL, mix well, and add the standard solution to form a 50ppm casein phosphopeptide dispersion solution;

[0058] (2) Deproteinization. The method is to take 5g of the pre-prepared sample in a 50mL plastic centrifuge tube, add 5g of water, mix well, then add 2.5mL of chloroform:n-butanol solution (pre-prepared to a volume ratio of 4:1 mixture), vortex and mix well. Sonicate for 15 minutes, then centrifuge at 4800 rpm for 5 minutes, take out the supernatant and repeat the above operation for 3 times. Take the supernatant and filter it with filter paper for later use;

[0059] (3) Decalcification by passing through a Rohm and Haas AMBERJET UP6040 cation exchange resin column. Put th...

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Abstract

The invention discloses a liquid-phase detection method for casein phosphopeptides in milk. The liquid-phase detection method comprises the following steps of adding casein phosphopeptides to milk to be detected to prepare a 10-100ppm casein phosphopeptides dispersion solution, carrying out decalcification through cation exchange resin after deproteinization, performing impurity removal and enrichment on an eluant obtained after the deproteinization through the cation exchange resin, carrying out vacuum rotary evaporation on a collecting liquid at the temperature of 60-70 DEG C at the revolving speed of 50-60rpm, and filtering through a membrane to take the collecting liquid as a liquid-phase detection sample; and carrying out liquid-phase detection on the liquid-phase detection sample, and calculating the concentration of the casein phosphopeptides added to the milk by using a peak area method according to an external standard method. The liquid-phase detection method provided by the invention has the advantages that the capital investments of reagents, instruments and the like at the initial stage of the experiment are reduced, the relative simple efficient purification steps are performed, the separation of the casein phosphopeptides is realized, the quantitative accuracy is guaranteed, the interference of impurities such as other proteins and lipids is effectively removed, the final detective sensitivity is improved, and the detection of a sample with low casein phosphopeptides content is beneficial.

Description

technical field [0001] The invention belongs to the technical field of food detection, and in particular relates to a liquid-phase detection method of casein phosphopeptide (CPP). Background technique [0002] Casein phosphopeptide (CPP) is a natural biologically active polypeptide containing phosphoserine, which is obtained by biotechnology from cow's milk casein, and then separated and purified. It can be used in various nutrition and health foods. Effectively promote the body's absorption and utilization of calcium, iron, zinc and other divalent mineral nutrients. Germany and Japan have classified casein phosphopeptide (CPP) as a functional food. In my country, casein phosphopeptide (CPP) has been widely used as a food additive in infant milk powder and health food. [0003] Although people have conducted extensive and in-depth research on the functional components, chemical characteristics, and physiological effects of casein phosphopeptide (CPP), its use in food and h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 曹庸李双祁刘飞彭维郭斌吴成顺
Owner GREENCREAM BIOTECH
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