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A method for extracting low-molecular-weight peptides in milk and dairy products

A low-molecular-weight peptide and extraction method technology, which is applied in the field of low-molecular-weight peptide extraction in milk and dairy products, can solve the problems of many impurities, unfavorable research, and poor extraction effect, and achieve high purity, good purification effect, and less impurities. Effect

Inactive Publication Date: 2015-09-16
HANGZHOU QUALITY TECH SUPERVISION INSPECTION INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

What these extraction methods obtain is the crude extract, the extraction effect is poor, and there are many impurities; and it is unfavorable for the development of imitation breast milk infant formula milk powder and the research of bioactive peptides in dairy products

Method used

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  • A method for extracting low-molecular-weight peptides in milk and dairy products
  • A method for extracting low-molecular-weight peptides in milk and dairy products
  • A method for extracting low-molecular-weight peptides in milk and dairy products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Add 20mL of n-hexane to the raw milk and liquid milk sample solution, shake and mix well, centrifuge at 6000rpm / min for 15 minutes, the upper layer is n-hexane-lipid solution, the middle layer is phosphate buffer solution, and the lowest layer is protein precipitation- For the particle debris layer, take the middle layer solution and pass it through an ultrafiltration membrane with a molecular weight cut-off of 5K, and the filtrate is the initial extraction solution.

[0063] Activation (conventional treatment procedure): Take a reverse-phase solid-phase extraction column (HLP column) and activate it with 10mL methanol and 10mL water in sequence.

[0064] Loading and washing: Add the primary extract to the activated HLP column, wash with 10mL PBS and 20mL 5% methanol solution in turn.

[0065] Elution: Use 10mL of 60% methanol elution solution to elute, and collect the elution solution.

[0066] Rotary evaporation: the eluate solution was concentrated to about 4 mL wit...

Embodiment 2

[0072] Add 20mL n-hexane to the milk powder and powdered dairy product solution, shake and mix well, centrifuge at 6000rpm / min for 15 minutes, the upper layer is n-hexane-lipid solution, the middle layer is the citric acid buffer solution extraction layer, and the lowest layer is protein Precipitation—particle impurity layer, take the middle layer solution, and pass it through an ultrafiltration membrane with a molecular weight cut-off of 5K, and the filtrate is the initial extraction solution.

[0073] Activation (conventional treatment procedure): Take a reverse-phase solid-phase extraction column (HLP column) and activate it with 10mL methanol and 10mL water in sequence.

[0074] Loading and washing: Add the primary extract to the activated HLP column, wash with 10mL PBS and 20mL 5% methanol solution in turn.

[0075] Elution: elute with 10mL 60% methanol elution solution, and collect the elution solution.

[0076] Rotary evaporation: the eluate solution was concentrated t...

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Abstract

The invention relates to an extraction method of low molecular peptide in milk and milk products, and has the advantages of good extraction effect and good purification performance. The technical scheme adopts the following steps: firstly, primary extraction: diluting fresh milk or milk powder by one to five times through buffer solution, and removing lipid, high molecular protein in milk and milk products, and other particulate matters so as to obtain low molecular peptide primary extracting solution; secondly, primary purification: performing extraction and decontamination to the primary extracting solution, then eluting low molecular peptide on an extraction column, collecting eluant, and finally performing rotary evaporation to remove methyl alcohol or methyl cyanide in the eluent so as to obtain low molecular peptide primary purification liquid; thirdly, repurification: adjusting the pH value of the obtained primary purification liquid to be 2 to 4, then absorbing and enriching low molecular peptide onto solid-phase resin, washing out small organic molecule matter without charge and impurities, then eluting the solid-phase resin through ammonia methanol solution, and collecting eluant; and fourthly, obtaining high-purity low molecular peptide mixture.

Description

technical field [0001] The invention relates to a method based on molecular sieve extraction and solid phase extraction to purify small peptides, oligopeptides and polypeptides (less than 50 amino acids) in milk and dairy products, which can be applied to low molecular weight peptides in human milk, cow milk, goat milk and dairy products In the field of peptide extraction, purification, and detection, it also has good application value in the detection of peptide-containing products and the research and development of active peptide functional products. Background technique [0002] The low-molecular-weight peptide derived from protein is a multifunctional factor, which consists of small peptides with short peptide chains of several amino acid residues, oligopeptides (5-10 amino acids) and polypeptides (11-50 amino acids); this Peptoids can be directly absorbed and utilized by the body, and have the characteristics of high absorption efficiency and low immunogenicity. In ad...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14
Inventor 肖海龙屠海云林赛君王红青
Owner HANGZHOU QUALITY TECH SUPERVISION INSPECTION INST