CN107383187B - Method for combined extraction of IgY, phosvitin, lecithin, egg oil and defatted egg powder from egg yolk - Google Patents
Method for combined extraction of IgY, phosvitin, lecithin, egg oil and defatted egg powder from egg yolk Download PDFInfo
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- CN107383187B CN107383187B CN201710557186.4A CN201710557186A CN107383187B CN 107383187 B CN107383187 B CN 107383187B CN 201710557186 A CN201710557186 A CN 201710557186A CN 107383187 B CN107383187 B CN 107383187B
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- 210000002969 egg yolk Anatomy 0.000 title claims abstract description 102
- 102000002322 Egg Proteins Human genes 0.000 title claims abstract description 33
- 108010000912 Egg Proteins Proteins 0.000 title claims abstract description 33
- 235000013345 egg yolk Nutrition 0.000 title claims abstract description 33
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 title claims abstract description 21
- 102100027992 Casein kinase II subunit beta Human genes 0.000 title claims abstract description 21
- 101710158100 Casein kinase II subunit beta Proteins 0.000 title claims abstract description 21
- 235000010445 lecithin Nutrition 0.000 title claims abstract description 21
- 229940067606 lecithin Drugs 0.000 title claims abstract description 21
- 239000000787 lecithin Substances 0.000 title claims abstract description 21
- 239000000843 powder Substances 0.000 title claims abstract description 17
- 235000013601 eggs Nutrition 0.000 title claims abstract description 16
- 238000000034 method Methods 0.000 title claims abstract description 10
- 238000000605 extraction Methods 0.000 title claims description 13
- 150000002632 lipids Chemical class 0.000 claims abstract description 41
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 239000006228 supernatant Substances 0.000 claims abstract description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 13
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 13
- 239000011780 sodium chloride Substances 0.000 claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 8
- 238000007865 diluting Methods 0.000 claims abstract description 6
- 239000012046 mixed solvent Substances 0.000 claims abstract description 6
- 239000002244 precipitate Substances 0.000 claims description 43
- 239000007864 aqueous solution Substances 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 18
- 239000000706 filtrate Substances 0.000 claims description 16
- 238000001291 vacuum drying Methods 0.000 claims description 16
- 239000008118 PEG 6000 Substances 0.000 claims description 13
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 claims description 13
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 13
- 239000012153 distilled water Substances 0.000 claims description 13
- 238000004108 freeze drying Methods 0.000 claims description 11
- 238000002390 rotary evaporation Methods 0.000 claims description 10
- 235000019441 ethanol Nutrition 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 238000007605 air drying Methods 0.000 claims description 5
- LJQKCYFTNDAAPC-UHFFFAOYSA-N ethanol;ethyl acetate Chemical compound CCO.CCOC(C)=O LJQKCYFTNDAAPC-UHFFFAOYSA-N 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- 238000002844 melting Methods 0.000 claims description 4
- 230000008018 melting Effects 0.000 claims description 4
- 238000002425 crystallisation Methods 0.000 claims description 2
- 230000008025 crystallization Effects 0.000 claims description 2
- 239000012535 impurity Substances 0.000 claims description 2
- 235000011837 pasties Nutrition 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 15
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 229920001223 polyethylene glycol Polymers 0.000 abstract description 2
- 239000002202 Polyethylene glycol Substances 0.000 abstract 1
- 230000007613 environmental effect Effects 0.000 abstract 1
- 231100000956 nontoxicity Toxicity 0.000 abstract 1
- 238000001556 precipitation Methods 0.000 abstract 1
- 238000011112 process operation Methods 0.000 abstract 1
- 238000005185 salting out Methods 0.000 abstract 1
- 238000001035 drying Methods 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 238000005119 centrifugation Methods 0.000 description 4
- 238000000967 suction filtration Methods 0.000 description 4
- 230000000975 bioactive effect Effects 0.000 description 3
- 238000011033 desalting Methods 0.000 description 3
- 238000010025 steaming Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 102000015779 HDL Lipoproteins Human genes 0.000 description 2
- 108010010234 HDL Lipoproteins Proteins 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 102000007330 LDL Lipoproteins Human genes 0.000 description 2
- 108010007622 LDL Lipoproteins Proteins 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000133 brain stem Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 235000020778 linoleic acid Nutrition 0.000 description 1
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/02—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from eggs
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/10—Phosphatides, e.g. lecithin
- C07F9/103—Extraction or purification by physical or chemical treatment of natural phosphatides; Preparation of compositions containing phosphatides of unknown structure
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/465—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from birds
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/108—Production of fats or fatty oils from raw materials by extracting after-treatment, e.g. of miscellae
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Abstract
The invention discloses a method for jointly extracting IgY, phosvitin, lecithin, egg oil and degreased egg powder from egg yolk. Firstly, diluting yolk liquid with water and centrifuging, and extracting IgY from obtained supernatant by salting out and polyethylene glycol precipitation; the lower layer is yolk plasma, firstly, ethanol is adopted to remove water in the yolk plasma, then mixed solvent of ethanol and ethyl acetate is utilized to extract total lipid of yolk, and then ethyl acetate is utilized to process so as to obtain two products of lecithin and yolk oil; finally, the protein after lipid removal is treated by sodium chloride solution to obtain the phosvitin. The invention has simple and convenient process operation, environmental protection, no toxicity, low production cost, high product purity and high yield, and provides technical guarantee for comprehensive utilization of the yolk.
Description
Technical Field
The invention belongs to the field of egg product deep processing, and particularly relates to a method for jointly extracting IgY, phosvitin, lecithin, egg oil and degreased egg yolk powder from egg yolk.
Background
The egg yolk is the most abundant part of the nutritive value in the egg, the protein content is high, the amino acid composition of the egg yolk is very similar to that of the human body protein, the egg yolk is high-quality complete protein second to cow milk, and the biological value of the egg yolk is as high as 96%; secondly, the lipids enriched in the egg yolk not only contain rich unsaturated fatty acids and proper linoleic acid content, but also contain phospholipid components which are indispensable to human beings in brain and nervous system, so that the development of bioactive components such as protein and lipid in the egg yolk is always a research hotspot of scholars at home and abroad.
Due to the complex composition of egg yolk, it is difficult to make the best use of the egg yolk. At present, research mainly focuses on the separation and extraction of single bioactive components in egg yolk, and the comprehensive utilization of the egg yolk is rarely researched. The invention adopts reagents allowed to be used on food to extract a target product, greatly reduces the potential safety hazard of food, simultaneously realizes the coproduction of various yolk bioactive components such as yolk immunoglobulin, yolk oil, lecithin, yolk hyperphosphatein and defatted yolk powder, establishes a set of egg product deep processing production process with high extraction efficiency, high purity, strong operability, high value and full utilization, and provides technical support for comprehensive utilization of yolk.
Disclosure of Invention
The invention aims to realize the co-production of various functional components of yolk and establish a set of egg deep processing production process with high extraction efficiency, high purity, simple operation, high value and full utilization.
The method of the invention comprises the following steps:
(1) and (3) extracting IgY: diluting fresh egg yolk liquid and water according to the volume ratio of 1: 1-12, adjusting the pH value to 4-7, uniformly stirring, placing in a refrigerator, standing overnight, centrifuging to obtain a supernatant A, wherein the lower layer is egg yolk serous substance (mainly containing high-density lipoprotein, low-density lipoprotein and water), then adding ammonium sulfate and sodium chloride into the supernatant A to salt out and precipitate the IgY, dissolving the centrifugally obtained crude IgY precipitate with distilled water, dialyzing to remove the salt to obtain a crude IgY aqueous solution, then adding PEG6000 into the crude IgY aqueous solution to ensure that the weight concentration reaches 2-10%, adjusting the pH value to 3-8, placing in a refrigerator, standing, centrifuging to obtain a purified IgY precipitate, finally dissolving the purified IgY precipitate again with distilled water, dialyzing to remove the PEG6000 to obtain a purified IgY aqueous solution, and freeze-drying to obtain an IgY solid;
(2) and (3) extracting total lipid of egg yolk: adding absolute ethanol into the yolk plasma obtained in the step (1), fully stirring, centrifuging to obtain supernatant B (containing water, ethanol, water-soluble protein and partial yolk lipid) and precipitate A (mainly containing low-density lipoprotein and high-density lipoprotein), placing the supernatant B into a refrigerator at-20 ℃ overnight to crystallize and separate out precipitate B, wherein the precipitate B mainly contains yolk lipid 2 and water-soluble protein, melting the solidified lipid yolk 2 by warm water to obtain oily liquid, and centrifuging to separate the yolk lipid 2 from the water-soluble protein, wherein the lipid is at the upper layer and the protein is at the lower layer; carefully pouring out the supernatant C (mainly ethanol, water and a part of the yolk lipid 1 which is not crystallized and precipitated at low temperature) and removing the ethanol by rotary evaporation, and then separating the liquid to obtain the yolk lipid 1; adding an ethanol-ethyl acetate mixed solvent into the obtained precipitate A for extraction, filtering to obtain a filtrate 1 and a filter residue 1, performing rotary evaporation on the filtrate 1 to remove the solvent to obtain yolk lipid 3, and performing vacuum drying on the mixed yolk lipids 1,2 and 3 to obtain yolk total lipid;
(3) extraction of lecithin: adding ethyl acetate into the total lipid of the egg yolk obtained in the step (2), crystallizing at low temperature, centrifuging to obtain a precipitate, and performing vacuum drying to obtain pasty lecithin;
(4) extracting egg yolk oil: performing rotary evaporation on the supernatant D obtained by centrifuging in the step (3) to remove the solvent, and performing vacuum drying to obtain egg yolk oil;
(5) extracting phosvitin and defatted yolk powder: naturally drying the filter residue 1 obtained in the step (2), adding a sodium chloride solution with the weight concentration of 10-20%, magnetically stirring for 0.5-2h to fully dissolve the water-soluble phosvitin, then preserving the heat at the high temperature of 50-100 ℃ for 10-30min to denature and precipitate other water-soluble impurity proteins, finally filtering to obtain a filtrate 2 and a filter residue 2, dialyzing the filtrate 2, desalting, freeze-drying to obtain the phosvitin, and naturally air-drying the filter residue 2 to obtain the defatted yolk powder.
Preferably, in step (1), the final concentrations of ammonium sulfate and sodium chloride in supernatant A are 20-35% and 0.1-1%, respectively, both by weight.
Preferably, in the step (2), the volume ratio of ethanol to ethyl acetate in the ethanol-ethyl acetate mixed solvent is 1: 1-6.
The invention has the beneficial effects that:
(1) the method has low requirement on equipment, simple and convenient operation, low operation cost and high extraction rate, does not cause toxic solvent residue, and can be used for industrial production.
(2) When the yolk oil and the yolk lecithin are extracted, the conditions are mild, the temperature is controlled below 50 ℃, the oxidation of the yolk oil and the yolk lecithin is avoided, and the functional activity of the yolk oil and the yolk lecithin is ensured to the greatest extent.
(3) The invention simultaneously obtains 5 products of yolk immunoglobulin, phosvitin, lecithin, yolk oil and degreased yolk powder, and realizes high-value full utilization of yolk.
(4) Compared with the single ethanol solvent, the ethanol-ethyl acetate mixed solvent has better extraction effect and higher oil extraction rate.
(5) The invention uses absolute ethyl alcohol to remove water in the yolk plasma, and does not adopt methods such as freeze-drying and drying, thereby saving energy, simplifying steps, providing good conditions for subsequent extraction of water-soluble phosvitin, greatly increasing the yield of the phosvitin, and simultaneously obtaining the yolk total lipid containing less water-soluble protein and having good fluidity.
Drawings
FIG. 1 is a process flow diagram of the present invention.
Detailed Description
In order to better illustrate the present invention, the present invention is described in detail below by way of examples.
Example 1
(1) Diluting 500ml of yolk stock solution with 9 times of distilled water, adjusting pH to 5.5, stirring uniformly, standing overnight in a refrigerator at 4 ℃, taking out, and centrifuging at 10000rpm for 15min to obtain supernatant A and yolk plasma; then adding ammonium sulfate and sodium chloride into the supernatant A to precipitate the IgY to ensure that the final weight concentration of the IgY reaches 28 percent and 0.5 percent respectively, completely dissolving the precipitate obtained by centrifugation with distilled water, dialyzing and desalting to obtain a crude IgY aqueous solution, then adding PEG6000 into the crude IgY aqueous solution, adjusting the pH to be 5.0 and the final weight concentration of the PEG6000 to be 8 percent, placing the crude IgY aqueous solution into a refrigerator for 1h, centrifuging to obtain a purified IgY precipitate, finally adding distilled water into the purified IgY precipitate to dissolve again, dialyzing to remove the PEG6000 to obtain a purified IgY aqueous solution, and freeze-drying to obtain an IgY finished product;
(2) adding 400ml of absolute ethyl alcohol into the yolk plasma in the step (1), stirring at 10000rpm for 10min, centrifuging to obtain a precipitate A and a supernatant B, then placing the supernatant B into a refrigerator at-20 ℃ for standing overnight, pouring out a supernatant C and a precipitate B, removing the ethyl alcohol from the supernatant C by rotary evaporation at the temperature of 45 ℃, obtaining yolk lipid 1 through liquid separation, and drying in vacuum (weighing 0.52 g); the precipitate B was prepared by melting the coagulated yolk lipid 2 with warm water at 30 ℃ and centrifuging to obtain yolk lipid 2 and vacuum-drying (weighing 14.01 g);
(3) adding 1335ml of absolute ethyl alcohol and 2670ml of ethyl acetate into the precipitate A, magnetically stirring for 90min at 40 ℃, carrying out vacuum filtration, carrying out rotary evaporation on the filtrate 1 at 45 ℃, and carrying out vacuum drying to obtain yolk lipid 3 (weighing 148.03 g); naturally air drying the filter residue 1, and adding 805ml of 10% NaCl2Magnetically stirring the solution at 4 ℃ for 1h, carrying out water bath at 90 ℃ for 20min, carrying out suction filtration to obtain filtrate 2, dialyzing for 24h, and freeze-drying to obtain phosvitin; naturally drying the filter residue 2 to obtain the defatted yolk powder;
(4) mixing yolk lipid 1,2 and 3, adding 960ml ethyl acetate, standing at 0 deg.C for crystallization for 2 hr, vacuum filtering, and vacuum drying to obtain refined lecithin; and rotatably steaming the supernatant at the temperature of D45 ℃, and drying in vacuum to obtain the egg yolk oil.
The yield and purity of each product are as follows:
| product(s) | IgY | Phosvitin | Lecithin | Egg oil | Defatted egg yolk powder |
| Yield/g | 3.2750 | 5.2505 | 53.8 | 107.5 | 42.18 |
| Purity/%) | 99.38 | 80.13 | 88.7 |
Example 2
(1) Diluting 500ml of yolk stock solution with distilled water with volume 3 times, adjusting pH to 4, stirring uniformly, standing overnight in a refrigerator at 4 ℃, rotating at 10000rpm, and centrifuging for 15min to obtain supernatant A and yolk plasma; adding ammonium sulfate and sodium chloride into the supernatant A to precipitate the IgY to enable the final concentration of the IgY to reach 20% and 1% respectively, completely dissolving the precipitate obtained by centrifugation with distilled water, dialyzing to remove salt to obtain a crude IgY aqueous solution, adding PEG6000 into the crude IgY aqueous solution to adjust the pH value to 7.0 and the final concentration of PEG6000 to be 4%, placing the crude IgY aqueous solution into a refrigerator for 1h, centrifuging to obtain a purified IgY precipitate, finally adding distilled water into the purified IgY precipitate to dissolve again, dialyzing to remove the PEG6000 to obtain the purified IgY aqueous solution, and freeze-drying to obtain an IgY finished product;
(2) adding 400ml of absolute ethanol into yolk plasma, stirring for 10min, centrifuging at 10000rpm for 10min to obtain a precipitate A and a supernatant B, then placing the supernatant B into a refrigerator at-20 ℃, standing overnight, pouring out a supernatant C and a precipitate B, performing rotary evaporation on the supernatant C at 45 ℃ to obtain yolk lipid 1, performing vacuum drying, and weighing 0.56 g. The precipitate B is melted by warm water to solidify lipid, and then is centrifuged to obtain lipid 2, and the obtained lipid 2 is dried in vacuum and weighed to 12.12 g;
(3) adding anhydrous ethanol 1322ml and ethyl acetate 2644ml into the precipitate A, magnetically stirring at 40 deg.C for 90min, vacuum filtering, rotary steaming the filtrate 1 at 45 deg.C, and vacuum drying to obtain yolk lipid 3, weighing 150.5 g; naturally air drying the filter residue 1, and adding 824ml of 15% NaCl2Magnetically stirring the solution at 4 ℃ for 0.5h, carrying out water bath at 50 ℃ for 30min, carrying out suction filtration to obtain filtrate 2, dialyzing for 24h, and freeze-drying to obtain phosvitin; naturally drying the filter residue 2 to obtain the defatted yolk powder;
(4) adding 970ml ethyl acetate into the mixed lipids 1,2 and 3, standing at 0 ℃ for 2 hours, then carrying out suction filtration, and carrying out vacuum drying on the obtained precipitate to obtain refined lecithin; the filtrate is rotary evaporated at 45 ℃ and dried in vacuum to obtain the egg yolk oil.
The yield and purity of each product are as follows:
| product(s) | IgY | Phosvitin | Lecithin | Egg oil | Defatted egg yolk powder |
| Yield/g | 3.1311 | 5.1750 | 51.2 | 109.6 | 41.67 |
| Purity/%) | 98.75 | 79.45 | 86.9 |
Example 3
(1) Diluting 500ml of yolk stock solution with 12 times of distilled water, adjusting pH to 7.0, stirring uniformly, standing overnight at 4 ℃, rotating at 10000rpm, and centrifuging for 15min to obtain supernatant A and yolk plasma; adding ammonium sulfate and sodium chloride into the supernatant A to precipitate the IgY to enable the concentration of the IgY to reach 35% and 0.1% respectively, completely dissolving the precipitate obtained by centrifugation with distilled water, dialyzing and desalting to obtain a crude IgY aqueous solution, adding PEG6000 into the crude IgY aqueous solution, adjusting the pH value to be 3.0 and the PEG concentration to be 10%, placing the crude IgY aqueous solution into a refrigerator for 1h, centrifuging to obtain a purified IgY precipitate, finally adding distilled water into the purified IgY precipitate for redissolving, dialyzing to remove the PEG6000 to obtain the purified IgY aqueous solution, and freeze-drying to obtain an IgY finished product;
(2) adding 400ml of absolute ethyl alcohol into the yolk plasma, stirring for 10min, performing centrifugation for 10min at 10000rpm to obtain a precipitate A and a supernatant B, then placing the supernatant B into a refrigerator at-20 ℃ for standing overnight, pouring out a supernatant C, collecting the precipitate B, performing rotary evaporation on the supernatant C at 45 ℃ to obtain yolk lipid 1, performing vacuum drying, and weighing 0.45 g; melting precipitate B, centrifuging to obtain lipid 2, vacuum drying, and weighing 13.20 g;
(3) adding anhydrous ethanol 1338ml and ethyl acetate 2676ml into the precipitate A, magnetically stirring at 40 ℃ for 90min, vacuum filtering, rotary steaming the filtrate 1 at 45 ℃ and then vacuum drying to obtain the yolk lipid 3, weighing 143.76 g. Naturally drying the filter residue 1, and adding 814ml of 20% NaCl2Magnetically stirring the solution at 4 ℃ for 2h, carrying out water bath at 100 ℃ for 10min, carrying out suction filtration to obtain filtrate 2, dialyzing for 24h, and freeze-drying to obtain phosvitin; naturally drying the filter residue 2 to obtain the defatted yolk powder;
(4) mixing yolk lipid 1,2 and 3, adding 960ml ethyl acetate, standing at 0 deg.C for 2 hr, vacuum filtering, collecting precipitate, and vacuum drying to obtain refined lecithin; the filtrate is rotary evaporated at 45 ℃ and dried in vacuum to obtain the egg yolk oil.
The yield and purity of each product are as follows:
| product(s) | IgY | Phosvitin | Lecithin | Egg oil | Defatted egg yolk powder |
| Yield/g | 3.0224 | 5.0753 | 52.1 | 104.1 | 41.68 |
| Purity/%) | 99.12 | 79.86 | 87.06 |
Claims (1)
1. A method for extracting IgY, phosvitin, lecithin, egg oil and degreased egg powder from egg yolk in a combined way is characterized by comprising the following steps:
(1) and (3) extracting IgY: diluting fresh egg yolk liquid and water according to the volume ratio of 1: 1-12, adjusting the pH value to 4-7, uniformly stirring, placing in a refrigerator, standing overnight, centrifuging to obtain a supernatant A, adding ammonium sulfate and sodium chloride into the supernatant A to salt out and precipitate the IgY, dissolving the coarse IgY precipitate obtained by centrifuging with distilled water, dialyzing to remove the salt to obtain a coarse IgY aqueous solution, adding PEG6000 into the coarse IgY aqueous solution to ensure that the weight concentration of the PEG6000 reaches 2-10%, adjusting the pH value to 3-8, placing in the refrigerator, standing, centrifuging to obtain a purified IgY precipitate, finally dissolving the purified IgY precipitate again with distilled water, dialyzing to remove the PEG6000 to obtain the purified IgY aqueous solution, and freeze-drying to obtain an IgY solid;
(2) and (3) extracting total lipid of egg yolk: adding absolute ethyl alcohol into the yolk plasma obtained in the step (1), fully stirring and then centrifuging to obtain a supernatant B and a precipitate A, putting the supernatant B into a refrigerator at the temperature of-20 ℃ for overnight crystallization to separate out the precipitate B, wherein the precipitate B mainly comprises yolk lipid 2 and water-soluble protein, melting the coagulated yolk lipid 2 by warm water to form oily liquid, and centrifuging to separate the yolk lipid 2 from the water-soluble protein; pouring out the supernatant C, removing the ethanol by rotary evaporation, and separating liquid to obtain yolk lipid 1; adding an ethanol-ethyl acetate mixed solvent into the obtained precipitate A for extraction, filtering to obtain a filtrate 1 and a filter residue 1, performing rotary evaporation on the filtrate 1 to remove the solvent to obtain yolk lipid 3, and performing vacuum drying on the mixed yolk lipids 1,2 and 3 to obtain yolk total lipid;
(3) extraction of lecithin: adding ethyl acetate into the total lipid of the egg yolk obtained in the step (2), crystallizing at low temperature, centrifuging to obtain a precipitate, and performing vacuum drying to obtain pasty lecithin;
(4) extracting egg yolk oil: performing rotary evaporation on the supernatant D obtained by centrifuging in the step (3) to remove the solvent, and performing vacuum drying to obtain egg yolk oil;
(5) extracting phosvitin and defatted yolk powder: naturally air-drying the filter residue 1 obtained in the step (2), adding 10-20% sodium chloride solution, magnetically stirring for 0.5-2h to fully dissolve the water-soluble phosvitin, then preserving heat at 50-100 ℃ for 10-30min to denature and precipitate other water-soluble impurity proteins, finally filtering to obtain filtrate 2 and filter residue 2, dialyzing the filtrate 2 to remove salt, freeze-drying to obtain phosvitin, naturally air-drying the filter residue 2 to obtain defatted yolk powder,
in the step (1), the final concentrations of the ammonium sulfate and the sodium chloride in the supernatant A are respectively 20-35% and 0.1-1%, which are weight concentrations,
in the step (2), the volume ratio of ethanol to ethyl acetate in the ethanol-ethyl acetate mixed solvent is 1: 1-6.
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