CN111588651A - Preparation method of selenium-rich collagen solution based on collagen artificial product - Google Patents
Preparation method of selenium-rich collagen solution based on collagen artificial product Download PDFInfo
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- CN111588651A CN111588651A CN202010404475.2A CN202010404475A CN111588651A CN 111588651 A CN111588651 A CN 111588651A CN 202010404475 A CN202010404475 A CN 202010404475A CN 111588651 A CN111588651 A CN 111588651A
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- Prior art keywords
- collagen
- solution
- acid
- selenium
- aqueous solution
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 36
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 36
- 239000011669 selenium Substances 0.000 title claims abstract description 36
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- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 claims description 8
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- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 6
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- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 4
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 4
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Images
Classifications
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/02—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from meat
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
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- A61L27/3637—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the origin of the biological material other than human or animal, e.g. plant extracts, algae
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Abstract
The invention discloses a preparation method of a selenium-rich collagen solution based on a collagen artificial product, and relates to the technical field of collagen preparation; in order to solve the problem of poor mouthfeel; the method specifically comprises the following steps: weighing collagen powder, placing the weighed collagen powder in a clean container, soaking the collagen powder in an acetic acid aqueous solution, and performing vortex or slight ultrasonic acceleration on the collagen powder to dissolve the collagen powder to prepare a collagen aqueous solution; weighing the polymer, placing the polymer in a clean container, dissolving the polymer by double distilled water, and performing vortex or stirring to accelerate the dissolution to prepare a biological polymer aqueous solution. The invention adds high-quality extracted selenium element, prepares jelly-like collagen products by the processes of mixing, glue boiling, filtering, blending, filling, cup sealing, sterilizing, cooling, packaging and the like, has rich raw materials, unique flavor and moderate elasticity and toughness, effectively solves the defects of over-fast degradation and poor taste of collagen, and meets the application requirements of beauty treatment, skin care, biological medicines and the like.
Description
Technical Field
The invention relates to the technical field of collagen preparation, in particular to a preparation method of a selenium-rich collagen solution based on a collagen artificial product.
Background
Collagen is a kind of scleroprotein constituting animal bones, cartilage, teeth, tendons, skin and fish scales, is a fibrous solid, and is in a tangled periodic structure with cross-stripes as observed under an electron microscope, is a structural protein, is often present in various mammals, and constitutes 30% of the total weight of all proteins of the body, and there are currently known 20 kinds of collagen, of which type I collagen is the largest, and is formed by transversely linking monomeric proteins having a molecular weight of about 300kDa through specific site covalent bonds, mature collagen assumes a typical fibrous shape, is insoluble in water, has high tensile strength, is composed of constitutive amino acids, has a structure in which three polypeptide chains are twisted spirally around another polypeptide chain through hydrogen bonds, collagen is not degraded in water, weak acid and weak base, but it is changed into gelatin of single chain structure after boiling, is soluble, has a certain viscosity without heating unlike gelatin, is easily gelated, and is more suitable for biological tissues and has high bioactivity since the molecular weight of collagen is larger than that of gelatin.
Through retrieval, the Chinese patent with the application number of CN200910248823.5 discloses a chitosan collagen gel and application thereof, which comprises 1 to 15 percent of chitosan, 1 to 20 percent of collagen, 0.1 to 1 percent of gel matrix and the balance of water; the chitosan is water-soluble chitosan, and the molecular weight of the chitosan is 1500-40 ten thousand daltons; the molecular weight of the collagen is 5000 daltons to 300 ten thousand daltons; the gel matrix is one or two of carbomer, polyethylene and cellulose derivative. The chitosan collagen gel and the application thereof in the patent have the following defects: the obtained collagen product has good thickening property, but has poor taste.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a preparation method of a selenium-rich collagen solution based on a collagen artificial product.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a selenium-rich collagen solution based on a collagen artificial product comprises the following steps:
s1: weighing collagen powder, placing the weighed collagen powder in a clean container, soaking the collagen powder in an acetic acid aqueous solution, and performing vortex or slight ultrasonic acceleration on the collagen powder to dissolve the collagen powder to prepare a collagen aqueous solution;
s2: weighing the macromolecules, placing the macromolecules into a clean container, dissolving the macromolecules by using double distilled water, and performing vortex or stirring to accelerate the dissolution of the macromolecules to prepare a biopolymer aqueous solution;
s3: mixing the collagen aqueous solution obtained in the step S1 with the biopolymer aqueous solution obtained in the step S2, and uniformly stirring to obtain a mixed solution A;
s4: adding carrageenan into the mixed solution A, mixing, stirring uniformly, slowly adding the obtained mixture into water with the temperature of 30-50 ℃ to disperse the mixture to prevent agglomeration, and soaking for 10-20min to ensure that the carrageenan fully absorbs water and swells;
s5: slowly heating to boil, keeping the micro-boiling state for 5-8min to obtain hot gel solution, standing for a moment, and removing surface foam to obtain mixed feed liquid;
s6: filtering while hot to remove impurities and some possible colloidal particles;
s7: cooling the temperature of the mixed material liquid to below 70 ℃, and aging the mixed material liquid at a low temperature of-9-12 ℃ for a certain time, wherein the aging time is 4-72 h, and the low temperature is-4-10 ℃; the aging time is 12-24 h;
s8: and filling, sealing the cup, sterilizing and cooling to obtain the jelly-like collagen product.
Preferably: the volume mass fraction of collagen in the acetic acid aqueous solution in the S1 is 0.1-100 mg-kg-1The volume mass fraction is 1-10 mg/kg-1。
Preferably: the volume mass fraction of the biopolymer in the aqueous solution in the S2 is 0.1-50mg kg-1The volume mass fraction is 0.1-20 mg/kg-1。
Preferably: the pH value of the mixed solution A in the S3 is adjusted to 6.0-7.3, and is adjusted by adding an alkaline substance or an acidic substance, wherein the alkaline substance is any one or a mixture of more than two of sodium hydroxide, potassium hydroxide and sodium carbonate; the acidic substance is any one or mixture of more than two of hydrochloric acid, sulfuric acid and nitric acid.
Preferably: the collagen product in the S8 is at a temperature of between 20 ℃ below zero and 5 ℃, the mass contents of the collagen and the biopolymer in the collagen product are respectively 0.1 to 1 percent and 1 to 15 percent, the mass content of the organic selenium is more than or equal to 150 mu g/kg, and the balance is water.
Preferably: the preparation method of the collagen powder in the S1 comprises the following steps:
s11: selecting connective tissues on an animal body, wherein the selected surface area of the connective tissues is 20 square millimeters to 2 square meters;
s12: immersing the connective tissue in a first acid solution to expand the connective tissue by at least one time to form an expanded connective tissue;
s13: cleaning the expanded connective tissue with a cleaning solution to form a collagen matrix;
s14: extracting collagen from the collagen matrix by using the extraction liquid to form a solution containing the collagen;
s15: purifying;
s16: concentrating and drying to obtain collagen powder.
Preferably: the animal in S11 is selected from cattle, pig, horse, sheep, chicken, duck, turkey, goose, whale or shark, and specifically comprises dermal tissue, subcutaneous tissue, ligament, tendon, aponeurosis, cartilage and bone tissue, wherein the pig is selected from connective tissue treated with organic solvent such as alcohol, ketone, acetone, acetonitrile, chloroform, N-dimethylformamide, dimethyl sulfoxide or mixture of organic solvent and water, and the volume ratio is 1: 1 to 5.
Preferably: the first acid solution in the S12 is organic acid, such as formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid or a mixture thereof, and the concentration of the first acid solution is 0.6-3.2 mol/L.
Preferably: the cleaning liquid in the S13 comprises the following raw materials in parts by weight: 100-150 parts of water, 1-10 parts of surfactant, 1-10 parts of chelating agent, 1-10 parts of proteolytic enzyme, 0.1-1 part of 1-methyl-3-ethylimidazole chloronium salt and 1-5 parts of sodium chloride.
Preferably: the preparation method of the organic selenium in the S3 comprises the following steps:
s31: screening qualified cardamine hupingshanensis plants, cleaning, and drying by drying or naturally drying in the sun;
s32: crushing the dried cardamine hupingshanesis plant, adding pure clear water into the crushed cardamine hupingshanesis plant, soaking for 30 minutes, and controlling the material-liquid ratio to be 1: 20;
s33: slowly heating the feed liquid to 90 ℃, stirring and extracting for 1 hour at constant temperature, and controlling the ratio of the feed liquid to be 1: 20, adding the stirred and extracted feed liquid into a filtering device, and filtering to obtain liquid;
s34: and carrying out spray drying or reduced pressure distillation drying on the obtained liquid to obtain the organic selenium.
The invention has the beneficial effects that: the method has the advantages that the high-purity collagen can be obtained, the extraction time is shortened, the extraction rate is improved, the performance is uniform and stable, the jelly-shaped collagen product is prepared by adding the high-quality extracted selenium element and carrying out the processes of mixing, glue boiling, filtering, blending, filling, cup sealing, sterilizing, cooling, packaging and the like, the raw materials are rich, the flavor is unique, the elasticity and the toughness are moderate, the defect that the collagen is degraded too fast and the taste is not good is effectively overcome, the safety performance is high, and the method can be applied to beauty skin care, acne removal, anti-inflammation, tissue engineering, biological medicines and the like.
Drawings
Fig. 1 is a schematic flow structure diagram of a preparation method of a selenium-rich collagen solution based on a collagen artificial product provided by the invention.
Detailed Description
The technical solution of the present patent will be described in further detail with reference to the following embodiments.
Reference will now be made in detail to embodiments of the present patent, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are exemplary only for the purpose of explaining the present patent and are not to be construed as limiting the present patent.
In the description of this patent, it is to be understood that the terms "center," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in the orientations and positional relationships indicated in the drawings for the convenience of describing the patent and for the simplicity of description, and are not intended to indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the patent.
In the description of this patent, it is noted that unless otherwise specifically stated or limited, the terms "mounted," "connected," and "disposed" are to be construed broadly and can include, for example, fixedly connected, disposed, detachably connected, disposed, or integrally connected and disposed. The specific meaning of the above terms in this patent may be understood by those of ordinary skill in the art as appropriate.
Example 1:
a method for preparing selenium-rich collagen solution based on collagen artificial product, as shown in figure 1, comprises the following steps:
s1: weighing collagen powder, placing the weighed collagen powder in a clean container, soaking the collagen powder in an acetic acid aqueous solution, and performing vortex or slight ultrasonic acceleration on the collagen powder to dissolve the collagen powder to prepare a collagen aqueous solution;
s2: weighing the macromolecules, placing the macromolecules into a clean container, dissolving the macromolecules by using double distilled water, and performing vortex or stirring to accelerate the dissolution of the macromolecules to prepare a biopolymer aqueous solution;
s3: mixing the collagen aqueous solution obtained in the step S1 with the biopolymer aqueous solution obtained in the step S2, uniformly stirring, adding organic selenium, and stirring and mixing to obtain a mixed solution A;
s4: adding carrageenan into the mixed solution A, mixing, stirring uniformly, slowly adding the obtained mixture into water with the temperature of 30-50 ℃ to disperse the mixture to prevent agglomeration, and soaking for 10-20min to ensure that the carrageenan fully absorbs water and swells;
s5: slowly heating to boil, keeping the micro-boiling state for 5-8min to obtain hot gel solution, standing for a moment, and removing surface foam to obtain mixed feed liquid;
s6: filtering while hot to remove impurities and some possible colloidal particles;
s7: cooling the temperature of the mixed solution to below 70 ℃, aging the mixed solution at a low temperature of-9-12 ℃ for a certain time, wherein the aging time is 4-72 h, preferably, the low temperature in the embodiment is-4-10 ℃; the aging time is 12-24 h;
s8: and filling, sealing the cup, sterilizing and cooling to obtain the jelly-like collagen product.
The volume mass fraction of collagen in the acetic acid aqueous solution in the S1 is 0.1-100 mg-kg-1Preferably, this is done hereIn the case, the volume mass fraction is 1-10 mg/kg-1。
The volume mass fraction of the biopolymer in the aqueous solution in the S2 is 0.1-50mg kg-1Preferably, the volume mass fraction in the embodiment is 0.1-20 mg-kg-1。
The pH value of the mixed solution A in the S3 is adjusted to 6.0-7.3, and is adjusted by adding alkaline substances or acidic substances, wherein the alkaline substances are any one or a mixture of more than two of sodium hydroxide, potassium hydroxide, sodium carbonate and the like; the acidic substance is any one or mixture of more than two of hydrochloric acid, sulfuric acid, nitric acid, etc.
The collagen product in the S8 is at a temperature of between 20 ℃ below zero and 5 ℃, the mass contents of the collagen and the biopolymer in the collagen product are respectively 0.1 to 1 percent and 1 to 15 percent, the mass content of the organic selenium is more than or equal to 150 mu g/kg, and the balance is water.
The preparation method of the collagen powder in the S1 comprises the following steps:
s11: selecting connective tissues on an animal body, wherein the selected surface area of the connective tissues is 20 square millimeters to 2 square meters;
s12: immersing the connective tissue in a first acid solution to expand the connective tissue by at least one time to form an expanded connective tissue;
s13: cleaning the expanded connective tissue with a cleaning solution to form a collagen matrix;
s14: extracting collagen from the collagen matrix by using the extraction liquid to form a solution containing the collagen;
s15: purifying;
s16: concentrating and drying to obtain collagen powder.
In S11, the animal such as cattle, pig, horse, sheep, chicken, duck, turkey, goose, whale or shark is domestic animal or fish, the specific parts are dermal tissue, subcutaneous tissue, ligament, tendon, aponeurosis, cartilage and bone tissue, preferably, the pig is used in this embodiment, the pig is rich in selenium, the connective tissue can be treated by suitable organic solvent such as alcohol, ketone, acetone, acetonitrile, chloroform, N-dimethylformamide, dimethyl sulfoxide or mixture of organic solvent and water, and the volume ratio is 1: 1-5, so that the organic solvent can permeate into the connective tissue.
The first acid solution in the S12 is an organic acid, such as formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid or a mixture thereof, and the concentration of the first acid solution is 0.6-3.2 mol/L.
The cleaning liquid in the S13 comprises the following raw materials in parts by weight: 100-150 parts of water, 1-10 parts of surfactant, 1-10 parts of chelating agent, 1-10 parts of proteolytic enzyme, 0.1-1 part of 1-methyl-3-ethylimidazole chloronium salt and 1-5 parts of sodium chloride to obtain a high-purity collagen matrix, wherein the surfactant is one of sodium dodecyl sulfate, Tego compound, cetylpyridinium chloride and cetyltrimethylammonium bromide; the chelating agent is one of ethylenediamine tetraacetic acid, 1, 4, 7, 10-tetraazacyclododecyl-1, 4, 7, 10 '-tetraacetic acid and 1, 4, 7, 10-tetraazacyclododecyl-1, 4, 7, 10' -tetramethylene phosphoric acid; the proteolytic enzyme may be ficin, pepsin, trypsin, dispase, and hemolysin to remove proteins attached to the extracellular matrix, other non-collagenous proteins, and terminal peptides of collagen molecules.
Grinding, vibrating, stirring, homogenizing, mincing, tearing, cutting, grinding, cutting and the like of the collagen matrix in the S14, and then putting the collagen matrix into an extraction liquid, stirring and mixing, wherein the extraction liquid contains a solution of organic weak acid or inorganic salt, the pH value of the solution is 3-7, and the organic weak acid is formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid, boric acid, phosphoric acid or a mixture of the organic weak acid and the inorganic salt; the inorganic salt comprises sodium chloride or potassium chloride, and the concentration of the inorganic salt is 0.2-2.0 mol/L.
The purification treatment in the S15 comprises the following steps:
s21: cooling the solution containing the collagen to 40-60 ℃, and removing solidified grease on the surface of the solution containing the collagen to obtain a solution of the degreased collagen;
s22: filtering the collagen solution to obtain a filtered solution of the degreased collagen, and adopting a 40-mesh double-layer filter screen;
s23: cooling the filtered solution of the degreased collagen to 15-25 ℃, and then performing cold storage treatment to obtain a collagen frozen initial product, wherein the temperature of the cold storage treatment is 0-4 ℃ and the time is 1-3 hours;
s24: removing impurities from the primary product to obtain collagen jelly, and storing at 0-4 deg.C by removing oil layer on the surface and a little precipitate at the bottom of the primary product.
The preparation method of the organic selenium in the S3 comprises the following steps:
s31: screening qualified cardamine hupingshanensis plants, cleaning, and drying by drying or naturally drying in the sun;
s32: crushing the dried cardamine hupingshanesis plant, adding pure clear water into the crushed cardamine hupingshanesis plant, soaking for 30 minutes, and controlling the material-liquid ratio to be 1: 20;
s33: slowly heating the feed liquid to 90 ℃, stirring and extracting for 1 hour at constant temperature, and controlling the ratio of the feed liquid to be 1: 20, adding the stirred and extracted feed liquid into a filtering device, and filtering to obtain liquid;
s34: and carrying out spray drying or reduced pressure distillation drying on the obtained liquid to obtain the organic selenium.
Example 2:
a method for preparing selenium-rich collagen solution based on collagen artificial product, as shown in figure 1, comprises the following steps:
s1: weighing collagen powder, placing the weighed collagen powder in a clean container, soaking the collagen powder in an acetic acid aqueous solution, and performing vortex or slight ultrasonic acceleration on the collagen powder to dissolve the collagen powder to prepare a collagen aqueous solution;
s2: weighing the macromolecules, placing the macromolecules into a clean container, dissolving the macromolecules by using double distilled water, and performing vortex or stirring to accelerate the dissolution of the macromolecules to prepare a biopolymer aqueous solution;
s3: mixing the collagen aqueous solution obtained in the step S1 with the biopolymer aqueous solution obtained in the step S2, and uniformly stirring to obtain a mixed solution A;
s4: adding carrageenan into the mixed solution A, mixing, stirring uniformly, slowly adding the obtained mixture into water with the temperature of 30-50 ℃ to disperse the mixture to prevent agglomeration, and soaking for 10-20min to ensure that the carrageenan fully absorbs water and swells;
s5: slowly heating to boil, keeping the micro-boiling state for 5-8min to obtain hot gel solution, standing for a moment, and removing surface foam to obtain mixed feed liquid;
s6: filtering while hot to remove impurities and some possible colloidal particles;
s7: cooling the temperature of the mixed solution to below 70 ℃, aging the mixed solution at a low temperature of-9-12 ℃ for a certain time, wherein the aging time is 4-72 h, preferably, the low temperature in the embodiment is-4-10 ℃; the aging time is 12-24 h;
s8: and filling, sealing the cup, sterilizing and cooling to obtain the jelly-like collagen product.
The volume mass fraction of collagen in the acetic acid aqueous solution in the S1 is 0.1-100 mg-kg-1Preferably, the volume mass fraction in the embodiment is 1-10 mg-kg-1。
The volume mass fraction of the biopolymer in the aqueous solution in the S2 is 0.1-50mg kg-1Preferably, the volume mass fraction in the embodiment is 0.1-20 mg-kg-1。
The pH value of the mixed solution A in the S3 is adjusted to 6.0-7.3, and is adjusted by adding alkaline substances or acidic substances, wherein the alkaline substances are any one or a mixture of more than two of sodium hydroxide, potassium hydroxide, sodium carbonate and the like; the acidic substance is any one or mixture of more than two of hydrochloric acid, sulfuric acid, nitric acid, etc.
The collagen product in the S8 is at a temperature of between 20 ℃ below zero and 5 ℃, the mass contents of the collagen and the biopolymer in the collagen product are respectively 0.1 to 1 percent and 1 to 15 percent, the mass content of the organic selenium is more than or equal to 150 mu g/kg, and the balance is water.
The preparation method of the collagen powder in the S1 comprises the following steps:
s11: selecting connective tissues on an animal body, wherein the selected surface area of the connective tissues is 20 square millimeters to 2 square meters;
s12: immersing the connective tissue in a first acid solution to expand the connective tissue by at least one time to form an expanded connective tissue;
s13: cleaning the expanded connective tissue with a cleaning solution to form a collagen matrix;
s14: extracting collagen from the collagen matrix by using the extraction liquid to form a solution containing the collagen;
s15: pouring the solution containing the collagen into a salt solution to obtain collagen precipitate;
s16: and (3) putting the collagen precipitate into a dialysis bag, putting the dialysis bag into clear water for dialysis, taking out and drying.
In S11, the animal such as cattle, pig, horse, sheep, chicken, duck, turkey, goose, whale or shark is domestic animal or fish, the specific parts are dermal tissue, subcutaneous tissue, ligament, tendon, aponeurosis, cartilage and bone tissue, preferably, the pig is used in this embodiment, the pig is rich in selenium, the connective tissue can be treated by suitable organic solvent such as alcohol, ketone, acetone, acetonitrile, chloroform, N-dimethylformamide, dimethyl sulfoxide or mixture of organic solvent and water, and the volume ratio is 1: 1-5, so that the organic solvent can permeate into the connective tissue.
The first acid solution in the S12 is an organic acid, such as formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid or a mixture thereof, and the concentration of the first acid solution is 0.6-3.2 mol/L.
The cleaning liquid in the S13 comprises the following raw materials in parts by weight: 100-150 parts of water, 1-10 parts of surfactant, 1-10 parts of chelating agent, 1-10 parts of proteolytic enzyme, 0.1-1 part of 1-methyl-3-ethylimidazole chloronium salt and 1-5 parts of sodium chloride to obtain a high-purity collagen matrix, wherein the surfactant is one of sodium dodecyl sulfate, Tego compound, cetylpyridinium chloride and cetyltrimethylammonium bromide; the chelating agent is one of ethylenediamine tetraacetic acid, 1, 4, 7, 10-tetraazacyclododecyl-1, 4, 7, 10 '-tetraacetic acid and 1, 4, 7, 10-tetraazacyclododecyl-1, 4, 7, 10' -tetramethylene phosphoric acid; the proteolytic enzyme may be ficin, pepsin, trypsin, dispase, and hemolysin to remove proteins attached to the extracellular matrix, other non-collagenous proteins, and terminal peptides of collagen molecules.
Grinding, vibrating, stirring, homogenizing, mincing, tearing, cutting, grinding, cutting and the like of the collagen matrix in the S14, and then putting the collagen matrix into an extraction liquid, stirring and mixing, wherein the extraction liquid contains a solution of organic weak acid or inorganic salt, the pH value of the solution is 3-7, and the organic weak acid is formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid, boric acid, phosphoric acid or a mixture of the organic weak acid and the inorganic salt; the inorganic salt comprises sodium chloride or potassium chloride, and the concentration of the inorganic salt is 0.2-2.0 mol/L.
The concentration of the salt solution in the S15 is 0.3-2 mol/L, and the volume ratio of the salt solution to the solution containing the collagen is 5-10: 1.
the dialysis time in the S16 is 3-5 hours, and the clear water is replaced every 30-60 minutes.
The preparation method of the organic selenium in the S3 comprises the following steps:
s31: screening qualified cardamine hupingshanensis plants, cleaning, and drying by drying or naturally drying in the sun;
s32: crushing the dried cardamine hupingshanesis plant, adding pure clear water into the crushed cardamine hupingshanesis plant, soaking for 30 minutes, and controlling the material-liquid ratio to be 1: 20;
s33: slowly heating the feed liquid to 90 ℃, stirring and extracting for 1 hour at constant temperature, and controlling the ratio of the feed liquid to be 1: 20, adding the stirred and extracted feed liquid into a filtering device, and filtering to obtain liquid;
s34: and carrying out spray drying or reduced pressure distillation drying on the obtained liquid to obtain the organic selenium.
When the collagen preparation method is used, high-purity collagen can be obtained, the extraction time is shortened, the extraction rate is improved, the performance is uniform and stable, a jelly-shaped collagen product is prepared by adding high-quality extracted selenium element and through the processes of mixing, glue boiling, filtering, blending, filling, cup sealing, sterilizing, cooling, packaging and the like, the raw materials are rich, the flavor is unique, the elasticity and the toughness are moderate, the defect that the collagen is degraded too fast and the taste is not good is effectively overcome, the safety performance is high, and the collagen preparation method can be applied to beauty skin care, acne removal and inflammation resistance, tissue engineering, biological medicines and the like.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A preparation method of a selenium-rich collagen solution based on a collagen artificial product is characterized by comprising the following steps:
s1: weighing collagen powder, placing the weighed collagen powder in a clean container, soaking the collagen powder in an acetic acid aqueous solution, and performing vortex or slight ultrasonic acceleration on the collagen powder to dissolve the collagen powder to prepare a collagen aqueous solution;
s2: weighing the macromolecules, placing the macromolecules into a clean container, dissolving the macromolecules by using double distilled water, and performing vortex or stirring to accelerate the dissolution of the macromolecules to prepare a biopolymer aqueous solution;
s3: mixing the collagen aqueous solution obtained in the step S1 with the biopolymer aqueous solution obtained in the step S2, and uniformly stirring to obtain a mixed solution A;
s4: adding carrageenan into the mixed solution A, mixing, stirring uniformly, slowly adding the obtained mixture into water with the temperature of 30-50 ℃ to disperse the mixture to prevent agglomeration, and soaking for 10-20min to ensure that the carrageenan fully absorbs water and swells;
s5: slowly heating to boil, keeping the micro-boiling state for 5-8min to obtain hot gel solution, standing for a moment, and removing surface foam to obtain mixed feed liquid;
s6: filtering while hot to remove impurities and some possible colloidal particles;
s7: cooling the temperature of the mixed material liquid to below 70 ℃, and aging the mixed material liquid at a low temperature of-9-12 ℃ for a certain time, wherein the aging time is 4-72 h, and the low temperature is-4-10 ℃; the aging time is 12-24 h;
s8: and filling, sealing the cup, sterilizing and cooling to obtain the jelly-like collagen product.
2. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 1, wherein the volume mass fraction of collagen in acetic acid aqueous solution in S1 is 0.1-100 mg-kg-1The volume mass fraction is 1-10 mg/kg-1。
3. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 2, wherein the volume mass fraction of biopolymer in the aqueous solution of S2 is 0.1-50 mg-kg-1The volume mass fraction is 0.1-20 mg/kg-1。
4. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 3, wherein the pH of the mixed solution A in S3 is adjusted to 6.0-7.3 by adding alkaline substance or acidic substance, wherein the alkaline substance is one or mixture of more than two of sodium hydroxide, potassium hydroxide and sodium carbonate; the acidic substance is any one or mixture of more than two of hydrochloric acid, sulfuric acid and nitric acid.
5. The method for preparing selenium-enriched collagen solution based on artificial collagen product as claimed in claim 4, wherein the collagen product in S8 is at-20 deg.C to 5 deg.C, the collagen and biopolymer content in the collagen product are 0.1-1% and 1-15% respectively, the organic selenium content is not less than 150 μ g/kg, and the rest is water.
6. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 1, wherein the method for preparing collagen powder in S1 comprises the following steps:
s11: selecting connective tissues on an animal body, wherein the selected surface area of the connective tissues is 20 square millimeters to 2 square meters;
s12: immersing the connective tissue in a first acid solution to expand the connective tissue by at least one time to form an expanded connective tissue;
s13: cleaning the expanded connective tissue with a cleaning solution to form a collagen matrix;
s14: extracting collagen from the collagen matrix by using the extraction liquid to form a solution containing the collagen;
s15: purifying;
s16: concentrating and drying to obtain collagen powder.
7. The method for preparing selenium-rich collagen solution based on collagen artificial product as claimed in claim 6, wherein the animal such as cattle, pig, horse, sheep, chicken, duck, turkey, goose, whale or shark at S11 is selected from dermal tissue, subcutaneous tissue, ligament, tendon, aponeurosis, cartilage and bone tissue, and the pig is treated with organic solvent such as alcohol, ketone, acetone, acetonitrile, chloroform, N-dimethylformamide, dimethyl sulfoxide or mixture of organic solvent and water at a volume ratio of 1: 1 to 5.
8. The method as claimed in claim 7, wherein the first acidic solution in S12 is an organic acid, such as formic acid, oxalic acid, acetic acid, citric acid, lactic acid, malic acid or their mixture, and has a concentration of 0.6-3.2 mol/L.
9. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 8, wherein the cleaning solution in S13 comprises the following raw materials (by weight portion): 100-150 parts of water, 1-10 parts of surfactant, 1-10 parts of chelating agent, 1-10 parts of proteolytic enzyme, 0.1-1 part of 1-methyl-3-ethylimidazole chloronium salt and 1-5 parts of sodium chloride.
10. The method for preparing selenium-enriched collagen solution based on collagen artificial product as claimed in claim 5, wherein the method for preparing organic selenium in S3 comprises the following steps:
s31: screening qualified cardamine hupingshanensis plants, cleaning, and drying by drying or naturally drying in the sun;
s32: crushing the dried cardamine hupingshanesis plant, adding pure clear water into the crushed cardamine hupingshanesis plant, soaking for 30 minutes, and controlling the material-liquid ratio to be 1: 20;
s33: slowly heating the feed liquid to 90 ℃, stirring and extracting for 1 hour at constant temperature, and controlling the ratio of the feed liquid to be 1: 20, adding the stirred and extracted feed liquid into a filtering device, and filtering to obtain liquid;
s34: and carrying out spray drying or reduced pressure distillation drying on the obtained liquid to obtain the organic selenium.
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| CN102131403A (en) * | 2009-07-27 | 2011-07-20 | 成功大学 | Preparation of high purity collagen |
| CN102391315A (en) * | 2011-08-10 | 2012-03-28 | 恩施盛硒生物科技有限公司 | Method for extracting purely natural plant organic selenium |
| CN105613939A (en) * | 2014-11-04 | 2016-06-01 | 恩施盛硒生物工程有限公司 | Pure natural plant selenoprotein preparation method |
| CN110713727A (en) * | 2018-06-27 | 2020-01-21 | 中国科学院过程工程研究所 | Collagen hydrogel prepared at low temperature, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022226731A1 (en) * | 2021-04-26 | 2022-11-03 | 北京盛美诺生物技术有限公司 | Pretreatment method for quantitative detection of undenatured type ii collagen in collagen product or cartilage, and application |
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Application publication date: 20200828 |