CN110144373A - Extract the method for chondroitin sulfate and collagen peptide respectively using animal cartilage - Google Patents
Extract the method for chondroitin sulfate and collagen peptide respectively using animal cartilage Download PDFInfo
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- CN110144373A CN110144373A CN201910457319.XA CN201910457319A CN110144373A CN 110144373 A CN110144373 A CN 110144373A CN 201910457319 A CN201910457319 A CN 201910457319A CN 110144373 A CN110144373 A CN 110144373A
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- chondroitin sulfate
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- cartilage
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- collagen peptide
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- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 title claims abstract description 74
- 229920001287 Chondroitin sulfate Polymers 0.000 title claims abstract description 74
- 229940059329 chondroitin sulfate Drugs 0.000 title claims abstract description 74
- 210000000845 cartilage Anatomy 0.000 title claims abstract description 62
- 102000008186 Collagen Human genes 0.000 title claims abstract description 58
- 108010035532 Collagen Proteins 0.000 title claims abstract description 58
- 229920001436 collagen Polymers 0.000 title claims abstract description 58
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 54
- 239000000284 extract Substances 0.000 title claims abstract description 39
- 241001465754 Metazoa Species 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims description 49
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 104
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 45
- 238000001728 nano-filtration Methods 0.000 claims abstract description 28
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000006243 chemical reaction Methods 0.000 claims abstract description 19
- 239000012141 concentrate Substances 0.000 claims abstract description 14
- 108091005804 Peptidases Proteins 0.000 claims abstract description 12
- 239000004365 Protease Substances 0.000 claims abstract description 12
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 10
- 230000000694 effects Effects 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims description 77
- 235000019441 ethanol Nutrition 0.000 claims description 69
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 28
- 239000000706 filtrate Substances 0.000 claims description 24
- 230000008569 process Effects 0.000 claims description 22
- 230000003068 static effect Effects 0.000 claims description 22
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Inorganic materials [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 235000013305 food Nutrition 0.000 claims description 17
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 16
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 16
- 239000000463 material Substances 0.000 claims description 15
- 239000008213 purified water Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 14
- 102000004190 Enzymes Human genes 0.000 claims description 11
- 108090000790 Enzymes Proteins 0.000 claims description 11
- 229910052799 carbon Inorganic materials 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 11
- 239000000047 product Substances 0.000 claims description 10
- 238000004062 sedimentation Methods 0.000 claims description 10
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 8
- 238000010792 warming Methods 0.000 claims description 7
- 239000003513 alkali Substances 0.000 claims description 6
- 239000000796 flavoring agent Substances 0.000 claims description 6
- 235000019634 flavors Nutrition 0.000 claims description 6
- 238000009928 pasteurization Methods 0.000 claims description 6
- 238000005507 spraying Methods 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 4
- 238000007254 oxidation reaction Methods 0.000 claims description 3
- 229940006423 chondroitin sulfate sodium Drugs 0.000 claims description 2
- 238000012869 ethanol precipitation Methods 0.000 claims description 2
- 239000012263 liquid product Substances 0.000 claims 1
- 238000001556 precipitation Methods 0.000 abstract description 37
- 238000000605 extraction Methods 0.000 abstract description 7
- 230000018044 dehydration Effects 0.000 abstract description 4
- 238000006297 dehydration reaction Methods 0.000 abstract description 4
- 238000000746 purification Methods 0.000 abstract description 4
- 238000001179 sorption measurement Methods 0.000 abstract description 4
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 241000251730 Chondrichthyes Species 0.000 abstract description 2
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 2
- 229920001184 polypeptide Polymers 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 43
- 238000004519 manufacturing process Methods 0.000 description 14
- 235000013402 health food Nutrition 0.000 description 8
- 238000004064 recycling Methods 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- 238000002955 isolation Methods 0.000 description 7
- 238000003756 stirring Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 239000003814 drug Substances 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 210000000988 bone and bone Anatomy 0.000 description 4
- 230000009977 dual effect Effects 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 210000003491 skin Anatomy 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 3
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 3
- 230000003796 beauty Effects 0.000 description 3
- 239000011575 calcium Substances 0.000 description 3
- 229910052791 calcium Inorganic materials 0.000 description 3
- 239000002537 cosmetic Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 208000006820 Arthralgia Diseases 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 229920002567 Chondroitin Polymers 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 230000003712 anti-aging effect Effects 0.000 description 2
- 229940030225 antihemorrhagics Drugs 0.000 description 2
- 206010003246 arthritis Diseases 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 description 2
- KXKPYJOVDUMHGS-OSRGNVMNSA-N chondroitin sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](OS(O)(=O)=O)[C@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](C(O)=O)O1 KXKPYJOVDUMHGS-OSRGNVMNSA-N 0.000 description 2
- 230000007691 collagen metabolic process Effects 0.000 description 2
- 150000002016 disaccharides Chemical group 0.000 description 2
- 239000003889 eye drop Substances 0.000 description 2
- 229940012356 eye drops Drugs 0.000 description 2
- 230000000025 haemostatic effect Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 150000003333 secondary alcohols Chemical class 0.000 description 2
- 239000010865 sewage Substances 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 208000025747 Rheumatic disease Diseases 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000006196 drop Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000020510 functional beverage Nutrition 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000003648 hair appearance Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000000774 hypoallergenic effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000000867 larynx Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000537 nasal bone Anatomy 0.000 description 1
- 208000004296 neuralgia Diseases 0.000 description 1
- 210000001331 nose Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000000552 rheumatic effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 210000004894 snout Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 210000000996 thick albumen Anatomy 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
-
- 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
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Sustainable Development (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a kind of preparation methods for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage (ox, pig, shark etc.): animal cartilage raw material is extracted with acetic acid, extract is obtained into concentrate A by ultrafiltration concentration, concentrate A is by alcohol precipitation, purification, filtering, alcohol precipitation, dehydration, dry chondroitin sulfate.It is filtered after cartilaginous element remaining in reaction kettle is hydrolyzed with protease, activated carbon adsorption is added to filter, carry out being spray-dried obtained collagen polypeptide after nanofiltration concentration, double effect concentration.The present invention realizes the higher value application of cartilage resource, high efficiency extraction isolating chondroitin sulfate and collagen peptide, raising added value of product can reduce environmental pollution simultaneously.
Description
Technical field
The present invention relates to a kind of processing technologies of animal cartilage, extract chondroitin sulfate respectively in particular with animal cartilage
With the method for collagen peptide, and in particular to first extract chondroitin sulfate with the method that acetic acid extracts and extract collagen with enzymatic isolation method again
A kind of production technology of protein peptides.
Background technique
China is a populous nation, and than more developed, cartilage resource is relatively abundanter for agricultural, animal husbandry.Animal cartilage, larynx
Drug, health care have been widely used in rich in chondroitin sulfate and collagen isoreactivity ingredient in bone, nasal bone and tracheae
In the fields such as food, cosmetics and food.
Chondroitin sulfate element (Chondroitin Sulfate, CS) is one kind by D-Glucose aldehydic acid and N- acetyl ammonia
The acid mucopolysaccharide for the repetition disaccharide unit composition that base galactolipin is constituted.It contains 50-70 disaccharide basic structure, and molecular weight is
Between 10000-50000.Can be specifically divided into according to its chemical composition and structure: chondroitin sulfate A (CSA), B, C, D, E, F, H etc. are more
Kind isomer.Chondroitin sulfate is extracted from animal cartilage, mainly chondroitin sulfate A (CSA) and chondroitin sulfate C and its
The mixture of his chondroitin sulfate.Chondroitin sulfate A (CSA) and chondroitin sulfate C have D- aminoglucose uronic acid and N- acetyl-D-
Amine-galactose composition, only the position of sulfate is different.
Chondroitin sulfate is usually and collagen combines the form for forming glycoprotein to exist.Chondroitin sulfate production method
It is divided into alkaline hydrolysiss (yield is low, impurity is more, seriously polluted be eliminated) and enzymatic isolation method (chondroitin sulfate manufacturer master at present
Stream production method) it is all that chondroitin sulfate and collagen peptide are passed through the effect of enzyme not of the same race from cartilage simultaneously while being hydrolyzed
Into same solution, resin adsorption is then had stepped through, ultrafiltration, oxidation, alcohol precipitation, is dehydrated, is dried to obtain chondroitin sulfate；Resin
Waste liquid or ultrafiltration filter liquor after absorption obtain thick collagen peptide by nanofiltration, double effect concentration and spray drying.Part is raw
Produce enterprise selective extraction chondroitin sulfate and collagen peptide is slatterned with discharge of wastewater, part producing producer, which does, simply to be mentioned
It obtains thick albumen powder to sell in the form of feed or biological fermentation culture medium, added value is very low.Enzymatic isolation method and alkaline hydrolysiss phase
Than with high income, foreign protein content is relatively low, the higher advantage of content of chondroitin sulfate, but be also not optimal life
Production method.
Chondroitin sulfate is widely used in the fields such as drug, health food, cosmetics and food, because sulfuric acid is soft
Ossein is clinically mainly used for anti-curing arthritis, treatment neuralgia, arthralgia, arteriosclerosis, dysacousis and hepatitis etc.
Disease has good curative effect to reducing blood lipid, antithrombotic, antitumor and wound healing etc. to coronary heart disease, rheumatic inflammation.Also
Can come as eye drops using.Chondroitin sulfate as health food, food, food additives in addition to have effects that it is above with
Also have the effect of emulsification, the dispelling abnormal flavor of food outside.Chondroitin sulfate is used as the adjustable skin cell metabolism of cosmetics, rush
Into the absorption of nutriment, to skin moisture-keeping and improvement hair quality.
Collagen is a kind of the most abundant functional protein of content in animal body.Account for the 33% of protein total content
Left and right.Collagen is mainly distributed in the organs such as skin, cartilage, tendon, body of gland, bone.U.S. FDA returns collagen peptide
Class is in highest security level catalogue (GRAS).It can be seen that its safety is very high.Collagen peptide have it is extraordinary it is water-soluble,
Dispersibility, trophism, absorption easy to digest, hypoallergenic.Application of the collagen peptide in food: preventive health care's food, drop
Blood pressure food, supplementary calcium food, beauty, anti-aging class health food, collagen peptide are applied in calm health food.City
Middle and high end calcium supplementing product, beauty product treat in the health food of osteoarthritis and have collagen peptide.Go out in the market
Show the solid functional beverage based on collagen peptide, also adds collagen peptide inside dairy products.Collagen peptide promotees
It is food and medicine field into the physiological function of skin collagen metabolism, quick-acting haemostatic powder, reduction serum cholesterol levels etc.
Important additive.
As the bioactivity of chondroitin sulfate and collagen peptide is constantly found and in every field using not
Disconnected is extended, extraction, purifying in relation to chondroitin sulfate and collagen peptide, isolation technics have been achieved with very big research at
Fruit.It produces extractive technique and also needs improve and perfect.Especially chondroitin sulfate and collagen peptide are separated to obtain
It is not first to hydrolyze to same solution to carry out separation acquisition again.Chondroitin sulfate manufacturer uses enzymatic isolation method production cost at present
Relatively high, yield and purity increase with respect to alkaline hydrolysiss, but production method is less than optimal that (product yield is with respect to cartilage sheet
The content of body).
Summary of the invention
The present invention in order to overcome the drawbacks of the prior art, solve technical solution used by one of above-mentioned technical problem and be:
Extract the method for chondroitin sulfate and collagen peptide respectively using animal cartilage, it is characterised in that: the method are as follows: first use
The method of acetic acid extracting side extracts chondroitin sulfate from animal cartilage, then extracts collagen peptide from remaining cartilage.
Preferably, described method includes following steps:
(1) animal cartilage is put into reaction kettle, purified water is added；35-40 DEG C of feed liquid temperature is adjusted, adjusts feed liquid with acetic acid
PH value 4-6 be stripped 5-10 hours (in extractive process keep pH value constant)；By extract releasing plate and frame filter
Filter to obtain extract A；Purified water is added again into reaction kettle, adjusts 40-60 DEG C of temperature of feed liquid, adjusts material liquid pH with NaOH
Value 8-9 is added enzyme 1 and is digested and (keep pH value constant in enzymolysis process) 4-6 hours, feed liquid temperature is adjusted after enzymatic hydrolysis
50-60 DEG C, enzyme 2 is added with the pH value 5.5-6.5 that NaOH adjusts feed liquid and is digested and (keep pH value constant in enzymolysis process) 1-3
Hour, enzymatic hydrolysis terminates, and 70-90 DEG C of heating makes to digest enzyme-deactivating used after adjusting the pH value 4-6 of feed liquid；It is 1-5 hours static, it uses
Plate and frame filter is filtered to obtain enzymolysis liquid B；
(2) preparation method of chondroitin sulfate:
(2a) is precipitated to obtain primary sedimentation sulfuric acid soft with ethyl alcohol after gained extract A in step (1) is concentrated by ultrafiltration
Ossein；
(2b) dissolves gained primary sedimentation chondroitin sulfate sodium chloride (food grade) solution in step (2a), adjusts pH value of solution
Value 10-12 heats up solution 40-50 DEG C, and it is 4-6 hours static that hydrogen peroxide (concentration of hydrogen peroxide is 1-4%) is added；
Solution plate and frame filter filters after (2c) will be static in step (2b), and filtrate ethanol precipitation is dehydrated, is drying to obtain
Chondroitin sulfate finished product；
(3) preparation method of collagen peptide:
(3a) adjusts the temperature of gained enzymolysis liquid B in step (1) to 50-70 DEG C；Addition active carbon powder (active carbon weight:
Enzymolysis liquid volume=10-20g:1L), material liquid pH value 4-6 is adjusted with hydrochloric acid, is reacted 1-4 hours；
(3b) filters gained feed liquid plate and frame filter in step (3a), and filtrate collecting and filtering apparatus nanofiltration is double after nanofiltration
Effect inspissator is concentrated to give concentrate again；
(3c) is spray-dried (inlet air temperature of spraying apparatus after gained concentrate in step (3b) is sterilized with pasteurization
220 ± 5 DEG C, 100 ± 5 DEG C of leaving air temp) up to food-grade collagen peptide.
Preferably, animal cartilage is is put into reaction kettle by processing described in step (1), the ratio of cartilage and purified water
For 1:1Kg/L(animal fresh cartilage) or the dry cartilage of 1:8Kg/L(animal)；Adjust feed liquid temperature be 35-40 DEG C, preferably 37 ±
0.5℃；The pH value 4-6 of adjusting feed liquid, preferably 4.5 ± 0.2；Extracting 5-10 hours, preferably 8 hours.
Preferably, 40-60 DEG C of temperature of adjusting feed liquid after processing is extracting described in step (1), preferably 50 ± 2
At DEG C adjust solution pH value be 8-10, preferably 9.5 ± 0.2；Enzymolysis time is 4-6 hours, preferably 5 hours.
Preferably, the mass concentration of the NaOH solution be 10-50%, preferably 20%.
Preferably, 50-60 DEG C of feed liquid temperature is adjusted after processing is digests for the first time described in step (1), preferably
55±2℃；The pH value that feed liquid is adjusted with hydrochloric acid is 5.5-6.5, preferably 6 ± 0.2；Enzymatic hydrolysis 1-4 hours, preferably 1.5 hours；
The pH value 4-6 of adjusting feed liquid after enzymatic hydrolysis, preferably 4.5 ± 0.2.
Preferably, the mass concentration of the hydrochloric acid solution be 8-30%, preferably 15%.
Preferably, it is alkali protease that processing described in step (1), which is enzyme 1, and dosage is 1-5 ‰ (with the poidometer of cartilage
Calculate), preferably 2 ‰；It is 1-3 ‰ (being calculated with the weight of cartilage) that enzyme 2, which is flavor protease dosage, preferably 1.5 ‰.
Preferably, heating described in step (1) is to be warming up to 70-90 DEG C.Preferably 80 ± 2 DEG C.
Preferably, described static 1-5 hours, preferably 3 hours.
Preferably, it is handled described in (2a, 2c) step in step (2) are as follows: alcohol, which is sunk in stillpot, to carry out.
It preferably, the use of concentration of alcohol is >=90% when precipitating, preferably with the mode that ethyl alcohol is added while stirring.
Preferably, when precipitating alcohol mixeding liquid concentration of alcohol be 60-80%, preferably 68 ± 0.5%.
Preferably, it is handled described in (2b) step in step (2) are as follows: concentration of sodium chloride solution 1-3%, preferably 1.5%；It adjusts
The pH value 10-12 of section solution, preferably 10.5 ± 0.2；Solution temperature is 40-50 DEG C, preferably 48 ± 2 DEG C；Hydrogen peroxide it is dense
Spending is 1-4%, preferably 3%.Oxidization time is 4-8 hours, preferably 6 hours.
Preferably, it is handled described in (3a) step in step (2) are as follows: the temperature of enzymolysis liquid B is adjusted to 50-70 DEG C, preferably
60±2℃；Hydrochloric acid adjust material liquid pH value 4-6, preferably 4.5 ± 0.2；The amount of active carbon is added are as follows: active carbon weight: enzymolysis liquid
Volume=10-20g:1L, preferably 15g:1L are added active carbon and react 1-4 hours, preferably 1 hour.
The beneficial effects of the present invention are embodied in: the present invention provides a kind of completely new method (acetic acid extraction process-enzymatic isolation method,
That is: chondroitin sulfate is first first extracted with acetic acid extraction process, then extracts collagen peptide with enzymatic isolation method), it is extracted from animal cartilage
Chondroitin sulfate and collagen peptide can effectively reduce cost, improve the yield of product, purity (purity up to 99%, albumen
Content≤2%), quality, improve benefit；And it reduces environmental pollution.
The present invention is by technological innovation, using new production method.First temperature production chondroitin sulfate, then pass through two kinds of enzymes
Subsection enzymolysis technology is implemented to feed liquid to produce collagen peptide.Chondroitin sulfate and collagen peptide is realized separately to produce simultaneously
In conjunction with respective extraction process ultrafiltration, purification, alcohol precipitation, the advanced production technology such as nanofiltration, sterilizing, spray drying.By alone
Production method increase the yield of product, profit margin increases, and economic benefit increases, and energy consumption reduces, the discharge of sewage
The problem of amount reduces, and reduces sewage treatment expense and reduces environmental pollution.
The present invention is by technological improvement, and using the method for separated extraction, preparation method simple process is feasible, the sulfuric acid of production
Chondroitin color is pure white, and major impurity protein content is very low≤and 2%.The purity of chondroitin sulfate very up to 99% with
On；The collagen peptide mouthfeel of reproduction is very good, and color is pure white, and the content of collagen peptide is very high.Impurity content is very
Low, all kinds of heavy metals meet national drug food, standard.
Chondroitin sulfate prepared by the present invention can be used for treating cardiovascular and cerebrovascular disease, arthralgia, arthritis, eye drops and
In health food and food etc.；
Collagen peptide prepared by the present invention is mainly used for health food, blood pressure reducing food, supplementary calcium food, beauty, anti-aging
Class health food, collagen peptide promote the life of skin collagen metabolism, quick-acting haemostatic powder, reduction serum cholesterol levels etc.
Function is managed, is the important additive in food and medicine field.
Specific embodiment
To facilitate the understanding of the present invention, the present invention is further illustrated with comparative example in conjunction with the embodiments.This field skill
Art personnel are not it will be clearly understood that following the description is defined its content just for the sake of understanding, explaining the present invention.
The specific scheme is that
One: the preparation method of chondroitin sulfate
(1) acetic acid extracts
Animal cartilage is put into reaction kettle, add purified water (ratio of cartilage and purified water be the fresh cartilage of 1:1Kg/L(animal) or
The dry cartilage of 1:8Kg/L(animal)), the temperature for adjusting feed liquid is 37 ± 0.5 DEG C, and adjusting material liquid pH value with acetic acid is 4.5 ± 0.2,
Extract 8 hours (pH value 4.5 ± 0.2 of feed liquid is remained in extractive process).After extract plate and frame filter mistake
Filter, collection filtrate are extract A.
(2) alcohol precipitation is concentrated
Extract A in step (1) is concentrated by ultrafiltration with ultrafilter, filtrate is transferred in alcohol precipitation kettle, is added ethyl alcohol (concentration >=90%)
Alcohol precipitation is carried out, the concentration of last alcohol mixeding liquid is 68 ± 0.5%, alcohol precipitation system left undisturbed overnight.
(3) dissolution purification
Supernatant in step (2) in alcohol precipitation system is transferred at ethyl alcohol recycling, it is (dense that sodium chloride solution is added into sediment
Degree is 1.5%, m/v), sediment is completely dissolved and is shifted in exquisite kettle, is heated up 48 ± 2 DEG C, is adjusted with 20% NaOH solution
Hydrogen peroxide is added in the pH value 10.5 ± 0.2 of feed liquid (concentration of the concentration of hydrogen peroxide in the solution is 3%).Static 6 hours.
(4) secondary alcohol precipitation
The solution plate and frame filter that purification is completed in step (3) is filtered, filtrate is transferred in secondary alcohol precipitation kettle, and second is added
Alcohol (concentration >=90%) carries out alcohol precipitation, and the concentration of alcohol mixeding liquid is 68 ± 0.5%, alcohol precipitation system left undisturbed overnight.
(5) it is dehydrated, is dry
By supernatant is transferred at ethyl alcohol recycling in alcohol precipitation system in step (4), first sediment is dehydrated, then in 90 ± 2 DEG C of rings
Chondroitin sulfate is dried to obtain under border.
Two: the preparation method of collagen peptide
(6) it digests, filter
The purified water of specified amount is added in remaining cartilage in step (1), is warming up to 50 ± 2 DEG C, is adjusted with 20% NaOH solution
PH value 9.5 ± 0.2 is added 2 ‰ alkali proteases (with the weight calculating of cartilage) enzymatic hydrolysis and (keeps pH value in enzymolysis process in 5 hours
It is constant).Adjusting temperature after enzymatic hydrolysis again is 55 ± 2 DEG C, adjusts pH value 6.0 ± 0.2 with 20% NaOH, 1.5 ‰ wind are added
Taste protease (is calculated) with the weight of cartilage, and enzymatic hydrolysis 1.5 hours (keeping pH value constant in enzymolysis process).Enzymatic hydrolysis terminates to adjust pH
Value 4.5 ± 0.2 heats up 80 ± 2 DEG C, static 3 hours.Static end filters to obtain enzymolysis liquid B with plate and frame filter.
(7) activated carbon adsorption
Enzymolysis liquid B in step (6) is transferred in reaction kettle, adjust 60 ± 2 DEG C of temperature, with salt acid for adjusting pH value be 4.5 ±
0.2.Active carbon (active carbon weight: enzymolysis liquid volume=15g:1L) is added to react 1 hour.Flame filter press is used after reaction
It filters and collects filtrate.
(8) nanofiltration
Filtrate in step (7) is subjected to nanofiltration with collecting and filtering apparatus (nanofiltration membrane 300Da), as conductivity≤200 μ s/cm of filter liquor
When, nanofiltration terminates.
(9) economic benefits and social benefits, sterilizing
By the trapped fluid in step (8) by dual-effect concentrator be concentrated, be concentrated to feed concentration be >=350g/L stop economic benefits and social benefits it is dense
Contracting, obtains concentrate.
(10) it sterilizes
By the gained concentrate pasteurization sterilization treatment in step (10), no bacterium concentrate is obtained.
(11) dry
Gained in step (10) is spray-dried (220 DEG C of the inlet air temperature of spraying apparatus, 100 DEG C of leaving air temp) without bacterium concentrate
Both food-grade collagen peptide is obtained.
Embodiment 1
A kind of preparation method being produced chondroitin sulfate and collagen peptide respectively using animal cartilage, is included the following steps:
(1) cartilage is handled: the fresh nose of an ox bone of 4500Kg being put into the reaction kettle of 10000L, purified water is added, until 9000L, will expect
The temperature of liquid is adjusted to 36.9 DEG C.
(2) it with the pH value 4.5 of acetic acid adjustment feed liquid, is extracted at 36.9 DEG C 8 hours and (maintains pH with acetic acid in extractive process
Value is 4.5 ± 0.2), extracting terminates, and shifts extract and filters to obtain extract with plate and frame filter, extract enters next
A process.Remaining cartilage enters another process.
The processing of extract:
Stop being concentrated when (2a) extract is concentrated by ultrafiltration with ultrafilter to 1/4 that filtrate volume is extract volume, by filtrate
Liquid is transferred in alcohol precipitation kettle, and opening stirring and ethyl alcohol (concentration 92.6%) concentration of ethyl alcohol into alcohol precipitation system is added is 68.2%.
Left undisturbed overnight.Supernatant ethyl alcohol in alcohol precipitation mixed system is transferred at ethyl alcohol recycling, primary sedimentation chondroitin sulfate is obtained.
(2b) dissolves 1.5% sodium chloride solution of primary sedimentation chondroitin sulfate, by adjusting sodium chloride solution
Amount make the concentration of chondroitin sulfate 11%.It is 10.6 with the pH value that 20% NaOH solution adjusts solution, adjusts solution temperature
It is 48.5 DEG C, is added hydrogen peroxide (hydrogen peroxide concentration is the 3% of solution concentration), static 6 hours.
(2c) will be static after solution filtered with plate and frame filter, filtrate is transferred in alcohol precipitation kettle, open stirring,
It is 68.2% that ethyl alcohol (concentration 92.6%) concentration of ethyl alcohol into alcohol precipitation system, which is added,.Room temperature left undisturbed overnight.By alcohol precipitation mixture
Supernatant ethyl alcohol is transferred at ethyl alcohol recycling in system, obtains secondary precipitation chondroitin sulfate.
Secondary precipitate is dehydrated by (2d) with ethyl alcohol (concentration 92.6%), and dehydration is three times.By sediment centrifugal drying
Both chondroitin sulfate is obtained.
(3) purified water is added again and heats up 51.2 DEG C for remaining cartilage in step (2), and adjustment pH value is 9.6, is added
9Kg alkali protease digests 5 hours；Adjustment temperature is 56.5 DEG C again, and adjustment pH value is 6.0, and 6.75Kg flavor albumen is added
Enzyme digests 1.5 hours, adjusts pH value 4.5, is warming up to 80.6 DEG C, static 3 hours.It is filtered after static with plate and frame filter
Obtain enzymolysis liquid.
(4) enzymolysis liquid is transferred in reaction kettle, adjusting temperature is 60.8 DEG C, and adjusting pH value is 4.6, is added 135Kg's
Activated carbon adsorption is adsorbed 1 hour.It is filtered after absorption with plate and frame filter and filtrate is transferred in nanofiltration kettle.
Nanofiltration is carried out with collecting and filtering apparatus (nanofiltration membrane 300Da).When measuring the 186 μ s/cm of conductivity of nanofiltration filter liquor, nanofiltration terminates.
(5) nanofiltration filtrate is concentrated with dual-effect concentrator, the concentration for being concentrated to material is 376g/L.Stopping is concentrated to give concentration
Liquid.
(6) concentrate is sterilized with pasteurization, is opened and 223 DEG C of inlet air temperature of adjustable spraying equipment, out wind-warm syndrome
It spends 102 DEG C and is spray-dried to obtain collagen peptide.
(7) two kinds of production technology products obtained therefrom tables of comparisons (see the table below 1):
Embodiment 2
A kind of preparation method for being extracted chondroitin sulfate and collagen peptide respectively using animal cartilage, is included the following steps:
(1) cartilage is handled: the fresh hog snout bone of 4500Kg is put into the reaction kettle of 10000L, purified water is added, until 9000L,.
(2) feed liquid temperature is adjusted to 36.8 DEG C, adjusts material liquid pH value 4.4 with acetic acid, extract 8 hours (in extractive process
PH value 4.5 ± 0.2 is maintained with acetic acid), extracting terminates, and shifts extract with plate and frame filter and filters to obtain extract, and enters
Next process.Remaining cartilage enters another process.
The processing of extract:
Stop being concentrated when (2a) extract is concentrated by ultrafiltration with ultrafilter to 1/4 that filtrate volume is extract volume, it will
Filtrate is transferred in alcohol precipitation kettle, opens stirring addition ethyl alcohol (concentration 93.1%) concentration of ethyl alcohol into alcohol precipitation system and is
68.2%.Left undisturbed overnight.Supernatant ethyl alcohol in alcohol precipitation mixed system is transferred at ethyl alcohol recycling, primary sedimentation chondroitin sulfate is obtained
Element.
(2b) dissolves 1.5% sodium chloride solution of primary sedimentation chondroitin sulfate, by adjusting sodium chloride solution
Amount make the concentration 10% of chondroitin sulfate.Adjusting solution ph with 20% NaOH solution is 10.6, adjusts solution temperature and is
48.1 DEG C, hydrogen peroxide, which is added, makes the 3% of hydrogen peroxide concentration solution concentration, and static 6 hours.
Solution is filtered with plate and frame filter after (2c) will be static, and filtrate is transferred in alcohol precipitation kettle, is opened stirring, is added
The concentration for entering ethyl alcohol (concentration 93.1%) ethyl alcohol into alcohol precipitation system is 67.8%.Left undisturbed overnight.On in alcohol precipitation mixed system
Clear ethyl alcohol is transferred at ethyl alcohol recycling, obtains secondary precipitation chondroitin sulfate.
Secondary precipitation chondroitin sulfate is dehydrated by (2d) with ethyl alcohol (concentration 93.1%), and dehydration is three times.By sulfate precipitate
Chondroitin centrifugal drying is up to chondroitin sulfate.
(3) purified water is added again and is warming up to 51.0 DEG C for remaining cartilage in step (2), and adjusting pH value is 9.7, adds
Enter 9Kg alkali protease to digest 5 hours (keeping material liquid pH value in enzymolysis process is 9.5 ± 0.2)；Adjusting temperature again is 56.5
DEG C, adjusting pH value is 6.0, and 6.75Kg flavor protease enzymatic hydrolysis being added 1.5 hours, (it is 6.0 that material liquid pH value is kept in enzymolysis process
± 0.2) material liquid pH value 4.4, is adjusted, is warming up to 80.5 DEG C, static 3 hours.It is filtered after static with plate and frame filter
Enzymolysis liquid.
(4) enzymolysis liquid is transferred in reaction kettle, adjustment temperature is 61.0 DEG C, and adjustment pH value is 4.5, and it is living that 135Kg is added
Property charcoal react 1 hour.It is filtered after reaction with plate and frame filter and filtrate is transferred in nanofiltration kettle.It (is received with collecting and filtering apparatus
Filter membrane is 300Da) nanofiltration concentration.As the 192 μ s/cm of conductivity of nanofiltration filter liquor, nanofiltration terminates.
(5) nanofiltration filtrate is concentrated with dual-effect concentrator, the concentration for being concentrated to material is 367g/L.Stop concentration
Liquid.
(6) concentrate is sterilized with pasteurization, opens simultaneously 220 DEG C of adjustable spraying equipment inlet air temperature, leaving air temp
103 DEG C are spray-dried to obtain collagen peptide.
(7) two kinds of production technology products obtained therefrom tables of comparisons (see the table below 2):
Embodiment 3
A kind of preparation method for being extracted chondroitin sulfate and collagen peptide respectively using animal cartilage, is included the following steps:
(1) cartilage is handled: the fresh shark end bone of 4500Kg being put into the reaction kettle of 10000L, adds purified water, until 9000L.
(2) temperature of feed liquid is adjusted to 37.1 DEG C, the pH value 4.6 of feed liquid is adjusted with acetic acid, extracting (extracted for 8 hours
The pH value that feed liquid is kept in journey is 4.5 ± 0.2), extracting terminates, and shifts extract and filters to obtain extracting with plate and frame filter
Liquid, into next process.Remaining cartilage enters another process.
The processing of extract:
Stop being concentrated when (2a) extract is concentrated by ultrafiltration with ultrafilter to 1/4 that filtrate volume is extract volume, by filtrate
It is transferred in alcohol precipitation kettle, opening stirring and ethyl alcohol (concentration 92.2%) concentration of ethyl alcohol into alcohol precipitation system is added is 68.7%, quiet
Only overnight.Supernatant ethyl alcohol in alcohol precipitation mixed system is transferred at ethyl alcohol recycling, primary sedimentation chondroitin sulfate is obtained.
(2b) dissolves 1.5% sodium chloride solution of primary sedimentation chondroitin sulfate, by the amount for adjusting sodium chloride solution
Making chondroitin sulfate concentration is 11%.It is 10.7 with the pH value that 20% NaOH solution adjusts solution, adjusting solution temperature is 48.4
DEG C, it is added hydrogen peroxide (concentration of hydrogen peroxide is the 3% of solution concentration), static 6 hours.
(2c) will be static after solution filtered with plate and frame filter, filtrate is transferred in alcohol precipitation kettle, open stirring,
It is 67.0% that ethyl alcohol (concentration 92.2%) concentration of ethyl alcohol into alcohol precipitation system, which is added,.Left undisturbed overnight.It will be in alcohol precipitation mixed system
Supernatant ethyl alcohol is transferred at ethyl alcohol recycling, obtains secondary precipitation chondroitin sulfate.
Secondary precipitation chondroitin sulfate is dehydrated by (2d) with ethyl alcohol (concentration 92.2%), and dehydration is three times.By precipitated sulfur
Aching and limp ossein centrifugal drying is up to chondroitin sulfate.
(3) purified water is added again and heats up 51.4 DEG C for remaining cartilage in step (2), and adjusting pH value is 9.6, is added
9Kg alkali protease digests 5 hours (pH value that feed liquid is kept in enzymolysis process is 9.5 ± 0.2)；Adjusting temperature again is 56.0
DEG C, adjusting pH value is 5.9, and 6.75Kg flavor protease enzymatic hydrolysis being added 1.5 hours, (pH value of holding feed liquid is in enzymolysis process
6.0 ± 0.2), enzymatic hydrolysis terminates to adjust material liquid pH value 4.7, is warming up to 80.1 DEG C, static 3 hours.With plate and frame mistake after static
Filter filters to obtain enzymolysis liquid.
(4) enzymolysis liquid is transferred in reaction kettle, adjust temperature be 61.2 DEG C, adjust pH value be 4.6, be added active carbon into
Reaction 1 hour.It is filtered after reaction with plate and frame filter and filtrate is transferred in nanofiltration kettle.With collecting and filtering apparatus (nanofiltration membrane
Nanofiltration is carried out for 300Da).As the 189 μ s/cm of conductivity of nanofiltration filter liquor, nanofiltration terminates.
(5) nanofiltration filtrate is concentrated with dual-effect concentrator, the concentration for being concentrated to material is 370g/L.Stop concentrate.
(6) concentrate is sterilized with pasteurization, 218 DEG C of adjustable spraying equipment inlet air temperature, 101 DEG C of leaving air temp sprays
Mist it is dry collagen peptide.
(7) two kinds of production technology products obtained therefrom tables of comparisons (see the table below 3):
The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；Although referring to foregoing embodiments
Invention is explained in detail, those skilled in the art should understand that: it still can be to aforementioned each implementation
Technical solution documented by example is modified, or equivalent substitution of some or all of the technical features；And these
It modifies or replaces, the range for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution should all
Cover within the scope of the claims and the description of the invention；To those of ordinary skill in the art, to this hair
Any alternate modification or transformation that bright embodiment is made are fallen within the scope of protection of the present invention.
Place is not described in detail by the present invention, is the well-known technique of those skilled in the art of the present technique.
Claims (10)
1. extracting the method for chondroitin sulfate and collagen peptide respectively using animal cartilage, it is characterised in that: the method are as follows:
Chondroitin sulfate is first extracted from animal cartilage using the method for acetic acid extracting side, then extracts collagen from remaining cartilage
Peptide.
2. the method according to claim 1 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
Be characterized in that: described method includes following steps:
(1) animal cartilage is put into reaction kettle, purified water is added；35-40 DEG C of feed liquid temperature is adjusted, adjusts feed liquid with acetic acid
PH value 4-6 be stripped 5-10 hours (in extractive process keep pH value constant)；By extract releasing plate and frame filter
Filter to obtain extract A；Purified water is added again into reaction kettle, adjusts 40-60 DEG C of temperature of feed liquid, adjusts material liquid pH with NaOH
Value 8-9 is added enzyme 1 and is digested and (keep pH value constant in enzymolysis process) 4-6 hours, feed liquid temperature is adjusted after enzymatic hydrolysis
50-60 DEG C, enzyme 2 is added with the pH value 5.5-6.5 that NaOH adjusts feed liquid and is digested and (keep pH value constant in enzymolysis process) 1-3
Hour, enzymatic hydrolysis terminates, and 70-90 DEG C of heating makes to digest enzyme-deactivating used after adjusting the pH value 4-6 of feed liquid；It is 1-5 hours static, it uses
Plate and frame filter is filtered to obtain enzymolysis liquid B；
(2) preparation method of chondroitin sulfate:
(2a) is precipitated to obtain primary sedimentation sulfuric acid soft with ethyl alcohol after gained extract A in step (1) is concentrated by ultrafiltration
Ossein；
(2b) dissolves gained primary sedimentation chondroitin sulfate sodium chloride (food grade) solution in step (2a), adjusts pH value of solution
Value 10-12 heats up solution 40-50 DEG C, and it is 4-6 hours static that hydrogen peroxide (concentration of hydrogen peroxide is 1-4%) is added；
Solution plate and frame filter filters after (2c) will be static in step (2b), and filtrate ethanol precipitation is dehydrated, is drying to obtain
Chondroitin sulfate finished product；
(3) preparation method of collagen peptide:
(3a) adjusts the temperature of gained enzymolysis liquid B in step (1) to 50-70 DEG C；Addition active carbon powder (active carbon weight:
Enzymolysis liquid volume=10-20g:1L), material liquid pH value 4-6 is adjusted with hydrochloric acid, is reacted 1-4 hours；
(3b) filters gained feed liquid plate and frame filter in step (3a), and filtrate collecting and filtering apparatus nanofiltration is double after nanofiltration
Effect inspissator is concentrated to give concentrate again；
(3c) is spray-dried (inlet air temperature of spraying apparatus after gained concentrate in step (3b) is sterilized with pasteurization
220 ± 5 DEG C, 100 ± 5 DEG C of leaving air temp) up to food-grade collagen peptide.
3. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: animal cartilage is put into reaction kettle to, and the ratio of cartilage and purified water is 1 for processing described in step (1):
1Kg/L(animal fresh cartilage) or the dry cartilage of 1:8Kg/L(animal)；Adjusting feed liquid temperature is 35-40 DEG C, preferably 37 ± 0.5 DEG C；
The pH value 4-6 of adjusting feed liquid, preferably 4.5 ± 0.2；Extracting 5-10 hours, preferably 8 hours.
4. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: being adjusted at 40-60 DEG C of temperature, preferably 50 ± 2 DEG C of feed liquid after processing is extracting described in step (1)
Adjust solution pH value be 8-10, preferably 9.5 ± 0.2；Enzymolysis time is 4-6 hours, preferably 5 hours.
5. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: 50-60 DEG C of feed liquid temperature is adjusted after processing is digests for the first time described in step (1), preferably 55 ± 2
℃；The pH value that feed liquid is adjusted with hydrochloric acid is 5.5-6.5, preferably 6 ± 0.2；Enzymatic hydrolysis 1-4 hours, preferably 1.5 hours；Enzymatic hydrolysis
After adjust feed liquid pH value 4-6, preferably 4.5 ± 0.2.
6. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: it is alkali protease that processing described in step (1), which is enzyme 1, dosage is 1-5 ‰ (being calculated with the weight of cartilage),
Preferably 2 ‰；It is 1-3 ‰ (being calculated with the weight of cartilage) that enzyme 2, which is flavor protease dosage, preferably 1.5 ‰.
7. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: heating described in step (1) is to be warming up to 70-90 DEG C；Preferably 80 ± 2 DEG C.
8. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: being handled described in (2a, 2c) step in step (2) are as follows: alcohol, which is sunk in stillpot, to carry out.
9. the method according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage,
It is characterized by: being handled described in (2b) step in step (2) are as follows: concentration of sodium chloride solution 1-3%, preferably 1.5%；It adjusts molten
The pH value 10-12 of liquid, preferably 10.5 ± 0.2；Solution temperature is 40-50 DEG C, preferably 48 ± 2 DEG C；The concentration of hydrogen peroxide is
1-4%, preferably 3%；Oxidization time is 4-8 hours, preferably 6 hours.
10. the side according to claim 1 or 2 for extracting chondroitin sulfate and collagen peptide respectively using animal cartilage
Method, it is characterised in that: handled described in (3a) step in step (2) are as follows: the temperature of enzymolysis liquid B is adjusted to 50-70 DEG C, preferably 60
±2℃；Hydrochloric acid adjust material liquid pH value 4-6, preferably 4.5 ± 0.2；The amount of active carbon is added are as follows: active carbon weight: enzymatic hydrolysis liquid
Product=10-20g:1L, preferably 15g:1L are added active carbon and react 1-4 hours, preferably 1 hour.
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| CN113234181A (en) * | 2021-05-05 | 2021-08-10 | 山东广昊生物制品有限公司 | Preparation method of chondroitin sulfate |
| CN115340595A (en) * | 2022-05-09 | 2022-11-15 | 湖南伍星生物科技有限公司 | Production method for preparing bovine cartilage collagen peptide by novel degreasing process |
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