CN102839207A - Method for preparing collagen peptide from fish scales - Google Patents
Method for preparing collagen peptide from fish scales Download PDFInfo
- Publication number
- CN102839207A CN102839207A CN2012103434041A CN201210343404A CN102839207A CN 102839207 A CN102839207 A CN 102839207A CN 2012103434041 A CN2012103434041 A CN 2012103434041A CN 201210343404 A CN201210343404 A CN 201210343404A CN 102839207 A CN102839207 A CN 102839207A
- Authority
- CN
- China
- Prior art keywords
- fish scales
- decalcification
- protease
- fish
- collagen peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 88
- 102000008186 Collagen Human genes 0.000 title claims abstract description 83
- 108010035532 Collagen Proteins 0.000 title claims abstract description 83
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 81
- 229920001436 collagen Polymers 0.000 title claims abstract description 77
- 238000000034 method Methods 0.000 title claims abstract description 26
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 39
- 239000004365 Protease Substances 0.000 claims abstract description 37
- 108091005804 Peptidases Proteins 0.000 claims abstract description 33
- 238000001035 drying Methods 0.000 claims abstract description 30
- 235000019419 proteases Nutrition 0.000 claims abstract description 28
- 102000004190 Enzymes Human genes 0.000 claims abstract description 25
- 108090000790 Enzymes Proteins 0.000 claims abstract description 25
- 229940088598 enzyme Drugs 0.000 claims abstract description 22
- 239000000796 flavoring agent Substances 0.000 claims abstract description 22
- 235000019634 flavors Nutrition 0.000 claims abstract description 22
- 108090000145 Bacillolysin Proteins 0.000 claims abstract description 16
- 102000035092 Neutral proteases Human genes 0.000 claims abstract description 16
- 108091005507 Neutral proteases Proteins 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 108090000526 Papain Proteins 0.000 claims abstract description 8
- 235000019834 papain Nutrition 0.000 claims abstract description 8
- 229940055729 papain Drugs 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 8
- 230000007935 neutral effect Effects 0.000 claims abstract description 6
- 238000010298 pulverizing process Methods 0.000 claims abstract description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract 13
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 11
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract 2
- 239000012528 membrane Substances 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 7
- 239000013505 freshwater Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims 1
- 230000000415 inactivating effect Effects 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 35
- 239000007788 liquid Substances 0.000 abstract description 11
- 235000019658 bitter taste Nutrition 0.000 abstract description 6
- 230000002255 enzymatic effect Effects 0.000 abstract description 3
- 238000005406 washing Methods 0.000 abstract description 2
- 238000002791 soaking Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 25
- 102000035195 Peptidases Human genes 0.000 description 22
- 230000007062 hydrolysis Effects 0.000 description 20
- 238000006460 hydrolysis reaction Methods 0.000 description 20
- 235000001014 amino acid Nutrition 0.000 description 14
- 150000001413 amino acids Chemical class 0.000 description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 239000000284 extract Substances 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 9
- 238000002156 mixing Methods 0.000 description 8
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 7
- 239000011575 calcium Substances 0.000 description 7
- 229910052791 calcium Inorganic materials 0.000 description 7
- 239000012141 concentrate Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000012360 testing method Methods 0.000 description 4
- 102000012422 Collagen Type I Human genes 0.000 description 3
- 108010022452 Collagen Type I Proteins 0.000 description 3
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 210000000988 bone and bone Anatomy 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 3
- 229960002591 hydroxyproline Drugs 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 3
- QDHHCQZDFGDHMP-UHFFFAOYSA-N Chloramine Chemical class ClN QDHHCQZDFGDHMP-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 description 1
- 102100034866 Kallikrein-6 Human genes 0.000 description 1
- 101710093813 Scale keratin Proteins 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 239000003519 biomedical and dental material Substances 0.000 description 1
- 230000003570 biosynthesizing effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- -1 pharmacy Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种从鱼鳞制备胶原蛋白肽的方法，其特征在于，包括如下步骤：1）取鱼鳞，用盐酸浸泡脱钙后，清洗至中性，干燥粉碎；2）取步骤1）得到的脱钙干燥粉碎后的鱼鳞加入酶反应器中，再加入水和混合蛋白酶以提取胶原蛋白肽，提取结束后，灭酶，离心，得酶解液，其中混合蛋白酶为中性蛋白酶、木瓜蛋白酶和风味蛋白酶的混合物；3）步骤2）得到的酶解液进行膜浓缩，浓缩液冷冻干燥后得到鱼鳞胶原蛋白肽粉。本发明用中性蛋白酶、木瓜蛋白酶和风味蛋白酶一次性酶法提取鱼鳞胶原蛋白肽，不仅降低酶解时间（酶解时间9h）、提高胶原蛋白肽提取率（72.72%），而且解决胶原蛋白肽的苦味等风味问题。The invention discloses a method for preparing collagen peptides from fish scales, which is characterized in that it comprises the following steps: 1) taking fish scales, soaking in hydrochloric acid for decalcification, washing until neutral, drying and pulverizing; 2) taking step 1) to obtain The decalcified, dried and pulverized fish scales are added to the enzyme reactor, and then water and mixed protease are added to extract collagen peptides. After the extraction is completed, the enzyme is inactivated and centrifuged to obtain an enzymatic solution, wherein the mixed protease is neutral protease and papain and flavor protease; 3) The enzymolysis liquid obtained in step 2) is membrane-concentrated, and the concentrated liquid is freeze-dried to obtain fish scale collagen peptide powder. The invention uses neutral protease, papain and flavor protease to extract collagen peptides from fish scales one-time enzymatically, which not only reduces the enzymatic hydrolysis time (enzymolysis time 9h), improves the extraction rate of collagen peptides (72.72%), but also solves the problem of collagen peptides Bitterness and other flavor issues.
Description
Technical field
The invention belongs to the collagen peptide preparation field, be specifically related to a kind of method for preparing collagen peptide from fish scale.
Background technology
Collagen protein extensively is present in the animal body, accounts for 30% of the interior total protein of animal body, for the mechanicalness protection of tissue and organ and the adjusting of cell micro-environment very important effect is arranged.The type more complicated of collagen protein, that has reported just has about 27 kinds, and comparatively common have I type, II type, III type, IV type, a V type, and wherein type i collagen albumen has using value most.Type i collagen albumen all has in fields such as food-processing and packing, biomedical material, pharmacy, leather, papermaking, image, biosynthesizing and cosmetics and skincare product very to be used widely, particularly rapid at beauty and skin care and Application as Medical Material development.
Fish scale accounts for the 1-5% of fish body weight, contains rich in protein, lecithin and several mineral materials; Wherein organism accounts for 41%～55%; And protein accounts for 70% of total organic matter; Be mainly collagen protein and fish scale Keratin sulfate; Wherein collagen protein is main with type i collagen albumen, and its proterties at aspects such as immunogenicity, biocompatibility, mechanical property, biodegradability, the property digested and assimilated, moisture retention, bacterinertness, antioxygen property, securities is good; The fish scale ash oontent is about 30%, and staple is a Win 40350, and the overwhelming majority concentrates on bone bed.
China is maximum in the world fresh-water fishes producing country, and the annual according to estimates fish scale that produces reaches 50,000 tons approximately, and major part is only treated as domestic refuse at present, produces the fish scale collagen product and can solve problem of environmental pollution effectively, can make full use of Biological resources again.Simultaneously, the outburst of BSE, TSE, FMD etc. is on the hazard the security of the collagen protein that from ox-hide, pigskin, extracts traditionally.Therefore from fish scale, extract collagen protein and become more promising.
The main method of extracting fish scale collagen has acid extraction method, alkali extraction method, enzyme extraction method, associating extraction method etc.The soda acid extraction method is a traditional method, and disclosed the earliest patent was exactly that acid system extracts fish scale collagen (JP5155900) in 1993 in Japan.But because of its production cycle long, environment is polluted, so enzyme extraction method comes into one's own.
The fish scale bone bed contains the ash content that mainly exists with the Win 40350 form, need at first destroy the bone bed ash content during with the Enzymatic Extraction collagen peptide, generally all uses hydrochloric acid decomposition method.Be reducing the pollution of hydrochloric acid to environment, have report directly to extract with proteolytic enzyme yet, is not the proteolytic enzyme of using always and be difficult to industrialized problem but there is the low or used proteolytic enzyme of extraction efficiency.Extract collagen peptide and generally all use the grease and the foreign protein of alkaline purification fish scale.The collagen peptide that extracts with proteolytic enzyme generally all has bitter taste, has limited it in the application such as aspects such as food, and the debitterize of collagen peptide is the comparison difficulty.Enzymolysis product is concentrated, and also being one with the purity that improves collagen peptide has problem to be solved.
Summary of the invention
In order to solve the problems of the technologies described above, the present invention provides a kind of and prepares the novel method of collagen peptide from the fresh water fish scale, and its principal feature is for to carry out the immersion decalcification pre-treatment of disposable short period of time with hydrochloric acid to fish scale; Mix a step with neutral protease, papoid and flavor protease and extract collagen peptide, not only reduce extraction time, improve the collagen peptide extraction yield, and solve local flavor problem such as bitter taste; With peptide content and the purity of membrane-concentrated zyme extract with raising collagen peptide product.
Provided by the inventionly prepare the method for collagen peptide, comprise the steps: from fish scale
1) gets fish scale, after the decalcification of salt soak, clean to neutral drying and crushing;
2) fish scale of getting after the decalcification drying and crushing that step 1) obtains adds in the enzyme reactor; Add entry and mixing protease again with the enzymolysis and extraction collagen peptide; After extracting end, the enzyme that goes out, centrifugal; Get enzymolysis solution, wherein mixing protease is the mixture of neutral protease, papoid and flavor protease;
3) enzymolysis solution that step 2) obtains carries out membrane-concentrated, obtains the collagen peptide powder after the liquid concentrator lyophilize.
As optimal technical scheme, fish scale is with concentration 0.3 ~ 1 molL in the step 1)
-1The decalcification of salt soak, fish scale is 10:100 ~ 40:100 g/mL with the ratio of hydrochloric acid, decalcification time 5 ~ 20 h.
More preferably, in the step 1) fish scale with concentration 0.5 molL
-1The decalcification of salt soak, fish scale is 1:5 g/mL with the ratio of hydrochloric acid, decalcification time 5h.
As optimal technical scheme, step 2) described in fish scale and the ratio of water after the decalcification drying and crushing be 1:100 ~ 4:100 g/mL; In the mixing protease: neutral protease be after the decalcification drying and crushing fish scale weight 1 ~ 6%, papoid be after the decalcification drying and crushing fish scale weight 0.5 ~ 4%, flavor protease be after the decalcification drying and crushing fish scale weight 0.25 ~ 2%; Enzymolysis time 9 ~ 20 h; 35 ~ 50 ℃ of hydrolysis temperatures, pH 6.5 ~ 7.5.
More preferably, the fish scale step 2) after the decalcification drying and crushing and the ratio of water are 3:100 g/mL.
More preferably, said neutral protease 200,000 U/g, said papoid is 980,000 U/g, said flavor protease is 20 U/mg.
Further preferably, said neutral protease be after the decalcification drying and crushing fish scale weight 4%, said papoid be after the decalcification drying and crushing fish scale weight 2%, said flavor protease be after the decalcification drying and crushing fish scale weight 1%.
As optimal technical scheme, the enzyme step of going out is: enzymolysis solution is heated to more than 90 ℃.
As optimal technical scheme; Supernatant carries out the secondary membrane-concentrated in the step 3): supernatant concentrates in combined films equipment; The one-level film is selected the rolled film of molecular weight 2000Da for use; Under normal temperature, 1.0 ~ 1.5Mpa, concentrate, the secondary film is selected the rolled film of molecular weight 400Da for use, under normal temperature, 2.5-3.0Mpa, concentrates.
Preferably, said fish scale is the fresh water fish scale.
Of the present inventionly can reach following effect: handle (0.5moLL with the disposable short period of time of hydrochloric acid
-1HCL soaks 5 h) the fish scale decalcification, omit the degreasing of fish scale and taken off foreign protein technology; With the disposable Enzymatic Extraction collagen peptide of neutral protease, papoid and flavor protease; Not only reduce enzymolysis time (enzymolysis time 9 h), improve collagen peptide extraction yield (72.72%), and solve the local flavor problems such as bitter taste of collagen peptide; Concentrate collagen peptide to improve the peptide content and the purity of collagen peptide product with the two-stage membrane sepn, the peptide content of product>=96%, molecular weight are 73.5% for the 400-1000Da peptide content.
Description of drawings
Fig. 1 is the hydrochloric acid decalcification process of fish scale.
Fig. 2 is that mixed enzyme of the present invention is extracted the collagen peptide process.
Fig. 3 is a neutral protein enzyme extraction collagen peptide process.
Fig. 4 is that papoid extracts the collagen peptide process.
Fig. 5 is that flavor protease extracts the collagen peptide process.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is described further so that those skilled in the art can better understand the present invention and implementing, but the embodiment that lifts not conduct to qualification of the present invention.
Embodiment one: the fish scale pre-treatment
Fish scale for collect from the food market be master's mixing of fresh water fish scale with the crucian fish scale, clean up the back with tap water and soak decalcification with hydrochloric acid, after the decalcification with the tap water washing to neutral, after 60 ℃ down dry (24 ~ 48 h), pulverizing then with kibbler.
Analyze suitable solid-liquid ratio, concentration of hydrochloric acid and the decalcification time (time that calcium concn tends towards stability in the soak solution) of hydrochloric acid decalcification through detecting the soak solution calcium concn, calcium concn adopts the EDTA titration measuring.
Experimental result such as following table 1 under the different experimental conditions:
Fish scale decalcification experimental result under table 1 different experimental conditions
| Experiment numbers | 1 | 2 | 3 | 4 |
| The ratio of fish scale and hydrochloric acid (g/mL) | 10:100 | 20:100 | 30:100 | 40:100 |
| Concentration of hydrochloric acid (mol/L) | 0.3 | 0.5 | 0.7 | 1.0 |
| Decalcification time (h) | 20 | 5 | 10 | 15 |
| Soak solution calcium concn (mol/L) | 0.1283 | 0.3208 | 0.4985 | 0.6233 |
Under experiment 2 solid-liquid ratio 20:100 (W/V), concentration of hydrochloric acid 0.5moL/L, soak decalcification, regularly measure the soak solution calcium concn, its calcium concn change curve in time is as shown in Figure 1.Can be known that by Fig. 1 the soak solution calcium concn tends towards stability behind 3 h, its suitable time is 3-7 h, preferred 5 h.
Embodiment two: mixed enzyme is extracted collagen peptide
The fish scale of getting after the decalcification drying and crushing that the experiment 2 of embodiment one makes adds in the enzyme reactor; Add entry and mixing protease again and carry out mixed enzyme extraction collagen peptide; Enzyme goes out after mixed enzyme is extracted and finished; Behind the centrifugal 10min of 4000rpm, get enzymolysis solution, wherein mixing protease is the mixture of neutral protease, papoid and flavor protease.The enzyme concrete steps of going out are: enzymolysis solution is heated to more than 90 ℃, keeps 10min.
With extraction yield, degree of hydrolysis and the enzymolysis solution total free aminoacids amount of collagen peptide as the check and analysis index.In the leaching process, when the extraction yield of the collagen peptide in the solution tends towards stability peak, promptly be judged as to extract and finish, be recorded as enzymolysis time.
The oxyproline amount (g) * 100 that the THP amount (g) of collagen peptide extraction yield (%)=every g fish scale enzymolysis solution/every g fish scale is total.
The hydroxyproline determination that fish scale is total: accurately take by weighing raw material fish scale 1g (the fish scale powder after decalcification and the drying and crushing); Add 50mL 6mol/L HCl in 120～130 ℃ of following hydrolysis 8h; Get supernatant behind the centrifugal 10min of 4000rpm and be settled to 100mL; Get 2mL and transfer pH to 6.0, be settled to 100mL again as sample liquid.Sample thief liquid (dilution on demand) 4.0mL (4mL zero(ppm) water is used in contrast) adds chloramines T solution (time spent joins at present) 2.0mL and shakes up back normal temperature held 20min in the 25ml color-comparison tube, adds the abundant mixing of developer 2.0mL then; Behind 60 ℃ of water-bath 20min, take out cooling; Measure the OD value down in 558nm, try to achieve the oxyproline amount from typical curve and (open the person of outstanding talent, Huaihai Institute of Technology journal (natural science edition); 2004,13 (1): 53-55).
Enzymolysis solution hydroxyproline determination: get enzymolysis supernatant (dilution on demand) 4.0ml (4mL zero(ppm) water is used in contrast) after centrifugal in the 25ml color-comparison tube; Add chloramines T solution (time spent joins at present) 2.0mL and shake up back normal temperature held 20min; Add the abundant mixing of developer 2.0mL then, behind 60 ℃ of water-bath 20min, take out cooling, measure the OD value down in 558nm; Try to achieve the oxyproline amount from typical curve and (open the person of outstanding talent; Huaihai Institute of Technology journal (natural science edition), 2004,13 (1): 53-55).
Degree of hydrolysis (DH value)/%=enzymolysis solution amino acid nitrogen content/fish scale total nitrogen content * 100
The enzymolysis solution amino acid nitrogen content is measured and is adopted FTM.
The fish scale total nitrogen content is measured and is adopted nitrogen determination.
Spectrophotometry is adopted in the acidimetric estimation of enzymolysis solution free amine group.
Experimental result under various conditions and the proportioning such as following table 2:
Mixed protein enzyme extraction collagen peptide experimental result under table 2 different condition and the proportioning
| Experiment numbers | 1 | 2 | 3 | 4 |
| Fish scale after the decalcification drying and crushing and the ratio of water (g/mL) | 1:100 | 2:100 | 3:100 | 4:100 |
| Neutral protease addition (accounting for the per-cent of the fish scale weight after the decalcification drying and crushing) | 1% | 2% | 4% | 6% |
| Papain dosage (accounting for the per-cent of the fish scale weight after the decalcification drying and crushing) | 0.5% | 1% | 2% | 4% |
| Flavor protease addition (accounting for the per-cent of the fish scale weight after the decalcification drying and crushing) | 0.25% | 0.5% | 1% | 2% |
| Hydrolysis temperature (℃) | 35 | 40 | 45 | 50 |
| pH | 6.0 | 6.5 | 7.0 | 7.5 |
| Enzymolysis time (h) | 20 | 10 | 9 | 15 |
| Collagen peptide extraction yield (%) | 15.28 | 71.64 | 82.32 | 83.65 |
| Degree of hydrolysis (%) | 25.64 | 58.74 | 62.52 | 64.79 |
| Enzymolysis solution total free aminoacids amount (mg/g) | 36.57 | 80.54 | 88.56 | 89.65 |
In the table 2, comprehensive extraction effect and cost are considered, the best results of experiment 3.Test fish scale and water ratio (solid-liquid ratio) 3:100 after the 3 decalcification drying and crushing (W/V, g/mL), the neutral protease addition be after the decalcification drying and crushing fish scale weight 4%, papain dosage be after the decalcification drying and crushing fish scale weight 2%, the flavor protease addition be after the decalcification drying and crushing fish scale weight 1%, the result is as shown in Figure 2 under the condition of 45 ℃ of hydrolysis temperatures, pH 7.0.
Can know that by Fig. 2 collagen peptide extraction yield, degree of hydrolysis and enzymolysis solution total free aminoacids amount tend towards stability gradually behind the enzymolysis 6h, near reaching maximum, reach maximum behind the enzymolysis 12h behind the enzymolysis 9h; Therefore suitable enzymolysis time is 6-12h, preferred 9h.Behind the enzymolysis 9h collagen peptide extraction yield, degree of hydrolysis and enzymolysis solution total free aminoacids amount be respectively 82.32%, 62.52%, 88.56mg/g.
The used neutral protease of present embodiment is precious along Science and Technology Ltd. available from Beijing letter profit, and enzyme activity is 200,000 U/g.Used papoid is precious along Science and Technology Ltd. available from Beijing letter profit, and enzyme activity is 980,000 U/g.Used flavor protease is precious along Science and Technology Ltd. available from Beijing letter profit, and enzyme activity is 20 U/mg.
Comparative Examples one neutral protein enzyme extraction collagen peptide
Under the condition of the fish scale after the decalcification drying and crushing that the experiment 2 of embodiment one makes and water ratio (solid-liquid ratio) 3:100 (W/V), enzyme addition 4%, 45 ℃ of hydrolysis temperatures, pH 7.0, carry out proof test; Regularly measure the extraction yield of collagen peptide; Collagen peptide degree of hydrolysis and total free aminoacids amount, the result is as shown in Figure 3.Find out that by Fig. 3 behind the enzymolysis 12h, extraction yield tends towards stability, and collagen peptide degree of hydrolysis and total free aminoacids amount raise slightly.Enzymolysis 12 h collagen peptide extraction yields, degree of hydrolysis and total free aminoacids amount are respectively 48.86%, 45.74% and 40.26mg/g.
Comparative Examples two papoids extract collagen peptide
Fish scale and water ratio (solid-liquid ratio) 3:100 (W/V), enzyme addition 2%, 45 ℃ of hydrolysis temperatures, pH after the decalcification drying and crushing that the experiment 2 of embodiment one makes carry out proof test 7.0 times; Regularly measure collagen peptide extraction yield, degree of hydrolysis and enzymolysis solution total free aminoacids amount, the result is as shown in Figure 4.Find out that by Fig. 4 collagen peptide extraction yield, degree of hydrolysis and total free aminoacids amount all tend towards stability behind the enzymolysis 24h, are respectively: 20.03%, 37.41% and 28.42mg/g.
Comparative Examples three flavor proteases extract collagen peptide
Fish scale and water ratio (solid-liquid ratio) 3:100 (W/V), enzyme addition 10%, 45 ℃ of hydrolysis temperatures, pH after the decalcification drying and crushing that the experiment 2 of embodiment one makes carry out proof test 7.0 times; Regularly measure collagen peptide extraction yield, degree of hydrolysis and enzymolysis solution total free aminoacids amount, the result is as shown in Figure 5.Find out that by Fig. 5 extraction yield, degree of hydrolysis tend towards stability behind the enzymolysis 12h, and the total free aminoacids amount tends towards stability behind enzymolysis 18h.Collagen peptide extraction yield, degree of hydrolysis and total free aminoacids amount are respectively 4.86%, 52.03% and 3.14mg/g during enzymolysis 12h.Therefore to extract the extraction yield of collagen peptide separately very low for flavor protease.
Embodiment three: the membrane-concentrated collagen peptide
The enzymolysis solution centrifugal 10min under 4000rpm that makes by the experiment 3 of embodiment two gets supernatant, and will filtrate then concentrates in combined films equipment.The one-level film is selected the rolled film of molecular weight 2000Da for use, under normal temperature, 1.0-1.5Mpa, carries out.The secondary film is selected the rolled film of molecular weight 400Da for use, under normal temperature, 2.5 ~ 3.0Mpa, carries out.Carry out lyophilize behind the secondary membrane-concentrated and obtain the collagen peptide powder.
Table 3 is the collagen peptide yield of secondary membrane-concentrated, and table 4 is peptide content, MWD and the bitter taste of collagen peptide powder.
The collagen peptide yield (%) of membrane-concentrated=THP after concentrating/THP * 100 before concentrating.
Hydroxyproline determination is with embodiment two, and collagen peptide powder peptide content is measured and adopted nitrogen determination, and the collagen peptide molecular weight distribution determination adopts HPLC.
Table 3 membrane-concentrated collagen peptide yield
| Membrane-concentrated | The one-level membrane-concentrated | The secondary membrane-concentrated |
| Collagen peptide yield/% | 95.42 | 92.58 |
Table 4 collagen peptide powder peptide content, MWD and bitter taste
The above embodiment is the preferred embodiment that proves absolutely that the present invention lifts, and protection scope of the present invention is not limited thereto.Being equal to that the technician in present technique field is done on basis of the present invention substitutes or conversion, all within protection scope of the present invention.Protection scope of the present invention is as the criterion with claims.
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103434041A CN102839207A (en) | 2012-09-17 | 2012-09-17 | Method for preparing collagen peptide from fish scales |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103434041A CN102839207A (en) | 2012-09-17 | 2012-09-17 | Method for preparing collagen peptide from fish scales |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102839207A true CN102839207A (en) | 2012-12-26 |
Family
ID=47366925
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103434041A Pending CN102839207A (en) | 2012-09-17 | 2012-09-17 | Method for preparing collagen peptide from fish scales |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102839207A (en) |
Cited By (35)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103082080A (en) * | 2013-02-07 | 2013-05-08 | 广东海洋大学 | Fish skin and fish scale protein enzymolysis powder with function of relieving fatigue |
| CN103834464A (en) * | 2014-03-05 | 2014-06-04 | 青岛金海源食品有限公司 | Method for extracting omega-3 polyunsaturated fatty acid, bone collagen and active calcium from cod bone |
| CN103993061A (en) * | 2014-05-20 | 2014-08-20 | 海南美合泰生物科技有限公司 | Production method of fish collagen |
| CN104140992A (en) * | 2013-11-26 | 2014-11-12 | 国家海洋局第三海洋研究所 | Large-scale preparation method of fish scale type I collagen peptides |
| CN104232713A (en) * | 2013-06-24 | 2014-12-24 | 诸城绿维食品有限公司 | Sewage treatment process for extracting collagen corpus fibrosum from aquatic product leftovers by virtue of enzymolysis method |
| CN104419741A (en) * | 2013-08-22 | 2015-03-18 | 新肌饮(上海)投资管理有限公司 | Preparation method for nano level collagen peptide essence by taking fish scale as raw material |
| CN104479995A (en) * | 2014-12-30 | 2015-04-01 | 广州康洛信生物科技有限公司 | Sheepskin collagen extraction system and method |
| CN104945502A (en) * | 2015-06-30 | 2015-09-30 | 石狮海星食品有限公司 | ACE (angiotensin converting enzyme) inhibitory pentapeptide |
| CN105039483A (en) * | 2015-09-21 | 2015-11-11 | 广东海洋大学 | Method for preparing fishskin fish-scale collagen protein |
| CN106170213A (en) * | 2014-04-10 | 2016-11-30 | 三得利控股株式会社 | Bitterness concealing method containing collagen protein peptide combinations |
| CN106834396A (en) * | 2015-12-03 | 2017-06-13 | 威海市宇王集团有限公司 | The preparation method of fish scale collagen active peptide |
| WO2017209587A1 (en) | 2016-06-01 | 2017-12-07 | Institut National De Recherche Halieutique | Method for the extraction and purification of native marine collagen from sardina pilchardus sardine scales |
| CN107475335A (en) * | 2017-08-04 | 2017-12-15 | 青岛金典生化器材有限公司 | A kind of method that collagen is prepared using fish scale as raw material |
| CN107522781A (en) * | 2017-09-01 | 2017-12-29 | 浦江县欧立生物技术有限公司 | The method that collagen peptide is extracted from grass carp scales |
| CN107712686A (en) * | 2017-11-01 | 2018-02-23 | 安徽省林锦记食品工业有限公司 | A kind of processing method of collagen peptide jelly |
| CN108004288A (en) * | 2017-12-08 | 2018-05-08 | 厦门医学院 | One kind is without fishy smell milkfish low molecular weight fish scale collagen and preparation method thereof |
| CN108096054A (en) * | 2017-12-08 | 2018-06-01 | 厦门医学院 | The purposes of milkfish fish scale collagen |
| CN108203728A (en) * | 2018-01-19 | 2018-06-26 | 四川大学 | A kind of preparation method of collagen peptide |
| CN109170698A (en) * | 2018-09-12 | 2019-01-11 | 沈阳师范大学 | A kind of low-fat hemp nutrition powder and preparation method thereof |
| CN109371082A (en) * | 2018-10-29 | 2019-02-22 | 浙江海洋大学 | A kind of preparation method of tilapia fish scale immunomodulatory peptide |
| CN109943612A (en) * | 2017-12-21 | 2019-06-28 | 长沙理工大学 | Method for producing high-activity minced fillet and product antifreeze agent by enzymolysis of silver carp scales |
| CN111484893A (en) * | 2020-04-24 | 2020-08-04 | 中国科学院兰州化学物理研究所 | A kind of method for simultaneously preparing fish roe essential oil and fish roe functional peptide |
| CN111621537A (en) * | 2020-05-09 | 2020-09-04 | 华中农业大学 | Preparation method of freshwater fish scale collagen peptide powder |
| CN111748599A (en) * | 2020-07-28 | 2020-10-09 | 卫青健康科技(苏州)有限公司 | Preparation method of fish scale collagen peptide |
| CN112493359A (en) * | 2020-11-19 | 2021-03-16 | 郑州轻工业大学 | Preparation method of fish scale collagen peptide glue powder |
| CN112956583A (en) * | 2021-02-24 | 2021-06-15 | 北京金华雅轩健康科技有限公司 | Extraction process of fish collagen peptide |
| CN113105539A (en) * | 2021-04-13 | 2021-07-13 | 海南三元星生物科技股份有限公司 | Preparation method of silver carp scale collagen peptide |
| CN113461806A (en) * | 2021-06-17 | 2021-10-01 | 湖北瑞邦生物科技有限公司 | Preparation method of fish scale collagen and application of fish scale collagen as antioxidant and carbohydrase inhibitor |
| CN113754759A (en) * | 2021-09-16 | 2021-12-07 | 润科生物工程（福建）有限公司 | Process for extracting multiple nutritional ingredients from fish scales |
| CN114560927A (en) * | 2022-03-15 | 2022-05-31 | 厦门元之道生物科技有限公司 | Method for producing collagen peptide by subcritical water extraction assisted fish scales and application of method |
| CN115754092A (en) * | 2022-12-16 | 2023-03-07 | 江西师范大学 | Identification method of gelatin of cyprinus carpio |
| CN116262938A (en) * | 2022-12-28 | 2023-06-16 | 衢州鲟龙水产食品科技开发有限公司 | A method for preparing collagen peptides from sturgeon dark fish intestines |
| RU2810511C1 (en) * | 2022-12-14 | 2023-12-27 | Федеральное государственное автономное образовательное учреждение высшего образования "Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского" | Method of obtaining fish collagen homogenizate |
| CN117441868A (en) * | 2023-11-27 | 2024-01-26 | 山东美佳集团有限公司 | Preparation method of seafood enzymatic seasoning liquid |
| CN118994371A (en) * | 2024-08-14 | 2024-11-22 | 广州崇真生物科技有限公司 | Preparation method and application of anti-aging collagen peptide |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN102154424A (en) * | 2011-01-14 | 2011-08-17 | 国家海洋局第三海洋研究所 | Process for preparing micro-molecular fish scale collagen peptides |
-
2012
- 2012-09-17 CN CN2012103434041A patent/CN102839207A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101899491A (en) * | 2010-05-18 | 2010-12-01 | 武汉艾诗妮娜生物科技有限公司 | Method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme |
| CN102154424A (en) * | 2011-01-14 | 2011-08-17 | 国家海洋局第三海洋研究所 | Process for preparing micro-molecular fish scale collagen peptides |
Non-Patent Citations (3)
| Title |
|---|
| 刘庆慧等: "鱼鳞酶解工艺的研究", 《海洋水产研究》 * |
| 刘艳: "草鱼鳞胶原蛋白肽的制备、性质及应用研究", 《中国优秀硕士学位论文全文数据库》 * |
| 廉志清: "酶法耦合膜技术制备鱼鳞胶原蛋白低聚肽的研究", 《中国优秀硕士论文全文数据库》 * |
Cited By (43)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103082080A (en) * | 2013-02-07 | 2013-05-08 | 广东海洋大学 | Fish skin and fish scale protein enzymolysis powder with function of relieving fatigue |
| CN104232713A (en) * | 2013-06-24 | 2014-12-24 | 诸城绿维食品有限公司 | Sewage treatment process for extracting collagen corpus fibrosum from aquatic product leftovers by virtue of enzymolysis method |
| CN104419741A (en) * | 2013-08-22 | 2015-03-18 | 新肌饮(上海)投资管理有限公司 | Preparation method for nano level collagen peptide essence by taking fish scale as raw material |
| CN104140992A (en) * | 2013-11-26 | 2014-11-12 | 国家海洋局第三海洋研究所 | Large-scale preparation method of fish scale type I collagen peptides |
| CN104140992B (en) * | 2013-11-26 | 2018-10-09 | 国家海洋局第三海洋研究所 | A kind of large-scale preparation method of fish scale Type I collagen protein peptides |
| CN103834464A (en) * | 2014-03-05 | 2014-06-04 | 青岛金海源食品有限公司 | Method for extracting omega-3 polyunsaturated fatty acid, bone collagen and active calcium from cod bone |
| CN103834464B (en) * | 2014-03-05 | 2016-02-17 | 青岛中仁日用化妆品有限公司 | A kind of method extracting omega-3 polyunsaturated fatty acids and bone collagen and Calcium of intensifiens from cod bone |
| CN106170213B (en) * | 2014-04-10 | 2020-04-21 | 三得利控股株式会社 | Bitterness masking method containing collagen peptide composition |
| CN106170213A (en) * | 2014-04-10 | 2016-11-30 | 三得利控股株式会社 | Bitterness concealing method containing collagen protein peptide combinations |
| CN103993061A (en) * | 2014-05-20 | 2014-08-20 | 海南美合泰生物科技有限公司 | Production method of fish collagen |
| CN104479995B (en) * | 2014-12-30 | 2016-08-24 | 广州康洛信生物科技有限公司 | Corii Caprae seu Ovis collagen protein extraction system and extracting method |
| CN104479995A (en) * | 2014-12-30 | 2015-04-01 | 广州康洛信生物科技有限公司 | Sheepskin collagen extraction system and method |
| CN104945502A (en) * | 2015-06-30 | 2015-09-30 | 石狮海星食品有限公司 | ACE (angiotensin converting enzyme) inhibitory pentapeptide |
| CN105039483A (en) * | 2015-09-21 | 2015-11-11 | 广东海洋大学 | Method for preparing fishskin fish-scale collagen protein |
| CN106834396A (en) * | 2015-12-03 | 2017-06-13 | 威海市宇王集团有限公司 | The preparation method of fish scale collagen active peptide |
| WO2017209587A1 (en) | 2016-06-01 | 2017-12-07 | Institut National De Recherche Halieutique | Method for the extraction and purification of native marine collagen from sardina pilchardus sardine scales |
| CN107475335A (en) * | 2017-08-04 | 2017-12-15 | 青岛金典生化器材有限公司 | A kind of method that collagen is prepared using fish scale as raw material |
| CN107522781A (en) * | 2017-09-01 | 2017-12-29 | 浦江县欧立生物技术有限公司 | The method that collagen peptide is extracted from grass carp scales |
| CN107712686A (en) * | 2017-11-01 | 2018-02-23 | 安徽省林锦记食品工业有限公司 | A kind of processing method of collagen peptide jelly |
| CN108004288A (en) * | 2017-12-08 | 2018-05-08 | 厦门医学院 | One kind is without fishy smell milkfish low molecular weight fish scale collagen and preparation method thereof |
| CN108096054A (en) * | 2017-12-08 | 2018-06-01 | 厦门医学院 | The purposes of milkfish fish scale collagen |
| CN109943612A (en) * | 2017-12-21 | 2019-06-28 | 长沙理工大学 | Method for producing high-activity minced fillet and product antifreeze agent by enzymolysis of silver carp scales |
| CN109943612B (en) * | 2017-12-21 | 2022-07-19 | 长沙理工大学 | Method for producing high-activity minced fillet and product antifreeze agent by enzymolysis of silver carp scales |
| CN108203728A (en) * | 2018-01-19 | 2018-06-26 | 四川大学 | A kind of preparation method of collagen peptide |
| CN109170698A (en) * | 2018-09-12 | 2019-01-11 | 沈阳师范大学 | A kind of low-fat hemp nutrition powder and preparation method thereof |
| CN109371082A (en) * | 2018-10-29 | 2019-02-22 | 浙江海洋大学 | A kind of preparation method of tilapia fish scale immunomodulatory peptide |
| CN111484893A (en) * | 2020-04-24 | 2020-08-04 | 中国科学院兰州化学物理研究所 | A kind of method for simultaneously preparing fish roe essential oil and fish roe functional peptide |
| CN111621537B (en) * | 2020-05-09 | 2022-06-14 | 华中农业大学 | A kind of preparation method of freshwater fish scale collagen peptide powder |
| CN111621537A (en) * | 2020-05-09 | 2020-09-04 | 华中农业大学 | Preparation method of freshwater fish scale collagen peptide powder |
| CN111748599A (en) * | 2020-07-28 | 2020-10-09 | 卫青健康科技(苏州)有限公司 | Preparation method of fish scale collagen peptide |
| CN112493359A (en) * | 2020-11-19 | 2021-03-16 | 郑州轻工业大学 | Preparation method of fish scale collagen peptide glue powder |
| CN112956583A (en) * | 2021-02-24 | 2021-06-15 | 北京金华雅轩健康科技有限公司 | Extraction process of fish collagen peptide |
| CN113105539A (en) * | 2021-04-13 | 2021-07-13 | 海南三元星生物科技股份有限公司 | Preparation method of silver carp scale collagen peptide |
| CN113461806A (en) * | 2021-06-17 | 2021-10-01 | 湖北瑞邦生物科技有限公司 | Preparation method of fish scale collagen and application of fish scale collagen as antioxidant and carbohydrase inhibitor |
| CN113754759A (en) * | 2021-09-16 | 2021-12-07 | 润科生物工程（福建）有限公司 | Process for extracting multiple nutritional ingredients from fish scales |
| CN114560927A (en) * | 2022-03-15 | 2022-05-31 | 厦门元之道生物科技有限公司 | Method for producing collagen peptide by subcritical water extraction assisted fish scales and application of method |
| CN114560927B (en) * | 2022-03-15 | 2024-06-11 | 厦门元之道生物科技有限公司 | Method for producing collagen peptide by sub-critical water extraction assisted fish scales and application thereof |
| RU2810511C1 (en) * | 2022-12-14 | 2023-12-27 | Федеральное государственное автономное образовательное учреждение высшего образования "Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского" | Method of obtaining fish collagen homogenizate |
| CN115754092A (en) * | 2022-12-16 | 2023-03-07 | 江西师范大学 | Identification method of gelatin of cyprinus carpio |
| CN116262938A (en) * | 2022-12-28 | 2023-06-16 | 衢州鲟龙水产食品科技开发有限公司 | A method for preparing collagen peptides from sturgeon dark fish intestines |
| CN117441868A (en) * | 2023-11-27 | 2024-01-26 | 山东美佳集团有限公司 | Preparation method of seafood enzymatic seasoning liquid |
| CN118994371A (en) * | 2024-08-14 | 2024-11-22 | 广州崇真生物科技有限公司 | Preparation method and application of anti-aging collagen peptide |
| CN118994371B (en) * | 2024-08-14 | 2025-06-17 | 德州蓝力生物技术有限公司 | Preparation method and application of anti-aging collagen peptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102839207A (en) | Method for preparing collagen peptide from fish scales | |
| CN102154424B (en) | Process for preparing micro-molecular fish scale collagen peptides | |
| CN103058749B (en) | Method for preparing amino acid liquid fertilizer by hydrolyzing animal proteins with multiple acids and application of amino acid liquid fertilizer | |
| CN101787078B (en) | A kind of collagen polypeptide and its preparation method and application | |
| CN102690853B (en) | A kind of preparation method of squid skin low molecular weight collagen peptide | |
| CN101407834A (en) | Method for preparing collagen peptide by using cod skin | |
| CN103040002A (en) | Tortoise herbal jelly with skin modifying effect and preparation technique thereof | |
| CN105039483A (en) | Method for preparing fishskin fish-scale collagen protein | |
| JP2008206470A (en) | Method for producing fish scale-derived collagen peptide | |
| CN104622782B (en) | A kind of fermented tcm detergent and preparation method thereof | |
| CN102475266A (en) | Preparation method of natural zinc-rich oyster powder | |
| CN104490732B (en) | A kind of anti-aging scale collagen polypeptide facial mask and preparation method thereof | |
| CN104906555A (en) | Preparation method of swallow nest extractive, and obtained swallow nest extractive and application thereof | |
| CN103343153A (en) | Method for preparing forest frog oil peptides by enzymatic hydrolysis and forest frog oil peptides | |
| CN102154421A (en) | Method for extracting collagen from fish processing waste including skin, bones and scale | |
| CN108402471A (en) | A kind of preparation method of poultry bone collagen polypeptide chelated calcium | |
| CN103467621A (en) | Preparation method of high-purity sturgeon chondroitin sulfate | |
| CN102732592A (en) | Method for preparing freshwater fish bone gelatin by enzyme process | |
| CN101649341A (en) | Method for extracting protein peptide from membranes of fowl eggshells | |
| CN103082282A (en) | Method for producing fish collagen calcium chelate by using fishbone | |
| CN102475634A (en) | Hibiscus syriacus leaf facial cleanser | |
| CN104381990A (en) | Preparation method of acete chinensis sauce rich in peptide calcium | |
| CN102492562A (en) | Mosquito repellent laundry detergent | |
| CN106187798A (en) | A kind of chelated calcium preparation method of aminoacid nanometer | |
| CN102702340A (en) | Feather fibrin and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121226 |