CN1682765A - Method for refining sheep placenta active component using membrane separating technology - Google Patents
Method for refining sheep placenta active component using membrane separating technology Download PDFInfo
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Abstract
The present invention relates to the extracting and separating method of active matters in sheep placenta, and is especially process of extracting active matters from sheep placenta with microfiltering film and nanofiltering film. The extracting process includes material collection, sorting, pulping, crushing, enzymolysis, film separation, and freeze drying. The film separating includes using inorganic ceramic film of aluminum oxide, zirconium oxide or antimony oxide or organic film of polysulfone, cellulose acetate, PTFE or polypropylene with intercepting particle range of 0.05-1 micron to eliminate hybrid protein, rib segment and residual degrading enzyme; and using nanofiltering film of polyamide, polypeperazine amide or polyether sulfone to intercept molecular weight 150-1000 components while eliminating free amino acid and water.
Description
Technical field
The present invention is a kind of method that adopts membrane separation technique extraction separation Foetus Caprae seu Ovis active substance, especially with micro-filtration membrane and the refining method of extracting the Foetus Caprae seu Ovis active substance of NF membrane.
Background technology
Placenta Hominis is the organ that fetus and parent carry out mass exchange, it is the physiological product of supplying with the fetus fully nutrient, protein content reaches more than 70%, amino acid content is abundant, A wide selection of colours and designs, reasonable mixture ratio, about fat content 15%, wherein contain abundant phospholipid, lipopolysaccharide, and contain multivitamin and needed by human constant, trace element, and panimmunity globulin, multiple antibody, also have with the closely-related interferon of immunologic function and with the body function relevant active polypeptide that runs well.Especially somatomedin and the positive growth factor in the sheep embryo different organs, these mammals birth backs be because organ or tissue degenerates no longer excretory " the magical factor ", can be equal to good fortune the effect of body.They can make the nervus centralis reparative regeneration of damage, can make the organs and tissues of pathological changes rebuild function; Can make bone marrow hematogenesis, epidermis more living.Can impel the gonad secretion, the endocrine regulation balance can be replenished the anti-ageing anti-old factors such as thymosin, to promote the metabolism of body, recovers incretion balance, strengthens the function of each internal organs, shines vitality.Active polypeptide constituents wherein can all absorb by intestinal, directly be transported to the different organ of whole body and tissue and bring into play bioactive functions through blood circulation, but protein component then can not directly be absorbed and used, must just can be absorbed and used by being degraded into polypeptide or aminoacid, and micromolecule polypeptide has higher biological activity and nutritive value than aminoacid, the cell active substance that from Foetus Caprae seu Ovis, extracts at present, also the clear and definite quality inspection standard of neither one now adopts the monitor control index of polypeptide as assay mostly.
At present existing public technology Chinese patent: publication number is that " a kind of method of making frozen dry extracts of sheep embryo " patent documentation of CN1193515A adopts the direct lyophilizing of Foetus Caprae seu Ovis raw material pulverize at low temperature, and carries out the method that secondary is pulverized under the liquid nitrogen cooling protection; Publication number is that CN1203794A's " freeze-dried sheep placenta extract powder and production method thereof " patent documentation adopts sheep embryo quick-freezing, and cryodesiccated method is back and forth freezed in the low temperature making beating.Though above-mentioned two kinds of methods have kept biological most of activity, also have a lot of invalid impurity components in the finished product, dna purity is not high.Publication number is that CN1307871A's " goat fetus essence injecta and preparation technology " patent documentation adopts the Foetus Caprae seu Ovis fragmentation, homogenate, freeze thawing, supernatant ultrafiltration after centrifugal, collection is less than the method for 30000 molecular weight filtered solutions, the active component that wherein utilizes freeze thawing technology to extract is many far away from the active substance of enzymolysis process output, utilizes Production by Enzymes Foetus Caprae seu Ovis active substance to have characteristics such as content height, production capacity are strong, is direct, the most economic a kind of production method.Publication number is to regulate pH value after CN1238207A's " tonic liquid of black ewe's placenta extract and production method thereof " patent documentation adopts the Foetus Caprae seu Ovis raw material powder to be broken into pasty state, adds enzyme hydrolysis, through coarse filtration, fine straining, obtains the method for extract; Publication number is that CN12730%A's " process for extracting sheep placenta extract " patent documentation adopts Foetus Caprae seu Ovis and Placenta Hominis making beating, enzymolysis, the method for ultrafiltration membrance filter; Publication number is that CN1432388A's " Foetus Caprae seu Ovis peptide oral liquid and preparation method thereof " patent documentation adopts Foetus Caprae seu Ovis and Placenta Hominis homogenate, micronized pulverization, Proteolytic enzyme, centrifugal ultrafiltration, the method for collecting molecular weight 10000 following ultrafiltrates; China's biochemical drug magazine, Liu Shishan etc. adopt the sheep embryonic disc to rub in " functional test of sheep embryonic disc peptide " non-patent literature that 2002 the 23rd the 5th phases of volume delivered, homogenate, freeze thawing, through microfiltration, ultrafiltration, nanofiltration makes the method for finished product after the lyophilizing.Above-mentioned these methods are in the extracting solution that obtains with enzymolysis or freeze thawing technology, except that containing peptide active component, protein molecule, aminoacid, nucleic acid etc., the plurality of impurities compositions such as residue that also have insoluble protein molecule, Foetus Caprae seu Ovis fascia fragment, digestive enzyme, and the material of holding back less than 30,000 or 10,000 molecular weight with ultrafilter membrane reaches the purpose of extracting purification, can lose some activated proteins, the yield of extraction reduces; Though existing bibliographical information has adopted membrane separation technique, does not indicate used membrane material and molecular cut off.Therefore aspect operating procedure and dna purity, all there is defective in above method.So it is higher to seek a kind of yield, purity is better, and the more extracting method of peptide active component content realizes that to the extraction that promotes the Foetus Caprae seu Ovis active substance suitability for industrialized production seems particularly important.
Summary of the invention
The objective of the invention is to improve the content of yield, purity and the active component peptide of product, provide a kind of simple to operate, pollute little, easily preserve, be suitable for the big refining separation method of producing, adopt inorganic ceramic film or organic membrane microfiltration and composite membrane nanofiltration to carry out separation and purification and extract the Foetus Caprae seu Ovis active substance.
The extracting method of existing Foetus Caprae seu Ovis active ingredient is: sheep embryo and Placenta Hominis are cleaned clean, fragmentation adds the water low-temperature homogenate, behind enzymolysis, obtains enzymolysis solution, utilizes ultrafiltration to hold back the active ingredient of certain molecular weight.
The technical solution adopted in the present invention is:
In enzymolysis solution, utilize inorganic ceramic film or organic membrane microfiltration to remove the fragment of the foreign protein of 100,000 above molecular weight in the enzymolysis solution, fascia and the residue of digestive enzyme, obtain clarifying the Foetus Caprae seu Ovis extracting solution, carry out purification, concentrate through organic hybrid films nanofiltration removal free amino acid, most of moisture content again, lyophilization obtains the Foetus Caprae seu Ovis lyophilized powder.
Membrane material involved in the present invention can be: the inorganic ceramic membrane material is oxides such as aluminium oxide, zirconium oxide or stibium oxide; The organic membrane material is high-molecular organic materials such as polysulfones, acetate fiber, politef or polypropylene type; The organic hybrid films material is aromatic polyamides class, poly-piperazine acidamide or polyether sulfone.It is 0.05～1um that related microfiltration is held back particle range, and nanofiltration molecular cut off scope is 150～1000.
Optimized technical scheme of the present invention is:
Described inorganic ceramic membrane material is a zirconium oxide, and holding back particle range is 0.08～0.2um.
Described organic membrane is a polysulfone membrane, and holding back particle range is 0.08～0.2um.
Described organic hybrid films is aromatic polyamides class or polyether sulfone, and its molecular weight is 150-1000.
Described NF membrane, separating spissated flow process all is to carry out at normal temperatures.
The present invention compared with prior art has following characteristics:
1. this extraction process adopts enzymolysis-membrance separation associating, and is simple to operate, extracts the yield height, and the active component that product keeps is more, has removed most of invalid impurity.
2. do to extract solvent with water, in the process of membrane separation purification, reaction temperature and, avoided losing of active component, and the permeate water after nanofiltration concentrates can be recycled.
3. adopt inorganic ceramic film separation efficiency height, resistance tocrocking is strong, is easy to regeneration and cleans.
4. adopt the NF membrane filtering and concentrating, reduced original volume 3-4 amount doubly, shortened the time of concentrate drying, saved the energy, extraction cost greatly reduces.
5. simple to operate, the efficient height, energy savings is suitable for industrialized great production.
The specific embodiment
Embodiment 1
With fresh or refrigerated sheep embryo and Placenta Hominis through quarantining qualified, rub into meat pulp, the low temperature making beating, taking by weighing said extracted serosity 2000 grams drops in the enzymatic vessel, starting agitator stirs, be warmed up to 50 ℃, take by weighing 3 gram neutral protease, hydrolysis 40 minutes was carried out enzyme-deactivating 5 minutes at 75 ℃, be that the zirconia ceramics film of 0.1um carries out microfiltration with said extracted liquid with interception immediately, filtration pressure is 0.1Mpa, and the collection permeate carries out nanofiltration again and concentrates, and the nanofiltration device adopts the polyether sulfone composite membrane, the molecular cut off scope is 150-180, filtration pressure is 0.1-0.2Mpa, removes most of moisture and obtains 500 milliliters of Foetus Caprae seu Ovis concentrated solutions, carries out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.5%, and the peptide content of surveying is 33.8%.
Embodiment 2
Sheep embryo and Placenta Hominis pretreatment, low-temperature homogenate, enzymolysis process is with embodiment 1, centrifugal filtration immediately, rotating speed 4000-5000 rev/min, time is 20 minutes, the polysulfone membrane that with interception is 0.1um is carried out microfiltration with above-mentioned centrifuged supernatant, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts the polyether sulfone composite membrane, and the molecular cut off scope is 150-180, and filtration pressure is 0.1-0.2Mpa, remove most of moisture and obtain 520 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.1%, and the peptide content of surveying is 32.4%.
Embodiment 3
Sheep embryo and Placenta Hominis pretreatment, homogenate, enzymolysis process is with embodiment 1, centrifugal filtration immediately, rotating speed 4000-5000 rev/min, time is 20 minutes, the polysulfone membrane that with interception is 0.08um is carried out microfiltration with above-mentioned centrifuged supernatant, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts polyamide-based composite membrane, and the molecular cut off scope is 300-1000, and filtration pressure is 0.1-0.4Mpa, remove most of moisture and obtain 700 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization, the yield of Foetus Caprae seu Ovis extract is 5.9%, and the peptide content of surveying is 30.8%.
Embodiment 4
Sheep embryo and Placenta Hominis pretreatment, homogenate, enzymolysis process are with embodiment 1, with interception is that the zirconia ceramics film of 0.08um carries out microfiltration with above-mentioned enzymolysis solution, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts polyamide-based composite membrane, the molecular cut off scope is 300-1000, filtration pressure is 0.1-0.4Mpa, remove most of moisture and obtain 650 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization, the yield of Foetus Caprae seu Ovis extract is 5.9%, and the peptide content of surveying is 32.5%.
Embodiment 5
Sheep embryo and Placenta Hominis pretreatment, low-temperature homogenate, enzymolysis process is with embodiment 1, centrifugal filtration immediately, rotating speed 4000-5000 rev/min, time is 20 minutes, the polysulfone membrane that with interception is 0.2um is carried out microfiltration with above-mentioned centrifuged supernatant, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts the polyether sulfone composite membrane, and the molecular cut off scope is 150-180, and filtration pressure is 0.1-0.2Mpa, remove most of moisture and obtain 660 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.1%, and the peptide content of surveying is 31.6%.
Embodiment 6
Sheep embryo and Placenta Hominis pretreatment, low-temperature homogenate, enzymolysis process are with embodiment 1, the alumina ceramic membrane that with interception is 0.2um carries out microfiltration with above-mentioned enzymolysis solution liquid, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts the polyether sulfone composite membrane, the molecular cut off scope is 150-180, filtration pressure is 0.1-0.2Mpa, remove most of moisture and obtain 640 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.7%, and the peptide content of surveying is 33.4%.
Embodiment 7
Sheep embryo and Placenta Hominis pretreatment, low-temperature homogenate, enzymolysis process are with embodiment 1, with interception is that the zirconia ceramics film of 0.1um carries out microfiltration with enzymolysis solution, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts polyamide-based composite membrane, the molecular cut off scope is 300-1000, filtration pressure is 0.1-0.4Mpa, remove most of moisture and obtain 590 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.4%, and the peptide content of surveying is 32.9%.
Embodiment 8
Sheep embryo and Placenta Hominis pretreatment, low-temperature homogenate, enzymolysis process are with embodiment 1, the alumina ceramic membrane that with interception is 0.08um carries out microfiltration with enzymolysis solution, filtration pressure is 0.1Mpa, the collection permeate carries out nanofiltration again and concentrates, the nanofiltration device adopts polyamide-based composite membrane, the molecular cut off scope is 300-1000, filtration pressure is 0.1-0.4Mpa, remove most of moisture and obtain 720 milliliters of Foetus Caprae seu Ovis concentrated solutions, carry out lyophilization again, the yield of Foetus Caprae seu Ovis extract is 6.3%, and the peptide content of surveying is 31.6%.
Claims (7)
1. the membrane technology separation and refining method of a sheep placenta active component, comprise feedstock capture, clear Xian's sorting, making beating, fragmentation, enzymolysis, use membrane technology separation and purification, lyophilization, it is characterized in that: in enzymolysis solution, utilize inorganic ceramic film or organic membrane microfiltration to remove foreigh protein removing, the fragment of fascia, the residue of digestive enzyme, remove free amino acid, most of moisture content through the organic hybrid films nanofiltration again, concentrate, lyophilization obtains the Foetus Caprae seu Ovis lyophilized powder.
2. the membrane technology separation and refining method of sheep placenta active component according to claim 1, it is characterized in that: the inorganic ceramic membrane material is aluminium oxide, zirconium oxide or stibium oxide, holding back particle range is 0.05～1um.
3. the membrane technology separation and refining method of sheep placenta active component according to claim 2, it is characterized in that: the inorganic ceramic membrane material is a zirconium oxide, holding back particle range is 0.08～0.2um.
4. the membrane technology separation and refining method of sheep placenta active component according to claim 1, it is characterized in that: the organic membrane material is polysulfones, acetate fiber, politef or polypropylene type, holding back particle range is 0.05～1um.
5. the membrane technology separation and refining method of sheep placenta active component according to claim 4, it is characterized in that: organic membrane is a polysulfone membrane, holding back particle range is 0.08～0.2um.
6. the membrane technology separation and refining method of sheep placenta active component according to claim 1 is characterized in that: the organic hybrid films material is aromatic polyamides class, poly-piperazine acidamide or polyether sulfone, and nanofiltration molecular cut off scope is 150～1000.
7. the membrane technology separation and refining method of sheep placenta active component according to claim 6, it is characterized in that: the organic hybrid films material is aromatic polyamides class or polyether sulfone, nanofiltration molecular cut off scope is 150～1000.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2005100539588A CN1300169C (en) | 2005-03-15 | 2005-03-15 | Method for refining sheep placenta active component using membrane separating technology |
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| CNB2005100539588A CN1300169C (en) | 2005-03-15 | 2005-03-15 | Method for refining sheep placenta active component using membrane separating technology |
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| CN1300169C CN1300169C (en) | 2007-02-14 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102485903A (en) * | 2011-08-18 | 2012-06-06 | 内蒙古健元鹿业有限责任公司 | Method for preparing biological active protein oligopeptide powder from deer placenta |
| CN102703535A (en) * | 2012-06-19 | 2012-10-03 | 江苏久吾高科技股份有限公司 | New technology for producing acrylamide by using ceramic membrane bioreactor |
| CN105998076A (en) * | 2016-07-20 | 2016-10-12 | 雁杰 | Submandibular gland active molecule separation and extraction process and active molecule product thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1110128A (en) * | 1994-04-14 | 1995-10-18 | 浙江康恩贝集团研究开发中心 | Method for preparation of active placenta powder |
| CN1054513C (en) * | 1998-04-14 | 2000-07-19 | 姚炳坤 | Method for production of frozen dry extracts of sheep embryo |
| CN1072955C (en) * | 1999-05-21 | 2001-10-17 | 海南椰风企业有限公司 | Tonic liquid of black ewe's placenta extract and its preparing method |
| CN1235026A (en) * | 1999-05-28 | 1999-11-17 | 孙月英 | Preparing method for injection of placenta peptide and products thereof |
| CN1273096A (en) * | 2000-04-07 | 2000-11-15 | 吴文惠 | Process for extracting sheep placenta extract |
-
2005
- 2005-03-15 CN CNB2005100539588A patent/CN1300169C/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102485903A (en) * | 2011-08-18 | 2012-06-06 | 内蒙古健元鹿业有限责任公司 | Method for preparing biological active protein oligopeptide powder from deer placenta |
| CN102703535A (en) * | 2012-06-19 | 2012-10-03 | 江苏久吾高科技股份有限公司 | New technology for producing acrylamide by using ceramic membrane bioreactor |
| CN102703535B (en) * | 2012-06-19 | 2014-07-30 | 江苏久吾高科技股份有限公司 | New technology for producing acrylamide by using ceramic membrane bioreactor |
| CN105998076A (en) * | 2016-07-20 | 2016-10-12 | 雁杰 | Submandibular gland active molecule separation and extraction process and active molecule product thereof |
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| CN1300169C (en) | 2007-02-14 |
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