CN1298709A - Process for preparing polypeptide-nucleic acid nutritive liquid of sheep placenta - Google Patents
Process for preparing polypeptide-nucleic acid nutritive liquid of sheep placenta Download PDFInfo
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- CN1298709A CN1298709A CN00132315A CN00132315A CN1298709A CN 1298709 A CN1298709 A CN 1298709A CN 00132315 A CN00132315 A CN 00132315A CN 00132315 A CN00132315 A CN 00132315A CN 1298709 A CN1298709 A CN 1298709A
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Abstract
A preparation method of sheep embryo polypeptide nucleic acid nutritive liquid is as follows: a whole sheep embryo after quick-frozen is washed to remove connective tissue and umbilical cord, then homogenized in a homogenizer, quick-frozen at minus 60 to minus 70 deg.C, thawed at 15-30 deg.C, repeat frozen and thawing several times, then centrifuged, the supernatant is dialyzed or ultrafiltered to obtain placenta transfer factor; the foresaid centrifuge precipitant after enzymolysis to obtain protein hydrolysate containing different peptide segments and nucleic acid; the said vacuum filtered precipitant is ultrasonic-treated, then through removing residues and grease, and adding saturated phenol for centrifugation after inc ubatation to obtain nucleic acid solution. Said method is complete in effective components and nutrient extraction, the obtained nutritive liquid can be used to prepare health-care product, cosmetics and immune regulating agent.
Description
The present invention relates to a kind of is raw material with the Foetus Caprae seu Ovis, the preparation method of the nutritional solution of rich polypeptide-nucleic acid.
The Placenta Hominis of humans and animals is a Chinese medicine, contains multiple bioactive substance with immunoregulation effect, and rich protein, polypeptide, aminoacid and nucleic acid.Fetus is a complete life, except that containing rich in protein, polypeptide, aminoacid and nucleic acid, also contain a large amount of Lecithins, unsaturated fatty acid, multivitamin and various trace elements, the more important thing is and contain great deal of bioactive substances in the animal embryo, growth promoter, immunomodulating, metabolism to biology have crucial effect, and the nutrient substance in the fetus and bioactive substance have easy absorption, advantage that utilization rate is high.
In animal product, some pays attention to its biological activity, and only dialysis obtains to have the immunocompetence placenta pepton, and does not utilize in the Placenta Hominis very rich nutrient substances.More is to extract nutrition again after adopting high temperature sterilize, and traditional traditional Chinese medical science also is to boil the back medication, thereby causes the high temperature of part nutritional labeling (as protein, vitamin etc.) to destroy and the bioactive substance loss of activity.Though the appearance of full Placenta Hominis frozen drying goods is arranged, bioactive substance and nutritional labeling are kept, its absorbance and utilization rate are not high, and can only be used for orally, and can not make injection and cosmetics.Simultaneously, most placental articles have been ignored the high nucleic acid resource of nutrition in Placenta Hominis and the fetus.
The object of the present invention is to provide a kind of preparation method of polypeptide-nucleic acid nutritive liquid of sheep placenta.It can effectively extract and preserve the bioactive substance in the Placenta Hominis, can fully extract nutrient substance such as protein nutritious in the Placenta Hominis and nucleic acid again.Mainly solve prior art to the extraction of active princlple and nutrition not exclusively, many, the absorbance of loss shortcoming such as less.Simultaneously, provide a kind of polypeptide gene tonic and immune formulation.
Purpose of the present invention can realize by following technology:
Nutritional solution comprises that Placenta Hominis dialysis solution, protein hydrolyzate and nucleic acids for preparation technological process are as follows: the extracting and proteolysis and the nucleic acid extracting three big steps that comprise bioactive substance.
1, the extracting of active substance.
(1) raw material is handled: get 1～3 month the full Placenta Hominis of sheep (comprising fetus and Placenta Hominis) at-20 ℃～-40 ℃ rapid freezing, clean, cleanings and removal connective tissue and umbilical cord.
(2) embryotomic dish tissue: tissue is cut into small pieces, and homogenate in homogenizer is in-60 ℃～-80 ℃ quick-freezings, afterwards in 15 ℃～30 ℃ thawings, multigelation.
(3) preparation of active substance: above-mentioned freeze thawing product is centrifugal, get above-mentioned clear liquid through dialysis or ultrafiltration, the degerming of reuse filtering with microporous membrane promptly gets and contains abundant bioactive substance placental transfer factor.
2, the preparation of protein hydrolyzate, the precipitation part of getting above-mentioned centrifugal gained adopts enzyme-sour associating hydrolysis.
(1) the precipitation adding distil water is got in enzyme hydrolysis, scalding, and aseptic condition downward modulation PH to 7～8 add pancreatin, and temperature is controlled at 36 ℃～42 ℃, enzymolysis.
(2) said hydrolyzed thing, heat boil after, transfer PH to 3 ± 0.5, high temperature and high pressure hydrolysis transfers pH value to 6+0.5 sucking filtration to remove precipitation afterwards, collecting clear liquid is protein hydrolyzate, wherein contains the peptide section and the various nucleic acid of all size.
3, further extracting nucleic acid:
(1) in the precipitation of above-mentioned sucking filtration, adds distilled water, fully be mixed, adopt the supersound process instrument that remaining genomic DNA is sheared, hatch for 50 ℃ ± 5 ℃, filter and remove residue and oils and fats.
(2) saturated phenol extracting adds the saturated phenol with volume, mixing, centrifugalize.
(3) above-mentioned gained sucks in another clean window, promptly gets nucleic acid solution.
More than each extracting solution stand-by in low temperature storage.
Advantage that the present invention compared with prior art has and good effect:
1, the placental preparations of Huo Deing has kept in the Placenta Hominis 96% natural activity composition.Wherein have with the extremely close SOD of health (seeing Table 1)
2, method for hydrolysis is united in enzyme-acid, increased substantially the content of materials such as aminoacid, small peptide, and wherein contained abundant micromolecule nutrient is absorbed by the body easily.And the percentage ratio that 8 kinds of essential amino acids account for total amount is 42.48% (seeing Table 2).And contain abundant trace element (seeing Table 3).
3, the placental preparations of this acquisition contains nutritive nucleic acid materials such as nucleotide that the abundant utmost point is beneficial to absorption, all size nucleic acid fragment, is a kind of nutrient and healthcare products of gene preferably.(seeing Table 1)
4, the method is scientific and reasonable, and all raw material is a natural materials, wide material sources, and flow process is short, and is simple to operate, is suitable for suitability for industrialized production, and application and development prospect is preferably arranged.
The character of table 1 nutritional solution and Main Ingredients and Appearance
Project content
The brown transparency liquid of character
PH value 6.8
Density 1.21g/cm
3
Aminoacid 33.68mg/ml
Protein 27mg/ml
Superoxide dismutase 639u/ml
Nucleic acid 1.63mg/ml
(1) measures superoxide dismutase activity with pyrogallol autoxidation method.
(2) fixing phosphorus method is measured nucleic acid content.
(3) protein and peptide concentration are measured and are adopted: the Lowry method.
The various amino acid contents of table 2 nutritional solution (mg/ml)
The amount of amino acid amount of amino acid
Ser 1.15 Tyr 0.31
ASP 3.69 Ile 2.66
Glu 4.36 Phe 2.11
Val 1.48 Pro 2.69
Ala 2.47 Lys 3.43
Met 0.90 Trp 0.25
Cys 0.32 His 0.62
Thr 1.54 Leu 1.94
Gly 2.23 Arg 1.53
Amino acid content: measure with Bechman 121MB automatic amino acid analyzer.
Part micronutrient levels (ug/ml) in table 3 preparation
The constituent content constituent content
Manganese 0.179 ferrum 0.85
Calcium 0.92 bronze medal 0.18
Zinc 0.16 germanium 0.0047
Selenium 0.0052 lithium 0.0016
Cobalt 0.0067 micronutrient levels: adopt aas determination.
Further specify below in conjunction with embodiment:
Embodiment 1:
From the full Placenta Hominis of sheep (comprising fetus and Placenta Hominis) extracting method: A, first freeze thawing broken cell and low temperature dialysis, keep bioactive substance and easy nutritional labeling such as ruined little living element in Foetus Caprae seu Ovis and the Placenta Hominis to greatest extent.B, enzyme-sour pyrohydrolysis protein and further extract aminoacid, peptide class and nucleic acid extracting three big steps more afterwards.
1, the extracting of active substance.
(1) raw material is handled: get 1～3 month the full Placenta Hominis of sheep (comprising fetal placenta)-40 ℃ rapid freezing, 4 ℃ of normal saline are cleaned, cleaning and remove connective tissue and umbilical cord.
(2) embryotomic dish tissue: tissue is cut into small pieces, uses homogenizer, 12000 change homogenate in 5 minutes 5 times; In-70 ℃ of quick-freezings, afterwards in 25 ℃ of thawings, multigelation 9 times.
(3) preparation of active substance: above-mentioned freeze thawing product centrifugal 20 minutes of 0～4 ℃ of 10000rpm, is got supernatant through dialysis or ultrafiltration, and the filtering with microporous membrane degerming of reuse 0.22 or 0.3ul promptly gets and contains abundant bioactive substance placental transfer factor.
2, the preparation of protein hydrolyzate, the precipitation part of getting above-mentioned centrifugal gained adopts enzyme-sour associating hydrolysis.
(1) enzyme hydrolysis is got precipitation and is added 6 times of weight distilled water, boils sterilization in 10 minutes, and aseptic condition downward modulation PH to 7.5 adds pancreatin (trypsin or chymase), and enzyme dosage is 1: 250, and temperature is controlled at about 36 ℃, enzymolysis 6h.
(2) said hydrolyzed thing, heat boil after, add HCI and transfer under the condition of PH to 3 left and right sides.High temperature and high pressure hydrolysis 3h reduces to room temperature, again after the high temperature and high pressure hydrolysis 3h, transfers about ph to 6 with NaOH afterwards.Sucking filtration is removed precipitation, and collecting clear liquid is protein hydrolyzate, wherein also contains the peptide section and the various nucleic acid of all size.
3, further extracting nucleic acid:
(1) distilled water of 2 times of volumes of adding in the precipitation of above-mentioned sucking filtration fully is mixed, and employing has the supersound process instrument that immerses tip remaining genomic DNA is sheared, peak power was handled 1 minute, repeat 5 times, hatched 10 minutes for 50 ℃, filter and remove residue and oils and fats.
(2) saturated phenol extracting adds the saturated phenol with volume, mixing, centrifugal (12000rpm, 4 ℃) 15 minutes.
(3) supernatant promptly gets and sucks in another clean window, promptly gets nucleic acid solution.
More than each extracting solution stand-by in-2 ℃ of storages.
Products obtained therefrom of the present invention can be used as health product, cosmetics and immunomodulator.
Embodiment 2
Polypeptide-nucleic acid nutritive liquid of sheep placenta: the product that three steps were extracted mixes, and filtration sterilization below is aseptic subpackaged: every 50ml adds 5g sucrose, 0.1ml spice, is oral storage liquid.Effect: be conventional Chinese medicine, it is warm in nature, and sweet-salty is gone into lung, liver, kidney three warps, and the merit of benefiting qi and nourishing blood benefit essence is arranged, and the effect of building up resistance with immunity is arranged on immunology.
Embodiment 3:
The close cosmetics of Foetus Caprae seu Ovis polypeptide-nucleic acid nutrition: the above-mentioned three step products of carrying are mixed, and filtration sterilization below is aseptic subpackaged: every 20ml adds in the 80ml emulsifying agent, and mix homogeneously can obtain milky paste.Have antioxidation, whiten, improve skin metabolism, multiple nutrient nourishes rough skin, defying age, the effect of the skin care of preserving youthful looks.
Embodiment 4:
The placenta pepton injection: 5ml/ props up cryopreservation with the aseptic subpackaged one-tenth of first step product.Promptly can be used for clinically through being up to the standards, these product are immunomodulator, and indication is widely arranged, and are usually used in weakness due to chronic disease, resistance difference and treatment chronic disease.
Claims (1)
1, the preparation method of polypeptide-nucleic acid nutritive liquid of sheep placenta is characterized in that being made up of following processing step:
(1) active substance extracting:
(a) raw material is handled: get the full tire of sheep, comprise fetus and Placenta Hominis, at-20 ℃-40 ℃ rapid freezing, clean, cleanings and removal connective tissue and umbilical cord.
(b) broken placenta tissue: will be organized in homogenate in the homogenizer, in-60 ℃ of-80 ℃ of quick-freezings, afterwards in 15-30 ℃ of thawing, multigelation.
(c) preparation of active substance: with the centrifugalize of above-mentioned freeze thawing product, get supernatant, promptly get with the filtering with microporous membrane degerming and contain abundant bioactive substance placental transfer factor through dialysis or ultrafiltration.
(2) preparation of protein hydrolyzate:
(a) enzyme hydrolysis, the precipitation of getting above-mentioned centrifugal gained adds the water boil sterilization, transfers pH value 7-8 to add 36 ℃ of-42 ℃ of enzymolysis of pancreatin;
(b) after the said hydrolyzed thing is heated and boiled, transfer pH value 3 ± 0.5, high temperature and high pressure hydrolysis transfers pH value to 6 ± 0.5 sucking filtration to go precipitation afterwards, and collecting clear liquid is protein hydrolyzate, wherein contains various peptide sections and nucleic acid;
(3) further extracting nucleic acid:
(a) in the precipitation of above-mentioned sucking filtration, add entry and be mixed, adopt the supersound process instrument that remaining genomic DNA is sheared, hatch for 50 ℃ ± 5 ℃, filter and remove residue and oils and fats.
(b) saturated phenol extracting adds the saturated phenol with volume, centrifugalize.Supernatant promptly gets and sucks in another clean window, promptly gets nucleic acid solution; More than each extracting solution stand-by in low temperature storage.
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| CNB001323156A CN1178672C (en) | 2000-11-03 | 2000-11-03 | Process for preparing polypeptide-nucleic acid nutritive liquid of sheep placenta |
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| CNB001323156A CN1178672C (en) | 2000-11-03 | 2000-11-03 | Process for preparing polypeptide-nucleic acid nutritive liquid of sheep placenta |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310939C (en) * | 2002-09-11 | 2007-04-18 | 北京陆桥技术有限责任公司 | Method for extracting deoxyribonucleic acid from edible oil |
| CN101703259B (en) * | 2009-11-05 | 2012-01-04 | 辽宁大学 | Method for deeply processing deer placenta |
| CN102329841A (en) * | 2011-07-12 | 2012-01-25 | 湖州康海斯生物科技有限公司 | Lamb placenta protein and preparation method thereof |
| CN103463131A (en) * | 2013-09-23 | 2013-12-25 | 河南牧翔动物药业有限公司 | Preparation method of sheep placenta transfer factor solution |
| CN103897015A (en) * | 2012-12-26 | 2014-07-02 | 重庆禾影生物科技发展有限公司 | Placenta active extraction method |
| CN104745664A (en) * | 2015-04-17 | 2015-07-01 | 浙江华尔成生物药业股份有限公司 | Preparation process of animal placenta extract |
| CN106318929A (en) * | 2016-09-21 | 2017-01-11 | 百源科创分子医学研究所（南通）有限公司 | Process for producing oligonucleotides through enzymolysis method |
| CN113088549A (en) * | 2021-05-06 | 2021-07-09 | 内蒙古银宏干细胞生命科技投资有限公司 | Method for extracting antioxidant polypeptide from human placental blood |
| CN114652664A (en) * | 2020-12-23 | 2022-06-24 | 科索瑞生物科技(天津)有限公司 | Repairing gel and preparation method thereof |
| CN115125193A (en) * | 2021-07-09 | 2022-09-30 | 广州赛莱拉生物基因工程有限公司 | Animal umbilical cord exosome and extraction method and application thereof |
| CN115708798A (en) * | 2023-01-10 | 2023-02-24 | 昆明时光肌生物技术有限公司 | One-step hydrolysis preparation method of sheep placenta extract |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100402548C (en) * | 2006-02-22 | 2008-07-16 | 郑彬 | Method for preparing physiological active polypeptide of deer placenta |
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2000
- 2000-11-03 CN CNB001323156A patent/CN1178672C/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310939C (en) * | 2002-09-11 | 2007-04-18 | 北京陆桥技术有限责任公司 | Method for extracting deoxyribonucleic acid from edible oil |
| CN101703259B (en) * | 2009-11-05 | 2012-01-04 | 辽宁大学 | Method for deeply processing deer placenta |
| CN102329841A (en) * | 2011-07-12 | 2012-01-25 | 湖州康海斯生物科技有限公司 | Lamb placenta protein and preparation method thereof |
| CN103897015A (en) * | 2012-12-26 | 2014-07-02 | 重庆禾影生物科技发展有限公司 | Placenta active extraction method |
| CN103463131B (en) * | 2013-09-23 | 2015-09-30 | 河南牧翔动物药业有限公司 | A kind of preparation method of sheep placenta transfer factor solution |
| CN103463131A (en) * | 2013-09-23 | 2013-12-25 | 河南牧翔动物药业有限公司 | Preparation method of sheep placenta transfer factor solution |
| CN104745664A (en) * | 2015-04-17 | 2015-07-01 | 浙江华尔成生物药业股份有限公司 | Preparation process of animal placenta extract |
| CN104745664B (en) * | 2015-04-17 | 2018-08-07 | 浙江华尔成生物药业股份有限公司 | A kind of preparation process of animal placenta extract |
| CN106318929A (en) * | 2016-09-21 | 2017-01-11 | 百源科创分子医学研究所（南通）有限公司 | Process for producing oligonucleotides through enzymolysis method |
| CN114652664A (en) * | 2020-12-23 | 2022-06-24 | 科索瑞生物科技(天津)有限公司 | Repairing gel and preparation method thereof |
| CN113088549A (en) * | 2021-05-06 | 2021-07-09 | 内蒙古银宏干细胞生命科技投资有限公司 | Method for extracting antioxidant polypeptide from human placental blood |
| CN115125193A (en) * | 2021-07-09 | 2022-09-30 | 广州赛莱拉生物基因工程有限公司 | Animal umbilical cord exosome and extraction method and application thereof |
| CN115125193B (en) * | 2021-07-09 | 2024-04-26 | 广州赛莱拉生物基因工程有限公司 | Animal umbilical cord extract and extraction method and application thereof |
| CN115708798A (en) * | 2023-01-10 | 2023-02-24 | 昆明时光肌生物技术有限公司 | One-step hydrolysis preparation method of sheep placenta extract |
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| CN1178672C (en) | 2004-12-08 |
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