CN102327291B - Stock solution of sheep embryo and sheep placental peptide and preparation method thereof - Google Patents
Stock solution of sheep embryo and sheep placental peptide and preparation method thereof Download PDFInfo
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- CN102327291B CN102327291B CN201110193930XA CN201110193930A CN102327291B CN 102327291 B CN102327291 B CN 102327291B CN 201110193930X A CN201110193930X A CN 201110193930XA CN 201110193930 A CN201110193930 A CN 201110193930A CN 102327291 B CN102327291 B CN 102327291B
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Abstract
The invention relates to a sheep embryo and sheep placenta biological product and a preparation method thereof. The preparation method of a stock solution of the sheep embryo and sheep placental peptide comprises the following steps of: (1) preliminarily treating raw materials of the sheep embryo and sheep placenta; (2) carrying out ultra-fine pulverization and cell-wall breaking on the sheep embryo and sheep placenta; and (3) carrying out extraction and enzymolysis on the sheep embryo and sheep placenta: adding 3-10 volumes by weight of distilled water in sheep embryo and sheep placenta slurry subjected to cell-wall breaking, carrying out warm immersion and extraction and then filtering to obtain filtrate A and precipitation A; carrying out protease enzymolysis on the precipitation A and suction filtration to obtain hydrolyzate B; and combining the filtrate A and the hydrolyzate B to obtain the stock solution of the sheep embryo and sheep placental peptide. The stock solution of the sheep embryo and sheep placental peptide by adopting the method not only retains natural active substances in the placenta, but also increases the contents of polypeptides and amino acid in the placenta.
Description
Technical field
The present invention relates to a kind of sheep embryo and Placenta caprae seu ovis biological product and preparation method thereof.
Background technology
China utilizes that various organs, tissue and the metabolite of animal are cured the disease, diseases prevention and health care with a long history, existing complete pharmacy ancient books and records---" legendary god of farming's book on Chinese herbal medicine opinion " just has the record of the many animals medicines such as Calculus Bovis, Cornu rhinocerotis, Cornu Cervi Pantotrichum, Colla Corii Asini.Illustrated that China utilizes already animal pharmaceuticals to process, extract, refiningly forms comparatively pure medicament on pharmaceutical technology.
Placenta Hominis (Placenta) has another name called " Placenta Hominis ", be conventional Chinese medicine, modern medicine study shows: Placenta Hominis contains the bioactive substance such as panimmunity globulin, bioactive peptide and aminoacid, mineral matter and other components (general designation living cells element), is a kind of regulator of body's immunity and nutrition, cosmetic health product.Scientist finds that the cell tissue structure of Placenta caprae seu ovis is the most similar to people's Placenta Hominis after the many animals Placenta Hominis is compared experiment, is expected to become with the human placenta to have the substitute that equal health preserving is worth.
In recent years, in succession occur again animal Placenta Hominis hydrolyzed solution and two kinds of Novel sheep Placenta Hominiss of Placenta Hominis lixiviating solution biological product at home and abroad, because of its unique usefulness, caused gazing at of people.But we think that above-mentioned two kinds of preparations respectively have drawback, and the former destroys the natural active matter in the Placenta Hominis in hydrolytic process, loses the meaning of utilizing Placenta Hominis; Although the latter has multiple natural active matter, polypeptide and amino acid content are very little, if the two is combined together, have both kept the natural active matter in the Placenta Hominis, increase again its polypeptide and amino acid content.This is a new breakthrough in Foetus Caprae seu Ovis and the developmental research of Placenta caprae seu ovis biological product undoubtedly.
Summary of the invention
In order to solve above-mentioned technical problem, an object of the present invention is to provide the preparation method of a kind of sheep embryo and Placenta caprae seu ovis element stock solution, the method had both kept the natural active matter in the Placenta Hominis, increased again its polypeptide and amino acid content.Another object of the present invention provides above-mentioned sheep embryo and Placenta caprae seu ovis element stock solution.
In order to realize first above-mentioned purpose, the present invention has adopted following technical scheme:
The preparation method of a kind of sheep embryo and Placenta caprae seu ovis element stock solution, the method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: getting sheep embryo and Placenta caprae seu ovis is raw material, cleans, and removes impurity, is ground into coarse powder under 0～5 ℃ of low temperature;
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: behind the freezing 1～3h of sheep embryo and Placenta caprae seu ovis coarse powder low temperature-40 ℃～-50 ℃ ,-30～-10 ℃ of micronizing, cell wall breaking reaches 0.1～10 μ m granularity;
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: the sheep embryo after the cell wall breaking and Placenta caprae seu ovis serosity add filter to get filtrate after 3～10 times of distilled water warm macerating extract A and precipitate A, sucking filtration got hydrolyzed solution B after precipitate A adopted protease hydrolyzed, merging filtrate A and hydrolyzed solution B get sheep embryo and Placenta caprae seu ovis element stock solution.
As preferably, in the above-mentioned step 1) take the sheep embryo at 2～3 monthly ages and Placenta caprae seu ovis as raw material.
As preferably, protease adopts that one or both mix in papain and the trypsin in the above-mentioned step 3).
As preferably, the use amount of protease is 0.1%～2% of precipitate A weight in the above-mentioned step 3).
Use amount as protease in preferred again, the above-mentioned step 3) is 0.3%～1% of precipitate A weight.
As preferably, protease adopts papain to mix with two kinds of trypsin in the above-mentioned step 3), and the temperature of enzyme digestion reaction is 40～50 ℃, and pH is 7.0～8.0, papain and tryptic enzyme proportioning 1:0.5～2.
As preferred again, the temperature of enzyme digestion reaction is 45 ℃ in the step 3), and pH is 7.5, papain and tryptic enzyme proportioning 1:1.
In order to realize second above-mentioned purpose, sheep embryo and Placenta caprae seu ovis element stock solution that the present invention also provides above-mentioned any one technical scheme to prepare.
The present invention is owing to adopted above-mentioned technical scheme, the process of manufacture of this technology can fully extract the effective ingredient in Foetus Caprae seu Ovis and the Placenta caprae seu ovis, and the active substance of Foetus Caprae seu Ovis and Placenta caprae seu ovis is maximized the retention, and is convenient to again human body and absorbs comprehensively, technique is advanced, reasonable, quality controllable.The sheep embryo and the Placenta caprae seu ovis element stock solution that adopt said method to make had both kept the natural active matter in the Placenta Hominis, increased again its polypeptide and amino acid content.
Description of drawings
Fig. 1 is the GPC-ELSD reference colour spectrogram of sheep embryo of the present invention and Placenta caprae seu ovis element stock solution.
The specific embodiment
Embodiment 1
The preparation method of a kind of sheep embryo and Placenta caprae seu ovis element stock solution, the method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: take the sheep embryo at 2-3 monthly age and Placenta caprae seu ovis as raw material, clean, remove impurity, be ground into coarse powder under the 0-5 ℃ of low temperature.
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: with low temperature (20 ℃) micronizing (the medicinal super micron mill of WZJ100) behind sheep embryo and Placenta caprae seu ovis coarse powder freezing (40 ℃～-45 ℃) 2h, cell wall breaking reaches 0.1-10 μ m granularity.
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: add filter to get filtrate after 6 times of distilled water warm macerating extract A and precipitate A through the sheep embryo after micronizing, the cell wall breaking and Placenta caprae seu ovis serosity, precipitate A with papain and trypsin digestion after sucking filtration get hydrolyzed solution B, 45 ℃ of hydrolysis temperatures, enzyme proportioning 1g:1g, pH7.5, enzyme-to-substrate are 20mg:3g; Merging filtrate A and hydrolyzed solution B get sheep embryo and Placenta caprae seu ovis element stock solution.
Adopt gel permeation chromatography-evaporat light scattering (GPC-ELSD) method to measure the molecular weight distribution of above-mentioned Foetus Caprae seu Ovis and Placenta caprae seu ovis biological product, formulate its quality standard.Step is as follows:
1. get sheep embryo and Placenta caprae seu ovis stock solution, add an amount of pure water heating or ultrasonic dissolution, let cool, standardize solution filters, and gets subsequent filtrate as need testing solution.
2. liquid-phase condition: mobile phase is pure water, flow velocity 1ml/min, 30 ℃ of column temperatures.
3. immobile phase: gel filtration chromatography post (GPC) chromatographic column Shodex OHpak SB-806 HQ, Shodex OHpak SB-805 HQ, Shodex OHpak SB-804 HQ, Shodex OHpak SB-803 HQ series connection.
4. detector: evaporative light scattering detector (ELSD), evaporating temperature: 100 ℃, atomization temperature: 80 ℃, nitrogen flow rate: 1.5L/min.
The GPC-ELSD reference colour spectrogram of sheep embryo and Placenta caprae seu ovis element stock solution as shown in Figure 1.
Embodiment 2
The preparation method of a kind of sheep embryo and Placenta caprae seu ovis element stock solution, the method enzymolysis in step 3) adopts the papain, and the other technologies scheme is as described in Example 1.
Claims (4)
1. the preparation method of a sheep embryo and Placenta caprae seu ovis element stock solution is characterized in that the method comprises the steps:
1) sheep embryo and the pre-treatment of Placenta caprae seu ovis raw material: getting sheep embryo and Placenta caprae seu ovis is raw material, cleans, and removes impurity, is ground into coarse powder under 0～5 ℃ of low temperature;
2) sheep embryo and Placenta caprae seu ovis micronizing, cell wall breaking: behind the freezing 1～3h of sheep embryo and Placenta caprae seu ovis coarse powder low temperature-40 ℃～-50 ℃ ,-30～-10 ℃ of micronizing, cell wall breaking reaches 0.1～10 μ m granularity;
3) extraction of sheep embryo and Placenta caprae seu ovis and enzymolysis: the sheep embryo after the cell wall breaking and Placenta caprae seu ovis serosity add filter to get filtrate after 3～10 times of distilled water warm macerating extract A and precipitate A, sucking filtration got hydrolyzed solution B after precipitate A adopted protease hydrolyzed, merging filtrate A and hydrolyzed solution B get sheep embryo and Placenta caprae seu ovis element stock solution; The use amount of described protease is 0.1%～2% of precipitate A weight, protease adopts papain to mix with two kinds of trypsin, the temperature of enzyme digestion reaction is 40～50 ℃, and pH is 7.0～8.0, papain and tryptic enzyme proportioning 1:0.5～2.
2. the preparation method of described a kind of sheep embryo and Placenta caprae seu ovis element stock solution according to claim 1 is characterized in that: in the step 1) take the sheep embryo at 2～3 monthly ages and Placenta caprae seu ovis as raw material.
According to claim 1 described a kind of sheep embryo and Placenta caprae seu ovis the element stock solution preparation method, it is characterized in that: the use amount of protease is 0.3%～1% of precipitate A weight in the step 3).
A kind of sheep embryo according to claim 1 and Placenta caprae seu ovis the element stock solution preparation method, it is characterized in that: the temperature of enzyme digestion reaction is 45 ℃ in the step 3), pH is 7.5, papain and tryptic enzyme proportioning 1:1.
5According to claim 1 or the sheep embryo for preparing of 2 or 3 or 4 described methods and Placenta caprae seu ovis element stock solution.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110193930XA CN102327291B (en) | 2011-07-12 | 2011-07-12 | Stock solution of sheep embryo and sheep placental peptide and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110193930XA CN102327291B (en) | 2011-07-12 | 2011-07-12 | Stock solution of sheep embryo and sheep placental peptide and preparation method thereof |
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| CN102327291B true CN102327291B (en) | 2013-02-13 |
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| JP6282243B2 (en) * | 2015-03-11 | 2018-02-21 | 株式会社ホルス | Method for producing fermentation-aged placenta solution |
| CN107184609A (en) * | 2017-06-09 | 2017-09-22 | 郑州源创吉因实业有限公司 | A kind of lossless sterile bulk preparation method of Human plactnta |
| CN107216366A (en) * | 2017-07-26 | 2017-09-29 | 西藏天慈生物科技有限公司 | A kind of preparation method and applications of Goat Placenta small-molecular peptides |
| CN108498568A (en) * | 2018-06-04 | 2018-09-07 | 沈旸 | Chinese patent drug, medical food and the preparation method of autoimmune and immune related diseases are treated with Goat Placenta or embryo |
| CN111888322B (en) * | 2020-09-18 | 2022-09-13 | 成都清科生物科技有限公司 | Sheep placenta extract with whitening function and extraction method and application thereof |
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| CH684741A5 (en) * | 1992-06-11 | 1994-12-15 | Lucchini Lab Sa | A process for the preparation of an extract. |
| CN1273096A (en) * | 2000-04-07 | 2000-11-15 | 吴文惠 | Process for extracting sheep placenta extract |
| CN1836669A (en) * | 2005-03-24 | 2006-09-27 | 杭州天奇医药科技有限公司 | Method for preparing sheep placenta extract |
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Application publication date: 20120125 Assignee: Anhui Hehua Biological Medical Technology Co., Ltd. Assignor: Huzhou Kanghaisi Biological Technology Co., Ltd. Contract record no.: 2013340000043 Denomination of invention: Stock solution of sheep embryo and sheep placental peptide and preparation method thereof Granted publication date: 20130213 License type: Exclusive License Record date: 20130408 |
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