CN108685828A - A kind of extracting method of placental hormone - Google Patents
A kind of extracting method of placental hormone Download PDFInfo
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- CN108685828A CN108685828A CN201810615770.5A CN201810615770A CN108685828A CN 108685828 A CN108685828 A CN 108685828A CN 201810615770 A CN201810615770 A CN 201810615770A CN 108685828 A CN108685828 A CN 108685828A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
- A61K8/982—Reproductive organs; Embryos, Eggs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a kind of preparation methods of placental hormone, include the following steps:1) placenta is taken, water is added, is homogenized, centrifugation takes and resets and add trypsin hydrolysis, obtains enzymolysis liquid;2) trichloroacetic acid is added into enzymolysis liquid obtained by step 1), centrifuges, obtains supernatant;3) activated carbon, deodorant are added in supernatant obtained by step (2), filtering obtains filtrate;4) filtrate filtering with microporous membrane obtained by step 3), obtains clarified solution;5) acetone, mixing are added into clarified solution obtained by step 4), centrifugation takes supernatant;6) dry, sterilizing.The method of the present invention can effectively extract placental hormone, and application prospect is excellent.
Description
Technical field
The present invention relates to the preparation methods of placental hormone, belong to field of medicaments.
Background technology
With advancing age and the influence of ambient enviroment, human body can gradual agings.And aging can show that human body is each
A position, people seeking beauty are intended to that the aging of face can be slowed down, and reduce the aging of skin, relaxation, dark yellow, pigmentation, wrinkle
The problems such as line.But high-grade expensive skin care item, which are not for everyone, has the ability purchase or the purchase of duration, so people
Begin look for more economical mode nursing skin.
Have much about research of the animal placenta element in terms of beauty and skin care, existing many is using sheep placenta as the activity of raw material
Cell health products, health and beauty significant effect, but placental hormone are beauty costly.It is more suitable in order to be developed using placental hormone
The cost-effective skin care item that consumers in general use are closed, we have started the research to mouse placental hormone, it has been investigated that mouse tire
The similarity of disk element and human body is higher compared with sheep placenta, and the cost of mouse placental hormone is also low compared with sheep placenta, and raw material is just
In extraction, in terms of ingredient function, mouse placental hormone is rich in multiple proteins, it is necessary to which amino acid, nucleic acid, hormone is former, and collagen is exempted from
The nutriments such as the epidemic disease factor and calcium iron zinc.
It not only has wrinkle unfolds effect well, in whitening, eliminates color spot, enhances the effect of the elasticity aspect of skin
Also very well.Mouse placental hormone in all respects the advantages of, make it that can become a kind of new selection of the placental hormone in terms of beauty and skin care.
Placental hormone is usually extracted from placenta, but the yield of existing extracting method is too low, needs to improve.
Invention content
In view of the above-mentioned problems, the present invention provides a kind of extracting methods of placental hormone
The preparation method of placental hormone of the present invention, includes the following steps:
1) placenta is taken, water is added, is homogenized, centrifugation takes and resets and add trypsin hydrolysis, obtains enzymolysis liquid;
2) trichloroacetic acid is added into enzymolysis liquid obtained by step 1), centrifuges, obtains supernatant;
3) activated carbon, deodorant are added in supernatant obtained by step (2), filtering obtains filtrate;
4) filtrate filtering with microporous membrane obtained by step 3), obtains clarified solution;
5) acetone, mixing are added into clarified solution obtained by step 4), centrifugation takes supernatant;
6) it sterilizes.
In step 1), the volume of the water of addition is 0.5~1.5 times of placental weight, and preferably the volume of water is placental weight
1 times;And/or the condition of the centrifugation is that 10000~14000r/min centrifuges 10~20min, preferably 12000r/min centrifugations
15min。
In step 1) when trypsin hydrolysis, 0.01~0.03 times of the amount of application placental weight of trypsase, preferably
0.02 times, hydrolysis time is 1~3h, preferably 2h.
In step 1), the placenta is the placenta of mouse.
In step 2), a concentration of 1~8% (g/ml) of trichloroacetic acid in trichloroacetic acid to solution is added in enzymolysis liquid;
And/or the condition of the centrifugation is that 10000~14000r/min centrifuges 10~20min, preferably 12000r/min centrifuges 15min.
In step 3), the amount of the activated carbon of addition is 1.5~2% (g/g) of supernatant weight;And/or the deodorant is
Mode is 1~1.5h of stirring, and the rotating speed of stirring is 40~80r/min, preferably 60r/min.
In step 4), the aperture of institute's miillpore filter is 4~7 microns;After the nanofiltration, using normal saline flushing, close
And clarified solution.
In step 5), the volume that acetone is added is 8~10 times of clarified solution volume, preferably 10 times;The mixing be 0~
1~3h is stirred at 4 DEG C, the rotating speed of preferably 2h, stirring are 40~80r/min, preferably 60r/min.
In step 5), centrifugation is that 10000~14000r/min centrifuges 10~20min, preferably 12000r/min centrifugations
15min。
In step 6), the drying is spray drying;The sterilizing is to use Co-60 radiation sterilization.
The purity for the placental hormone product that the method for the present invention is produced is high, and the peptide of peptide molecular weight 300-1000 dalton contains
Amount reaches 98% or more, makes one physical efficiency and preferably absorbs, does not generate any poisonous and harmful substance in production process, safety, nontoxic
Side effect, product free from extraneous odour can be widely applied to the fields such as skin care item, food, health products, drug, the life of the method for the present invention
The production period is short, at low cost, and high income, prospects for commercial application are good.
Obviously, the above according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific implementation mode of form by the following examples remakes further specifically the above of the present invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention.
Specific implementation mode
The method of the present invention is raw materials used can be commercially available by purchase, wherein miillpore filter is the micro- of model G4
Hole filter membrane, aperture are 4~7 microns.
The preparation of 1 placental hormone of the present invention of embodiment
1, preparation method
(1) take gestational age tire mouse on the 15 to 20th with placenta out of female rat abdomen rapidly take out (avoiding bacterium as possible), remove blood,
Umbilical cord and fat freeze after cleaning, and Preservation in sterile condition is spare;
(2) aseptic condition calls rounding placenta (mouse containing tire) 100g in the following text, by 1:1 ratio (i.e. 100ml water is added in 100g placentas)
Sterile water is added, is homogenized with refiner.15min is centrifuged with 12000r/min with high speed low temperature centrifugal machine under aseptic condition, is added
Until 2g trypsin hydrolysis 1h to protease complete hydrolysis, enzymolysis liquid is obtained;
(3) trichloroacetic acid is added in the enzymolysis liquid of step (2) so that the mass concentration of trichloroacetic acid is in the solution
1% (g/ml) centrifuges 15min with 12000r/min again, removes foreigh protein removing, obtains supernatant;
(4) activated carbon (the 1.5% of supernatant weight) deodorant is added in the supernatant of step (3), stirs (60r/min)
Then 1~1.5h is filtered to remove activated carbon, obtain filtrate;
(5) filtrate of step (4) is detached with miillpore filter, normal saline flushing, clarified solution is used in combination to reach
200ml;
(6) 1 is pressed under aseptic condition:(0-4 DEG C) stirring (60r/min) 2h of acetone, high-speed low temperature centrifugation is added in 10 ratio
Machine centrifuges 15min with 12000r/min, takes supernatant;
(7) spray drying (uses BUCHI spray-dried instruments, pump speed 5-10,60 degrees Celsius of inlet temperature, air pressure
500, Aspiration 25-40), placental hormone is obtained, using Co-60 radiation sterilization.
2, property and yield detection
By detection, the yield of placental hormone of the present invention is 1.3%.The calculation formula of yield is:Yield=placental hormone weight/
Negotiate weight × 100%.
Property is as follows:
The preparation of 2 placental hormone of the present invention of embodiment
1, preparation method
(1) take gestational age tire mouse on the 15 to 20th with placenta out of female rat abdomen rapidly take out (avoiding bacterium as possible), remove blood,
Umbilical cord and fat freeze after cleaning, and Preservation in sterile condition is spare;
(2) aseptic condition calls rounding placenta (mouse containing tire) 100g in the following text, by 1:Sterile water is added in 2 ratios, is homogenized with refiner.
15min is centrifuged with 12000r/min with high speed low temperature centrifugal machine under aseptic condition, 2g trypsin hydrolysis 1h is added to protease
Until complete hydrolysis, enzymolysis liquid is obtained;
(3) trichloroacetic acid is added in the enzymolysis liquid of step (2) so that the mass concentration of trichloroacetic acid is in the solution
4% (g/ml) centrifuges 15min with 12000r/min again, removes foreigh protein removing, obtains supernatant;
(4) activated carbon (the 1.7% of supernatant weight) deodorant is added in the clear liquid of step (3), stirs (60r/min) 1
~1.5h, is then filtered to remove activated carbon, obtains filtrate;
(5) filtrate of step (4) is detached with miillpore filter, normal saline flushing, clarified solution is used in combination to reach
200ml;
(6) 1 is pressed under aseptic condition:(0-4 DEG C) stirring (60r/min) 2h of acetone, high-speed low temperature centrifugation is added in 10 ratio
Machine centrifuges 15min with 12000r/min, takes supernatant;
(7) spray drying (uses BUCHI spray-dried instruments, pump speed 5-10,60 degrees Celsius of inlet temperature, air pressure
500, Aspiration 25-40), placental hormone is obtained, using Co-60 radiation sterilization.
2, property and yield detection
By detection, the yield of placental hormone of the present invention is 0.8%.The calculation formula of yield is:Yield=placental hormone weight/
Negotiate weight × 100%.
The preparation of 3 placental hormone of the present invention of embodiment
1, preparation method
(1) take gestational age tire mouse on the 15 to 20th with placenta out of female rat abdomen rapidly take out (avoiding bacterium as possible), remove blood,
Umbilical cord and fat freeze after cleaning, and Preservation in sterile condition is spare;
(2) aseptic condition calls rounding placenta (mouse containing tire) 100g in the following text, by 2:Sterile water is added in 1 ratio, is homogenized with refiner.
15min is centrifuged with 12000r/min with high speed low temperature centrifugal machine under aseptic condition, 2g trypsin hydrolysis 1h is added to protease
Until complete hydrolysis, enzymolysis liquid is obtained;
(3) trichloroacetic acid is added in the enzymolysis liquid of step (2) so that the mass concentration of trichloroacetic acid is in the solution
8% (g/ml) centrifuges 15min with 12000r/min again, removes foreigh protein removing, obtains supernatant;
(4) activated carbon (the 2.0% of supernatant weight) deodorant is added in the supernatant of step (3), stirs (60r/min)
Then 1~1.5h is filtered to remove activated carbon, obtain filtrate;
(5) filtrate of step (4) is detached with miillpore filter, normal saline flushing, clarified solution is used in combination to reach
200ml;
(6) 1 is pressed under aseptic condition:(0-4 DEG C) stirring (60r/min) 2h of acetone, high-speed low temperature centrifugation is added in 10 ratio
Machine centrifuges 15min with 12000r/min, and precipitation separates acetone solution, takes supernatant;
(7) spray drying (uses BUCHI spray-dried instruments, pump speed 5-10,60 degrees Celsius of inlet temperature, air pressure
500, Aspiration 25-40), placental hormone is obtained, using Co-60 radiation sterilization.
2, property and yield detection
By detection, the yield of placental hormone of the present invention is 1.5%.The calculation formula of yield is:Yield=placental hormone weight/
Negotiate weight × 100%.
In conclusion the method for the present invention can effectively extract placental hormone, high income, and preparation method is easy to operate, fits
For industrialized production, application prospect is had a vast market.
Claims (10)
1. a kind of preparation method of placental hormone, which is characterized in that include the following steps:
1) placenta is taken, water is added, is homogenized, centrifugation takes and resets and add trypsin hydrolysis, obtains enzymolysis liquid;
2) trichloroacetic acid is added into enzymolysis liquid obtained by step 1), centrifuges, obtains supernatant;
3) activated carbon, deodorant are added in supernatant obtained by step (2), filtering obtains filtrate;
4) filtrate filtering with microporous membrane obtained by step 3), obtains clarified solution;
5) acetone, mixing are added into clarified solution obtained by step 4), centrifugation takes supernatant;
6) dry, sterilizing.
2. preparation method according to claim 1, it is characterised in that:In step 1), the volume of the water of addition is Placental Weight
0.5~1.5 times of amount, the preferably volume of water are 1 times of placental weight;And/or the condition of the centrifugation be 10000~
14000r/min centrifuges 10~20min, and preferably 12000r/min centrifuges 15min.
3. preparation method according to claim 1, it is characterised in that:In step 1) when trypsin hydrolysis, trypsase
0.01~0.03 times of amount of application placental weight, preferably 0.02 times, hydrolysis time is 1~3h, preferably 2h.
4. preparation method according to claim 1, it is characterised in that:In step 1), the placenta is the placenta of mouse.
5. preparation method according to claim 1, it is characterised in that:In step 2), trichloroacetic acid is added in enzymolysis liquid
To a concentration of 1~8% (g/ml) of trichloroacetic acid in solution;And/or the condition of the centrifugation is 10000~14000r/min
10~20min is centrifuged, preferably 12000r/min centrifuges 15min.
6. preparation method according to claim 1, it is characterised in that:In step 3), the amount of the activated carbon of addition is supernatant
1.5~2% (g/g) of liquid weight;And/or it is 1~1.5h of stirring that the deodorant, which is mode, the rotating speed of stirring is 40~80r/
Min, preferably 60r/min.
7. preparation method according to claim 1, it is characterised in that:In step 4), the aperture of institute's miillpore filter is 4~7
Micron;After the nanofiltration, using normal saline flushing, merge clarified solution.
8. preparation method according to claim 1, it is characterised in that:In step 5), the volume that acetone is added is clarified solution
8~10 times of volume, preferably 10 times;The mixing is 1~3h of stirring at 0~4 DEG C, preferably 2h, the rotating speed of stirring is 40~
80r/min, preferably 60r/min.
9. preparation method according to claim 1, it is characterised in that:In step 5), centrifugation is 10000~14000r/min
10~20min is centrifuged, preferably 12000r/min centrifuges 15min.
10. preparation method according to claim 1, it is characterised in that:In step 6), the drying is spray drying;Institute
It is to use Co-60 radiation sterilization to state sterilizing.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810615770.5A CN108685828A (en) | 2018-06-14 | 2018-06-14 | A kind of extracting method of placental hormone |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810615770.5A CN108685828A (en) | 2018-06-14 | 2018-06-14 | A kind of extracting method of placental hormone |
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ID=63848401
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109172607A (en) * | 2018-11-13 | 2019-01-11 | 哈哈驴（天津）国际贸易有限公司 | Process and method for extracting placenta from fresh donkey placenta |
| CN111544363A (en) * | 2020-04-26 | 2020-08-18 | 无锡可尚生物科技有限公司 | Preparation method of bovine placenta extract |
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| CN1440770A (en) * | 2002-12-26 | 2003-09-10 | 牛朋斌 | Sheep fetus peptide capsule and its prepn process |
| CN101028286A (en) * | 2006-03-01 | 2007-09-05 | 红原牦牛肉食品有限责任公司 | Sheep placenta extracts and preparation thereof |
| CN101837005A (en) * | 2010-04-12 | 2010-09-22 | 湖南圣雅凯生物科技有限公司 | Process for preparing pig placenta extract |
| CN104372057A (en) * | 2014-11-17 | 2015-02-25 | 得利斯集团有限公司 | Extracting method of placenta |
| CN105950693A (en) * | 2016-07-20 | 2016-09-21 | 白城天福粮油食品集团有限公司 | Extraction method for bioactive polypeptides of bovine placenta |
| CN106727709A (en) * | 2016-11-24 | 2017-05-31 | 湖州珍露生物制品有限公司 | A kind of production technology of sheep placenta extract solution |
-
2018
- 2018-06-14 CN CN201810615770.5A patent/CN108685828A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1440770A (en) * | 2002-12-26 | 2003-09-10 | 牛朋斌 | Sheep fetus peptide capsule and its prepn process |
| CN101028286A (en) * | 2006-03-01 | 2007-09-05 | 红原牦牛肉食品有限责任公司 | Sheep placenta extracts and preparation thereof |
| CN101837005A (en) * | 2010-04-12 | 2010-09-22 | 湖南圣雅凯生物科技有限公司 | Process for preparing pig placenta extract |
| CN104372057A (en) * | 2014-11-17 | 2015-02-25 | 得利斯集团有限公司 | Extracting method of placenta |
| CN105950693A (en) * | 2016-07-20 | 2016-09-21 | 白城天福粮油食品集团有限公司 | Extraction method for bioactive polypeptides of bovine placenta |
| CN106727709A (en) * | 2016-11-24 | 2017-05-31 | 湖州珍露生物制品有限公司 | A kind of production technology of sheep placenta extract solution |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109172607A (en) * | 2018-11-13 | 2019-01-11 | 哈哈驴（天津）国际贸易有限公司 | Process and method for extracting placenta from fresh donkey placenta |
| CN109172607B (en) * | 2018-11-13 | 2020-09-22 | 哈哈驴（天津）国际贸易有限公司 | Process and method for extracting placenta from fresh donkey placenta |
| CN111544363A (en) * | 2020-04-26 | 2020-08-18 | 无锡可尚生物科技有限公司 | Preparation method of bovine placenta extract |
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Application publication date: 20181023 |