CN103462864B - Embryo water extract extracted from animal embryo internal organs, and extraction method and applications thereof - Google Patents
Embryo water extract extracted from animal embryo internal organs, and extraction method and applications thereof Download PDFInfo
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Abstract
The invention discloses an embryo water extract extracted from animal embryo internal organs, and an extraction method and applications thereof, and belongs to the field of extraction technology of embryo extract from animal embryos and applications of the extraction technology. The extraction method is capable of extracting the water soluble embryo extract without enzymatic hydrolysis by proteolytic enzyme, wherein the molecular weight of the water soluble embryo extract is less than 4000D. The extraction method is capable of increasing production efficiency and raw material utilization ratio, reducing production cost, meeting the requirements of cosmetic and food production, and ensuring easy absorption of the substances in the extract by human skin and mucosal tissues, and is beneficial to industrialized large-scaled production.
Description
Technical field
The present invention relates to from animal embryo, particularly from animal embryo internal viscera such as sheep, pig and Canis familiaris L.s, comprise the Technology field of extracting eggembryosin in the internal organs such as the liver of embryo, heart, kidney, intestinal and blood.
Background technology
Switzerland scientist Ka Er in 1912 teaches because there being a kind of magical material that human body cell can be made to rejuvenate in Late Cambrian sheep embryonic cell, i.e. sheep embryo extract and obtain Nobel Prize in medicine.The mankind find a kind of can the pure natural animal product of slow down aging, i.e. sheep embryo extract, thus make people pursue dream that is young and life lengthening to become a reality.
According to science textual criticism, the number of times of biological cell division and there is certain dependency between the life-span, such as, the life-span is the chicken of 30 years, and cell division is 25 times; And the life-span is only the white mice of 3 years, only division 12 times.And cause the basic reason of human senility to be form human body life the most basic unit cell to occur aging; Along with the increase at age, cell division is more and more slower, and aged cells makes organ function decline, and even causes pathological changes, and people is also thereupon old and feeble.Ka Er teaches the sheep embryo extract found in sheep embryo can make animal body frequency dividing cell increase, and can make to have failed or aging cell reactivation or be eliminated, thus aging function is restored.
1931, Switzerland Borrow. the mode that Ni Han teaches first passage injection the parathyroidin of lamb is used for parathyroid gland impaired be dying patient successfully saved its life, this have milestone formula meaning be found to be the mankind in the future sheep embryo extract lay a good foundation in application that is biomedical and defying age field, the Pope, in order to represent the respect to him, awards Feng Qiwei the Vatican academy of science academician.Nineteen fifty, Switzerland Dr.Alfred Pfister and professor Ni Han, through cooperation for many years and effort, find that the extract of sheep embryonic cell is the optimum carrier of competent cell material.
Nineteen fifty-two, Dr.Alfred Pfister has set up Switzerland's Lei Manna health medical treatment center, promotes sheep embryo extract living cells therapy.Nineteen fifty-five, the damage very easily caused when he solves cell extraction under the graduate support of Switzerland Batelle and deteriorating course, and the technological deficiency of preserving, this achievement is named as Cellvital.1956, achievement in research Cellvital got back and once obtained the Otto of Nobel Prize in medicine in 1931. the demonstration of Heinrich. Wa Erbao cell biological scholar.
Over more than 70 year, whole world hundreds of thousands of them carried out the associated treatment of sheep embryo extract, comprising front Pope Borrow 12 generation, Queen of England and royalty member, Philippine president Marx, Saudi Arabia and Yemen king, Japanese Emperor Hirohito, mound gill prime minister, French gaullist president, Charlie Chaplin, and Norma Jean Remix, Dai Deli, Garbo, soccer star's Maradona etc.
Sheep embryo extract contains the multiple active component that can play mediator soma organ dysfunction, and its Concentrations of Epidermal Growth Factor (EGF), Hyaluronic acid-stimulating factor (HASF) and basic fibroblast growth factor (bFGF) etc. have the effect of beautifying skin.EGF particularly in sheep embryo extract, just becomes fashion as a kind of emerging beauty and skin care factor.Research shows, the corneocyte of people and fibrocyte etc. all have specificity EGF receptor to exist, and provides theoretical foundation for human skin cell has the ability absorbing EGF.
In addition, also containing multiple small active peptides, nucleic acid, vitamin, several kinds of mineral elements and abundant aminoacid isoreactivity composition in sheep embryo extract, while promotion skin cell renewal, the content of its Endogenous Growth Factors can also be increased, promote emiocytosis hyaluronic acid and glycoprotein, there is adjustment skin texture, slow down aging, reduce wrinkle, suppress the effect of acne and the growth of control comedo.Therefore, sheep embryo extract will have more and more stronger vitality and good development prospect as a kind of cosmetics additive with unique biological effect.
Through century-old research, sheep placental extract is affirmative in the effect in body aging that delays; And the effect in raising immunity of organisms of the small-molecule active substance contained by it is also very significant; In addition, at adjustment body hormonal readiness, improve anemia, strengthen metabolism and promote also there is remarkable result in body energy etc.Sheep embryo extract be one both can within use, also can the active small molecular polypeptides matter of external, it is absorbed by body and the mode that utilizes also is diversified, and it will be played a greater and greater role to the beauty treatment of the mankind and healthy cause.
The leaching process of current sheep placental extract approximately can be divided into three phases: first from lamb foetus organ cell, extract active component, removes impurity, thermo-responsive protein and allergenic factor; Secondly by isolation technics, obtain the active substance with different efficacies, bioactive peptide as dissimilar in embryonic thymus element, multiple somatomedin etc., finally adopt freeze drying technology to carry out lyophilization, ensure that it is active.It should be noted that the extraction of sheep placental extract is a very complicated biotechnology processes, nowadays most leaching process can only complete in the lab, is difficult to carry out a large amount of suitability for industrialized production and preserves for a long time.Laboratory is had to adopt the method for proteolytic enzyme enzymolysis to extract sheep embryo extract in China, although this method can increase substantially total protein content in hydrolyzed solution, but to find through current research in the gross protein detected that main what increase is do not have the aminoacid of bio-regulating activity and the fragment of bioactive peptide, the eggembryosin content that really can play regulating action is then lower, illustrate that the sheep embryo extract extracted by the enzyme solution of proteolytic enzyme is active and content is all lower, proteolytic enzyme has certain destruction to sheep embryo extract, present this extracting method gradually eliminate by new extracting method.
Summary of the invention
The present invention first object is to propose a kind of eggembryosin water extract extracted from animal embryo internal viscera.
Water extract of the present invention is the water solublity eggembryosin that the molecular weight extracted from the internal organs of animal embryo inside is less than 4000D.
Indication animal of the present invention is not sheep, can also be extended to the animal such as pig and Canis familiaris L., and animal embryo internal organs can as internal organs such as liver, heart, kidney, intestinal and blood, and the available embryo's commercial weight of the present invention is extensive, is easy to get, is beneficial to industrialized mass.The molecular weight of water solublity eggembryosin of the present invention is less than 4000D and can guarantees that in extracting solution, material is easily absorbed by human body skin and mucosal tissue.
The present invention second object is that invention is a kind ofly beneficial to large-scale production, the method from animal embryo internal viscera extract at low temperature eggembryosin of production cost of increasing operation rate, reduce.
The present invention includes following steps:
1) the animal embryo distilled water of freezen protective under-15 ~-25 DEG C of temperature conditions is cleaned, isolate internal organs in embryo;
2) in atmospheric conditions, adopt temperature be the distilled water of 0 ~ 5 DEG C steep for mechanical activation comminution embryo in internal organs 20 ~ 30 minutes, the mixed weight of described distilled water and internal organs in described embryo is than 1 ~ 4 ︰ 1;
3) mixed liquor be mixed to form by internal organs in distilled water and embryo is repeatedly freezing and thaw 2 ~ 4 times, at every turn freezing ambient temperature is-15 ~-30 DEG C, each cooling time is 24 ~ 48 hours, thaws to 5 DEG C at every turn, adopts low-frequency ultrasonic waves process 20 ~ 40 minutes in course of defrosting;
4) with the refrigerated centrifuger of 4000 ~ 15000 revs/min to above-mentioned mixed liquor process 10 ~ 15 minutes, removing solid impurity, obtain water extraction solution A;
5) the Ph value of water extraction solution A is adjusted to 1.5 ~ 3.5, adopts the method for frozen centrifugation to remove precipitate, get supernatant and adjust Ph value to 3.5 ~ 6.5 again, then remove precipitate through the method for frozen centrifugation, obtain water extraction solution B;
6) under the ambient temperature of 1 ~ 5 DEG C, ultrafiltration is carried out to water extraction solution B, isolate the water solublity eggembryosin that molecular weight is below 4000D.
Raw material of the present invention adopts the animal embryo of freezen protective, farthest keeps the activity of eggembryosin in embryo; The object adopting distilled water to soak improves purity when extracting eggembryosin, reduces the pollution that impurities in water brings, and distilled water temperature keeps 0 ~ 5 DEG C, is conducive to the activity protecting eggembryosin in extracting solution; Through the freezing and processing method of thawing repeatedly, make mixed liquor be meat paste shape, object is all to improve extraction efficiency, increases the content of eggembryosin in extracting solution, and farthest ensures the activity of eggembryosin in leaching process, is beneficial to follow-up separation; The Ph value adopting first water transfer to put forward solution A is adjusted to 1.5 ~ 3.5, adjusts twice frozen centrifugation method of Ph value to 3.5 ~ 6.5 again, efficiently can remove the large protein in Aqueous extracts A, is beneficial to follow-up ultrafiltration and extracts; Under the ambient temperature of 1 ~ 5 DEG C, carry out ultrafiltration to water extraction solution B, object guarantees to extract the activity of eggembryosin not by the impact of temperature, is beneficial to and extracts active ingredients efficiently.The natural conditions that final products pH value and eggembryosin exist are close, fully can ensure the natural activity of the eggembryosin extracted.
Invent the high activity eggembryosin that just can extract natural structure without the need to the enzymolysis of proteolytic enzyme, improve eggembryosin content, be beneficial to industrial-scale production, enhance productivity, improve the utilization rate of raw material, to reduce production cost, cosmetics and food production can be met.
The animal embryo age in days of step 1) of the present invention is 30 ~ 145 days, and object makes the content of eggembryosin in extracted embryo more.
Separately, in described step 5), the water extraction solution A of 0 ~ 5 DEG C being carried out the adjustment of Ph value, is also the activity of the effective ingredient more advantageously kept in water extraction solution A.
3rd object of the present invention is the purposes that the above molecular weight of proposition is less than the water solublity eggembryosin of 4000D.
One of purposes: the purposes of eggembryosin water extract in Cosmetic Manufacture extracted from animal embryo internal viscera, feature is: when production cosmetic, and the ratio that water solublity eggembryosin molecular weight being less than 4000D accounts for 5 ~ 50% in the quality that feeds intake replaces original water in cosmetics.
Purposes two: the purposes of eggembryosin water extract in Cosmetic Manufacture extracted from animal embryo internal viscera, feature is: when production cosmetic, water solublity eggembryosin molecular weight being less than 4000D, after lyophilization, packs and makes cosmetics.
Through on probation, above two class cosmetics are remarkable to cosmetic result, and have no side effect, and low cost, simple.
Purposes three: the eggembryosin water extract extracted from animal embryo internal viscera is in the purposes of food production, and feature is: molecular weight is less than the water solublity eggembryosin of 4000D for the preparation of health care oral liquid and capsule.
The present invention is easy to use, good absorbing effect, cheap as the advantage of health care oral liquid and capsule, can be used as general health product and immunostimulant uses.
Detailed description of the invention
One, molecular weight is less than the extracting method of the water solublity eggembryosin of 4000D:
1, get under 30 ~ 145 age in days fresh animal embryos are placed in-15 ~-25 DEG C of cryogenic conditions and store.
2, the animal embryo distilled water of freezen protective is cleaned, and be separated the internal organs such as fetal liver, heart, kidney, intestinal and blood, then the internal organs after being separated are adopted mechanical activation comminution, stand-by.
3, in atmospheric conditions, adopt temperature be the distilled water of 0 ~ 5 DEG C steep for mechanical activation comminution embryo in internal organs 20 ~ 30 minutes.The mixed weight of distilled water and internal organs in embryo is than 1 ~ 4 ︰ 1.
4, by freezing for the mixed liquor iterative cycles be mixed to form by internal organs in distilled water and embryo and process 4 times of thawing, at every turn freezing ambient temperature is-15 ~-30 DEG C, each cooling time is 24 ~ 48 hours, thaws to 5 DEG C at every turn, adopts low-frequency ultrasonic waves process 20 ~ 40 minutes in course of defrosting.
5, with the refrigerated centrifuger of 4000 ~ 15000 revs/min to above-mentioned freezing through iterative cycles and the mixed liquor process of thawing 10 ~ 15 minutes, removing solid impurity, obtains water extraction solution A.
6, the water extraction solution A of 0 ~ 5 DEG C is adjusted Ph value to 1.5 ~ 3.5 with 1 ~ 2N hydrochloric acid, adopt the method for frozen centrifugation to remove precipitate, get supernatant.By the supernatant of 0 ~ 5 DEG C with 1 ~ 2N sodium hydroxide adjustment Ph value to 3.5 ~ 6.5, then remove precipitate through the method for frozen centrifugation, obtain water extraction solution B.
7, under the ambient temperature of 1 ~ 5 DEG C, adopt the ultrafilter of 4000D to carry out ultrafiltration to water extraction solution B, isolate the water solublity eggembryosin that molecular weight is below 4000D.
Two, molecular weight is less than the qualification of the water solublity eggembryosin of 4000D:
Eggembryosin content in supernatant after adopting nanodrop2000 ultraviolet spectrometry to measure ultrafiltration, eggembryosin content of the present invention can be reached for 0.4% ~ 4% after measured, be 2 ~ 10 times of traditional eggembryosin extracting method content, extraction efficiency and the activity of eggembryosin can be increased substantially.
Three, cosmetics are prepared:
1, cosmetics one:
Factory formula as astringent:
Take molecular weight respectively and be less than the water solublity eggembryosin 5kg of 4000D, the 95kg that purifies waste water by above raw material mix homogeneously, carry out bottled, namely make astringent.
2, cosmetics two:
Freeze-dried preparation as eggembryosin water extract:
Water solublity eggembryosin molecular weight being less than 4000D, after lyophilization, packs and makes freeze-dried preparation.
Above astringent is directly used in respectively clean after face, by freeze-dried preparation directly or adopt introducer be used for clean after face, absorb faster than mill run.
Be applied to skin aging crowd, it is remarkable that skin recovers elastic effect, and have no side effect.
Four, health product are prepared:
1, eggembryosin oral formulations A is prepared:
Be that the water solublity eggembryosin of below 4000D and Mel or other food additive are mixed well by molecular weight, after ultrafiltration and pasteurization sterilizing, the bottled oral liquid of fill.
2, eggembryosin oral formulations B is prepared:
Be obtain lyophilized powder after the water solublity eggembryosin lyophilization of below 4000D as the raw material producing various capsule using molecular weight.
3, eggembryosin capsule is prepared:
Be obtain lyophilized powder 5 kilograms, the lyophilized powder after Flos Camelliae Japonicae powder lyophilization 95 kilograms after the water solublity eggembryosin lyophilization of below 4000D by molecular weight, granulate after the two mixing, the hard capsule that fill every net content is 250 milligrams.
4, apply:
Take object: physical weakness person.
Taking dose: according to 4 ~ 10 milliliters for each person every day when oral liquid uses, connects and takes 20 ~ 30 days.Oral capsule, if capsule content is lyophilized powder 10 ~ 50 milligrams/, everyone takes 20 ~ 30 days continuously according to 1 ~ 4/day.
Frequent easy catching a cold person and hyperlipemia, take product of the present invention about 20 days, body immunity improves not easy catching a cold, and hyperlipidemia index can recover normal.
Claims (5)
1. the eggembryosin water extract extracted from animal embryo internal viscera, it is characterized in that described water extract is the water solublity eggembryosin that the molecular weight extracted from the internal organs of animal embryo inside is less than 4000D, the extracting method of described water extract comprises the following steps:
1) cleaned by the animal embryo distilled water of freezen protective under-15 ~-25 DEG C of temperature conditions, isolate internal organs in embryo, the age in days of described animal embryo is 30 ~ 145 days;
2) in atmospheric conditions, adopt temperature be the distilled water of 0 ~ 5 DEG C steep for mechanical activation comminution embryo in internal organs 20 ~ 30 minutes, the mixed weight of described distilled water and internal organs in described embryo is than 1 ~ 4 ︰ 1;
3) mixed liquor be mixed to form by internal organs in distilled water and embryo is repeatedly freezing and thaw 2 ~ 4 times, at every turn freezing ambient temperature is-15 ~-30 DEG C, each cooling time is 24 ~ 48 hours, thaws to 5 DEG C at every turn, adopts low-frequency ultrasonic waves process 20 ~ 40 minutes in course of defrosting;
4) with the refrigerated centrifuger of 4000 ~ 15000 revs/min to above-mentioned mixed liquor process 10 ~ 15 minutes, removing solid impurity, obtain water extraction solution A;
5) pH value of the water extraction solution A of 0 ~ 5 DEG C is adjusted to 1.5 ~ 3.5, adopts the method for frozen centrifugation to remove precipitate, get supernatant adjust pH to 3.5 ~ 6.5 again, then remove precipitate through the method for frozen centrifugation, obtain water extraction solution B;
6) under the ambient temperature of 1 ~ 5 DEG C, ultrafiltration is carried out to water extraction solution B, isolate the water solublity eggembryosin that molecular weight is below 4000D.
2. the preparation method of the eggembryosin water extract extracted from animal embryo internal viscera as claimed in claim 1, is characterized in that comprising the following steps:
1) cleaned by the animal embryo distilled water of freezen protective under-15 ~-25 DEG C of temperature conditions, isolate internal organs in embryo, the age in days of described animal embryo is 30 ~ 145 days;
2) in atmospheric conditions, adopt temperature be the distilled water of 0 ~ 5 DEG C steep for mechanical activation comminution embryo in internal organs 20 ~ 30 minutes, the mixed weight of described distilled water and internal organs in described embryo is than 1 ~ 4 ︰ 1;
3) mixed liquor be mixed to form by internal organs in distilled water and embryo is repeatedly freezing and thaw 2 ~ 4 times, at every turn freezing ambient temperature is-15 ~-30 DEG C, each cooling time is 24 ~ 48 hours, thaws to 5 DEG C at every turn, adopts low-frequency ultrasonic waves process 20 ~ 40 minutes in course of defrosting;
4) with the refrigerated centrifuger of 4000 ~ 15000 revs/min to above-mentioned mixed liquor process 10 ~ 15 minutes, removing solid impurity, obtain water extraction solution A;
5) pH value of the water extraction solution A of 0 ~ 5 DEG C is adjusted to 1.5 ~ 3.5, adopts the method for frozen centrifugation to remove precipitate, get supernatant adjust pH to 3.5 ~ 6.5 again, then remove precipitate through the method for frozen centrifugation, obtain water extraction solution B;
6) under the ambient temperature of 1 ~ 5 DEG C, ultrafiltration is carried out to water extraction solution B, isolate the water solublity eggembryosin that molecular weight is below 4000D.
3. the purposes of eggembryosin water extract in Cosmetic Manufacture extracted from animal embryo internal viscera as claimed in claim 1, it is characterized in that, when production cosmetic, the ratio that water solublity eggembryosin molecular weight being less than 4000D accounts for 5 ~ 50% in the quality that feeds intake replaces original water in cosmetics.
4. the purposes of eggembryosin water extract in Cosmetic Manufacture extracted from animal embryo internal viscera as claimed in claim 1, it is characterized in that, when production cosmetic, water solublity eggembryosin molecular weight being less than 4000D, after lyophilization, packs and makes cosmetics.
5. the eggembryosin water extract extracted from animal embryo internal viscera as claimed in claim 1, in the purposes of food production, is characterized in that, molecular weight is less than the water solublity eggembryosin of 4000D for the preparation of health care oral liquid and capsule.
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| CN105646647A (en) * | 2015-12-30 | 2016-06-08 | 耿威 | Chick embryo polypeptide as well as preparation method and application of chick embryo polypeptide |
| CN106562993A (en) * | 2016-10-26 | 2017-04-19 | 沈会强 | Preparation method and application of sika deer embryo factors |
| CN106581467A (en) * | 2016-10-26 | 2017-04-26 | 沈会强 | Corn embryonic factor manufacturing method and application |
| CN106581466A (en) * | 2016-10-26 | 2017-04-26 | 沈会强 | Manufacturing method and applications of rye embryo extract |
| CN106562995A (en) * | 2016-10-26 | 2017-04-19 | 沈会强 | Preparation method for preparing swan embryos and application of swan embryos |
| CN108323750A (en) * | 2017-01-20 | 2018-07-27 | 内蒙古草原鑫河食品有限公司 | A kind of active constituent and its extracting method and application with liver protection |
| CN106821939A (en) * | 2017-03-19 | 2017-06-13 | 湖南海济药业有限公司 | The extracting method of sheep embryo extract and the preparation containing its extract |
| CN109908181A (en) * | 2019-04-23 | 2019-06-21 | 独角兽生命科学股份公司 | The preparation method of anti-aging Porcine HGF |
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