CN105534849A - Chicken embryo peptide as well as preparation method and application thereof in preparation of anti-inflammation, acne removal and black hair generation products - Google Patents
Chicken embryo peptide as well as preparation method and application thereof in preparation of anti-inflammation, acne removal and black hair generation products Download PDFInfo
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- 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
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- A61K38/00—Medicinal preparations containing peptides
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- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/02—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings containing insect repellants
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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Abstract
The invention relates to the field of biotechnology and particularly relates to a chicken embryo peptide as well as a preparation method and application thereof. The chicken embryo peptide provided by the invention comprises chicken embryo peptide and free amino acid, wherein the molecular weight of the chicken embryo peptide is less than 50kDa. The chicken embryo peptide provided by the invention can realize the anti-inflammation, acne removal and black hair generation functions; and survey result indicates that the chicken embryo peptide provided by the invention has a good effect on local inflammation and detumescence of human body epidermis and particularly realizes a perfect curative effect on mosquito bites. Moreover, the chicken embryo peptide provided by the invention can realize a good effect on the acnes on human faces and also can promote the regeneration of black hair. The preparation method of the chicken embryo peptide provided by the invention is easy and convenient to operate; and through homogenization, freeze thawing and ultrasonication, the cytomembrane is broken to the greatest extent so that intracellular bioactive peptides are dissolved out well and the extraction efficiency is improved.
Description
Technical field
The present invention relates to biological technical field, particularly relate to a kind of Embryo Gallus domesticus peptide and preparation method thereof and the application preparing antiinflammatory, in product that anti-acne, hair color regenerate.
Background technology
Aminoacid is the common name of the class organic compound containing amino and carboxyl.Wherein, glycine (GLY) has raising muscle vitality, human body is had to the Nutritions such as tonification; Leucine (LEU) has the effect promoting skin, wound healing; Methionine (MET) has the function of degrease, can promote the synthesis of skin protein; Tyrosine (TYR) has treatment Neurogenic inflammatory effect, participates in the inflammation of repairing skin.Relation is had with chromogenesis, can sharp albinism during shortage; Histidine (HIS) has repairing effect to allergic skin; Threonine (THR) needed by human, can make people become thin during shortage; Alanine (ALA) can promote that metabolism (decomposition) effect of ethanol in blood strengthens liver function; Isoleucine (ILE) can maintain organism balance, has the effect of appetitive increase; Tryptophan (TRY) promotes the synthesis of hemoglobin, prevents and treats dermatosis, Promote cell's growth; Cystine (CYS) has the damage of repairing skin, effective in cure to alopecia; Lysine (LYB) participates in the formation of connective tissue, microvascular endothelial cells interstitial, and keeps normal permeability; Aspartic acid (ASP) has protective effect to muscle and skin; Valine (VAL) impels nervous function normal, if when lacking, tactile sensing degree can be caused to improve especially; Phenylalanine (PHE) changes tyrosine in body, promotes thyroxine and adrenergic synthesis; Proline (PRO) is to the effective in cure effect of hypertension; Serine (SER) can reduce the cholesterol concentration in blood, has antibacterial action; Glutamic acid (GLU) has protection skin wet, as the cosmetics of preparation, has good effect to skin, mucosa unit and cell recovery, is suitable for child and dermatosis patient use; Arginine (ARG) can increase muscle vitality, keeps the normal function of body.
Peptide is that a-amino acid links together with peptide bond and the compound formed, and it is also the intermediate product of proteolysis.There is no obvious boundary between polypeptide and protein, usually, according to the quantity of amino acid residue, the peptide of 2 ~ 10 aminoacid compositions is called oligopeptide (or claiming small-molecular peptides); The peptide of 10 ~ 50 aminoacid compositions is called polypeptide; The peptide be made up of the aminoacid of more than 50 is large peptide.
The embryo of Embryo Gallus domesticus and chicken is after fertilized eggs hatching to certain natural law, the edible egg of also non-broken shell.Research shows, the nutritional labeling comprised in Embryo Gallus domesticus comprises protein, polypeptide, aminoacid, carbohydrate (glucose, fructose, galactose, arabinose, furanose etc.), lipid, fatty acid etc.In addition, infection composition and bioactive substance is also comprised in Embryo Gallus domesticus.Described infection composition has globulin, interferon, fetoprotein, lysozyme and phosphodiesterase; Described bioactive substance has sialic acid, youngster's phenol amine, insulin like growth factor-1, insulin, kininase, histaminase, choline, erythropoietin, deoxyribonuclease and ribonuclease etc.Relative to unhatched egg, the content of Embryo Gallus domesticus Middle nutrition composition, particularly small molecular protein or polypeptide is abundanter, and the fat paid no attention to human body, cholesterol equal size then decline to a great extent.
Embryo Gallus domesticus is nutritious, has potential skin protection, health care and medical value.But also less at present to the research of peptide in Embryo Gallus domesticus at present, tackle it and carry out further development and utilization in the research of cosmetics, health product, medicine.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of Embryo Gallus domesticus peptide and preparation method thereof and the application preparing antiinflammatory, in product that anti-acne, hair color regenerate.Embryo Gallus domesticus peptide provided by the invention has good UVResistant effect, can Cell protection normal growth under ultra-vioket radiation, and cell growth has facilitation.
Embryo Gallus domesticus peptide provided by the invention, comprises Embryo Gallus domesticus peptide and free amino acid; Wherein, the molecular weight of Embryo Gallus domesticus peptide is less than 50kDa.
In the Embryo Gallus domesticus peptide that the present invention obtains, bioactive peptide 404 detected altogether, respectively from 98 kinds of different protein.According to PSMs (Peptide-SpectralMatch), these protein are mainly: Ovalbumin (accession number P01012), ApolipoproteinB (accession number Q197X2), Ovotransferrin (accession number E1BQC2), HistoneH3.2 (accession number P84229), Vitellogenin-2 (accession number P02845), accession number is the albumen of E1BTF4, Apovitellenin-1 (accession number P02659), OvoglobulinG2typeAB (accession number I0J174), OvoglobulinG2 (accession number I0J170) and Alpha-1-acidglycoprotein (accession number A7UEB0).
In Embryo Gallus domesticus peptide of the present invention, the concentration of Embryo Gallus domesticus peptide is 0.1g/ μ L.
The present invention extracts the Embryo Gallus domesticus peptide obtained and comprises 18 kinds of free amino acids, comprising 7 kinds of essential amino acids, is respectively threonine, valine, isoleucine, leucine, phenylalanine, lysine and tryptophan.Also comprise 11 kinds of non essential amino acid in addition, be respectively aspartic acid, serine, glutamic acid, glycine, alanine, methionine, tyrosine, histidine, arginine, cystine and proline.
In Embryo Gallus domesticus peptide of the present invention, the concentration of free amino acid is 14.3mg/mL.
The preparation method of Embryo Gallus domesticus peptide provided by the invention comprises: shelled by Embryo Gallus domesticus, after homogenate, smudge cells; After papain enzymolysis, collected by centrifugation supernatant, molecular cut off is less than the component of 50kDa.
Embryo Gallus domesticus of the present invention is the Embryo Gallus domesticus of hatching to 14 ~ 19 days.
Concrete, the obtain manner of Embryo Gallus domesticus is, gets SPF level fertilized eggs, puts into incubator and hatch, incubation temperature controls at 37.5 DEG C, relative humidity 60%, and every 2 hours automatic egg turnings once, treat that hatching was to 14 ~ 19 days, as preferably, adopt the Embryo Gallus domesticus of hatching to 16 ~ 17 days.
The embryo of the Embryo Gallus domesticus of fertilization hatching to 14 day ~ 19 days is in the stem cell height rise period, and various histoorgan basically forms.In the hatching process of Embryo Gallus domesticus, the metabolism of protein is very violent, and between the incubation period, the content of proteins and peptides is in continuing propradation.Research shows, at the 14th day ~ 19 days of hatching, in Embryo Gallus domesticus, the increasing degree of proteins and peptides was maximum, and cytokine class polypeptide is maximum, and the content of free amino acid also reaches the highest during this period.
Before preparing Embryo Gallus domesticus polypeptide, hatching chicken embryo stops hatching.Be specially, hatching chicken embryo be positioned over-20 DEG C and stop hatching.
In the present invention, described homogenate adopts tissue refiner.
Be specially: the egg surface sterilization to 14d ~ 19d will be hatched, with tissue refiner homogenate after removal eggshell.As preferably, the temperature of homogenate is 4 DEG C, and the time is 20min rotating speed is 2500 revs/min.
Through tissue homogenate, chicken embryo tissue is pulverized, and cell is preliminary broken under the mechanism of refiner.In order to further smudge cells, carry out freeze thawing and ultrasonication.
In embodiments of the present invention, smudge cells comprises the step of freeze thawing and ultrasonication;
The cryogenic temperature of freeze thawing is-20 DEG C ~-40 DEG C, and the time is 60h ~ 80h; Solution temperature is 4 DEG C ~ 8 DEG C;
The temperature of ultrasonication is 4 DEG C ~ 8 DEG C, and power is 800W ~ 1200W, and the time is 15min ~ 25min.
In certain embodiments, smudge cells comprises the step of freeze thawing and ultrasonication;
The cryogenic temperature of freeze thawing is-30 DEG C, and the time is 72h; Solution temperature is 4 DEG C;
The temperature of ultrasonication is 4 DEG C, and power is 1000W, and the time is 20min.
Freeze thawing makes cell in the process of freeze thaw, makes the salinity of residue cytosol increase and cause cell breakage owing to forming ice pellets in cell.In the present invention, the temperature of dissolving is 4 DEG C, is dissolved to cell homogenates and melts.After freeze thawing, cell membrane further breaks, then carries out ultrasonic disruption, thus makes the degree of crushing of cell membrane more complete, improves the extraction efficiency of bioactive peptide.
In the present invention, after the tissue homogenate of ultrasonication mixes with isopyknic water, enzymolysis is carried out.
Described enzymolysis refers to jointly hatch at tissue homogenate and pancreatin, makes protein that the process of decomposing occur.
In the present invention, during enzymolysis, the mass fraction of pancreatin is 2.5%.Adding tissue homogenate to mass fraction by pancreatin is 2.5%.
In the present invention, the enzyme activity of pancreatin is 37000u/g.
In the present invention, the temperature of enzymolysis is 45 DEG C ~ 55 DEG C, and pH value is 7.3 ~ 7.7, and enzymolysis time is 40min ~ 80min.
In certain embodiments, the temperature of enzymolysis is 50 DEG C, and pH value is 7.5, and enzymolysis time is 1h.
After centrifugal, reject fragment of tissue, collects supernatant.Rich in proteins, bioactive peptide and free amino acid in this supernatant.
In an embodiment of the present invention, centrifugal rotating speed is 4500 turns/min, and the centrifugal time is 20min, and centrifugal temperature is 4 DEG C.
Retain the filter membrane of specifically instigating supernatant through 50kDa, collect the ultrafiltrate within 50kDa.
Those skilled in the art can also carry out lyophilization according to known method to the component that tissue homogenate, supernatant and ultrafiltration are collected.
As preferably, after retaining, also comprise the step that adjust ph is 6.8 ~ 7.2.
In the scope that pH value is 6.8 ~ 7.2, the stable in properties of Embryo Gallus domesticus peptide, activity is higher.
Described lyophilization is that Embryo Gallus domesticus bioactive peptide mixes with lyophilized preparation, obtained by freeze drying lyophilized powder.Described lyophilization excipient is mannitol.Concrete, adding mannitol to mass fraction in the component that tissue homogenate, supernatant and ultrafiltration are collected is 8% carry out lyophilization.Preferably, adding mannitol to mass fraction in the component of collecting in ultrafiltration is 8% carry out lyophilization.
The dermatitis that mosquito bite causes in insect dermatitis is modal.Owing to containing plurality of antigens composition in the saliva of mosquito or the leachate of poison gland, these antigens can produce atopic reaction with antibody and cause inflammation after entering human body skin.Comedo is chronic inflammation hair follicle sebum gland disease, is one of modal disease of department of dermatologry.The factors such as it occurs main and hypersteatosis, pilosebaceous duct block, bacteriological infection and inflammatory reaction are closely related.
The present invention investigates experiment and proves, Embryo Gallus domesticus peptide has significant antiphlogistic effects, can treat mosquito bite or comedo.For mosquito bite, its effective percentage is 100%, and cure rate can reach 95%, and for comedo, anti-acne effect can reach 99%, and cure rate can reach 95%.The cycle of curing is 7 ~ 14 days.
Therefore, the invention provides the application of Embryo Gallus domesticus peptide in the product of preparation antiinflammatory, anti-acne.
A product for antiinflammatory, anti-acne, comprises Embryo Gallus domesticus peptide, hyaluronic acid, xanthan gum, Jojoba oil, arbutin, glycyrrhizic acid dipotassium and glycerol.
Wherein, the concentration of Embryo Gallus domesticus peptide is 0.5mg/mL ~ 3.0mg/mL, hyaluronic mass fraction is 0.3%, xanthan gum mass fraction 0.3%, Jojoba oil mass fraction 1%, arbutin mass fraction 0.5%, the mass fraction of glycerol is 3%, glycyrrhizic acid dipotassium mass fraction is 0.1%, and surplus is deionized water.
The dosage form of the product of antiinflammatory provided by the invention, anti-acne is unguentum, spray or patch.
As preferably, in the unguentum of antiinflammatory provided by the invention, anti-acne, hair color regeneration, the mass fraction of Embryo Gallus domesticus peptide is 3mg/ml, also comprises the acceptable adjuvant of unguentum.
As preferably, in the spray of antiinflammatory provided by the invention, anti-acne, hair color regeneration, the mass fraction of Embryo Gallus domesticus peptide is 3mg/ml, also comprises the acceptable adjuvant of spray.
As preferably, during antiinflammatory mosquito bite provided by the invention pastes, the mass fraction of Embryo Gallus domesticus peptide is 2.0mg/ml, also comprises the acceptable adjuvant of patch.
Under normal circumstances, have abundant blood vessel in hair papilla, for hair papilla, ball top portion provide sufficient nutrition, melanin granule just synthesizes smoothly.When melanin granule is at the formation generation obstacle in hair papilla, ball top portion, although or formed because of certain factor, can not be transported in hair and go, thus the melanin granule of medulla of hair, cortical section is reduced, when disappearing, just there will be poliosis.
The present invention investigates experiment and proves, Embryo Gallus domesticus peptide has the significant effect promoting hair color regeneration.Embryo Gallus domesticus peptide is allowed fully to enter into basal layer and the skin corium of hair follicle tissue and epidermis, make the cell activation of its stratum germinativum, activate the cell of stratum germinativum and the cell of skin base layer, produce and utilize melanic ability to strengthen, make hair, at the initial stage of regeneration, basic change just occur, thus make hair blackening.Dark hair regeneration is large about about 30%.
Therefore, the invention provides the application of Embryo Gallus domesticus peptide in the product preparing hair color regeneration.
A product for hair color regeneration, comprises Embryo Gallus domesticus peptide, hyaluronic acid, xanthan gum, Jojoba oil, arbutin, glycyrrhizic acid dipotassium and glycerol.
Wherein, the concentration of Embryo Gallus domesticus peptide is 3.0mg/mL, and hyaluronic mass fraction is 0.3%, xanthan gum mass fraction 0.3%, Jojoba oil mass fraction 1%, arbutin mass fraction 0.5%, the mass fraction of glycerol is 3%, and glycyrrhizic acid dipotassium mass fraction is 0.1%, and surplus is deionized water.
The product of promotion hair color regeneration provided by the invention is shampoo, hair oil, pomade, suppurative mastitis and hair dye.
Embryo Gallus domesticus peptide provided by the invention, comprises Embryo Gallus domesticus peptide and free amino acid; Wherein, the molecular weight of Embryo Gallus domesticus peptide is less than 50kDa.Embryo Gallus domesticus peptide provided by the invention can play the effect that antiinflammatory, anti-acne and hair color regenerate, and the survey showed that, and Embryo Gallus domesticus peptide provided by the invention has good effect to human epidermal local inflammation and detumescence, particularly has good curative effect to mosquito bite.Further, Embryo Gallus domesticus peptide provided by the invention can have good effect to human body face comedo.Further, Embryo Gallus domesticus peptide provided by the invention can promote the regeneration of hair color.The preparation method of Embryo Gallus domesticus peptide provided by the invention is easy and simple to handle, and by homogenate, freeze thawing and ultrasonication, broken to the full extent cell membrane, makes the bioactive peptide in born of the same parents obtain good stripping, thus improve extraction efficiency.
Accompanying drawing explanation
Fig. 1 shows that embodiment 1 obtains the antiinflammatory mosquito bite effect of Embryo Gallus domesticus peptide;
Fig. 2 shows that embodiment 1 obtains the anti-acne effect of Embryo Gallus domesticus peptide.
Detailed description of the invention
The invention provides a kind of Embryo Gallus domesticus peptide and preparation method thereof and the application preparing antiinflammatory, in product that anti-acne, hair color regenerate, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can change methods and applications herein or suitably changes and combination not departing from content of the present invention, spirit and scope, realizes and applies the technology of the present invention.
The material that the present invention adopts, instrument or reagent are all common commercially available product, all can buy in market.
Below in conjunction with embodiment, set forth the present invention further:
Embodiment 1
(1) hatching chicken embryo: get SPF level fertilized eggs, put into incubator and hatch, incubation temperature controls at 37.5 DEG C, relative humidity 60%, every 2 hours automatic egg turnings once, treat that hatching was to 16 days, take out fertilized eggs, put in the freezer of-20 DEG C, stop hatching and saving backup.
(2) homogenate and cell breakage: surface sterilization is carried out to the egg of hatching, remove eggshell and take out Embryo Gallus domesticus, at 4 DEG C by Embryo Gallus domesticus with after tissue refiner homogenate, pour in the seal box after sterilizing, to be put in the freezer of-30 DEG C freezing 72 hours, in the environment of 4 DEG C, cell ultrasonication is carried out to it after melting, add the water for injection of equivalent, be positioned in the freezer of 4 DEG C for subsequent use.
(3) enzyme hydrolysis and centrifugal: get the cold preservation liquid heated water bath to 50 DEG C after cell breakage, adjust pH to 7.5, adding pancreatin to concentration is that 2.5% (mass fraction) is hydrolyzed water bath with thermostatic control 1 hour, at 4 DEG C, under 4500r/min condition centrifugal 20 minutes, receive supernatant to be cooled to 4 DEG C of cold preservations for subsequent use.
(4) ultrafiltration and tune pH value: by the filter membrane of Embryo Gallus domesticus serosity ultra filtration 50kDa, collect the ultrafiltrate within 50kDa, adjust pH to 6.8 ~ 7.2, obtained Embryo Gallus domesticus peptide.
Embodiment 2
(1) hatching chicken embryo: get SPF level fertilized eggs, put into incubator and hatch, incubation temperature controls at 37.5 DEG C, relative humidity 60%, every 2 hours automatic egg turnings once, treat that hatching was to 14 days, take out fertilized eggs, put in the freezer of-20 DEG C, stop hatching and saving backup.
(2) homogenate and cell breakage: surface sterilization is carried out to the egg of hatching, remove eggshell and take out Embryo Gallus domesticus, at 8 DEG C by Embryo Gallus domesticus with after tissue refiner homogenate, pour in the seal box after sterilizing, to be put in the freezer of-20 DEG C freezing 80 hours, in the environment of 4 DEG C, cell ultrasonication is carried out to it after melting, add the water for injection of equivalent, be positioned in the freezer of 4 DEG C for subsequent use.
(3) enzyme hydrolysis and centrifugal: get the cold preservation liquid heated water bath to 55 DEG C after cell breakage, adjust pH to 7.7, adding pancreatin to concentration is that 2.5% (mass fraction) is hydrolyzed water bath with thermostatic control 80min, at 4 DEG C, under 4500r/min condition centrifugal 20 minutes, receive supernatant to be cooled to 4 DEG C of cold preservations for subsequent use.
(4) ultrafiltration and tune pH value: by the filter membrane of Embryo Gallus domesticus serosity ultra filtration 50kDa, collect the ultrafiltrate within 50kDa, adjust pH to 6.8 ~ 7.2, obtained Embryo Gallus domesticus peptide.
Embodiment 3
(1) hatching chicken embryo: get SPF level fertilized eggs, put into incubator and hatch, incubation temperature controls at 37.5 DEG C, relative humidity 60%, every 2 hours automatic egg turnings once, treat that hatching was to 19 days, take out fertilized eggs, put in the freezer of-20 DEG C, stop hatching and saving backup.
(2) homogenate and cell breakage: surface sterilization is carried out to the egg of hatching, remove eggshell and take out Embryo Gallus domesticus, at 0 DEG C by Embryo Gallus domesticus with after tissue refiner homogenate, pour in the seal box after sterilizing, to be put in the freezer of-40 DEG C freezing 60 hours, in the environment of 0 DEG C, cell ultrasonication is carried out to it after melting, add the water for injection of equivalent, be positioned in the freezer of 0 DEG C for subsequent use.
(3) enzyme hydrolysis and centrifugal: get the cold preservation liquid heated water bath to 45 DEG C after cell breakage, adjust pH to 7.3, adding pancreatin to concentration is that 2.5% (mass fraction) is hydrolyzed water bath with thermostatic control 40min, at 4 DEG C, under 4500r/min condition centrifugal 20 minutes, receive supernatant to be cooled to 4 DEG C of cold preservations for subsequent use.
(4) ultrafiltration and tune pH value: by the filter membrane of Embryo Gallus domesticus serosity ultra filtration 50kDa, collect the ultrafiltrate within 50kDa, adjust pH to 6.8 ~ 7.2, obtained Embryo Gallus domesticus peptide.
Embodiment 4
Embodiment 1 obtains bioactive peptide and analyzes through shotgun, and the content of bioactive peptide is 0.1 μ g/ μ L; Peptide hop count amount amounts to 404, and respectively from 98 kinds of different albumen, concrete outcome is as table 1:
Table 1shotgun analysis result
Embodiment 5
The method adopting GB/T5009.124-2003 to provide, utilizes L-8800 type automatic amino acid analyzer, detects the Embryo Gallus domesticus peptide Free Amino Acids that embodiment 1 provides.Result is:
| Amino acid name | Content (mg/ml) |
| Aspartic acid | 1.2400 |
| Threonine | 0.4966 |
| Serine | 0.7433 |
| Glutamic acid | 2.1670 |
| Glycine | 0.6891 |
| Alanine | 0.8337 |
| Valine | 0.9657 |
| Methionine | 0.3995 |
| Isoleucine | 0.7108 |
| Leucine | 1.2117 |
| Tyrosine | 0.2629 |
| Phenylalanine | 0.5486 |
| Lysine | 1.1358 |
| Histidine | 0.1835 |
| Arginine | 0.7548 |
| Tryptophan | 0.9735 |
| Cystine | 0.6228 |
| Proline | 0.3891 |
| Add up to (mg/ml) | 14.3284 |
Embodiment 6
Whether can eliminate the local inflammation of people's normal skin surface to measure Embryo Gallus domesticus peptide, the curative effect having carried out eliminating the local inflammation of mosquito bite is observed, and research method is:
Embryo Gallus domesticus peptide prepared by the embodiment 1 first preparing variable concentrations, concentration is: 0.5mg/ml, 0.8mg/ml, 1.0mg/ml, 1.5mg/ml, 2.0mg/ml, 2.5mg/ml, 5ml/ bottle, each 60 bottles, totally 360 bottles, gives 360 people of energy unrecoverable data at random.Be divided into 6 groups according to the concentration of Embryo Gallus domesticus peptide, smear in time after requiring everyone mosquito bite and connect painting three times, observing effect.
Judgment criteria is: antipruritic in 30 minutes, and the normal condition returning to skin of subsiding a swelling in two hours is designated as healing, otherwise is only designated as effectively.Statistical result is as Fig. 1.
Showing efficiency according to the effect data regained is 100%, and optium concentration is 2.0mg/ml, and during optium concentration, cure rate is 95%.
There is good effect according to above result of study Embryo Gallus domesticus peptide to the local inflammation of human epidermal and detumescence, particularly have good curative effect to mosquito bite.Can reduce the misery of people in summer, range of application widely, can be used for the unguentum of external, spray, mosquito bite subsides etc.
Embodiment 7
In order to adolescence is not by the puzzlement of comedo, reduce comedo to study, work, the trouble brought of living, measure the comedo whether Embryo Gallus domesticus peptide can eliminate adolescence's normal skin surface for this reason, the curative effect simultaneously having carried out eliminating comedo is observed.Research method is:
Embryo Gallus domesticus peptide prepared by the embodiment 1 first preparing variable concentrations, concentration is: 2.0mg/ml, 2.5mg/ml, 3.0mg/ml, 3.5mg/ml, 4.0mg/ml, 5ml/ bottle, each 60 bottles, allot to the classmate of senior middle school's long comedo by school physician and the form master, smear consumption 2ml every day for three times carry out the effect observation of two weeks and respectively add up once result in the 7th day and 14 days, carry out Comprehensive Assessment to effective and healing, statistical result is as Fig. 2.
Judgment criteria is: everyone all show as comedo diminish, move back red recovery skin normal color and do not stay pox to cheat pox print for curing, otherwise be only designated as effectively.Showing efficiency according to the effect data regained is 99%, and optium concentration is 3.0mg/ml, and during optium concentration, cure rate is 95%.Majority just do not cure to the 7th day and the 14th day, everyone all show as comedo diminish, move back red recovery skin normal color and do not stay pox to cheat pox print, particularly face has formed classmate's successful of acne.Good effect, particularly face is had to form classmate's successful of acne according to above result of study Embryo Gallus domesticus peptide to human face comedo
Embodiment 8
In order to solve the worry of grey poll, reduce the trouble and worry of white originator, several poliosis, grey originator are tested.
Embryo Gallus domesticus peptide prepared by the embodiment 1 first preparing variable concentrations, concentration is 3.0mg/ml.First allow experimenter scrape shaven head send out, smeared Embryo Gallus domesticus peptide solution 5 times first 7th day every day, then morning every day, noon, evening smear Embryo Gallus domesticus peptide solution three times, 30 days and 60 days effect observations of comparing.Experimenter represents, the poliosis of about 30% changes hair color into.
Illustrate: allow Embryo Gallus domesticus peptide essence fully enter into basal layer and the skin corium of hair follicle tissue and epidermis, make the cell activation of its stratum germinativum, activate the cell of stratum germinativum and the cell of skin base layer, produce and utilize melanic ability to strengthen, make hair, at the initial stage of regeneration, basic change just occur, thus make hair blackening.
Below be only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. an Embryo Gallus domesticus peptide, is characterized in that, comprises Embryo Gallus domesticus peptide and free amino acid; Wherein, the molecular weight of described Embryo Gallus domesticus peptide is less than 50kDa.
2. the preparation method of Embryo Gallus domesticus peptide described in claim 1, is characterized in that, comprising: shelled by Embryo Gallus domesticus, after homogenate, smudge cells; After pancreatin enzymolysis, collected by centrifugation supernatant, ultrafiltration molecular cut off is less than the component of 50kDa.
3. preparation method according to claim 2, is characterized in that, described smudge cells comprises the step of freeze thawing and ultrasonication;
The cryogenic temperature of described freeze thawing is-20 DEG C ~-40 DEG C, and the time is 60h ~ 80h; Solution temperature is 0 DEG C ~ 8 DEG C;
4 DEG C ~ 8 DEG C of described ultrasonication, power is 800W ~ 1200W, and the time is 15min ~ 25min.
4. preparation method according to claim 2, is characterized in that, in the enzymolysis solution of described enzymolysis, the mass fraction of pancreatin is 2.5%.
5. preparation method according to claim 2, is characterized in that, the temperature of described enzymolysis is 45 DEG C ~ 55 DEG C, and pH value is 7.3 ~ 7.7, and enzymolysis time is 40min ~ 80min.
6. preparation method according to claim 2, is characterized in that, described centrifugal rotating speed is 4500 turns/min, and the centrifugal time is 20min, and centrifugal temperature is 4 DEG C.
7. the application of Embryo Gallus domesticus peptide in the product of preparation antiinflammatory, anti-acne prepared of preparation method described in Embryo Gallus domesticus peptide according to claim 1 or any one of claim 2 ~ 6.
8. a product for antiinflammatory, anti-acne, hair color regeneration, is characterized in that, comprise the Embryo Gallus domesticus peptide that preparation method described in Embryo Gallus domesticus peptide according to claim 1 or any one of claim 2 ~ 6 is obtained.
9. the Embryo Gallus domesticus peptide that prepared by preparation method described in Embryo Gallus domesticus peptide according to claim 1 or any one of claim 2 ~ 6 promotes the application in the product of hair color regeneration in preparation.
10. promote to it is characterized in that the product that hair color regenerates, comprise the Embryo Gallus domesticus peptide that preparation method described in Embryo Gallus domesticus peptide according to claim 1 or any one of claim 2 ~ 6 is obtained.
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