CN102525867B - Chick embryo essence freeze-dried powder and preparation method and application thereof - Google Patents
Chick embryo essence freeze-dried powder and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of cosmetics and in particular relates to a chick embryo essence freeze-dried powder and preparation method and application thereof. The chick embryo essence freeze-dried powder is obtained by the preparation method which comprises the following steps of: breaking the shell of an embryo egg which is hatched for 8-16 days and taking a chick embryo, blending with a food blender to obtain a homogenate, adding a phosphate buffer solution into the homogenate, stirring at 4-10 DEG C and leaching overnight, filtering the leaching liquid with medical gauze, centrifuging the filtrate, collecting the supernatant, filtering the supernatant with a hollow fiber ultrafiltration membrane, collecting the filtrate, adding trehalose into the filtrate and freeze-drying. The freeze-dried powder contains various nutrients, anti-infective components and bioactive substances, and can be re-dissolved to make an essence which is formulated to cosmetics having anti-wrinkle, anti-skin aging, skin-rejuvenating and whitening effects. The freeze-dried powder is used in cosmetic compositions which are applied twice daily in morning and evening to reduce skin small wrinkles, moisturize the skin and improve elasticity and resistance of the skin, and has no irritation to the skin.
Description
Technical field
The invention belongs to the cosmetics technical field, be specifically related to a kind of Embryo Gallus domesticus essence lyophilized powder and preparation method thereof and application in cosmetics.
Background technology
Skin aging is the skin aging phenomenon that is caused by natural cause or non-natural cause.The outward manifestation of skin aging has cutis laxa, attenuation, drying, squama is arranged, and tarnishes; Elasticity and consolidation descend thereupon; Wrinkle forms, and increases gradually, deepens; The sensitivity of skin increases, hyperpigmentation.The factor that causes skin aging mainly contains inside and outside two aspects, and endogenous cause of ill comprises that mainly the flowability of collagen protein and the decline of elastin laminin synthesis capability, Cytoplasm and intercellular substance in age ageing, physiological ageing, the skin reduces.Exopathogenic factor comprises that mainly collagen protein and elastin laminin degeneration, sebum secretion imbalance, oxygen-derived free radicals that ultraviolet (photoaging), extraneous factor cause increase the normal cell of injury.The activity of fighting against senium composition that is applied at present in the cosmetics mainly comprises following four aspects: the composition (cell growth factor, animals and plants extract) that 1) influences the histiocyte growth metabolism; 2) skin moisture-keeping and the composition of repairing skin barrier function; 3) composition of anti-oxidative damage; 4) resist ultraviolet composition.
But, the present report that also the Embryo Gallus domesticus essence is not used for cosmetics as the activity of fighting against senium composition.
Summary of the invention
The purpose of this invention is to provide that a kind of defying age is effective, the simple Embryo Gallus domesticus essence of preparation technology lyophilized powder and preparation method thereof, and Embryo Gallus domesticus essence lyophilized powder is applied in the cosmetics as a kind of defying age additive.
The concrete steps of Embryo Gallus domesticus essence method for preparing freeze-dried powder of the present invention are:
(1) gets 8～16 days healthy chicken embryo (preferably hatching 12～14 days healthy chicken embryo) of hatching, strike shell and get Embryo Gallus domesticus;
(2) with food masher Embryo Gallus domesticus is broken into homogenate, add the phosphate buffer (30～50 mM) of pH6.8～7.2, press mass ratio, Embryo Gallus domesticus: under ℃ condition of phosphate buffer=1:1～3,4～10, Embryo Gallus domesticus and phosphate buffer fully stirred to leach spend the night;
(3) with the Embryo Gallus domesticus leaching liquid in multilamellar (for example 4-6 layer) the hospital gauze filtration step (2), with filtrate with the centrifugal 15～30min of the rotating speed of 4000～6000 rpm;
(4) with the supernatant after centrifugal, be that the doughnut membrane ultrafiltration of 10000～20000 Da filters with molecular cut off, filter liquor, i.e. Embryo Gallus domesticus essence;
(5) add trehalose in the Embryo Gallus domesticus essence, pressing mass volume ratio (g/mL) is trehalose: Embryo Gallus domesticus essence=1:5～10 at-45～-85 ℃ of following lyophilization 12～36h, promptly obtain Embryo Gallus domesticus essence lyophilized powder.
The invention provides above-mentioned Embryo Gallus domesticus essence lyophilized powder as the application of defying age additive in cosmetics, the skin tool of aging is improved significantly.
In the present invention, the addition of described Embryo Gallus domesticus essence lyophilized powder is 0.1%～5% of a cosmetics gross weight, preferred 1%～3%.
In the present invention, the improvement effect of described skin comprises elimination, minimizes and/or prevents the flaw of skin visually or by sense of touch to detect, for example wrinkle of skin, elasticity, moisture, melanocyte etc.
The present invention also provides the cosmetic composition that contains Embryo Gallus domesticus essence lyophilized powder.But described cosmetic composition topical application produces the defying age effect on skin, and mechanism is synthesizing by the inhibition of tryrosinase and stimulation collagen.In addition, Embryo Gallus domesticus essence lyophilized powder has antioxidation.Whether Embryo Gallus domesticus essence lyophilized powder of the present invention can be used for any type of cosmetic composition, no matter be present in the cosmetic composition.The amount of lyophilized powder should be a safety and effective for the dosage form (for example Cleansing Foam, facial cream, emulsifiable paste or gel) of effect of wanting to reach and compositions.
Among the present invention, described cosmetic composition can be cream, frost, dew, water, gel, lotion or eluting type product etc.
Among the present invention, the Embryo Gallus domesticus essence extracts from 8～16 days Embryo Gallus domesticus of healthy hatching, the Embryo Gallus domesticus in this stage is especially hatched 12～14 days Embryo Gallus domesticus, contains the abundant various nutrient substance of keeping the new life sprouting, the active component with significant promotion growth and human body immunity improving ability.Bioactive substance contained in the Embryo Gallus domesticus can be divided into 3 classes substantially, i.e. nutritional labeling, infection composition and bioactive substance.Nutritional labeling has protein, aminoacid, carbohydrate (glucose, fructose, galactose, arabinose etc.), lipid, fatty acid, multivitamin (A, E, C etc.), mineral (Fe, Zn, Cu etc.) etc.; The infection composition has immunoglobulin, interferon, fetus albumen, lysozyme and phosphodiesterase (anti-trypanosomicide) etc.; Bioactive substance has sialic acid, catecholamine, insulin like growth factor (IGF-I), insulin, kininase, histaminase, erythropoietin, deoxyribonuclease and ribonuclease etc.These compositions mainly contain (1) nutritive maintenance skin (2) water conservation skin moistening crease-resistant (3) infection to the effect of skin.
For example, contain insulin like growth factor (IGFs in the Embryo Gallus domesticus essence, molecular weight 7650 Da), effect about IGFs, nineteen ninety authority's New England Journal of Medicine has been delivered the antidotal clinical experiment report about IGF-1, prove that fully IGF-1 can make application on human skin elasticity increase, ruddy, the deflation of the colour of skin, wrinkle minimizing etc.
Human insulin's growth factor I GF-1 is present best activity of fighting against senium material, and is present unique material that can reverse skin aging, therefore in time replenishes IGF-1(quasi-insulin growthing factor I type) be antidotal best method.Doctor Rudman is divided into two groups with the old man in 26 61-80 years, carry out IGF-I ﹠ HGH(human growth hormone) double blind experiment, the result shows: after 6 months, do not use the old man of the assembled growth factor, old and feeble situation continues to worsen, and used the old man of the assembled growth factor, the average young 10-20 of bodily tissue year.He emphasizes: " the various degenerations that health occurs with age growth are not that inevitably we recognize that now its some aspect can be prevented or reverse ".Human growth hormone HGH is present best antiaging agent, and is at present unique material that can reverse aging.Yet HGH must change into IGF-1 after in the human body could be absorbed by liver, and therefore in time supplying the quasi-insulin growthing factor I type is antidotal best method.
(the hot medical college professor of University of Wisconsin-Madison, neuroendocrine expert) carried out the once clinical research of large-scale IGF-I HGH with doctor Meng De (recovering physiotherapy expert) in the doctor of medicine 1994-1996 spy, treatment target has people more than 800, comprise some video display stars and famous enterpriser, that maximum is the doctor, accounts for 20% of sum.They extract the data of 202 objects at random out and analyze, and this 202 people's age, the women accounted for 15% from 39 years old to 74 years old, the mean treatment time is 180 days (half a year), the shortest 15 days, the longest 720 days (2 years), the result shows that the antidotal effect of IGF-I ﹠ HGH reaches 100%.
Inventor of the present invention has developed the Embryo Gallus domesticus essence lyophilized powder that skin aging is had better improvement effect through for many years trial and effort, and that this lyophilized powder can be widely used in is crease-resistant, in defying age, skin care and the cosmetics such as whiten.
The present invention relates to the organism extract, the hatching egg of the Embryo Gallus domesticus that is used to extract and hatching are country's demonstrationization ecologic breeding pattern, and safety and sanitation are used for cosmetic composition and improve skin and/or the defying age effect is provided.
The invention still further relates to the proteic synthetic and/or active beautifying use of restraint of tyrosinase of described Embryo Gallus domesticus essence chafe cell collagen.
In addition, the present invention relates to described Embryo Gallus domesticus essence provides antioxidant effect for skin beautifying use.
The invention still further relates to cosmetic composition, for example cream frost, gel, elite dew, emulsion or lotion, it comprises the Embryo Gallus domesticus essence lyophilized powder of the present invention (consumption is 0.1%～5%) of safety and effective dose.
As mentioned above, Embryo Gallus domesticus essence lyophilized powder of the present invention is effective as the active component in the cosmetic composition.Especially, compositions preferably is suitable for wrinkly, coarse drying and easily plays skin bits, responsive, old and feeble and/or the UV damage, thereby improves the situation of skin, and is used for healthy skin, thereby is used to prevent or delaying skin aging.
Embryo Gallus domesticus essence lyophilized powder of the present invention is used in the cosmetic composition, and said composition can also add used for cosmetic oiliness raw material.For the not special restriction of described oiliness raw material, can comprise synthesizing ester, be specially and comprise the stearic acid esters, for example glyceryl tristearate, stearic acid shark liver ester, oil with hydrogenated soybean fatty glyceride; The oleic acid esters, linoleic acid esters, for example Ceraphyl 140A, propylene glycol mono-oleate, linoleic acid isopropyl ester; Palmic acid esters, for example Palmic acid iso stearyl ester, Palmic acid isocetyl ester; Isostearic acid esters, for example isostearic acid isopropyl ester, isostearic acid sorbitan ester, isostearic acid propylene glycol ester; Hydroxy stearic acid esters, for example hydroxy stearic acid monoglyceride, monohydroxy tristerin; Myristic acid esters, for example myristic acid iso stearyl ester, butyl myristate; Different Palmic acid esters, for example different octyl palmitate, different Palmic acid diglycerol base ester; Lauric acid esters, for example glyceryl laurate ester, lauric acid propylene glycol ester, lauric acid iso stearyl ester; Sad, n-nonanoic acid, capric acid esters, for example iso stearyl caprylate, different isodecyl pelargonate; Polyglycereol esters, for example monostearate polyglycerin ester, distearyl acid polyglycerin ester; Lanolin derivative class, for example hydroxylation lanoline, lanosterol; Pentaerythritol esters, for example pentaerythritol tetramyristate; Ricinoleic acid ester, for example castor oil acid glyceride, castor oil hydrogenated; Citric acid, succinic acid esters, for example citric acid acetyl triethyl, succinic acid diethoxy ethyl ester; Phosphoric acid ester, for example polyxyethylated (12～16) ether phosphates (6EO), trioleyl phosphate; Other esters, for example mountain Yu acid glycerol base ester, gallateoctylester; Also have fatty acid, for example coco-nut oil fatty acid, palm-kernel oil fatty acid, linoleic acid; Aliphatic alcohols, aromatic alcohol, for example Cortex cocois radicis oleyl alcohol, arachidic alcohol, Fructus Canarii albi oleyl alcohol; The wax class, for example synthetic chloroflo, synthetic squalane.
When adding above-mentioned oiliness raw material, their additions in cosmetic composition are preferably 0.01%～45%(weight of total amount), 0.05%～25%(weight more preferably).
Embryo Gallus domesticus essence lyophilized powder of the present invention is used in the cosmetic composition, said composition can also add the used for cosmetic nutritive additive, described nutritive additive comprises: (1) vitamins, for example nicotinic acid, allantoin DL-pantothenylol, natural Vitamin E, d-Delta-Tocopherol; (2) aminoacid, for example L-hydroxyproline, L-tyrosine, L-histidine; (3) adenylate, for example Flavin adenine dinucleotide disodium salt two water salt, adenylic acid; (4) sterols, for example sitosterol, cholesteryl oleate, plant sterol oleate, n-nonanoic acid cholesterol ester; (5) hydrolyzed protein class, for example oleoyl hydrolytic collagen, hydrolytic collagen ethyl ester liquid, coco-nut oil fatty acid hydrolytic collagen, isostearoyl hydrolytic collagen; (6) other nutritional substance, for example umbilical cord extracting solution, oil-soluble placenta extracting liquid, gluconic acid sodium salt.
When adding above-mentioned nutritive additive, their additions in cosmetic composition are 0.01%～45%(weight of total amount), especially preferred addition is 0.05%～25%(weight), 0.1%～20%(weight more preferably).
Embryo Gallus domesticus essence lyophilized powder of the present invention is used in the cosmetic composition, also can add surfactant in the said composition.Described surfactant comprises: sorbitan fatty acid ester (Span-20, Span-40, Span-60, Span-80) and its ethylene oxide adduct (Tween-20, Tween-40, Tween-60, Tween-80), polyoxyethylene lauryl ether, cocoanut fatty acid diethanolamide, monoolein, polyoxyethylene castor oil, Wool wax alcohols,ethoxylated, alkyl phosphate, alkyl phosphoric acid polyethylene glycol oxide ester, fatty acid alkali metal salt, alkyl glycerol base ether.Can add a kind of in the above-mentioned surfactant or some kinds.
When adding above-mentioned surfactant, their additions in cosmetic composition are preferably 0.01%～45%(weight of total amount), 0.05%～25%(weight more preferably).
Embryo Gallus domesticus essence lyophilized powder of the present invention is used in the cosmetic composition, also can add water-soluble polyol in the said composition.The example of described water-soluble polyol comprises that intramolecularly contains the alcohol of two or more hydroxyls, for example ethylene glycol, Polyethylene Glycol, propylene glycol, glycerol, glycerol-ethylene oxide adduct, sorbitol, POE(20) sorbitol, 1,3 butylene glycol, glucose, maltose, maltose alcohol, sucrose, fructose, xylitol, maltotriose, threitol, erithritol and the alcohol that obtains by the reduction amylolysis.Can adopt a kind of in the above-mentioned alcohol or several.
When adding above-mentioned water-soluble polyol, can select suitable adding proportion according to actual needs.Usually, their addition is preferably 0.01%～60%(weight of total amount), 0.1%～25%(weight more preferably).
Embryo Gallus domesticus essence lyophilized powder of the present invention is used in the cosmetic composition, can also further add used for cosmetic powder class raw material in the said composition.Described powder class raw material comprises: (1) silicates, for example montorillonite clay, pumice, calcium silicates, glass dust; (2) aluminum salt, for example aluminium myristate, isostearic acid aluminum, aluctyl.; (3) mica group, for example phlogopite, sericite, synthetic phlogopite; (4) lamination powder, for example polyethylene terephthalate-aluminum-epoxy resin layer press-powder, polyethylene-polyester layer press-powder; (5) surface treatment thing powder, for example cellulose powder, sodium abietate are handled calcium carbonate; (6) other powder, for example salicylic acid titanium, cobalt aluminum, calcium hydroxy phosphate.These powder class raw materials can come pretreatment with following material before use: the polysiloxanes with methyl hydrogen methyl polysiloxane, trimethylsiloxy silicic acid, methyl cyclopolysiloxane or similar substance is handled; Fluorine with phosphoric acid perfluoroalkyl ester, perfluor alcohol or analog is handled; Getting aminoacid with N-acyl glutamic acid or analog handles; Lecithin is handled; Metallic soap is handled; Fatty acid treatment; Alkylphosphonate is handled or similar processing.
When adding above-mentioned used for cosmetic powder class raw material, can select suitable adding proportion according to the dosage form of cosmetic composition.Usually, their addition is preferably 0.01%～50%(weight of total amount), more preferably 0.05%～30%.
The cosmetic composition that contains Embryo Gallus domesticus essence lyophilized powder of the present invention can also comprise the multiple additive that other uses at cosmetic field.The example of this constituents type comprises: wetting agent, film former, thickening agent, antibiotic antiseptic, pigment, spice, sunscreen, chelating agen, natural material.
The pH value preferred 3～10 that contains the cosmetic composition of Embryo Gallus domesticus essence lyophilized powder of the present invention.Preferred pH value is 4～9, like this near the normal acid-base value of skin, helps keeping the normal physiological function of skin.
The general conventional method of preparation cosmetic composition known in the art that adopts prepares the compositions that contains Embryo Gallus domesticus essence lyophilized powder of the present invention.These class methods are generally mixed various compositions by one or more steps, make its state that becomes relative homogeneous, wherein relate to have or not heating, cooling, homogenizing, use vacuum etc.Compositions can be prepared into any required conventional form, for example Cleansing Foam, facial cream, elite dew, essence, emulsifiable paste, gel.
In the prescription of any cosmetic composition, the addition scope of Embryo Gallus domesticus essence lyophilized powder of the present invention all is based on cosmetic composition and wants the effect that reaches, other component, types of compositions, cost and practicality in the prescription.Usually, the addition of Embryo Gallus domesticus essence lyophilized powder of the present invention is preferably 0.1%～5% of gross weight, and more preferably 1%～3%.
Cosmetic composition described in the present invention is that effectively wherein, this based composition of dermal administration to this type of improvement of needs improves the required persistent period of this type of skin as a part that is used to improve the beauty method of the skin with old and feeble sign.Though the effect of this type of persistent period according to this type of cosmetic composition changes, at least use one time cosmetic composition to skin preferred every day.
Cosmetic composition of the present invention can also comprise other multiple used for cosmetic additive.
Description of drawings
Fig. 1 is consumer's skin effect test.
Fig. 2 is among the embodiment 7, compares the variation of hydration value with the numerical value of initial time record.
Fig. 3 is among the embodiment 10, and the numerical value during with the processing beginning is compared the variation of maximal roughness.
The specific embodiment
For understanding content of the present invention better, the invention will be further described below in conjunction with concrete embodiment.But the present invention is not subjected to the limitation of these embodiments.
The preparation of embodiment 1 Embryo Gallus domesticus essence lyophilized powder
Get 13 days normal free of contamination chicken embryo of hatching, strike shell and get Embryo Gallus domesticus.With food masher Embryo Gallus domesticus is broken into homogenate, by quality than Embryo Gallus domesticus: phosphate buffer=1:1 adds phosphate buffer, Embryo Gallus domesticus and phosphate buffer is fully stirred leach 30min.Filter the Embryo Gallus domesticus leaching liquids with 3 layers of hospital gauze, with filtrate with the centrifugal 20min of the rotating speed of 6000 rpm.With the supernatant after centrifugal, be that the doughnut membrane ultrafiltration of 10000 Da gets effluent with molecular cut off, be the Embryo Gallus domesticus essence.Press mass volume ratio (g/mL) trehalose: Embryo Gallus domesticus essence=1:2.5, in the Embryo Gallus domesticus essence, add trehalose, get Embryo Gallus domesticus essence lyophilized powder at-80 ℃ of following lyophilization 18h.
Below be in vitro tests
The antioxidation of embodiment 2 Embryo Gallus domesticus essences (removing free radical) effect
Free radical is one of principal element that causes skin aging, the material of antioxidation can or take place with combined with radical to substitute, and makes the quantity of [DPPH] free radical reduce, and solution colour shoals, the absorbance value that shows as 515 nm places constantly reduces, only to reaching stable.Therefore can calculate the ability of (judgement) its antioxidation or removing free radical by measuring [removings] ability of catching of polyphenoils confrontation free radical under 515 nm.Be used for antioxidation, defying age, whitening function to the functional cosmetics raw material.[DPPH] (diphenyl bitterness diazanyl free radical) be a class comparatively stable be the free radical at center with N.[DPPH] methanol solution is purple, at 515 nm wavelength places maximum light absorption is arranged, and the depth and the concentration of color are linear.[DPPH] free radical is had the antioxidant of strong reducing power directly to catch, and the variation of characteristic absorption can be known the removing ability of antioxidant to aryl biradical by inference after being caught by antioxidant according to DPPH, takes this senile-resistant efficacy of cosmetics is characterized.Detection is the important investigation index that prevents skin aging, particularly at photoaging to the ability of knowing of free radical.
Set up the DPPH experimental technique of standard, Embryo Gallus domesticus essence lyophilized powder of the present invention redissolves with the phosphate buffer of 10 mM pH7.0.Experiment is over against photograph with vitamin C, 0.1 mL takes a sample, add the standard DPPH methanol solution that 3.9 mL mass concentrations are 25 ug/mL, with methanol is the zeroing contrast, 515 nm wavelength are measured the light absorption value of different time down, be scaled the mass concentration of DPPH according to standard curve, calculate the residual rate of DPPH.Judge the removing free radical ability-oxidation resistance of Embryo Gallus domesticus essence according to the concentration of the pairing antioxidant of DPPH residual rate, the result shows, compares with vitamin C, and the Embryo Gallus domesticus essence of suggestion use amount has the ability of 40～60% removing free radical.
Embodiment 3 Embryo Gallus domesticus essences are to the inhibition of tyrosinase activity
Tryrosinase is a kind of copper bearing metalloenzyme, is prevalent in mammal, plant and the microorganism, mainly participates in the formation of the interior melanin of organism and other polyphenols.The bioactive substance that external tyrosinase activity suppresses has certain tyrosinase activity to suppress ability, can be used to pass judgment on the biological effectiveness of Pear Power active substance.
Embryo Gallus domesticus essence lyophilized powder of the present invention redissolves with the phosphate buffer of 10 mM pH7.0, with the DOPA is substrate, and Mushroom Tyrosinase is over against photograph, and phosphoric acid buffer is negative contrast, set up the tyrosinase activity detection architecture, be used to detect the inhibitory action of Embryo Gallus domesticus essence tryrosinase.Get 1.0 mL Mushroom Tyrosinase solution, add the phosphate buffer of 3 mL pH 6.0, add 2.0 mL DOPA solution, shake up.After reacting 5 min, 480 nm wavelength are measured OD value (using l cm cuvette) down, increase the vigor of expression reaction in 5 minutes endoenzyme with the OD value of unit interval.The Embryo Gallus domesticus essence suppresses active mensuration to tyrosinase activity, the Embryo Gallus domesticus essence that in the enzyme reaction system, adds use amount, measure OD480 after 5 minutes, the enzymatic activity after the negative increase of unit of account time OD is represented to suppress, with unconstrained dose relatively calculate to suppress efficient.The result show the Embryo Gallus domesticus essence to inhibition of tyrosinase activity 30～50%.
The active substance of whitening has the biological action that suppresses melanin formation or promote the melanin degraded, utilize murine melanoma B16 system, detect the Embryo Gallus domesticus essence to the synthetic inhibitory action of B16 cell melanin, estimate the active function of the active active component of skin-lightening cosmetic.
Melanocyte is cultivated, and adding the Embryo Gallus domesticus essence in the culture medium is processed group, and not adding the Embryo Gallus domesticus essence is the matched group cultured cell, and 420 nm absorbance values are measured in collecting cell, fragmentation, centrifugal, represent the melanic depth with the size of OD420.The result shows that compared with the control, the Embryo Gallus domesticus essence can will lack B16 cell melanin synthetic 35 ~ 55%.
Embodiment 5 Embryo Gallus domesticus essences are to the effect of Skin Cell collagen protein synthesis
Dermal matrix index components-collagen protein; Cell based counterdie index components-IV collagen type, skin is rebuild the mensuration of equivalent and skin histology functional component index-fibronectin and hyaluronic acid etc., to pass judgment on the effect of these functional materials to skin texture and reconstruction.
With the HDF(human skin fibroblast) and HEK(people's epidermal keratinocyte) be system, it is processed group that cultured cell adds the Embryo Gallus domesticus essence, cultured cell 48 hours, collecting cell is with total collagen protein of ELISA kit measurement and IV collagen type.Experimental result shows that the Embryo Gallus domesticus essence can improve HDF and the synthetic collagen protein of HEK cell.
Table 1 improves the percentage rate (%) of collagen protein synthesis
| Cell | Total collagen protein | The IV collagen type |
| HDF | 35～45 | 40～50 |
| |
30～40 | 30～40 |
Below be human trial
The skin effect test of embodiment 6 consumers
The elite dew that consumer's (51.2 years old mean age) test group of being made up of 40 objects adopts the elite that does not contain Embryo Gallus domesticus essence lyophilized powder to reveal and contain 2% Embryo Gallus domesticus essence lyophilized powder carries out the skin efficacy study.Subjects has been tested above-mentioned two kinds of prescriptions.
Test period comprises three days conditioning stage, three days and 30 days product operational phase.This is a Blind Test, the prescription picked at random.Three day trial employment period, subjects writes down operating position and fill in questionnaires every day, answers the sense organ that reveals about every kind of elite and 4 problems of handling characteristics; 30 day trial employment period, subjects writes down operating position and fill in questionnaires every day, answers the use feeling revealed about every kind of elite and 6 problems of effect, makes evaluation according to from 1 to 3 grade.Consumer's nearly all character that the elite that contains the Embryo Gallus domesticus essence is revealed has all been made the evaluation (Fig. 1) that is higher than blank sample as can be seen from the results.Think that the water humidity that uses skin behind the product contain the Embryo Gallus domesticus essence, brightness, roughness etc. have all had improvement in various degree.
The test of embodiment 7 moisture of skin
Each is organized laboratory sample and is tried the position random number, decision test specimen and the contrast order of being tried the position.Form test group by 8 volunteers.Before the test, the experimenter need unify to clean both hands forearm inboard with warm clear water.Clean the back and carry out measurement markers in experimenter's both hands forearm inboard.Two pilot regions of each labelling of right-hand man's forearm in this experiment.A kind of thing that tried is smeared in each zone, and is interregional every 1cm.Every place pilot region is 4cm * 4cm, and the test specimen amount is 0.2g.The experimenter sits quietly in constant environment behind the 30min, uses Corneometer CM825 PC to be tried the measurement of position blank value, and 5 points are measured according to certain fixed order in each zone, draw its meansigma methods.Be responsible for smearing sample by the special messenger then, and pick up counting.Measure the variation of MMV respectively at day part by Experimental design.The meansigma methods that at every turn detects is deducted the changing value that blank value is this period MMV, remove the rate of increase that blank value promptly draws MMV again, the results are shown in Figure 2.
Skin moisture content Growth Rate Calculation formula is as follows:
Skin moisture content rate of increase=(MMV
t-MMV
0)/MMV
0* 100%
In the formula, MMV
0Skin MMV before expression is smeared; MMV
tBack t period skin MMV is smeared in expression.
Experimental result shows that the amplitude of experimenter's skin hydration meansigma methods increase of using the elite dew that contains 2% Embryo Gallus domesticus essence lyophilized powder all greater than placebo, illustrates that the Embryo Gallus domesticus essence has moistening effect preferably.
Use illumination that Mexameter MX18 the measures specific wavelength volume reflection after on the human body skin, determine the content of melanocyte in the skin, can estimate the white-skinned face function of cosmetics by this.
Form test group by 20 female volunteers (38 years old mean age).The experimenter needs unified gentle cleaning agent washing test position (buccal position, the left and right sides) before the test, at first detect each test position (left and right sides buccal test position is symmetry as far as possible) with Corneometer CM 825PC tester, repeat respectively 5 times, draw its meansigma methods, this is basic value C
0Allow every experimenter use 0.3g placebo elite to reveal at the right side buccal then, usage quantity is identical in the left side, and the elite that contains the Embryo Gallus domesticus essence is revealed, and every day 2 times, continues 20 days altogether.After 20 days, carry out apparatus measures on buccal, each is tried the position and is respectively detected 5 times, draws meansigma methods, and is designated as C respectively
TAnd C
CKCalculate the variation of skin melanin content.The result shows to use and contains the skin melanin content that Embryo Gallus domesticus essence elite reveals and reduce very obvious, the range of decrease is all more than 20%, be up to 55.30%, and use the skin melanin content at placebo position to change not obvious, the range of decrease is between 3～25%, experimental result explanation Embryo Gallus domesticus essence can be used as effective active component, is applied in the cosmetics, can be effectively and carry significantly bright with brighten the colour of skin.
The test of embodiment 9 skin elasticitys
Skin elasticity reduces with skin aging, so skin elasticity is one of important symbol of judging skin aging, is that skin aging detects requisite project.Form test group by 20 female volunteers (38 years old mean age).The experimenter needs unified gentle cleaning agent washing test position (buccal position, the left and right sides) before the test, at first detect each test position (left and right sides buccal test position is symmetry as far as possible) with Corneometer CM 825PC tester, repeat respectively 5 times, draw its meansigma methods, this is basic value Ue
0Allow every experimenter use 0.3g placebo elite to reveal at the right side buccal then, usage quantity is identical in the left side, and the elite that contains the Embryo Gallus domesticus essence is revealed, and every day 2 times, continues 20 days altogether.After 20 days, carry out apparatus measures on buccal, each is tried the position and is respectively detected 5 times, draws meansigma methods, and is designated as Ue respectively
TAnd Ue
CKThe result shows that use contains all increases to some extent of skin elasticity that Embryo Gallus domesticus essence elite is revealed, and amplification is up to 30%, and uses the skin elasticity variation at placebo position not obvious, partly increase is arranged, but amplification is only up to about 10%.Experimental result explanation Embryo Gallus domesticus essence can be used as effective active component, is applied in the cosmetics, can be effectively and improve the elasticity of skin significantly.
The test of embodiment 10 wrinkles of skin
The variation that the elite that this test is intended to estimate single and uses the placebo elite to reveal and contain 2% Embryo Gallus domesticus essence lyophilized powder is for several times respectively revealed the back skin coarseness.Form test group by 20 female volunteers (38 years old mean age).The experimenter needs unified gentle cleaning agent washing test position before the test.Reveal in experimenter's the right side facial 0.3g of use placebo elite then, usage quantity elite identical, that contain the Embryo Gallus domesticus essence is revealed in the left side, every day 2 times, continues 20 days altogether.When on-test, use elite to reveal 2h first after, extract the skin die behind the 4h and during off-test, analyze by the software of special use then, can obtain the relevant parameter of different skin wrinkle.On buccal, carry out apparatus measures, as basic control level.
Analyze and Bonferroni checks by the variance repeated measure, the record numerical value at each time control point is compared.When Probability p≤0.05, can think that difference has statistical significance, the results are shown in Figure 3.
Experimental result shows that the experimenter who uses the elite dew that contains 2% Embryo Gallus domesticus essence lyophilized powder uses the maximal roughness at position obviously to descend, compare with the result who uses placebo and to have significant difference, explanation is under used concentration, the Embryo Gallus domesticus essence can fast and effeciently produce the wrinkle removal effect, can be used in ripe skin protection in age and the anti-ageing cosmetics of waiting for a long time.
In this description, the present invention is described with reference to its certain embodiments.But, still can make various modifications and conversion obviously and not deviate from the spirit and scope of the present invention.Therefore, description and accompanying drawing are regarded in an illustrative, rather than a restrictive.
Claims (10)
1. the preparation method of an Embryo Gallus domesticus essence lyophilized powder is characterized in that concrete steps are:
(1) gets 8～16 days healthy chicken embryo of hatching, strike shell and get Embryo Gallus domesticus;
(2) with food masher Embryo Gallus domesticus is broken into homogenate, add pH and be 6.8～7.2, concentration is the phosphate buffer of 30～50 mM, presses mass ratio, Embryo Gallus domesticus: phosphate buffer=1:1～3, under 4～10 ℃ of conditions, Embryo Gallus domesticus and phosphate buffer are fully stirred leaching spend the night;
(3) with the Embryo Gallus domesticus leaching liquid in hospital gauze filtration step (2), with filtrate with the centrifugal 15～30min of the rotating speed of 4000～6000 rpm;
(4) with the supernatant after centrifugal, be that the doughnut membrane ultrafiltration of 10000～20000 Da filters with molecular cut off, filter liquor, i.e. Embryo Gallus domesticus essence;
(5) adding trehalose in the Embryo Gallus domesticus essence, is trehalose by mass volume ratio: Embryo Gallus domesticus essence=1:5～10g/mL at-45～-85 ℃ of following lyophilization 12～36h, promptly obtains Embryo Gallus domesticus essence lyophilized powder.
2. Embryo Gallus domesticus essence lyophilized powder for preparing by the described method of claim 1.
3. the application of Embryo Gallus domesticus essence lyophilized powder as claimed in claim 2 in the preparation anti-aging cosmetics.
4. a cosmetic composition is characterized in that containing the described Embryo Gallus domesticus essence of claim 2 lyophilized powder, and the content of this Embryo Gallus domesticus essence lyophilized powder is 0.1%～5% of cosmetic composition gross weight.
5. cosmetic composition according to claim 4 is characterized in that compositions is cream, frost, dew, water, gel, lotion.
6. cosmetic composition according to claim 4 is characterized in that said composition also adds used for cosmetic oiliness raw material is arranged, and described oiliness raw material is the stearic acid esters, the oleic acid esters, linoleic acid esters, Palmic acid esters, the isostearic acid esters, hydroxy stearic acid esters, myristic acid esters; Different Palmic acid esters, lauric acid esters, polyglycereol esters, lanolin derivative class, pentaerythritol esters, ricinoleic acid ester, succinic acid esters, phosphoric acid ester, mountain Yu acid glycerol base ester, wax class; The addition of described oiliness raw material is 0.01%～45% of a composition total weight.
7. cosmetic composition according to claim 4, it is characterized in that said composition also adds the used for cosmetic nutritive additive is arranged, described nutritive additive is: vitamins, aminoacid, adenylate, sterols, hydrolyzed protein class, and other nutritional substance: one or more in umbilical cord extracting solution, oil-soluble placenta extracting liquid, the gluconic acid sodium salt, the addition of described nutritive additive are 0.01%～45% of composition total weight.
8. cosmetic composition according to claim 4, it is characterized in that said composition also adds surfactant is arranged, described surfactant is: a kind of in sorbitan fatty acid ester and its ethylene oxide adduct, polyoxyethylene lauryl ether, cocoanut fatty acid diethanolamide, monoolein, polyoxyethylene castor oil, Wool wax alcohols,ethoxylated, alkyl phosphate, alkyl phosphoric acid polyethylene glycol oxide ester, fatty acid alkali metal salt, the alkyl glycerol base ether or some kinds; The addition of described surfactant is 0.01%～45% of a composition total weight.
9. cosmetic composition according to claim 4, it is characterized in that said composition also adds water-soluble polyol is arranged, described water-soluble polyol is ethylene glycol, Polyethylene Glycol, propylene glycol, glycerol, glycerol-ethylene oxide adduct, sorbitol, POE(20) a kind of in sorbitol, 1,3 butylene glycol, maltose alcohol, xylitol, threitol, erithritol and the alcohol that obtains by the reduction amylolysis or several; The addition of described water-soluble polyol is 0.01%～60% of a composition total weight.
10. according to the described cosmetic composition of one of claim 4-9, it is characterized in that said composition also adds used for cosmetic powder class raw material is arranged, described powder class raw material is a silicates, aluminum salt, mica group, lamination powder or surface treatment thing powder; The addition of described used for cosmetic powder class raw material is 0.01%～50% of a composition total weight.
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| CN103262939A (en) * | 2013-05-23 | 2013-08-28 | 江南大学 | Preparation method of antioxidant activity mixed peptide originated from chick embryo |
| CN103445163A (en) * | 2013-09-02 | 2013-12-18 | 蒋应文 | Chick embryo nutrient and preparation method thereof |
| CN103766947B (en) * | 2014-02-17 | 2015-10-07 | 雷正新 | A kind of preparation technology of young fowl food |
| CN106360421A (en) * | 2016-10-25 | 2017-02-01 | 桂林淮安天然保健品开发有限公司 | Preparation process of embryo egg health product capable of improving antifatigue efficacy |
| CN106509680A (en) * | 2016-10-25 | 2017-03-22 | 桂林淮安天然保健品开发有限公司 | Manufacture technology of embryo egg health-care products |
| CN106509679A (en) * | 2016-10-25 | 2017-03-22 | 桂林淮安天然保健品开发有限公司 | Making technology of embryonated egg health-care product having efficacy of enhancing immunity |
| CN106490507A (en) * | 2016-10-25 | 2017-03-15 | 桂林淮安天然保健品开发有限公司 | Improve the embryo egg hatching technique of immunoglobulin content |
| CN109601695A (en) * | 2018-12-18 | 2019-04-12 | 长春大学 | A kind of method that activated protein exempts from digesting protein with soybean protein for raw material preparation |
| CN111568781A (en) * | 2020-06-23 | 2020-08-25 | 上海美月生物科技有限公司 | Skin care gel and preparation method thereof |
| CN115025124B (en) * | 2022-06-17 | 2024-04-30 | 济南万泉生物技术有限公司 | Hematopoietic anti-aging active component extracted from chick embryo tissue and application thereof |
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| CN1128144A (en) * | 1995-07-13 | 1996-08-07 | 来风战 | Chicken embryo extract oral liquid |
| CN1226401A (en) * | 1998-02-16 | 1999-08-25 | 沈双宇 | Health-care products 'Fenghuangtai' and production thereof |
| CN1324625A (en) * | 2000-05-19 | 2001-12-05 | 邓先明 | Chicken embryo essence preparing process |
| CN1342461A (en) * | 2000-09-08 | 2002-04-03 | 邹清雁 | Process for preparing embryo nutrient |
| KR100542266B1 (en) * | 2003-08-04 | 2006-01-11 | 한국식품연구원 | Face-pack Composition Comprising Sericin Protein Extracted from Cocoon |
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