CN102517363A - Production method and application of egg-shell membrane collagen peptides - Google Patents
Production method and application of egg-shell membrane collagen peptides Download PDFInfo
- Publication number
- CN102517363A CN102517363A CN2011103631331A CN201110363133A CN102517363A CN 102517363 A CN102517363 A CN 102517363A CN 2011103631331 A CN2011103631331 A CN 2011103631331A CN 201110363133 A CN201110363133 A CN 201110363133A CN 102517363 A CN102517363 A CN 102517363A
- Authority
- CN
- China
- Prior art keywords
- membrane
- polypeptide
- eggshell membrane
- egg
- collagen polypeptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a production method and application of egg-shell membrane collagen peptides. The method comprises the following steps of: (1) taking an egg membrane out of an egg shell, and smashing to over 40 meshes; (2) adding egg membrane powder obtained in the step (1)into a papain solution with the concentration of 800-1,000 U/L, and stirring at the temperature of 35-50 DEG C for 2-3 hours to obtain a mixed material, wherein the solid-liquid ratio is 1:(2-5); (3) adding an NaOH solution into the mixed material obtained in the step (2) till the mass concentration of NaOH is 2-4.5 percent, stirring at the temperature of 45-85 DEG C for 3-4 hours, standing, and taking a supernatant; and (4) adjusting the pH value of the supernatant obtained in the step (3) to 7-11 with dilute hydrochloric acid to obtain an egg-membrane collagen peptide liquid, desalting, concentrating and drying to obtain egg-shell membrane collagen peptides. The production method has simple operation steps and simple conditions, egg-shell membrane collagen peptides can be produced on a large scale, energy consumption is low, environmental pollution is avoided simultaneously, and two or more polypeptide surfactants can be produced simultaneously; and by changing operation conditions, the performance of the polypeptide surfactant can be adjusted and controlled.
Description
Technical field
The present invention relates to the polypeptide technical field, the especially working method of eggshell membrane collagen polypeptide, and application.
Background technology
Ovum crusta Gallus domesticus exists as human lives's sub product in a large number, and the eggshell membrane that adheres in it causes great contaminate environment especially easily.Eggshell membrane contains multiple proteins (content is about 90%) like collagen protein (I, V, X type), SPP1, BSA, sialoprotein etc.; Also contain CHS, mucinase, N-acetylglucosamine semi-lactosi, glucuronic acid etc.; Being mainly used in treatment and alleviating arthrodynia etc., is first-class raw materials such as beauty treatment, healthcare products, makeup, daily use chemicals washing.Egg membrane protein is with a wide range of applications with aspects such as dressing materials, enzyme immobilization carrier, cell tactophily matrix or drug controllable slow-release materials at heavy metal ion selective absorbent, artificial skin or skin treating.
But eggshell membrane is difficult for from Ovum crusta Gallus domesticus, isolating on a large scale, at present also not about produce the report of polypeptide surfactant from eggshell membrane.Simultaneously, eggshell membrane contains a large amount of cross-link bonds and is difficult to dissolving, but strong excessively inappropriate treatment condition can cause the polypeptide excessive degradation, and product is difficult to separate purifies.These characteristic limitations the production and the application of eggshell membrane.
Although prior art is quite understood proteinic enzymolysis process mechanism, from the requirement of different starting material restrictions, variant prodn, new treating processes often need develop to satisfy technology and economic requirement specially.As:
1, (Qiqihar College of Light Industry journal such as Yu Shujuan; 1994,10 (30): 30-35) prepare aminoacids complex, promptly after the acid effect, add proteolytic enzyme again and be hydrolyzed with acid-enzyme hydrolysis method hydrolysis corn protein; This method can reduce the drawback of acid system and two kinds of methods of enzyme process; Make that amino acid kind in the product is more complete, content is higher, have very strong solubleness and emulsifying capacity, in the utilization ratio that improves product, reduce the discharging of pollutent.
2, people such as Wang Cuihua (the synthetic and froth stability [J] of protein-based whipping agent. Nanjing University of Technology's journal, 2006,28 (4): 92-96) adopting the cow hoof angle is the major protein raw material, at an amount of calcium hydroxide and little amount of N aHSO
3Hydrolysis under the condition that exists, successful synthetic protein type foamed concrete is used whipping agent.
3, (the papain hydrolysis Semen phaseoli radiati albumen prepares the research [J] of soluble peptide to Lu Zhenhua etc.Food science and technology, 2005,11:56-58) the research papoid is to the hydrolytic action of Semen phaseoli radiati albumen.The top condition of enzymolysis is enzyme concn 8% (an E/S mass ratio), concentration of substrate 7% ratio of water (Semen phaseoli radiati albumen with), and 65 ℃ of temperature of reaction, reaction times 3h, pH6.5, this work has been opened up vegetable-protein and has newly been originated.
4, (enzymatic hydrolysis of gingko activated protein and anti-oxidant activity research [J] such as Deng Qianchun.EI, 2005,21 (11): 155-159) studied enzymatic hydrolysis and prepared the method for gingko peptide section and the variation of anti-oxidant activity thereof; Through single factor analysis, orthogonal test and to the analysis of test-results; The optimum hydrolysising condition of having confirmed hydrolysis by novo gingko activated protein is that concentration of substrate is 1%, pH value 8.5,45 ℃ of hydrolysis temperatures; Enzyme concn 7000U.g-1, reaction times 6h.Biological activity through discovering gingko peptide section obviously improves, for the exploitation of gingko deep processing research and product innovation thereof lays the foundation.
5, Pushkar Singh Bora (Functional properties of native and succinylated lentil (lens culiaris) globulins [J] .Food Chemistry; 2002; 77:171-176) grind made internal disorder or usurp behind the succinylation with the performance of the resulting root of Szemao crotalaria sphaeroprotein of acidylate not, wherein the iso-electric point of the root of Szemao crotalaria sphaeroprotein after the acidylate changes to 3.5 from 4.5, has improved the solubility greater than 4 o'clock at pH; Wetting ability; Viscosity has 100% raising, and emulsifying property and emulsifying stability also improve a lot, and whipability has reduction slightly.
6, people's (synthesizer performance [J] of aminoacids complex bay alcohol ester such as Feng Guang wick.Zhengzhou Engineering College's journal; 2003,24 (4): 42-45) having studied the aminoacids complex liquid concentrator that obtains with the rapeseed meal hydrolysis is raw material, makes aminoacids complex bay alcohol ester with the lauryl alcohol esterification; The top condition of having confirmed building-up reactions is that the mol ratio of lauryl alcohol and aminoacids complex is 1: 2; Toluene is made band aqua (110～122 ℃ of temperature of reaction), and the reaction times is 4h, the dense H of catalyzer
2SO
4Amount be 8% of aminoacids complex, a spot of polyoxyethylene glycol (PEG600) is made phase-transfer catalyst, the product that makes has good froth stability, and good characteristic such as pungency is little, toxicity is low, degradation property is good and degraded product toxicity is low.
These above-mentioned examples explain that clearly the polypeptide products of each type usually has its corresponding raw material; Corresponding processing conditions and technological process; That is to say that the current production process of polypeptide of having reported all is not suitable for from eggshell membrane produces polypeptide or polypeptide surfactant in batches.Except that receiving material performance restriction, prior art is at production quality control, operability, energy consumption, particularly realize that aspects such as mass-producing, environmental friendliness production all have problems.
Summary of the invention
The present invention is directed to deficiency, propose a kind of working method of eggshell membrane collagen polypeptide, ability large-scale production, processing ease.
In order to realize the foregoing invention purpose, the present invention provides following technical scheme: a kind of working method of eggshell membrane collagen polypeptide may further comprise the steps:
1., from Ovum crusta Gallus domesticus, take out eggshell membrane, be crushed to more than 40 orders;
2., with step 1. the eggshell membrane powder to add concentration be in the papoid solution of 800～1000U/L, solid-to-liquid ratio is 1: 2～5, in 35～50 ℃ stirred 2～3 hours compound;
3., 2. add NaOH solution in the compound to step, to the mass concentration of NaOH be 2～4.5%, stirred 3～4 hours in 45～85 ℃, leave standstill; Get supernatant;
4., the supernatant pH value of using the Hydrogen chloride regulating step 3. to obtain is 7～11, must eggshell membrane collagen polypeptide liquid; Through desalination, concentrate and drying after, obtain the eggshell membrane collagen polypeptide.
Preferably, step 4. desalination is that the employing membrane pore size is the film of 1k with concentrating, and drying is 180 ℃ of following spraying dryings.
Preferably, step 4. desalination be that pure water wash-out eggshell membrane collagen polypeptide liquid to pH value is 7; Concentrate is that the employing membrane pore size is the membrane filtration of 300k; Drying is 180 ℃ of following spraying dryings of liquid concentrator or lyophilize behind the membrane filtration.
Preferably, it is the membrane sepn of 30k that the filtered solution that membrane filtration obtains adopts membrane pore size, and liquid and secondary filtered solution must dam; The said liquid that dams is in 180 ℃ of following spraying dryings of temperature; It is the 1k membrane-concentrated that said secondary filtered solution adopts the aperture, gets liquid concentrator in 180 ℃ of following spraying dryings of temperature.
The eggshell membrane collagen polypeptide that a kind of working method as stated makes.
A kind of makeup include the eggshell membrane collagen polypeptide of 0.01wt%～20wt%.
Preferably, eggshell membrane collagen polypeptide content is 0.1wt%～10wt%.
Preferably, eggshell membrane collagen polypeptide content is 0.1wt%～5wt%.
Compared with prior art; The present invention utilizes high efficiency, the specificity characteristics of enzyme; Eggshell membrane to common means difficult degradation; In specific temperature and pH scope, carry out the high-performance bio degraded, adopt membrane sepn to concentrate and purifying to the eggshell membrane of degrading, the egg membrane protein polypeptide is produced in the implement scale chemical industry.
The present invention is because operation steps and condition are simple; The eggshell membrane collagen polypeptide that is produced on a large scale, energy consumption is low simultaneously free from environmental pollution, and the polypeptide surfactant that can produce two or more simultaneously; Through changing operational condition, the performance of polypeptide surfactant can adjustedly be controlled.
The present invention has reported the main surface and interface characteristic of these polypeptide surfactants first.These eggshell membrane polypeptide surfactants are that reduction surface tension ability is the strongest in current known protein matter and the polypeptide; Mixing system with conventional surfactant formation can further reduce surface tension; These mixing systems can form highly stable foam and emulsion, and its relative stability can and mix decorum composition and regulate through the polypeptide type.
As the natural polypeptides product; Cooperate the green bio degraded and the gentle membrane sepn purification production process that adopt; Utilize eggshell membrane to produce the alternative conventional surface active substance of producing from petroleum of resulting polypeptide surfactant (tensio-active agent and surface-active polymer or their mixture), be used to improve currently available products or compound product innovation.The natural polypeptides tensio-active agent to the action temperature of skin and hair with; Stronger avidity and wetting effect are arranged, and nontoxic, nonirritant, in washing, can also be beneficial to the absorption of skin or hair to polypeptide; Be particularly suitable for allergic person and baby and use (Li Yi; Zhu Haiyang, Li Huashan, etc.The application [J] of N-fatty acyl group glutamate family tensio-active agent in household chemicals. household chemicals science, 2003,26 (4): 20-23).
This research work shows, polypeptide surfactant has better washability with a spot of other tensio-active agent coordinative roles the time, and pure natural eggshell membrane polypeptide surfactant is nontoxic, be easy to biological degradation and absorption, meets human requirements of green environmental protection.The eggshell membrane polypeptide surfactant is enjoyed in biological medicine, pharmacy, biomaterial, makeup and protective foods etc. and being had broad application prospects.
Description of drawings
Fig. 1 is products C its surface tension variation diagram in time under different concentration;
Fig. 2 is product D its surface tension variation diagram in time under different concentration;
Fig. 3 is the logarithm concentration mapping of the co-adsorption surface tension of products C and SDS to SDS; Wherein the small peptide concentration fixed is in 0.1 and 1 mg/ml.The surface tension of pure SDS as a reference.All experiments are all carried out in pure water, and temperature is controlled at 22～23 ℃;
What Fig. 4 was products C and SDS at silicon oxide surface is co-adsorption.
Embodiment
Describe the present invention below in conjunction with specific embodiment and accompanying drawing, the description of this part only is exemplary and explanatory, should any restriction not arranged to protection scope of the present invention.
Embodiment 1
Get the fresh Ovum crusta Gallus domesticus of 1000 grams, foreign matter is removed in the water flushing, shell is pulverized the Ovum crusta Gallus domesticus of removing wherein; The protein enzyme solution that configures the eggshell membrane that contains a small amount of shell then (200ml, concentration is 800U/L), design temperature is at 60 ℃; Stir after 2 hours, slowly add NaOH solution, make that the mass concentration of the NaOH in the compound is 3%; 65 ℃ of temperature stirred 3 hours, left standstill.Get supernatant, regulate pH value to 7 with Hydrogen chloride and get membranin liquid, the volume of membranin liquid is 3 times of former supernatant.Adopt the film equipment desalination and concentration of 1k,, get eggshell membrane collagen polypeptide products A (about 37 grams) in 180 ℃ of following spraying dryings of temperature.
Get the fresh Ovum crusta Gallus domesticus of 1000 grams, foreign matter is removed in the water flushing, shell is pulverized the Ovum crusta Gallus domesticus of removing wherein; The protein enzyme solution that configures the eggshell membrane that contains a small amount of shell then (200ml, concentration is 1000U/L), design temperature is at 35 ℃; Stir after 3 hours, slowly add NaOH solution, make that the mass concentration of the NaOH in the compound is 3%; 50 ℃ of temperature stirred 4 hours, left standstill.Get supernatant, regulate pH value to 9 with Hydrogen chloride and get membranin liquid, the volume of membranin liquid is 3 times of former supernatant.Adopting membrane pore size is the concentrated membranin liquid of membrane filtration of 300k, and the salinity that adds in the pure water elute soln makes the pH value get back to 7, reclaims liquid concentrator and filtered solution.Liquid concentrator is adopted spraying drying or cryodesiccated mode in temperature below 180 ℃, get eggshell membrane collagen polypeptide products B (about 2 grams).
With the resulting filtered solution of embodiment 2 preparing product B, adopt the film of 30k to separate, reclaim dam liquid and secondary filtered solution.In 180 ℃ of following spraying dryings of temperature, the eggshell membrane collagen polypeptide gets products C (about 15 grams) with the liquid that dams that obtains.
With the resulting secondary filtered solution of embodiment 3 preparing product C, adopting pore size is the 1k membrane-concentrated, gets liquid concentrator and gets product D (about 22 grams) in 180 ℃ of following spraying dryings of temperature.
Embodiment 5 a large amount of preparation eggshell membrane collagen polypeptide products B, C and D
Get 200 kilograms of fresh Ovum crusta Gallus domesticuss, foreign matter is removed in the water flushing, sterilization; Crushing; With reference to as before described step shell pulverize is removed, add the protein enzyme solution that configures to the eggshell membrane that contains a small amount of shell in proportion then, carry out alkaline degree of depth dissolving again; Acid neutralization film to be used such as is left standstill and desalination and concentration is set is separated.The step that adopts is identical with foregoing description, but the increase of clear liquid amount increases through parallel connection in proportion and separates the blooming operating unit, and suitably regulation system is operated balance and reached desalination and concentration and separate targets.Through getting about 410 grams of product B in 180 ℃ of following spraying dryings of temperature, about 3100 grams of C, about 4300 grams of D.
The product compositional analysis:
Get product B, C and D carry out chemical composition analysis, and the result shows:
Product B contains 88% organism (amino acid), 7% inorganics (being mainly calcium salt), 5% water.
Products C contains 75% organism (amino acid), 16% inorganics (being mainly calcium salt), 7% water.
Product D contains 82% organism (amino acid), 12% inorganics (being mainly calcium salt), 6% water.
Analytical results has the limit of error about 3%.
The performance of products test experience
To the product B that the foregoing description 5 makes, C and D carry out Performance Detection, and the result is following:
1, water-soluble:
The solvability of eggshell membrane collagen polypeptide products in the aqueous solution is better, and wherein product D (5kDa) is better than products C (40kDa) solvability.Elevated temperature helps the solubilising of products C.
2, surface tension
2.1, the surface tension of eggshell membrane collagen polypeptide
Products C as illustrated in fig. 1 and 2 and D its surface tension under different concentration changes in time.Add the eggshell membrane polypeptide and in the aqueous solution, can reduce surface tension significantly.Along with peptide concentration increases, it is strong more to reduce the surface tension ability.
The result shows that products C had similar reduction surface tension ability with D in incipient 5～10 minutes.In subsequently 5～10 minutes, surface tension tends to be steady.Therefore, the surface tension 20 minutes the time can be considered equilibrium value.Even if can be clearly seen that surface tension is still much lower than water when minimum concentration (0.03 mg/ml).When concentration raise, surface tension descended gradually.When the maximum concentration of being studied was 10 mg/ml, surface tension had only 30mN/m.
As far as we know, this is the minimum surface tension that albumen and small peptide can reach, people such as Cooper [Adsorption of frog foam nest proteins at the air/water interface, Cooper, A.; Kennedy, M.W.; Fleming, R.I.; Wilson, E.H.; Videlerm H.; Wokosin, D.L.; Su, T.J.; Green, R.J.; Lu, J.R., Biophys.J.2005,88,2114-2125] once utilized surface tension instrument and neutron reflection to study different proteic surface propertieies.The result shows that why frog nest albumen can stablize the nest foam is because it is that reduction surface tension effect is best in the known protein (like N,O-Diacetylmuramidase, bovine serum albumin, beta casein).Frog nest albumen is when 1 mg/ml, and surface tension is about 50mN/m.Comparatively speaking, under identical solution condition, products C and D but can be reduced to 33mN/m and 40mN/m with surface tension, show that the eggshell membrane polypeptide through processing treatment of the present invention has the strongest reduction surface tension ability.
As far as most of tensio-active agents, the surface tension when micellar concentration is everlasting 30mN/m (most of nonionic alkyl surfactants are such as C12E5) to changing between 40mN/m (most of ionic surface active agents are such as C12TAB and SDS).The small peptide tensio-active agent has the capillary ability of the attenuating similar with conventional surfactants, therefore can be used as substitute products.The small peptide tensio-active agent also has the incomparable character of oil tensio-active agent such as bio-compatibility, degradability and environment friendly simultaneously.
In practical application, the aqueous solution often contains salt.In addition, pH and temperature also can change.Ionic surface active agent is different to the reaction of environment with nonionogenic tenside.Such as, nonionogenic tenside C12E5 is insensitive to salt, but ionic surface active agent SDS is responsive to salt, such as, in SDS, add sodium-chlor and can reduce micellar concentration and make tensio-active agent more effective.By contrast, ionic surface active agent is then to temperature-insensitive, but nonionogenic tenside is to temperature sensitive.Heat and to cause nonionogenic tenside C12E5 to reduce micellar concentration.Under extreme conditions even can be insoluble.Therefore, when using, to consider these problems.
PH value of solution is to the capillary influence of polypeptide when having investigated 1 mg/ml.For product D (5kDa), pH changes at 7 o'clock from 5, and surface tension is had only less influence.But be raised at 8 o'clock, surface tension can rise 5 to 10mN/m.Possibly cause because solubleness rises.For products C (40kDa), pH changes at 8 o'clock from 5, to capillary influence less than 5mN/m.Add the capillary obvious change that sodium-chlor does not cause products C and D simultaneously.From 15 to 40 ℃ of temperature are influential to the surface tension of products C, but product D is not had much affect.But temperature does not have tangible trend to both influences.On the whole, Influence of Temperature is in most cases less than 5mN/m.
Therefore, compare with conventional surfactants, eggshell membrane collagen polypeptide tensio-active agent is to temperature, and pH and salt are all relatively not too responsive.This SOLUTION PROPERTIES with albumen and polypeptide is similar, but different with the conventional surfactants SOLUTION PROPERTIES.So environmental stability has important meaning for its application.Such as, the insensitivity of temperature makes them reach stable (sour milk, ice cream) in the emulsification of food, there is important use the emulsification of care products (face cream) aspect.
2.2, the surface tension of two-component mixture (containing tensio-active agent and polypeptide active agent)
Care products, normal some tensio-active agents that add in food and the medicine, polymkeric substance and other compositions are the most optimized to reach interface performance.Therefore, polypeptide surfactant can become their substitute in this series products, or is used to develop new product.Technical problem is that can polypeptide and other products compatible and mixing.
As shown in Figure 3, with the surface tension of pure SDS as a reference, the result is consistent with bibliographical information, shows that the purity of SDS sample is very high.When products C existed, it is low and more smooth that surface tension becomes.Because pure polypeptide wants high in this concentration range surface tension, explain that both interactions play leading role in SDS and polypeptide two-component mixture.
When SDS concentration raise, although final effect is the surface tension that keeps lower, its molecule was still unclear with composition in the structure at interface.People such as Green once utilized surface tension instrument and neutron reflection to study interface structure and composition [Interaction of lysozyme and sodium dodecyl sulphate at the air-water interface, Green, the R.J. of SDS-N,O-Diacetylmuramidase; Su, T.J.; Joy, H.; Lu, J.R., Langmuir 2000,16,5797-5805.].They find a surface tension peak clearly below aggregate concentration.Be below or above under the situation at this peak, it is stable that surface tension keeps.Though surface tension is seen quasi-stable, the neutron reflection result of experiment shows that the structure at its interface but alters a great deal with forming.Therefore, at this moment surface tension can not give too much interface information.But two kinds of technical tie-ups are used and can be reflected more detailed interface change procedure.They point out that leading role is played in charge neutrality when low SDS concentration.Oleophilic function is also played a role at formation SDS-albumen micella, makes globular preteins structural distortion.The distortion of capillary rising and protein structure and dissolving are carried out simultaneously, and competitive adsorption plays a leading role relevant at the interface with SDS in surface tension reduction subsequently.People such as Taylor [Taylor, D.J.F.; Thomas, R.K.; Hines, J.D.et al., Langmuir 2002,8,9783-9791] once studied the surface adsorption of the tensio-active agent CnTAB and the electronegative polymer P SS two-component mixture of positively charged.With SDS-N,O-Diacetylmuramidase system similarity, they have equally also observed the appearance at micelle critical concentration lower surface tensammetric peak, mean in the formation of these condition settle things and the tension force of following to raise.
System compares with the SDS-N,O-Diacetylmuramidase, and the surface tension of membrane polypeptides promoting agent is quite steady, shows that its interface structure is better with the composition coupling.It should be noted the N,O-Diacetylmuramidase positively charged, Comparatively speaking, the iso-electric point of small peptide promoting agent is between pH4～5, and is identical with the SDS charging property between pH5～8.Therefore, charge neutralization caused in such system adhere to and subsequent precipitate the surface tension peak value that is brought can not occur.
When product D mixed with SDS, its two-component system surface tension was same more smooth with the variation of SDS concentration, and surface tension drops with the concentration rising of polypeptide active agent; The two-component mixture system of these fundamemtal phenomenas and SDS and polypeptide products C is substantially similar; Explain the interface behavior receive both interact control, in other words, in very big SDS and polypeptide fraction scope; The interface certainly is to contain two components, and both play leading role at interaction.
In view of above-mentioned analysis, investigated equally products C and D under the same conditions with the mixed surface tension situation of positively charged ion C12TAB.Our table of discovery surface tension result and SDS-membrane polypeptides system similarity.Just almost smooth curve below the C12TAB micellar concentration.Small peptide concentration is when 1 mg/ml, and products C and C12TAB mixture surface tension have only 30mN/m.The disappearance at surface tension peak and low surface tension also are its two key characters.These results show the surface tension peak that different electro ultrafiltration causes and the bioactive peptide system that is deposited in us followed in can not take place, show that electrostatic interaction does not play a major role.This provides particular performances for active polypeptide in stable foam and emulsion process.
3, froth stability
3.1, the froth stability of polypeptide active agent
Low surface tension and high foam stability property are associated, but not all low surface tension can both produce high foam stability property.After understanding polypeptide active agent system performance, depend on practical situations, can adjust system stability.In the test below, with different protein dissolutions in water.Concentration is 1 mg/ml.With the same concussion of reagent bottle 1 minute.After leaving standstill 2 minutes, polypeptide products D foam is still very stable, and is similar with the foam of beta casein (beta-casein), and is better than bovine serum albumin (BSA).Comparatively speaking, N,O-Diacetylmuramidase (chicken egg white lysozyme), the foam of products C reduces a lot, its stable relative mistake.
3.2, the polypeptide active agent contains the froth stability of SDS
The beta casein of 1 mg/ml, product D (5kDa), bovine serum albumin, products C (40kDa) adds 0.2mM SDS in the N,O-Diacetylmuramidase.
With the same concussion of reagent bottle 1 minute.After leaving standstill 2 minutes, take pictures.Products C forms unstable foam too, but has been become on the contrary by the foamy stability of bovine serum albumin and N,O-Diacetylmuramidase formed thereby.
The result shows that simultaneously change SDS and polypeptide active agent concentration separately can change foamy stability, and in other words, mixed system foamy stability and unstable can be controlled through selecting conditions such as different polypeptides promoting agent type, concentration.Not hard to imagine, froth stability and unstable also can be passed through other condition, adjustings such as the kind of promoting agent, salt, other additive, concentration.Explanation can be according to the product request for utilization in practical application; Come the stability and the unstable of control foam through component; As make clean water not need stable foam to occur, but hair-cream but generally needs a little stability, and the food foam usually requires good stability.
According to the principle of as above describing; Through with positively charged ion, nonionic and other biological or synthetic interfacial activity material (natural surface active agent, modified-cellulose, SEPIGEL 305) compound; Eggshell membrane polypeptide adjustable foamy stability and unstable depend on usage.
4, emulsifying stability
4.1, the emulsifying stability of polypeptide active agent
In the research emulsion's stability, aqueous solution composition is kept identical with above-mentioned foamite system, after normal hexane 1 to 1 mixes, with the same concussion of reagent bottle 1 minute.After leaving standstill 2 minutes, take pictures.Figure 11 results show that the emulsifying effectiveness of product D is better than beta casein (beta-casein), bovine serum albumin (BSA) and N,O-Diacetylmuramidase (lysozyme).The stability of products C also a little less than, this trend is identical with froth stability.
4.2, the emulsifying stability of polypeptide active agent after adding SDS
On the basis of true protein emulsion stability experiment, in each sample, add 0.2mMSDS, mix with normal hexane 1 to 1 again.With the same concussion of reagent bottle 1 minute.After leaving standstill 2 minutes, take pictures.The froth stability that result and polypeptide active agent add behind the SDS is similar, shows to add the certain adjustable surface tension of low quantity of surfactant, influences interface molecular structure and foamy stability.The relative stability of products C and D is with not add SDS identical, but adds SDS adjustable stability.Further experiment shows, the stability of emulsion can be controlled through regulating polypeptide and SDS concentration.Similarly, regulate the type of polypeptide surfactant and the purpose that solvent nature can both reach the control emulsion stability, this performance is that basic product stability has very strong directive significance with the emulsion to control.
5, selective adsorption
Below the purpose of experiment is to show the adsorptive power of polypeptide at solid surface.As shown in Figure 4, pH is between 6.5～7 in experiment, and temperature is at 22～23 ℃.Surface excess is measured by oval photometer.After 60 minutes, solution is removed, and puts into the water of same amount, and continues to measure.Because the reduction of strength of solution, the absorption of rise is caused by bioactive peptide.
The polypeptide active agent all has very strong absorption at many dielectric surfaces, and this variation from surface contact angle can be confirmed easily.Simultaneously, adsorbed polypeptide active agent is difficult for water or damping fluid rinses out, and shows the non-reversibility that it is very strong.
Hydrophilic and the weak negative electricity of band of silicon oxide surface, similar with hair.Therefore SDS does not adsorb on the surface.On the contrary, polypeptide since the effect meeting of entropy in surface adsorption.Therefore, increase SDS concentration and can suppress polypeptide in surface adsorption.When SDS concentration was 5mM, polypeptide no longer adsorbed.Yet when solution dilution or cleaning, because SDS concentration sharply descends, its influence power reduces.Comparatively speaking, polypeptide is little in surface adsorption and concentration relationship, causes polypeptide in surface adsorption.Therefore, polypeptide can be regulated through mixture ratio in the absorption on surface.The result shows that simultaneously polypeptide has stronger surfactivity, can be through flushing in surface adsorption when condition is suitable.
According to the performance analysis of the above-mentioned eggshell membrane collagen polypeptide products that the application's working method is made, explain that this product can reduce surface tension effectively.With tensio-active agent and other component excellent compatibility when mixing use.The eggshell membrane polypeptide is also very strong in the solid interface adsorptive power.After other surfactant mixed, system performance changed with its component flexibly, and these performances show through mixing with the model surface surfactant system.These unique function declaration eggshell membrane polypeptide can be combined into Different products with different surface active agents very neatly, at food, and personal care, pharmacy, biosensor and other biological technical elements are with a wide range of applications.
Therefore, the present invention is used as surfactant or polypeptide active agent to these collagen polypeptides, can be used for various products and production process, plays and reduces surface tension, wetting, cleaning, dispersion, emulsification, foaming, anti-bubble effect.For example, the collagen polypeptide tensio-active agent can be used for cleaning product and comprise sanitising agent, the fabric softener, and the agricultural-food protective material comprises weedicide, insecticide, the Personal hygiene nursing comprises shampoo, face cream, synthetic emulsion such as coating, tackiness agent, ink, medicine.
Because peculiar bio-compatibility, degradability, the eggshell membrane degradation product series product through aforesaid method production also can be used for cell cultures based raw material, artificial skin and skin treating and wash with raw material with solid support material, treatment and alleviation arthrodynia main raw material, healthy food material, cosmetic material (comprising the hair skin care), daily use chemicals etc. with subsidiary material, enzyme are fixing.Depend on different application processes and mechanism, one or several product in the eggshell membrane degradation product series polypeptide products can independently use or undertaken compoundly by diverse ways and component, and products obtained therefrom can be liquid, pulvis; Gel, foam, emulsion or butterfat type occur; Like face washing lotion, face cream, emulsion; Ointment, toothpaste, deodorant spray.
Production of the presently claimed invention and product comprise that all contain eggshell membrane degrading activity polypeptide or polypeptide active agent product; Wherein polypeptide active agent product content is usually between 0.01wt%～20wt%; Its content is between 0.1wt%～10wt% in most of product, or between 0.1wt%～5wt%.Often contain the content following material between 0.01wt%-15wt% usually in these production processes and the product (like face washing lotion, face cream, anti-skin aging frost, emulsion, ointment, toothpaste, deodorant spray):
1, VITAMINs comprises A, B, and C, D, E, K series and their growth, Vogan-Neu (vitamin A) compound comprises the C2-C22 retinol ester, Vitamin A Palmitate 1.7 M.I.U/Gram, vitamin A acetic acid, vitamin A propionate, vitamin A acid;
2, salicylic acid compound (ester class and salt); Comprise β-sitosterols, lupenol, Stigmasterol, plant sterol,
3, substituted and unsubstituted flavonoid compound comprises flavonoid, tonka bean camphor, chromanols (for example, daizeol (7,4 '-dihydroxy isoflavone)),
4, natural and substituted amino acid, like N-acyl group phenylalanine(Phe) (natural amino acid, acetylamino acid, methylamino acid, N-undecylenoyl-L-phenylalanine, N-tetradecyl Methionin),
5, the small peptide of design and bionical thing comprises dipeptides, tripeptides, tetrapeptide, pentapeptide, six peptides etc., like Val-Trp, and Asp-Phe; Ala-His, Arg-Lys-Arg, His-Gly-Gly, Gly-His-Lys; Gly-Gly-His, Gly-His-Gly, Lys-Phe-Lys; Gly-Gln-Pro-Arg, lipopeptides such as N-PalmitoylAla-His, N-Palmitoyl-GlyLys-His andN-Palmitoyl-Lys-Thr-Thr-Lys-Ser; N-Palmitoyl-Tyr-Gly-Gly-Phe-Xwith X denoting Met or Leu, N-Palmitoyl-Val-Gly-Val-Ala-Pro-Gly etc. (supplying with) by other company of Sederma and
6, the granulate material additive comprises coloured and leuco-pigment, organic and inorganic powder, and viscosity strengthens and fluorescent brightening particle and their combinations under different shapes and component, like glass; Zinc oxide, silicon oxide, silk, the microparticle material of porous or atresia; Other earth mineral are like aluminium sesquioxide and clay, siliceous copolymer; Polystyrene derivative, these particles perhaps have coating
7, sunscreen activeconstituents comprises organic and inorganic anti-UV-A and UV-B compound, like the para-amino benzoic acid verivate; The verivate of ethyl-PABA, cinnamon derivative, ethylhexyl Whitfield's ointment; The methoxydibenzoylmethane butyl ester; The ethylhexyl triazone, UVNUL MS-40-3 phenyl benzoglyoxaline
8, local anesthetic comprises Benzocaine, lignocaine, and bupivacaine, chlorprocaine, dibucaine, ketamine, PROCAINE HCL, PHARMA GRADE and Ding Ka,
9, the acne activeconstituents comprises Lucidol, UNISEPT DHA, and zinc, sulphur, Resorcinol, Oxacyclotetradecane,erythromycin deriv, Whitfield's ointment,
10, preserve moisture and amendment, comprise urea, guanidine, glycollate, alkylamine salt; Lactic acid, polyhydroxy-alcohol, like sorbyl alcohol, N.F,USP MANNITOL, Xylitol; Glycerine, Ucar 35, polyoxyethylene glycol, sugar, starch and verivate thereof; Mucinase, alkyl sugar and starch ester, the non-ionic type of synthesizing water-solubility and ionic polymer are like the polyoxyethylene glycol and the glycol polypropylene of modification
11, antiphlogistic drug comprises reflunomide, like HYDROCORTISONE INJECTIONS, and piroxicam, Frosst), Ibuprofen BP/EP and verivate, from this type of material of natural origin (plant, fungi, mikrobe) extraction,
12, tanned active substance comprises dihydroacetone (DHA or 1,3-dihydroxyl-2-acetone),
13, whitening agent comprises kojic acid, quinhydrones, p-aminophenol derivative (N-SUV oxygen carbonyl p-aminophenol etc.,
14, antibacterials comprise coal tar, sulphur, and aluminum chloride, trichlosan, TTO, clove leaf oil, thiobendazole, polyenoid, hydroxypyridone, Lucidol, the 3-hydroxy-benzoic acid, benzoglyoxaline,
15, thickening and jelling agent comprise carboxylic acid polyalcohol, cross-linked polyacrylate polymer, polyacrylamide polymers; Like the Tylose CH 50 polysaccharide, FM propionic acid carboxylic acid, hydroxypropylcellulose; Microcrystalline Cellulose, sodium cellulose sulfate and their mixture
16, tensio-active agent comprises negatively charged ion (dodecyl sulfate and verivate), positively charged ion (palmityl trimethyl ammonium chloride and verivate), and non-ionic type (Triton series) and amphoterics (natural or synthetic lipoid and verivate thereof),
17, positively charged ion, negatively charged ion, nonionic and amphiphilic polymers and multipolymer thereof,
18, hair and skin conditioning agent comprise any improvement hair, skin gloss, flexibility, and compatibility, health, antistatic property damages the oily material.These materials are water insoluble usually, but water dispersible, make and disperse or emulsion, comprise organosilicon (silicone oil, the positively charged ion organosilicon, the organosilicon gum, the high refractive index organosilicon, components such as hydrocarbon oils,
19, anti-dandruff activeconstituents comprises pyridinethione salt, azole, selenium chlorine and sulphur.
Use the eggshell membrane collagen polypeptide of the embodiment of the invention 1 to 4, make following various makeup respectively:
Embodiment 6 hair conditioner
Get the product D of 2wt% embodiment 4, mix power 3wt%Incroquat Behenyl TMC (Behentrionium chlorine, cetostearyl alcohol); 1% phenyl trimethiocone, the sanitas of 0.2wt%, 0.1% POTASSIUM SORBATE GRANULAR WHITE; 2wt%
(contains PPG-26-Butech-25, methyltrimethylene glycol, less water; The PEG-40 THIXCIN, apigenin, Oleanolic Acid and Biotinyl tripeptides-1); 0.1%; Perfume, 0.025wt%sarcolactic acid, water reaches 100wt%.Emulsion is wiping gently, can promote at the hair regeneration resting stage of corium.
Embodiment 7 anti-wrinkle creams and skin whitening
Get the product D of 2wt% embodiment 4, admixture 0.1wt%Ultrez 10 (carbomer), 3wt%transcutol; 8wt% glycerine, 0.1% POTASSIUM SORBATE GRANULAR WHITE, 0.6wt%Volpo S2 (Steareth 2); 4wt%Crodafos CES (cetostearyl alcohol, double hexadecyl phosphoric acid, phosphoric acid salt ceteth 10); 2wt%DC344 (cyclomethicone), 10wt%CRODAMOL GTCC (sad/last of the ten Heavenly stems triglyceride level), 1.6wt%CrU1 3 (sorbyl alcohol Triple Pressed Stearic Acid); 0.3wt% benzene mixed manthanoate, the NaOH of 0.3wt% (30%), 3000
3WT% (glycerine; Methyltrimethylene glycol, water, carbomer; Polysorbate 20, palmitoyl oligopeptide, palm tetrapeptide-3; 4wt%
(sad/last of the ten Heavenly stems triglyceride level, di-acetyl boldine), 0.1% delicate fragrance; Water reaches 100wt%.This peptide complex body anti-wrinkle cream is to be fit to the beautifying skin treatment, comprises crow's feet, wrinkle, and pigmentation spots stimulates the toxin expelling system.
Embodiment 8 body lotions
Get the product D of 2wt% embodiment 4, mix power 0.3wt% Hydrocerol A, 1.2wt% Hydrocerol A trisodique; 10wt%alcool, the methylbenzene of 0.2wt%, 1WT%Volpo G 26 (glycereth 26); Crillet 1 (polysorbate20), 0.1% perfume.This emulsion is moistened, and preserves moisture, and sensation and the youth outward appearance loosened are provided.
Embodiment 9 natural body washs
Prescription: 1 glass of (50 milliliters) water, 1 of W-Gum (5 milliliters), 2 soupspoon glycerine (20 milliliters); 1 teaspoons of sucrose; 2 soupspoon product D collagen polypeptide powder (2 gram), 5～10 natural flavor oil (lemon or orange), 1～2 redness or xanchromatic food dye (optional); Be placed in a bowl or the beaker, mix all the components.Pour into squeeze bottle into, in 48 hours, use.Need longer-term to use, add sanitas.
The foregoing description 6～9 uses natural egg polypeptide surfactant to replace conventional surfactant and multipolymer surface, in the hope of improving the bio-compatibility of product to skin.This example shows this advantage through the skin cells experiment.
(1) become human keratinocyte (HEKa,＞25population doublings, from Invitrogen) to implant 24 well culture plates; In the use-DMEM-LG of low dextrose cultivates medium (Dulbecco ' s modified Eagle ' s medium-low glucose) and augments 10% Ox blood serum (fetal bovine serum; FBS), 1wt% penicillium mould-Streptomycin sulphate (from Sigma), every porocyte number is approximately 5.0 * 104; Be placed in 37 ℃, wet air and 5%CO
2Under cultivated 3 days, remove old cultivation medium then.New cultivation medium branch adds and does not add FBS, inserts 0,0.5,1,2 and 5% polypeptide surfactant D respectively, SDS, C12TAB, C12E5, non-ionic polymers PluronicF-127.Use and be inverted phase microscope observation of cell growth population form, and with standard MTT laboratory examination cell viability.Every group comprises 3 repetitions.
Observations:
(i) add ionic surface active agent SDS and C12TAB and cause cell in 1 to several hours internal surface desorptions, death, ionic surface active agent concentration is high more, and this phenomenon appearance is fast more.As if by contrast, the nonionogenic tenside negative interaction is not obvious under lower concentration, but high density causes slow apoptosis.
(ii) add polypeptide surfactant and obviously promote cell enlargement, concentration is high more, increases fast more.2 with 5% polypeptide D under growth almost the same with 10%FBS.
(iii) under the situation that does not add polypeptide tensio-active agent or FBS, cell is grown up slowly or zero growth almost gradually.
(2) the adult skin inoblast (HDF,＞25 population doublings fromInvitrogen) are used for cell growth experiment above the repetition, observe and last basically identical.
(3) use polypeptide surfactant C to repeat top cell growth experiment, observe and last basically identical.
Above cell growth contrast experiment thereby obviously show: (1) polypeptide surfactant to the skin cells nontoxicity, have good bio-compatibility, (2) can promote the skin cells growth.By contrast, conventional surfactant has toxicity in various degree, can not be as effective skin cells nutrition.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (8)
1. the working method of an eggshell membrane collagen polypeptide may further comprise the steps:
1., from Ovum crusta Gallus domesticus, take out eggshell membrane, be crushed to more than 40 orders;
2., with step 1. the eggshell membrane powder to add concentration be in the papoid solution of 800～1000U/L, solid-to-liquid ratio is 1: 2～5, in 35～50 ℃ stirred 2～3 hours compound;
3., 2. add NaOH solution in the compound to step, to the mass concentration of NaOH be 2～4.5%, stirred 3～4 hours in 45～85 ℃, leave standstill; Get supernatant;
4., the supernatant pH value of using the Hydrogen chloride regulating step 3. to obtain is 7～11, must eggshell membrane collagen polypeptide liquid; Through desalination, concentrate and drying after, obtain the eggshell membrane collagen polypeptide.
2. working method as claimed in claim 1 is characterized in that: step 4. desalination is that the employing membrane pore size is the film of 1k with concentrating, and drying is 180 ℃ of following spraying dryings.
3. working method as claimed in claim 1 is characterized in that: step 4. desalination is that pure water wash-out eggshell membrane collagen polypeptide liquid to pH value is 7; Concentrate is that the employing membrane pore size is the membrane filtration of 300k; Drying is 180 ℃ of following spraying dryings of liquid concentrator or lyophilize behind the membrane filtration.
4. working method as claimed in claim 3 is characterized in that: it is the membrane sepn of 30k that the filtered solution that membrane filtration obtains adopts membrane pore size, and liquid and secondary filtered solution must dam; The said liquid that dams is in 180 ℃ of following spraying dryings of temperature; It is the 1k membrane-concentrated that said secondary filtered solution adopts the aperture, gets liquid concentrator in 180 ℃ of following spraying dryings of temperature.
5. eggshell membrane collagen polypeptide that makes like the arbitrary said working method of claim 1～4.
6. makeup include the eggshell membrane collagen polypeptide of 0.01wt%～20wt%.
7. makeup as claimed in claim 6 is characterized in that: eggshell membrane collagen polypeptide content is 0.1wt%～10wt%.
8. makeup as claimed in claim 7 is characterized in that: eggshell membrane collagen polypeptide content is 0.1wt%～5wt%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103631331A CN102517363A (en) | 2011-11-16 | 2011-11-16 | Production method and application of egg-shell membrane collagen peptides |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103631331A CN102517363A (en) | 2011-11-16 | 2011-11-16 | Production method and application of egg-shell membrane collagen peptides |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102517363A true CN102517363A (en) | 2012-06-27 |
Family
ID=46288428
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103631331A Pending CN102517363A (en) | 2011-11-16 | 2011-11-16 | Production method and application of egg-shell membrane collagen peptides |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102517363A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104292364A (en) * | 2014-09-30 | 2015-01-21 | 浙江大学 | Method for extracting bioactive substances from eggshell membrane |
| CN104436175A (en) * | 2013-09-17 | 2015-03-25 | 陈新宇 | Fowl-egg-inner-shell-membrane biological antibacterial preparation |
| CN104920782A (en) * | 2015-06-03 | 2015-09-23 | 四川大学 | polypeptide chelate calcium preparation and preparation method thereof |
| CN105296583A (en) * | 2014-07-11 | 2016-02-03 | 淮安鸿玛生物科技有限公司 | Application and production method of egg membrane protein polypeptide |
| CN106902787A (en) * | 2017-03-24 | 2017-06-30 | 孙志廷 | A kind of preparation method of the compound lithium battery Absorbent of egg shell waste material/activated carbon |
| US10932952B2 (en) | 2015-11-11 | 2021-03-02 | Biovotec As | Dry biocompatible disintegratable films for delivering particulate egg shell membrane to a wound |
| US11045578B2 (en) | 2015-06-24 | 2021-06-29 | Biovotec As | Tissue engineering scaffolds comprising particulate egg shell membrane |
| US11992508B2 (en) | 2014-10-28 | 2024-05-28 | Biovotec As | Micronized eggshell membrane particles and the use thereof to promote the healing of wounds |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805773A (en) * | 2010-03-25 | 2010-08-18 | 淮安鸿玛生物科技有限公司 | Method for producing egg membrane protein |
| CN101870995A (en) * | 2009-04-21 | 2010-10-27 | 陈栋梁 | Process for preparing collagen tripeptide |
| CN101921317A (en) * | 2010-09-26 | 2010-12-22 | 福州大学 | Anti-freeze polypeptide prepared by utilizing papain to carry out enzymolysis to pigskin collagen |
-
2011
- 2011-11-16 CN CN2011103631331A patent/CN102517363A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101870995A (en) * | 2009-04-21 | 2010-10-27 | 陈栋梁 | Process for preparing collagen tripeptide |
| CN101805773A (en) * | 2010-03-25 | 2010-08-18 | 淮安鸿玛生物科技有限公司 | Method for producing egg membrane protein |
| CN101921317A (en) * | 2010-09-26 | 2010-12-22 | 福州大学 | Anti-freeze polypeptide prepared by utilizing papain to carry out enzymolysis to pigskin collagen |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104436175A (en) * | 2013-09-17 | 2015-03-25 | 陈新宇 | Fowl-egg-inner-shell-membrane biological antibacterial preparation |
| CN105296583A (en) * | 2014-07-11 | 2016-02-03 | 淮安鸿玛生物科技有限公司 | Application and production method of egg membrane protein polypeptide |
| CN104292364A (en) * | 2014-09-30 | 2015-01-21 | 浙江大学 | Method for extracting bioactive substances from eggshell membrane |
| CN104292364B (en) * | 2014-09-30 | 2016-08-03 | 浙江大学 | A kind of method extracting bioactive substance from egg shell membrane |
| US11992508B2 (en) | 2014-10-28 | 2024-05-28 | Biovotec As | Micronized eggshell membrane particles and the use thereof to promote the healing of wounds |
| CN104920782A (en) * | 2015-06-03 | 2015-09-23 | 四川大学 | polypeptide chelate calcium preparation and preparation method thereof |
| CN104920782B (en) * | 2015-06-03 | 2018-04-13 | 四川大学 | A kind of polypeptide chelate calcium preparation and preparation method thereof |
| US11045578B2 (en) | 2015-06-24 | 2021-06-29 | Biovotec As | Tissue engineering scaffolds comprising particulate egg shell membrane |
| US10932952B2 (en) | 2015-11-11 | 2021-03-02 | Biovotec As | Dry biocompatible disintegratable films for delivering particulate egg shell membrane to a wound |
| CN106902787A (en) * | 2017-03-24 | 2017-06-30 | 孙志廷 | A kind of preparation method of the compound lithium battery Absorbent of egg shell waste material/activated carbon |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102517363A (en) | Production method and application of egg-shell membrane collagen peptides | |
| CN102112106B (en) | Cosmetic composition for anti-aging of the skin comprising phaseolus radiatus seed extracts by fermentation and enzyme treatment | |
| CN102178636B (en) | Marine biological function cosmetic for minimizing pores | |
| EP1433463B1 (en) | Use of algal proteins in cosmetics | |
| CN102015764A (en) | Cosmetic or pharmaceutical compositions comprising metalloproteinase inhibitors | |
| WO2008018448A1 (en) | Activator comprising biosurfactant as the active ingredient, mannosyl erythritol lipid and method of producing the same | |
| KR101685581B1 (en) | Compositions for improving skin wrinkle or moisture comprising Orostachys japonicus and composition prepared therefrom | |
| JP2001261570A (en) | Skin care preparation | |
| CN101953758A (en) | Series of skin care products with oat protein peptide as basic component and preparation method thereof | |
| JP4945692B2 (en) | Cell-utilized skin external preparation composition and method for producing the same | |
| JP2008110927A (en) | Cosmetic containing royal jelly protein hydrolyzate | |
| KR20160000747A (en) | Emulsifying agent comprising polysaccharide, cosmetic composition comprising the same, and method of producing thereof | |
| KR100837879B1 (en) | Cosmetic Composition Containing Collagen from Deer Extracts and Process for Preparation Thereof | |
| JP2012087091A (en) | Cosmetic | |
| CN103830119A (en) | Preparation method of toning lotion taking natural sericin as matrix | |
| CN110522709A (en) | Skin care compositions and methods and application thereof | |
| JP2015007031A (en) | Collagen production enhancer containing cellobiose lipid | |
| CN105296583A (en) | Application and production method of egg membrane protein polypeptide | |
| CN104473793A (en) | Moisturizing and lipid layer enhancing composition and application of same in face-cleansing product | |
| JP2002284632A (en) | Extinction substance for superoxide anion extracted from sake lees as effective component | |
| CN1138523C (en) | Association of phenol derivatives with extract of iridaceae family for lightening skin and superficial body growth and composition containing same | |
| JP2004026766A (en) | Skin care preparation for external use | |
| CN113543767A (en) | Skin care composition comprising non-hydrolyzed nucleic acid material extracted from tuna | |
| JP2015007032A (en) | Activating agent containing cellobiose lipid as active ingredient | |
| CN102093241A (en) | Method for preparing high-purity crystallization type N-acylglutamate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120627 |