CN1785160A - Skin-care product contg. tortoise skin collagen composition, and its prepn. method - Google Patents
Skin-care product contg. tortoise skin collagen composition, and its prepn. method Download PDFInfo
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Abstract
A skin-protecting protect for suppress lipid peroxide, delaying sanility and removing spots and wrinkles features that its additive contains turtle skin collagen which is prepared from turtle skin through extracting by enzymolyzing.
Description
Technical field
The present invention relates to a kind of skin care item, be specifically related to contain the skin care item of collagen compositions.
Background technology
Collagen is a kind of native protein, extensively is present in skin, bone, cartilage, tooth, tendon ligament and the blood vessel of animal, and be the epochmaking structural protein of connective tissue, play the function that supports organ, protection human body.Collagen generally is transparent, the branchiess fibril of white, is the substrate that is made of mucopolysaccharide and other protein around it.All contain hydroxyproline in oligopeptide that produces after the collagen decomposition and the aminoacid, this is rare in other protein,split.Thereby measure hydroxyproline in the urine or contain the content of the small peptide of hydroxyproline, can be used as the index of judging collagen decomposition in the body.Collagen has very strong biological activity and biological function, can participate in migration, differentiation and the propagation of cell, makes bone, tendon, cartilage and skin have certain mechanical strength.Collagen is because of antigenicity a little less than it and excellent biological compatibility, and is of many uses at pharmaceutical sanitary fields such as burn, wound, cornea disease, beauty treatment, orthopedic, hard tissue repair, wound surface hemostasis.Collagen protein is the main protein composition of skin histology, aboundresources, and mouthfeel is soft, taste is light, is easy to digestion, also receives the concern of field of food always, is used as functional mass and nutritional labeling in numerous food product.
With advancing age, the elasticity of skin reduces, metabolism slows down and the biologic garbage that produces is induced the generation of having quickened free radical down in many oxygen-derived free radicals inducible factors and factors such as diet inequality, food pollution and pressure index increase that occurring in natures such as the pollution of water quality, air, over-drastic exposure in sunshine exist.A large amount of free radicals is accumulated on internal organs of the body and the skin, be easy to destroy the decomposition and the synthetic metabolic balance of collagen, people's self-resistance, body repair in trauma are descended, collagen cross-linking is aging, can have influence on cardiovascular, lung tissue, kidney, liver etc. and exercise normal physiological function.The easy dehydration drying of while skin, colour of skin deepening, lackluster, wrinkle appears.
The applicant studies and tests discovery through for many years, collagen cross-linking, pigment produce also has a prior biochemical mechanism (Yin, D., Biochemical basis of lipofuscin, ceroid, and age pigment-like fluorophores.Free Radic.Biol.Med., 1996,21:871-888.), age pigment (mottle) fluorescent material is the end-product of multiple biochemical side reaction in the basic life process of human body, mainly contains relevant with fat peroxidating carbonyl and the crosslinked fluorescent material that forms of protein amino of passing through; And it is not aerobic and with the associated glycosylation of saccharide or Maillard reaction and fluorescence glycation product that forms etc.The formation of age pigment is dynamic change procedure continuously in the organism, the pale brown color age pigment material that each biochemical process produces or freely exist or be attached on the other biological molecule such as albumen.From in essence, biochemical side reaction is decreased and is become the biochemical essence (Yin that property in bad repair accumulation is the physiological aging course, D., Chen, K., Theessentiai mechanisms of aging:Irreparable damage accumulation of biochemicalside-reactions.Exp.Gerontol., 2005,40:455-465). be the broad sense causes of senescence that the applicant proposes.
The current research result of the relevant Colla Plastri Testudinis proteomics in this research department shows, the outstanding senile-resistant efficacy of the longevity of Testudinis and its bodily tissue is closely related.One of the important behaviour that it then is the Colla Plastri Testudinis anti-aging effects that the Testudinis collagen tissue does not take place with human skin and the similar aging crosslinked change of blood vessel in increasing rheological processes (Wu Yaosong, Liao Yanyang, print big-and-middle, application and the skin aging Mechanism Study progress of collagen protein in beauty treatment, the modern skin science magazine of China, 2004,1:219-224. and Xiang Dan, He Weiguang, print big-and-middle, Zhu Zerui, the SDS electrophoresis and the fluorescence analysis of soluble protein in hibernation Testudinis body fluid and the tissue, the Chinese science journal, 2004,1:47-53.).
Currently marketed skin care item are of a great variety, and many kinds are all having some effects aspect skin moisture-keeping, the wrinkle resistant and defying age, but moisture preserving time is short, wrinkle resistant and defying age DeGrain; The requirement of effect is more and more higher in this respect and people are to skin care item, and therefore developing long, the wrinkle resistant and more tangible skin care item of defying age effect of a kind of safety, moisture preserving time is pressing for of people.
Summary of the invention
The purpose of this invention is to provide a kind of skin care item that contain the turtle-skin collagen compositions, to solve the problems referred to above that current skin care item exist.
The technical scheme that realizes the object of the invention is: the skin care item that contain the turtle-skin collagen compositions, comprise base material of forming by oil phase raw material and water raw material and additive with certain effect, the present invention has added turtle-skin collagen in additive, the addition of turtle-skin collagen is 2～8% of skin care item by weight percentage.The preparation method that contains the skin care item of turtle-skin collagen compositions comprises the dissolving of oil phase raw material, the adding of water raw material, heating and sterilization, and emulsifying and dilution and the adding that contains the additive of turtle-skin collagen, the preparation method of turtle-skin collagen follows these steps to carry out successively:
1): pre-treatment: Testudinis skin priority water and deionized water are cleaned, the Testudinis skin of cleaning at room temperature soaked 1～5 day with 0.5%～10% dilute alkaline soln, and the consumption of dilute alkaline soln is 1～10 (liter/kilogram Testudinis skin), and the reuse deionized water washes the Testudinis skin of alkali treatment repeatedly, up to pH 6.5～7.0
2): the extraction of Colla Plastri Testudinis: the acetic acid or the neutral deionized water that above-mentioned clean Testudinis skin are added pH 2.5～5.0, addition gets final product to cover the Testudinis skin, and interpolation protease, the weight ratio of protease and Testudinis skin is 1: 10～1: 100, placed 4 ℃ of enzymolysis 2～3 days, and filtered, extract and repeat 3 times, merge extractive liquid,
3): the purification of Colla Plastri Testudinis: above-mentioned pH 2.5～5.0 or neutral filtrate is centrifugal with 3000～10000r/min, and the extracting centrifugal liquid that removes slag is put into bag filter to centrifugal back clear liquid and is dialysed with deionized water, changes water once every 3～4 hours, up to 10% AgNO
3The solution check does not have precipitation and generates, and this moment, pH value was about 6.5～7.0.
4) enzymolysis of Colla Plastri Testudinis: collagen protein is poured out from bag filter, regulate the collagen viscosity with suitable quantity of water, ultrasonic Treatment 10min with 20000 hertz, at pH=5, add 1.5% pepsin and papain hydrolysis 12h in water bath with thermostatic control under 37 ℃ of conditions of temperature, 5min makes enzyme deactivation under 60 ℃ of temperature then, and freezing back is in 0～15 ℃ of preservation, and is stand-by.
Above-mentioned protease comprises at least a in pepsin, papain, the trypsin etc.
Be described in further detail the present invention below in conjunction with accompanying drawing
Description of drawings
Fig. 1 is the electrophoresis contrast figure of Colla Plastri Testudinis and Corii Bovis seu Bubali collagen, pigskin collagen, standard collagen;
Number in the figure: 1-sample one is a bovine collagen albumen; 2-sample two is pigskin collagen albumen samples;
3-sample three is standard I type collagen protein; 4-sample four is collagen protein of Testudinis skin;
5-sample five is collagen protein of Testudinis skin
Fig. 2 is the moisture survival rate curve of 2 kinds of collagen polypeptide;
Line style among the figure :-◆--water
-propylene glycol
Fig. 3 is a hydroxyproline ultraviolet spectrometry absorption value change list.
Tortoise is long-lived animal, and tortoise mainly is to use tortoiseshell as Chinese medicine, and the tortoise whole body all is precious, along with sending out of aquaculture Exhibition and the taking of Chinese medicine tortoiseshell also can a large amount of tortoise skin raw materials of by-product, are for the fine raw material of making collagen.
Except adding aloe extract, PCA and vitamin A, also added 2～8% in the skin care item of the present invention Turtle-skin collagen, turtle-skin collagen is organized in to increase and does not take place in the rheological processes and the similarly aging crosslinked change of human skin and blood vessel, Therefore have anti-aging effects, also have long moistening effect. The compatible aloe of adding and turtle-skin collagen is carried simultaneously Get thing, PCA and vitamin A can strengthen skin care item whitening, remove free radical and moist effect.
Turtle-skin Collagen and to contain the preparation method of skin care item of Turtle-skin Collagen composition as follows:
1, pre-treatment
Tortoise skin successively water and deionized water are cleaned, and the tortoise skin of cleaning at room temperature soaks with 0.5%～10% dilute alkaline soln Processed 1～5 day, the consumption of dilute alkaline soln is 1～10 (rising/kilogram tortoise skin), repeatedly washes alkali treatment with deionized water again The tortoise skin, until pH 6.5～7.0.
2, the extraction of tortoise glue
General method for extracting protein is divided into alkaline process, acid system and enzyme process. Acid-hydrolysis method is rapid and thorough, but tryptophan is complete Section is destroyed, and serine and tyrosine part are destroyed, and the product yield is low. Alkali hydrolysis method is rapid and thorough, but contains hydroxyl The amino acid of base and sulfydryl, all destroyed and generation racemization (structure variation). For obtaining better more completely activity Albumen adopts enzymatic isolation method. Above-mentioned clean tortoise skin is added acetic acid or the neutral deionized water of pH 2.5～5.0, add Amount gets final product to cover the tortoise skin, and adds protease, and the weight ratio of protease and tortoise skin is 1: 10～1: 100, at 4 ℃ of enzymes Separated 2～3 days, and filtered, the extraction of tortoise glue repeats 3 times, merges extract.
3, the purifying of tortoise glue
With above-mentioned pH 2.5～5.0 or neutral filtrate, centrifugal with 3000～10000r/min centrifuge, extracting centrifugal liquid removes slag, centrifugal rear clear liquid is put into the bag filter dialysis, with deionized water dialysis, changed water once every 3～4 hours, until with 10% AgNO3The solution check does not have precipitation and generates, and this moment, the pH value was about 6.5～7.0.
4, the enzymolysis of Colla Plastri Testudinis
Collagen protein is poured out from bag filter, regulate the collagen viscosity with suitable quantity of water, ultrasonic Treatment 10min with 20000 hertz, at pH=5, add 1.5% pepsin and papain hydrolysis 12h in water bath with thermostatic control under 37 ℃ of conditions of temperature, 5min makes enzyme deactivation under 60 ℃ of temperature then, and freezing back is in 0～15 ℃ of preservation, and is stand-by.
But the Testudinis micromicro that the present invention adopts is to be fresh also stored frozen, earlier with tap water flushing, reuse deionized water rinsing.Used Testudinis skin has the skin of Chinemys such as Chelomia mydas (Linnaeus)., amphibian Testudinis, Testudo elongata, non-irrigated Testudinis.
Above-mentioned protease comprises in pepsin, papain, trypsin, the microbial enzyme at least a.
The purity of the Colla Plastri Testudinis of purification and Corii Bovis seu Bubali collagen, pigskin collagen and standard collagen is higher as can be seen from Figure 1, and molecular size, subunit constitute similar.
5, the composition skin-care product that contain turtle-skin collagen
As used typical additive or excipient in the cosmetics of the present invention, comprise oil phase raw material and water raw material, wherein the oil phase raw material has vegetable and animals oils, mineral oil, fatty oil, wax, higher alcohol, aliphatic alcohol, silicone oil, phospholipid, fatty alcohol ether, polyol resin etc., the water raw material has water-soluble polymer, clay mineral and purify waste water triethanolamine etc.Also can add pH regulator agent, surfactant, antioxidant, chelating agen, antiseptic, antifungus agent, pigment thing and spice etc. as required.In addition, also can with have other active component that brighten beautification function or skin cosmetic effect, as blood circulation accelerant, disinfectant, antiinflammatory, cell activator, vitamins, amino acids, wetting agent and keratolytic agent etc. is used in combination.
Product of the present invention has carried out zoopery, behind this product coating animal subject rat, and hydroxyproline content in the irritant reaction of observation rat skin, the skin.Experimental result is not all had red and swollen unusual the generation for trying rat skin, and the hydroxyproline content (see figure 3) experimental technique that is significantly improved in this product rat skin: 6 male and female half and half of rat cut off back Mus gross area 5cm
2, be applied to no fur skin secondary every day with 0.1 this product of gram, continuous 3 months of each 0.1 gram, observing has vacuum response.Bark fetching carries out the hydroxyproline content analysis with chlorine ammonia T method simultaneously.Laggard pedestrian's body tuerculoderma does not yet find to have any stimulation phenomenon.
Water conservation experiment: accurately take by weighing the 1.0g collagen protein polypeptide, be dissolved in a certain amount of distilled water, and be settled to 100ml, measure 1ml sample liquid then, it is tiled in the culture dish that diameter is 10cm, in the incubator as for constant temperature and humidity, surveys the moisture survival rate of a sample liquid every 10min.Do contrast with distilled water simultaneously.The result shows that the moisture survival rate of turtle-skin collagen polypeptide is apparently higher than pig glue polypeptide (see figure 2).
Cosmetics of the present invention can be mixed with cream, oil preparation, Emulsion, unguentum, moisturizing emulsion, facial film, milky lotion etc. on demand.
The skin care item of the present invention's development meet cosmetics health chemical standard and microbial standard after testing; Through animal experiment, to the irritated reaction of the non-stimulated nothing of rat skin.Product hydroxyproline content in rat skin is significantly improved, water-retaining property is also obviously than similar height, therefore the skin care item tool of the present invention's development obviously suppresses the effect of lipid peroxide and contains the anti-crosslinked aged factor of skin, thereby has speckle dispelling, wrinkle resistant, moisture-keeping functions; The Colla Plastri Testudinis purity height of the present invention's preparation, cost is low, has the prospect of Application and Development.
The specific embodiment
Two Application Examples of these skin care item invention.
Embodiment 3: the preparation of Brazilian turtle-skin collagen
1: the tap water of 15 ℃ of 1 kilogram of usefulness of Brazilian Testudinis skin of stored frozen is rinsed well, removed scale and subcutaneous tissue, the reuse deionized water rinsing is clean.
2: Na with 5%
2CO
3Testudinis skin after 10 liters of soaking flushing of solution are clean is about 5 days, and water fully cleans the Testudinis skin of alkali treatment again, up to being neutral.
3: add 5 liters of the acetic acid of pH 2.5 at the Testudinis flesh side of above-mentioned processing, and add 10 gram pepsin, in 4 ℃ of following enzymolysis 2 days, use filtered through gauze, residue is enzymolysis once more, handle the same, merging filtrate.
4: centrifugal with the centrifuge of 5000r/min, remove slag and get clear liquid, centrifugal back clear liquid is put into bag filter, dialyse with deionized water, changed one time water in per 3～4 hours, up to 10% AgNO
3The solution check does not have precipitation and generates, and this moment, pH value was about 6.5～7.0.
5: the enzymolysis of Colla Plastri Testudinis: collagen protein is poured out from bag filter, regulate the collagen viscosity with suitable quantity of water, ultrasonic Treatment 10min with 20000 hertz, at pH=5, add 1.5% pepsin and papain hydrolysis 12h in water bath with thermostatic control under 37 ℃ of conditions of temperature, 5min makes enzyme deactivation under 60 ℃ of temperature then, and freezing back is in 0～15 ℃ of preservation, and is stand-by.
Embodiment 4: the preparation of Testudinis collagen
1: fresh Testudinis skin 500 grams are rinsed well with 20 ℃ tap water, removed subcutaneous tissue, the reuse deionized water rinsing is clean.
2: the Testudinis skin after 1 liter of soaking flushing of the NaOH solution with 0.5% is clean is about 1 day, and water fully cleans the Testudinis skin of alkali treatment again, up to being neutral.
3: add 2.5 liters of neutral waters at the Testudinis flesh side of above-mentioned processing, and add 50 gram papains, in 4 ℃ of following enzymolysis 2 days, use filtered through gauze, residue is enzymolysis once more, handle the same, merging filtrate.
4: centrifugal with the centrifuge of 10000r/min, remove slag and get clear liquid, centrifugal back clear liquid is put into bag filter, dialyse with deionized water, changed one time water in per 3～4 hours, up to 10% AgNO
3The solution check does not have precipitation and generates, and this moment, pH value was about 6.5～7.0.
5: the enzymolysis of Colla Plastri Testudinis: collagen protein is poured out from bag filter, regulate the collagen viscosity with suitable quantity of water, ultrasonic Treatment 10min with 20000 hertz, at pH=5, add 1.5% pepsin and papain hydrolysis 12h in water bath with thermostatic control under 37 ℃ of conditions of temperature, 5min makes enzyme deactivation under 60 ℃ of temperature then, and freezing back is in 0～15 ℃ of preservation, and is stand-by.
Embodiment 5: the preparation of Colla Plastri Testudinis cosmetics
Oil phase: oil phase raw material in embodiment 1 or 2 joined melt in the pot, heat, under constantly stirring, be heated to 70～75 ℃, it is fully dissolved.Avoid superheated and long-time heating to prevent the material composition oxidation deterioration.Oil content, antiseptic and the emulsifying agent etc. of oxidation can add oil phase before emulsifying easily, and dissolving evenly can be carried out emulsifying.
Water: 30%～50% deionized water is joined melt in the pot earlier, add water raw material in embodiment 1 or 2 simultaneously, be heated to 90～100 ℃ under stirring, keep sterilization in 20 minutes, stand-by.As containing water-soluble polymer in the prescription, should prepare separately, it is dissolved in the water, at room temperature fully stir and make its even swelling, prevent conglomeration, also can carry out homogenizing if necessary, before emulsifying, add water.Avoid long-time heating, change in order to avoid cause viscosity.The moisture that vapors away during for supplementary heating and emulsifying can be added 3%～5% water by prescription.
With above-mentioned water is joined in the oil phase earlier, carry out emulsifying and stir, the beginning emulsified body is a w/o type, and along with the adding of water, modification becomes O/W type emulsified body, is called condensed emulsified body.
The deionized water through ultraviolet disinfection dilutes without heating to add residue 50%～70% again, because the foreign minister of condensed emulsified body is a water, so the dilution of emulsified body can be finished smoothly, in this process, the temperature of emulsified body descends very fast.After deionized water added, the temperature of emulsified body can drop to 50～60 ℃.
Add antiseptic, essence this moment, slowly stirs when being cooled to 40 ℃ of left and right sides, adds additives such as turtle-skin collagen albumen, vitamin, stirs to be cooled to 16 ℃ and to stop, bottled, and carry out performance rating.
Claims (3)
1, a kind of skin care item that contain the turtle-skin collagen compositions, comprise base material of forming by oil phase raw material and water raw material and additive with certain effect, the invention is characterized in to have added turtle-skin collagen in the additive that the addition of turtle-skin collagen is 2～8% of skin care item by weight percentage.
2, the preparation method that contains the skin care item of turtle-skin collagen compositions as claimed in claim 1, comprise the dissolving of oil phase raw material, the adding of water raw material, heating and sterilization, emulsifying and dilution and the adding that contains the additive of turtle-skin collagen compositions, the invention is characterized in the preparation method of turtle-skin collagen, follow these steps to successively carry out:
1), pre-treatment: Testudinis skin priority water and deionized water are cleaned, the Testudinis skin of cleaning at room temperature soaked 1～5 day with 0.5%～10% dilute alkaline soln, the consumption of dilute alkaline soln is 1～10 (liter/kilogram Testudinis skin), and the reuse deionized water washes the Testudinis skin of alkali treatment repeatedly, up to pH6.5～7.0;
2), the extraction of Colla Plastri Testudinis: the acetic acid or the neutral deionized water that above-mentioned clean Testudinis skin are added PH2.5～5.0, addition gets final product to cover the Testudinis skin, and interpolation protease, the weight ratio of protease and Testudinis skin is 1: 10～1: 100, placed 4 ℃ of enzymolysis 2～3 days, filter, the extraction of Colla Plastri Testudinis repeats 3 times, merge extractive liquid;
3), the purification of Colla Plastri Testudinis: above-mentioned PH2.5～5.0 or neutral filtrate is centrifugal with 3000～10000r/min, and the extracting centrifugal liquid that removes slag is put into bag filter to centrifugal back clear liquid and is dialysed with deionized water, changes water once every 3～4 hours, up to 10% AgNO
3The solution check does not have precipitation and generates, and this moment, pH value was about 6.5～7.0;
4), the enzymolysis of Colla Plastri Testudinis: collagen protein is poured out from bag filter, regulate the collagen viscosity with suitable quantity of water, ultrasonic Treatment 10min with 20000 hertz, at pH=5, add 1.5% pepsin and papain hydrolysis 12h in water bath with thermostatic control under 37 ℃ of conditions of temperature, 5min makes enzyme deactivation under 60 ℃ of temperature then, and freezing back is in 0～15 ℃ of preservation, and is stand-by.
3, the preparation method of a kind of Colla Plastri Testudinis as claimed in claim 2 is characterized in that described protease comprises at least a in pepsin, papain, the trypsin etc.
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| CN108531536A (en) * | 2018-05-04 | 2018-09-14 | 浙江万里学院 | The preparation method and applications of Shelled Turtle Trionyx Sinensis collagen peptide |
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| US6096324A (en) * | 1995-06-13 | 2000-08-01 | Laboratory Skin Care | Methods of delivering materials into the skin, and compositions used therein |
| CN1183271A (en) * | 1996-11-27 | 1998-06-03 | 北京市法码医药咨询有限公司 | Collagen E milk and preparing method thereof |
| JP2003520217A (en) * | 2000-01-19 | 2003-07-02 | ザ、プロクター、エンド、ギャンブル、カンパニー | Skin care composition |
| US20030147830A1 (en) * | 2002-01-30 | 2003-08-07 | The Procter & Gamble Company | Topical skin and/or hair compositions containing protein |
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| CN108531536A (en) * | 2018-05-04 | 2018-09-14 | 浙江万里学院 | The preparation method and applications of Shelled Turtle Trionyx Sinensis collagen peptide |
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