CN114886830A - Scalp/hair/skin repairing composition using seawater pearl as raw material, preparation method and application thereof - Google Patents
Scalp/hair/skin repairing composition using seawater pearl as raw material, preparation method and application thereof Download PDFInfo
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- CN114886830A CN114886830A CN202210235950.7A CN202210235950A CN114886830A CN 114886830 A CN114886830 A CN 114886830A CN 202210235950 A CN202210235950 A CN 202210235950A CN 114886830 A CN114886830 A CN 114886830A
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- seawater
- pearl
- pearl powder
- preparing
- scalp
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Abstract
The invention discloses a scalp/hair/skin repairing composition using seawater pearl as raw material, a preparation method and application thereof, comprising the steps of preparing pearl powder, preparing seawater pearl extract, preparing hydrolyzed seawater pearl, preparing hydrolyzed fibroin, and mixing the pearl powder: seawater pearl extract: hydrolyzing seawater pearls: the hydrolyzed fibroin is mixed according to the mass ratio of (0.01-2) to (8-12) to (4-6). According to the invention, the pearl powder, the seawater pearl extract, the hydrolyzed seawater pearl and the hydrolyzed fibroin are compounded, so that various functional components are compounded and complemented, the hair quality repairing function is strong, the scalp sebaceous gland activity is regulated, the 5 alpha-reductase is inhibited, the obvious antioxidant and anti-inflammatory effects are achieved, and meanwhile, the hydrolyzed seawater pearl has the effects of moisturizing and repairing skin barriers.
Description
Technical Field
The invention belongs to the technical field of cosmetic compositions, and particularly relates to a scalp/hair/skin repairing composition taking seawater pearls as raw materials, a preparation method and application thereof.
Background
The pearl medicine has more than 2000 years of history in China, the Ming dynasty Li Shizhen pays more attention to the pharmacological action of the pearl, and the efficacy of the pearl is considered to be skin-beautifying, so that the pearl medicine is particularly written in Ben Cao gang mu (compendium of materia Medica): the pearl flavor is sweet, cold and nontoxic, and can be used for calming heart and improving eyesight: the pearl is used for coating the surface, so that people can feel moist and have good color. Smearing on hands and feet, removing skin and adverse rising of skin: phlegm reduction, face speckle removal, diarrhea relief, infantile convulsion removal, soul calming: stopping nocturnal emission, removing acne and treating toxic materials. It also describes various methods for pearl medicine. The Ming Dynasty Chen Ru Chang Shen (Duyianzhi), in the case of Tangwuzong plums inflammation in place, the Lideyu takes the jewelry powder, realgar and cinnabar decoction as a thick soup, each cup consumes about thirty thousand money, and the residue is discarded after three times of decoction. When the pill refining technology was prevailed, people thought that pearl powder, realgar and other substances can grow and be old after being refined and taken, and the hairiness and youth can be raised.
The seawater pearl powder contains abundant fatty alcohol, and is easy to be absorbed by human epidermal cells, so as to enhance cell activity and promote cell metabolism. The pearl powder can promote the growth of collagen cells of a human body, can promote the growth of the collagen cells to fill gaps and connect tissues when traumas and wounds occur, can promote the regeneration of skin, keep the skin tender, moist and white, is smooth and complete, has certain absorption capacity for body fluid secretion when seawater pearl necklaces are tightly attached to the neck of a person, can absorb the medicinal components of the pearls, even influences local affected parts, and has the anti-inflammatory treatment effect.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
The invention provides a preparation method of a scalp/hair/skin repairing composition taking seawater pearls as raw materials, which comprises the following steps:
preparing pearl powder: putting seawater pearls into a superfine pulverizer to pulverize into coarse powder with the particle size of 10-15 microns, then performing air flow pulverization to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight of 15-25% of the powder, pressurizing, instantly reducing pressure, taking out, mixing the obtained pearl powder and water into pearl slurry, performing wet grinding to obtain pearl powder, filtering and drying to obtain bright white pearl powder;
preparing a seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 3-5%, soaking for 1-3 h, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a seawater pearl extract;
preparing hydrolyzed seawater pearls: preparing 0.1-0.4 mg/mL streptothricin protease solution with the pH value of 7.0-7.5, adding the pearl powder, wherein the streptothricin protease accounts for 0.1-0.2% of the mass of the pearl powder, performing enzymolysis at 40-45 ℃ for 1-3 h, heating to inactivate enzyme, adjusting the pH value to 5.5-6.5, adding complex enzyme accounting for 0.1-0.2% of the mass of the pearl powder, wherein the complex enzyme is debitterized protease and bromelain according to the ratio of 1-2: 1, performing enzymolysis at 50-55 ℃ for 2-3 hours, heating to inactivate enzyme, performing reduced pressure evaporation concentration, and freeze-drying to obtain hydrolyzed seawater pearls;
preparing hydrolyzed fibroin:
mixing pearl powder: seawater pearl extract: hydrolyzing seawater pearls: the hydrolyzed fibroin is mixed according to the mass ratio of (0.01-2) to (8-12) to (4-6).
The preferable scheme of the preparation method of the scalp/hair/skin repairing composition using seawater pearls as raw materials comprises the following steps: the pearl powder preparation method comprises the steps of putting seawater pearls into an ultrafine grinder, grinding the seawater pearls into coarse powder with the particle size of 10-15 microns, carrying out air flow grinding on the coarse powder to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight being 20% of the powder, pressurizing to 0.5MP, maintaining for 4-5 hours, instantly reducing pressure, taking out, and mixing the obtained pearl powder with the water according to the mass ratio of 1: 4-5, carrying out wet grinding to obtain pearl powder with the particle size of 10-100nm, filtering and drying to obtain bright white pearl powder.
The preferable scheme of the preparation method of the scalp/hair/skin repairing composition using seawater pearls as raw materials comprises the following steps: the preparation method of the seawater pearl extract comprises the steps of adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2-3 hours, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain the bright white seawater pearl extract.
The preferable scheme of the preparation method of the scalp/hair/skin repairing composition using seawater pearls as raw materials comprises the following steps: the seawater pearl extract is prepared by preparing 0.2mg/mL streptothricin protease solution from streptothricin protease with phosphate buffer solution with the pH value of 7.5, adding pearl powder, wherein the streptothricin protease accounts for 0.16 percent of the mass of the pearl powder, performing enzymolysis for 2-3 h at the temperature of 40 ℃, heating to inactivate enzyme, adjusting the pH value to 6, adding complex enzyme accounting for 0.2 percent of the mass of the pearl powder, and performing reaction on debitterizing protease and bromelain according to the ratio of 2: 1, carrying out enzymolysis at 55 ℃ for 3-4 h, heating to inactivate enzyme, carrying out reduced pressure evaporation concentration, and carrying out freeze drying to obtain the hydrolyzed seawater pearl.
The preferable scheme of the preparation method of the scalp/hair/skin repairing composition using seawater pearls as raw materials comprises the following steps: the preparation method of the hydrolyzed silk protein comprises the steps of putting silk into a sodium carbonate aqueous solution with the mass fraction of 1-2% to boil for 30-40 min, removing sericin, filtering and drying, and mixing the silk after the sericin is removed with 0.1mol/L phosphoric acid aqueous solution with the pH value of 6.5 according to the g: ml of 1: 20-1: 40, mixing, adding 800U/g neutral protease, hydrolyzing at 45 deg.C for 4h, filtering to adjust pH to 7 to obtain hydrolyzed fibroin, concentrating under reduced pressure, freeze drying, and hydrolyzing fibroin.
The preferable scheme of the preparation method of the scalp/hair/skin repairing composition using seawater pearls as raw materials comprises the following steps: mixing pearl powder: seawater pearl extract: hydrolyzing seawater pearls: the hydrolyzed fibroin is mixed according to the mass ratio of 0.02-1: 10:10: 5.
As another aspect of the invention, the invention also provides a scalp/hair/skin repairing composition which is prepared by the method and takes seawater pearls as raw materials.
As another aspect of the present invention, the present invention also provides the use of the scalp/hair/skin care composition using seawater pearls as raw materials in cosmetics: the composition is used for preparing cosmetics comprising one or more of shampoo, hair conditioner, scalp care solution, emulsion, cream, shower gel, facial mask and face cleaning product, has strong damaged hair repair function, can regulate the activity of scalp sebaceous glands and inhibit the effect of 5 alpha-reductase, and has obvious antioxidant and anti-inflammatory effects, and the effects of moisturizing and repairing skin barrier.
The invention has the beneficial effects that: according to the invention, the pearl powder, the seawater pearl extract, the hydrolyzed seawater pearl and the hydrolyzed fibroin are compounded, so that various functional components are compounded and complemented, the hair quality repairing function is strong, the scalp sebaceous gland activity is regulated, the 5 alpha-reductase is inhibited, the obvious antioxidant and anti-inflammatory effects are achieved, and meanwhile, the hydrolyzed seawater pearl has the effects of moisturizing and repairing skin barriers.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein:
FIG. 1 is a schematic representation of the change in moisture content of the scalp before and after use of the composition of example 1.
FIG. 2 is a schematic representation of the change in the percutaneous water loss of the scalp before and after use of example 1 (g/m) 2 h)。
FIG. 3 is a graph of scalp sensitivity before and after application of the composition of example 1 to a portion of volunteers.
FIG. 4 is a skin moisturizing efficacy test of the products of the examples
Fig. 5 is a scalp 5 α -reductase inhibition test for the products of each example.
FIG. 6 is a graph showing the result of DPPH clearance test.
FIG. 7 is a hair tensile strength test after using the products of each example 1.
FIG. 8 is a graph showing the expression level of IL-6.
Fig. 9 is a schematic view of the change in percutaneous water loss of comparative example 7.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1:
preparation of component A: pearl powder: selecting pollution-free seawater pearls, removing dead pearls and impurities, cleaning the pearls with clear water, filtering, drying at low temperature, putting the pearls into an ultrafine pulverizer to pulverize into coarse powder with the particle size of 10-15 microns, then performing air flow pulverization to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight of 20% of the powder, pressurizing to 0.5MP, maintaining for 4 hours, instantly reducing the pressure, taking out, mixing the obtained pearl powder with water according to the ratio of 1: 4(m/m), grinding by a wet method to obtain pearl powder with the particle size of 10-100nm, filtering and drying to obtain a component A: bright white pearl powder;
preparation of component B: seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2 hours, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a component B: bright white seawater pearl extract;
preparation of component C: hydrolyzing seawater pearls: preparing 0.2mg/mL of streptothricin solution by using phosphate buffer solution with pH of 7.5, adding pearl powder, wherein the streptothricin accounts for 0.16 percent of the mass of the pearl powder, performing enzymolysis for 2 hours at 40 ℃, heating to inactivate enzyme, adjusting the pH to 6, adding complex enzyme accounting for 0.2 percent of the mass of the pearl powder, and the complex enzyme is debitterizing protease and bromelain according to the ratio of 2: 1, carrying out enzymolysis at 55 ℃ for 3 hours, heating to inactivate enzyme, carrying out reduced pressure evaporation concentration, and carrying out freeze drying to obtain a component C: hydrolyzing seawater pearls;
preparation of component D: hydrolyzing the fibroin: placing silk into sodium carbonate aqueous solution with the mass fraction of 1% to boil for 30min, removing sericin, filtering and drying, and mixing the silk without sericin with 0.1mol/L phosphoric acid aqueous solution with the pH value of 6.5 according to the ratio of 1: 20-1: 40(g: ml), adding 800U/g neutral protease, hydrolyzing at 45 ℃ for 4h, filtering to adjust pH to 7 to obtain hydrolyzed fibroin, concentrating by evaporation under reduced pressure, and freeze drying to obtain component D: and (3) hydrolyzing the fibroin.
Mixing a component A, B, C, D according to a mass ratio of 1:10:10:5 mixing uniformly to obtain the scalp/hair/skin repairing composition. The scalp/hair/skin rejuvenation composition was formulated into a solution according to table 1.
TABLE 1 composition formula
| Starting materials | Mass fraction (wt%) |
| Deionized water | To 100 |
| Component A | 1 |
| |
10 |
| |
10 |
| |
5 |
| Carbomer 940 | 0.3 |
The test method comprises the following steps:
DPPH free radical scavenging experiments: preparing a DPPH solution: dissolving 1mg DPPH with small amount of anhydrous ethanol, diluting with distilled water to constant volume of 1L, storing in dark place, adding a certain amount of substance to be detected into 2.5mL of 50 μ g/mL
After standing in a mL of DPPH absolute ethanol solution at room temperature for 30min, the absorbance of the solution was measured at a wavelength of 515 nm. The scavenging effect was calculated from the following formula:
S(％)＝[A0-(A-A1)]×100％/A0
in the formula: s is DPPH radical scavenging ratio, A 0 Absorbance of absolute ethanol solution of DPPH at wavelength of 515 nm; a is the absorbance of the mixed solution after being clear at the wavelength of 515nm for 30 min; a. the 1 The absorbance of the sample itself at a wavelength of 515 nm.
5 α -reductase inhibition assay: 5 α -reductase is a membrane protein localized on the microsomes of target cells and plays an important role in the development of androgenic alopecia. After binding of androgens to receptors, complex enzymatic reactions occur that indirectly alter the hair growth process. 5 alpha-reductase has 3 isozymes, i.e., type I, II, III. The type I5 alpha-reductase is present in skin tissues such as skin sebaceous glands, sweat glands, hair papilla cells, fibroblasts, epidermal keratinocytes, hair follicle keratinocytes and the like. While type II 5 alpha-reductases are mainly present in tissues such as prostate, reproductive skin, epididymis, seminal vesicle, etc. The conversion of testosterone to Dihydrotestosterone (DHT) is dependent on reduced coenzyme II, high levels of dihydrotestosterone are likely to cause androgen-dependent diseases, and the down-regulation of androgen levels by inhibiting 5 alpha-reductase activity is an important approach for the treatment of androgenetic alopecia. The 5 alpha-reductase inhibitors have the advantage of selectively blocking dihydrotestosterone synthesis without affecting the normal level and physiological function of testosterone. Currently studied 5 α -reductase inhibitors can be classified into components derived from natural plants and chemically synthesized steroids, pyridines, quinolinones, and phenoxybutyric acid compounds. Can screen high-efficiency inhibitor by taking the activity of 5 alpha-reductase as a target.
Preparation of 5 α -reductase: the SPF level mouse/rat is killed after fasting overnight without water, the liver and epididymis (fat tissue peeling) are taken out quickly, weighed, rinsed by normal saline, sheared, and added with 5 times precooled homogenate [0.32mol/l sucrose, 1mmol/l DTT, 1mmol/l EDTA, 20mmol/l PBS (liver pH 7.5; epididymis pH 5.5 ]) to be homogenized in a glass homogenizer, the homogenate is centrifuged at 4 ℃ and 3000g for 10min, the supernatant is centrifuged at 4 ℃ and 10000g for 60min, the supernatant is taken as crude enzyme, subpackaged, and stored in an ultra-low temperature refrigerator at-80 ℃ and taken out before use, and repeated freeze thawing is avoided.
Protein content is determined by adopting a Lowry method protein content detection kit or quantitative determination is carried out by adopting a Coomassie brilliant blue experiment, the concentration of 5 alpha-reductase is expressed by the total protein content in crude enzyme, and the operation is carried out according to the kit instruction.
Preparation of Coomassie brilliant blue G250 solution: 100mg of Coomassie brilliant blue G250 was weighed out accurately and 50ml of 95% ethanol solution was added. The solution is blue, is stirred and dissolved, then is added with 100ml of 85% (W/V) phosphoric acid and is stirred, the solution is bloodred, finally deionized water is used for fixing the volume to 1000ml, the solution is changed into brown, the solution is stirred on a magnetic heating stirrer overnight, and is filtered by filter paper for standby. BSA protein standards were serially diluted with PBS to 0.0, 2.5, 5.0, 10.0, 20.0, 40.0, 50.0. mu.g/ml. Sequentially adding 50 μ l BSA solution with each concentration into a 96-well plate, sequentially adding 150 μ l Coomassie brilliant blue G250 solution into each well, placing into a multifunctional microplate reader, shaking for 5min, and measuring the light absorption value at 595 nm. The Bradford protein standard curve is plotted with BSA concentration as abscissa and absorbance as ordinate.
The crude enzyme extract was diluted to an appropriate ratio, and the protein concentration of the crude enzyme extract was measured by the Bradford method, and the protein concentration was regarded as the content of 5 α -reductase.
Study of the inhibitory Activity of the Activity on type II 5 α -reductase: adding sample according to the following reaction system, incubating at 37 ℃ for a certain time, adding 2ml of precooled methanol to stop reaction, mixing uniformly, centrifuging for 5min at 10000r/min, taking supernatant, filtering by using a 0.22 mu m filter membrane, and measuring the absorbance value of each sample hole by using a multifunctional microplate reader; or the residual concentration of testosterone in each sample.
TABLE 15 study of alpha-reductase inhibitory Activity
| Reagent | Blank control tube | Sample tube | Positive control tube | Negative control tube |
| Phosphate buffer/. mu.l | 300 | 300 | 300 | 300 |
| 5 alpha-reductase crude enzyme solution/. mu.l | 500 | 500 | 500 | 500 |
| Testosterone/. mu.l | 50 | 50 | 50 | 50 |
| 75% ethanol/. mu.l | 50 | / | 50 | / |
| Sample solution/. mu.l | / | 50 | / | / |
| Finasteride/. mu.l | / | / | 50 | / |
| NADPH/ |
100 | 100 | 100 | 100 |
| Duration of reaction/t | t | t | t | t |
| Methanol (precooling) | 2000 | 2000 | 2000 | 2000 |
Volunteer experiments: volunteer subjects were enrolled as required and signed written informed consent. Before the group entry, the subjects are asked a series of questions about disease history, health condition and the like according to the criteria of entry and exclusion and the like, and the subjects are subjected to evaluation of scalp conditions before the use of the product and image shooting and recorded.
The test included 30 scalp sensitive subjects aged 20-50 years, and the final effective number of subjects was 30. The volunteer enters constant temperature and humidity evaluation room for 30min for standing and testing, 0.2ml of Margarita stock solution is applied to scalp every day and fixed at 2 x 2cm 2 An area; the same evaluations and tests were again performed on the product for 14 days, with the experimental results: FIG. 1 is a schematic representation of the change in scalp moisture content before and after use, showing a significant difference, p, from that before use<0.05，***p<0.001. FIG. 2 is a schematic diagram showing the change of percutaneous water loss (g/m) before and after use 2 h) In that respect FIG. 3 is a graph of scalp sensitivity before and after application of the composition of the present invention to a portion of volunteers. After using the sample, 30 subjects can conclude the following compared to before using the sample: 1) after the sample is used for 14 days, the moisture content of the scalp cuticle is obviously improved, has obvious difference,the percutaneous dehydration condition of the scalp is obviously reduced, and the instant moisturizing effect is achieved. 2) The test sample had the effect of relieving scalp redness, and the phenomenon of scalp redness was significantly reduced in the volunteers 14 days after the use of the sample (see fig. 3).
Comparative example 1:
preparation of component A: pearl powder: selecting pollution-free seawater pearls, removing dead pearls and impurities, cleaning the pearls with clear water, filtering, drying at low temperature, putting the pearls into an ultrafine pulverizer to pulverize into coarse powder with the particle size of 10-15 microns, then performing air flow pulverization to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight of 20% of the powder, pressurizing to 0.5MP, maintaining for 4 hours, instantly reducing the pressure, taking out, mixing the obtained pearl powder with water according to the ratio of 1: 4(m/m), grinding by a wet method to obtain pearl powder with the particle size of 10-100nm, filtering and drying to obtain a component A: pearl powder;
preparation of component B: seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2h, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a component B: seawater pearl extract;
preparation of component C: hydrolyzing seawater pearls: preparing 0.2mg/mL of streptothricin solution by using phosphate buffer solution with the pH value of 7.5, adding pearl powder, wherein the streptothricin accounts for 0.16 percent of the mass of the pearl powder, performing enzymolysis for 2 hours at the temperature of 40 ℃, heating to inactivate enzyme, adjusting the pH value to 6, adding complex enzyme accounting for 0.2 percent of the mass of the pearl powder, and the complex enzyme is debitterizing protease and bromelain according to the ratio of 2: 1, carrying out enzymolysis at 55 ℃ for 3 hours, heating to inactivate enzyme, carrying out reduced pressure evaporation concentration, and carrying out freeze drying to obtain a component C: hydrolyzing seawater pearls;
preparation of component D: commercially available keratin.
Mixing a component A, B, C, D according to a mass ratio of 1:10:10:5, mix well and formulate solutions according to example 1, table 1.
Comparative example 2:
preparation of component A: pearl powder: selecting pollution-free seawater pearls, removing dead pearls and impurities, cleaning the pearls with clear water, filtering, drying at low temperature, putting the pearls into an ultrafine pulverizer to pulverize into coarse powder with the particle size of 10-15 microns, then performing air flow pulverization to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight of 20% of the powder, pressurizing to 0.5MP, maintaining for 4 hours, instantly reducing the pressure, taking out, mixing the obtained pearl powder with water according to the ratio of 1: 4(m/m), grinding by a wet method to obtain pearl powder with the particle size of 10-100nm, filtering and drying to obtain a component A: pearl powder;
preparation of component B: seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2 hours, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a component B: seawater pearl extract;
preparation of component C: hydrolyzing seawater pearls: preparing papain into a papain solution of 0.2mg/mL by using a phosphate buffer solution with the pH of 6.0, adding the pearl powder and papain, wherein the mass of the papain is 0.36 percent of that of the pearl powder, performing enzymolysis at 55 ℃ for 5 hours, heating to inactivate enzyme, performing reduced pressure evaporation and concentration, and performing freeze drying to obtain a component C: hydrolyzing seawater pearls;
preparation of component D: hydrolyzing the fibroin: placing silk into sodium carbonate aqueous solution with the mass fraction of 1% to boil for 30min, removing sericin, filtering and drying, and mixing the silk without sericin with 0.1mol/L phosphoric acid aqueous solution with the pH value of 6.5 according to the ratio of 1: 20-1: 40(g: ml), adding 800U/g neutral protease, hydrolyzing at 45 ℃ for 4h, filtering to adjust pH to 7 to obtain hydrolyzed fibroin, concentrating by evaporation under reduced pressure, and freeze drying to obtain component D: and (3) hydrolyzing the fibroin.
Comparative example 3:
the hydrolyzed seawater pearls of component C of example 1 were replaced with polyglutamic acid and the other preparation methods were the same as example 1.
Comparative example 4:
the seawater pearl extract of the component B in the example 1 is replaced by conch hard protein powder, and other preparation methods are the same as the example 1.
The skin moisturizing efficacy of the products of each example was tested: the test included 30 volunteers, and the final number of effective subjects was 30. The volunteers enter a constant temperature and humidity evaluation room to wash the arms for 30min for testing, 0.2ml of sample is taken from each person to be smeared on the area of 2 x 2cm2 of the left forearm, and the testing is respectively carried out after 1, 2, 4 and 8h of smearing. No adverse reactions were reported by 30 volunteers. As shown in fig. 4, the 5 samples all had moisturizing ability, and the moisturizing effect of example 1 was the most excellent among all the samples.
The results of the scalp 5 α -reductase inhibition test for each example product are shown in fig. 5. The 5 α -reductase inhibition of the composition of example 1 was 79.01%, and the 5 α -reductase inhibition of control 4 was 68.34% (P < 0.001).
DPPH clearance assay: as shown in FIG. 6, the DPPH clearance of the composition of example 1 reached 93.7%, the DPPH clearance of control 1 was 80.67% (P < 0.01) with a significant difference, and the DPPH clearance of control 3 was 76.17% (P < 0.01) with a significant difference.
Hair strength and toughness test:
experimental hair tresses: highly damaged Chinese (heavily damaged hair bundle, multiple bleaching), 18cmx 1g, Shanghai Yan Yu Co; human hair made wig pieces 40cm x 25g, Hangzhou Bingyufei trade Co.
Using equipment: XS (08) XT-3 carbon fiber Strength tester, Shanghai Asahi Instrument Co., Ltd; HF5 constant temperature and humidity unit, available from Kunlin Air precision air Conditioning System (Suzhou) Inc.
Environmental conditions: the test result observation should be carried out in the environment with the temperature of 21 +/-1 ℃ and the relative humidity of 50 +/-10% RH, and the visual evaluation should be carried out under the constant illumination condition (the illumination of a fluorescent tube or an LED with the color temperature of 5500-6500K).
The experimental method comprises the following steps: selecting the severely damaged hair strands, and storing under the conditions of constant temperature and humidity, wherein the temperature is 25 +/-2 ℃ and the humidity is 50 +/-5 percent. The test group is used for smearing the hair strands for three minutes by using a quantitative product, washing the hair strands with warm water, stroking the hair strands for ten times by using similar force by using hands each time of warm water washing, and then drying the hair strands under the conditions of constant temperature and constant humidity, wherein the operation is one time. Each hair was treated five times with the product. The control group was untreated tresses. The tensile test was performed after 12h storage under constant temperature and humidity conditions. The diameter of the hair is measured by using an SN-1200W high-definition camera, and the specific method is to take the average value of the diameters of 3 positions in the middle section. Taking 30 hairs with diameter difference of 10 μm in the hair bundle, using a fiber strength tester to perform single fiber strength test, and calculating and comparing the hair tensile strength of the control group and each sample group by the following formula:
σ＝Fb/So
where σ is the tensile strength, Fb is the maximum force to which the sample is subjected when it is broken, and So is the original cross-sectional area of the sample. As shown in FIG. 7, after the product of each example 1 is used, compared with the control group, the toughness of the hair is improved by 22.13%, and the P is less than 0.01, so that the composition has the effect of improving the toughness of the hair.
Comparative example 5:
preparation of component A: pearl powder: selecting pollution-free seawater pearl, removing dead pearl and impurities, cleaning pearl with clear water, filtering, drying at low temperature, pulverizing pearl into coarse powder with particle size of 10-15 μm in an ultrafine pulverizer, pulverizing with gas flow to obtain seawater pearl powder with average particle size of 2-3 μm, mixing the obtained powder with water according to a ratio of 1: 4(m/m), grinding by a wet method to obtain pearl powder, filtering and drying to obtain a component A: the light purple pearl powder has slight fishy smell;
preparation of component B: seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2h, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a component B: light purple seawater Margarita extract.
The pearl powder and the seawater pearl extract obtained in the comparative example 5 are light purple and have slight fishy smell, the technical scheme of the example 1 increases the specific surface area of the seawater pearl particles, the prepared pearl powder is fine and porous, the color of the seawater pearl powder is obviously improved, the fishy smell of the seawater pearl is favorably removed, and the pearl powder obtained in the example 1 has no fishy smell.
Comparative example 6:
the component A in the example 1 is replaced by taurine, the other preparation method is the same as the example 1, and the component A, B, C, D is mixed according to the mass ratio of 0.02: 10:10:5, mix well and formulate solutions according to table 1.
Comparative example 7:
the seawater pearl of example 1 was replaced with a fresh water pearl, and the preparation method was the same as example 1. Mixing a component A, B, C, D according to a mass ratio of 1:10:10:5, mix well and formulate solutions according to table 1.
LPS induces release of inflammatory factors by Hacat cells: and (3) giving a sample to be detected with 0.5 per mill of HaCat cell volume fraction to incubate cells for 24 hours, then adding 1 mu g/ml LPS to act on the cells for 6 hours, and collecting cell culture supernatant after incubation for 24 hours, and respectively determining the content of the IL-6 inflammatory factors in the supernatant by using a kit. As shown in FIG. 8, the sample of example 1 had an inhibitory effect on the inflammatory factor IL-6 induced by LPS. Compared with the model group, the IL-6 secretion of example 1 is reduced by 34% (P < 0.001), the IL-6 secretion of control example 6 is reduced by 21% (P < 0.01), and the IL-6 secretion of control example 7 is reduced by 19% (P < 0.01).
30 volunteers enter a constant temperature and humidity evaluation room to wash arms for 30min for testing, 0.2ml of sample is taken by each person to smear on the area of 2 x 2cm2 of the left forearm, and the test is carried out 1, 2, 4 and 8h after smearing. No adverse reactions were reported by 30 volunteers. The rate of change in the skin's transepidermal water loss is shown in FIG. 9.
Compared with fresh water pearls, the seawater pearls selected by the invention contain about 0.02 percent of functional components such as taurine and the like, and contain a plurality of proteins such as chitin, a plurality of amino acid active components such as tyrosine, arginine, aspartic acid, lysine, glycine, L-cysteine and the like, and calcium carbonate, mineral macro-elements such as sodium, potassium, phosphorus and the like, and a plurality of trace elements required by human bodies, such as iron, zinc, iodine, selenium, germanium and the like And (3) repairing skin barrier.
It should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (9)
1. A method for preparing a scalp/hair/skin care composition using seawater pearls as raw materials is characterized in that:
preparing pearl powder: putting seawater pearls into a superfine pulverizer to pulverize into coarse powder with the particle size of 10-15 microns, then performing air flow pulverization to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight of 15-25% of the powder, pressurizing, instantly reducing pressure, taking out, mixing the obtained pearl powder and water into pearl slurry, performing wet grinding to obtain pearl powder, filtering and drying to obtain bright white pearl powder;
preparing a seawater pearl extract: adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 3-5%, soaking for 1-3 h, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain a seawater pearl extract;
preparing hydrolyzed seawater pearls: preparing 0.1-0.4 mg/mL streptothricin protease solution with the pH value of 7.0-7.5, adding the pearl powder, wherein the streptothricin protease accounts for 0.1-0.2% of the mass of the pearl powder, performing enzymolysis at 40-45 ℃ for 1-3 h, heating to inactivate enzyme, adjusting the pH value to 5.5-6.5, adding complex enzyme accounting for 0.1-0.2% of the mass of the pearl powder, wherein the complex enzyme is debitterized protease and bromelain according to the ratio of 1-3: 1, performing enzymolysis at 50-55 ℃ for 2-3 hours, heating to inactivate enzyme, performing reduced pressure evaporation concentration, and freeze-drying to obtain hydrolyzed seawater pearls;
preparing hydrolyzed fibroin:
mixing pearl powder: seawater pearl extract: hydrolyzing seawater pearls: the hydrolyzed fibroin is mixed according to the mass ratio of (0.01-2) to (8-12) to (4-6).
2. The method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 1, wherein: the pearl powder preparation method comprises the steps of putting seawater pearls into an ultrafine grinder, grinding the seawater pearls into coarse powder with the particle size of 10-15 microns, carrying out air flow grinding on the coarse powder to obtain seawater pearl powder with the average particle size of 2-3 microns, putting the seawater pearl powder into a pressurizing device, adding water with the weight being 20% of the powder, pressurizing to 0.5MP, maintaining for 4-5 hours, instantly reducing pressure, taking out, and mixing the obtained pearl powder with the water according to the mass ratio of 1: 4-5, carrying out wet grinding to obtain pearl powder with the particle size of 10-100nm, filtering and drying to obtain bright white pearl powder.
3. A method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 1 or 2, which comprises: the preparation method of the seawater pearl extract comprises the steps of adding the pearl powder into a lactic acid aqueous solution with the mass concentration of 4%, soaking for 2-3 hours, filtering, collecting filter residues, repeatedly washing with distilled water, carrying out wet grinding, drying, and carrying out secondary grinding by a high-speed grinder to obtain the bright white seawater pearl extract.
4. A method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 1 or 2, which comprises: the seawater pearl extract is prepared by preparing 0.2mg/mL streptothricin protease solution from streptothricin protease with phosphate buffer solution with the pH value of 7.5, adding pearl powder, wherein the streptothricin protease accounts for 0.16% of the mass of the pearl powder, performing enzymolysis for 2-3 h at 40 ℃, heating to inactivate enzyme, adjusting the pH value to 6, adding complex enzyme accounting for 0.2% of the mass of the pearl powder, and performing 1-3: 1, carrying out enzymolysis at 55 ℃ for 3-4 h, heating to inactivate enzyme, carrying out reduced pressure evaporation concentration, and carrying out freeze drying to obtain the hydrolyzed seawater pearl.
5. A method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 1 or 2, which comprises: the preparation method of the hydrolyzed silk protein comprises the steps of putting silk into a sodium carbonate aqueous solution with the mass fraction of 1-2% to boil for 30-40 min, removing sericin, filtering and drying, and mixing the silk after the sericin is removed with 0.1mol/L phosphoric acid aqueous solution with the pH value of 6.5 according to the g: ml of 1: 20-1: 40, adding 800-900U/g neutral protease, hydrolyzing for 4h at 45 ℃, filtering and adjusting the pH value to 7 to obtain hydrolyzed fibroin, decompressing, evaporating, concentrating, freezing, drying and hydrolyzing the fibroin.
6. A method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 1 or 2, which comprises: mixing pearl powder: seawater pearl extract: hydrolyzing seawater pearls: the hydrolyzed fibroin is mixed according to the mass ratio of 0.02-1: 10:10: 5.
7. The method for preparing a scalp/hair/skin-rejuvenating composition using seawater pearls as a raw material according to claim 4, wherein: the complex enzyme is debitterized protease and bromelain according to the ratio of 2: 1 is compounded in proportion.
8. A scalp/hair/skin care composition using seawater pearls as a raw material prepared by the method of claim 1.
9. Use of a scalp/hair/skin-rejuvenating composition based on seawater pearls as claimed in claim 1 in cosmetics, characterized in that: the composition is used for preparing cosmetics comprising one or more of shampoo, hair conditioner, scalp care solution, emulsion, cream, shower gel, facial mask and face cleaning product, has strong damaged hair repair function, can regulate the activity of scalp sebaceous glands and inhibit the effect of 5 alpha-reductase, and has obvious antioxidant and anti-inflammatory effects, and the effects of moisturizing and repairing skin barrier.
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