CN113925821A - Application of spirulina H11 strain extract as antioxidant and skin care product thereof - Google Patents
Application of spirulina H11 strain extract as antioxidant and skin care product thereof Download PDFInfo
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- CN113925821A CN113925821A CN202010611631.2A CN202010611631A CN113925821A CN 113925821 A CN113925821 A CN 113925821A CN 202010611631 A CN202010611631 A CN 202010611631A CN 113925821 A CN113925821 A CN 113925821A
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- spirulina
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
- A61K8/375—Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- Cosmetics (AREA)
Abstract
The invention relates to the field of spirulina application, and discloses application of a spirulina H11 strain extract as an antioxidant, wherein the spirulina H11 strain is preserved in a number of: CCTCC M2017772. The spirulina H11 strain extract selected by the invention has strong free radical scavenging capacity, has development value as an antioxidant production application, and the preparation method thereof obviously reduces the pigment and the smell of the extract. The invention also provides a skin care product, which takes the extract of the spirulina H11 strain as an antioxidant, can effectively remove free radicals, and reduces and avoids the damage of the free radicals to the skin.
Description
Technical Field
The invention relates to the field of spirulina application, in particular to application of a spirulina H11 strain extract as an antioxidant and a skin care product containing the spirulina H11 strain extract.
Background
Space breeding of microalgae is a relatively efficient artificial breeding mode, the mutagenic action of factors such as cosmic radiation, microgravity, complex electromagnetic environment and the like on microalgae is utilized to cause the genetic variation of microalgae cells, a large number of mutant strains are obtained at one time, and then the microalgae strains with high growth rate, high biomass, stable genetic character and development value are screened from the mutagenized strains. Compared with the traditional breeding, the space mutation breeding has the greatest advantages of high mutation probability, wide mutation range and short breeding period, and can create high-quality germplasm resources in a relatively short time.
For application No.: 201711461143.2, (Spirulina platensis) strain H11, deposited under the number: CCTCC M2017772. The Spirulina (Spirulina platensis) H11 strain is obtained by space breeding and multiple screening, the growth rate of the strain is fast, and the strain has development value in high content of phycocyanin, Spirulina polysaccharide and beta-carotene.
In natural antioxidants, phenols have a radical scavenger structure and can supply hydrogen from a hydroxyl group located on an aromatic ring to terminate radical oxidation of lipids and other compounds, thereby having a remarkable antioxidant effect. Polyphenol components play a key role in resisting chronic diseases and aging caused by free radicals (especially active oxygen and active nitrogen), while antioxidant components of spirulina are considered as main causes for these effects, but polyphenol compounds in microalgae do not cause much attention due to their low concentration, and the range of use is also limited.
Disclosure of Invention
The invention aims to provide the application of spirulina H11 strain extract in the aspect of antioxidation and provide a skin care product using the spirulina H11 strain extract as an antioxidant.
As a first aspect of the present invention, the technical solution adopted by the present invention to solve the technical problem is: provides the application of the spirulina H11 strain extract, the spirulina H11 strain is preserved as follows: CCTCC M2017772, and the application of the spirulina H11 strain extract as an antioxidant.
The Spirulina (Spirulina platensis) H11 strain of the invention is deposited in the Chinese type culture Collection on 12 months and 8 days in 2017, and the address is as follows: wuhan university in Wuchang Lojia mountain in Wuhan City of Hubei China; the strain preservation number is as follows: CCTCC M2017772. The classification of the spirulina H11 strain is named as spirulina platensis SCSIO-44012-H11.
Phenols have the structure of radical scavengers, capable of supplying hydrogen from hydroxyl groups located on aromatic rings to terminate radical oxidation of lipids and other compounds, have a significant antioxidant effect, and play a key role in combating chronic diseases and aging caused by free radicals, in particular reactive oxygen and reactive nitrogen. Polyphenolic compounds, including simple phenols, flavonoids, phenylpropionaldehyde, tannins, lignins, phenolic acids and their derivatives, are important components of complex defense mechanisms against a variety of plant stresses.
The spirulina H11 strain extract provided by the invention carries a 'practical ten' return satellite, returns to the ground after operating on orbit for 12 days, has the characteristics of strong free radical scavenging capacity and stable heredity, and is remarkably improved in polyphenol yield and remarkably enhanced in oxidation resistance. The spirulina H11 strain extract has strong free radical scavenging capacity, potential for extracting polyphenol and antioxidant capacity, and lays a foundation for industrial application.
The further scheme of the invention is as follows: use of extract of strain Spirulina H11 as antioxidant in cosmetics.
With age, the body protection mechanisms decline. Physiological damage in vivo is especially the induced lipid peroxidation, i.e. endogenous superoxide radicals act on unsaturated fatty acids in the body to produce unstable lipid peroxides which in turn decompose to produce aldehydes, especially malondialdehyde, which react to form lipid protein complexes by attacking phospholipids and proteins, with the deposition of lanipofuscin inside the cells, which is a hallmark of cell aging. Therefore, the antioxidation is always a powerful means for preventing and treating skin aging and is an important way for other compounds to achieve the effect of resisting the skin aging.
By extracting polyphenol from Spirulina H11 strain, the antioxidant effect of the polyphenol can reduce the generation of hydrogen peroxide, superoxide anion and hydroxyl radical which are harmful to living cells, protect cells from oxidative stress damage, recover skin elasticity, and reduce wrinkle.
The further scheme of the invention is as follows: the spirulina H11 strain extract is prepared by the following method:
culturing and collecting spirulina H11 strain, drying and pulverizing to obtain spirulina powder, adding ethanol into the spirulina powder, shaking by a shaking table, and filtering to obtain extractive solution;
evaporating the extracting solution by using a rotary evaporator, and dissolving the extracting solution by using deionized water to obtain a crude extract;
adsorbing the crude extract with pretreated AB-8 macroporous resin column, sequentially eluting with deionized water, 20-40% ethanol, 50-70% ethanol, and 95% ethanol to obtain ethanol solution, and concentrating the ethanol solution by evaporation to obtain extract.
In the invention, a spirulina H11 strain is cultured, and the cultured spirulina is collected, freeze-dried and pulverized to obtain spirulina powder. Then adding ethanol into the spirulina powder to leach active substances, and shaking and extracting by using a shaking table overnight. Filtering the obtained solution to remove residue, collecting liquid, rotary evaporating at 40 deg.C to remove ethanol, adding deionized water for several times to dissolve active substances to obtain crude extract.
And (3) carrying out adsorption purification on the obtained crude extract by using a pretreated AB-8 macroporous resin column, and then sequentially eluting with deionized water, 20-40% ethanol, 50-70% ethanol and 95% ethanol. And (4) carrying out rotary evaporation on the collected ethanol analysis liquid to remove ethanol and water to obtain the extract. The preparation method can remarkably reduce the pigment and smell of the extract, is more suitable for being applied to cosmetics and ensures the quality of the cosmetics.
As a second aspect of the present invention, the present invention also provides a skin care product comprising, by mass percent,
penetration enhancer: 0.01 to 5 percent;
humectant: 0.01-20%;
thickening agent: 0.02-0.8%;
pH regulators: 0.01-1%;
preservative: 0.01 to 0.15 percent;
skin conditioner: 0.01 to 5 percent;
solubilizer: 0.01 to 0.5 percent;
skin whitening agent: 0.01 to 5 percent;
aromatic agent: 0.005-0.5%;
antioxidant: 0.01-2.5%, wherein the antioxidant is extract of spirulina H11 strain, and the spirulina H11 strain has a preservation number of: CCTCC M2017772;
water: and (4) the balance.
In this example, the antioxidant used was extract of Spirulina H11 strain, strain H11 (Spirulina platensis) deposited in 2017 at 12/8 th day, china center for type culture collection, address: wuchang Lojia mountain in Wuhan, Hubei; the strain preservation number is as follows: CCTCC M2017772. The classification of the spirulina H11 strain is named as spirulina platensis SCSIO-44012-H11. Bis-diethoxydiol cyclohexane 1, 4-dicarboxylate is used as a penetration enhancer; the bis-diethoxydiol cyclohexane 1, 4-dicarboxylate is dissolved in water-soluble oil, and can promote penetration of effective components into hair and skin, and enhance product moisture retention.
In one embodiment, the penetration enhancer is bis-diethoxydiethylene glycol cyclohexane 1, 4-dicarboxylate.
In one embodiment, the moisturizer comprises one or a combination of two or more of dipropylene glycol, panthenol, butylene glycol, glycerin, PEG/PPG-17/6 copolymer, glycerin polyacrylate, glyceryl polyether-26, sodium hyaluronate.
In one embodiment, the thickener comprises one or a combination of two or more of acrylic/C10-30 alkanol acrylate crosspolymer, hydroxyethyl cellulose, xanthan gum, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, carbomer, and ammonium acryloyldimethyl taurate/VP copolymer.
In one embodiment, the solubilizer comprises one or more of polysorbate-20, PEG-40 hydrogenated castor oil, PPG-26-Butanethol-26, and glyceryl ether-25 PCA isostearate.
In one embodiment, the pH adjusting agent comprises one or a combination of two or more of aminomethyl propanol, citric acid, sodium citrate, potassium hydroxide, sodium hydroxide, arginine, and the like.
In one embodiment, the skin conditioning agent comprises one or a combination of two or more of allantoin, bisabolol, ulva extract, oat bran extract, phaeodactylum tricornutum extract, macroalgae extract, fucus extract, chlorella fermentation product, hydrolyzed collagen, ceramide 2, beta-glucan, trehalose, brown algae extract, hamamelis virginiana water, mistletoe ginkgo leaf extract, lalang grass rhizome extract, cactus extract, lactobacillus/soybean fermentation product extract.
By adding the skin conditioning agent, the skin is further supplemented with water, and the generation of wrinkles can be reduced. The effective components in the skin conditioner can penetrate into the deep part of skin and be absorbed by skin, thereby improving the state of skin.
In one embodiment, the skin whitening agent comprises one or a combination of more than two of niacinamide, magnolia sieboldii extract, kojic acid and derivatives thereof, arbutin and derivatives thereof.
By adding the skin whitening agent, the activity of tyrosinase can be inhibited, thereby achieving an excellent whitening effect.
In one embodiment, the antioxidant is added in an amount of 1% to 2.5%.
In the present invention, the antioxidant spirulina H11 strain extract is added in an amount of 0.01-2.5%, preferably in an amount of 1-2.5%, and more preferably in an amount of 2%. Wherein, the antioxidant performance is increased along with the increase of the dosage, and the acceleration is slowed down after the dosage is increased to 2 percent, and the content of the added antioxidant spirulina H11 strain extract is 2 percent based on the consideration of the cost and the effect.
In the invention, the addition amount of the preservative is 0.01-1.5%, and the preservative can comprise one or the combination of more than two of phenoxyethanol, methyl hydroxybenzoate, benzoic acid and the salt thereof, and propyl hydroxybenzoate.
In the present invention, the chelating agent is added in an amount of 0-1%, and the chelating agent may be EDTA-2Na and/or EDTA-4Na, etc.
The invention can also add aromatic, which can be essence.
The skin care product can be skin water, lotion, skin penetrating water and the like, the selected spirulina H11 strain extract is used as an antioxidant, the polyphenol in the extract is utilized, free radicals can be effectively eliminated, the damage of the free radicals to the skin is reduced and avoided, and the bis-diethoxydiol cyclohexane 1, 4-dicarboxylate is further matched as a penetration enhancer, so that the absorption of the skin to the skin care product is effectively promoted, the oxidation resistance of the skin care product is enhanced, and the skin elasticity is effectively improved.
The invention has the beneficial effects that:
the spirulina H11 strain extract selected by the invention has strong free radical scavenging capacity and development value as an antioxidant for production and application, and meanwhile, in the preparation process, AB-8 macroporous resin is adopted for adsorption and purification, and then deionized water and ethanol with different concentrations are used for washing and resolving, so that the pigment and smell of the extract can be obviously reduced, and the extract is more beneficial to being used as a cosmetic oxidant.
The skin care product of the invention takes the extract of spirulina H11 strain as the antioxidant, can effectively remove free radicals, and reduces and avoids the damage of the free radicals to the skin.
Drawings
FIG. 1 is a standard graph of gallic acid;
FIG. 2 is a graph of the clearance of free radicals from a test subject to DPPH;
FIG. 3 shows the DPPH radical scavenging IC of a test substance50Comparing the graphs;
FIG. 4 is a graph of test subject versus ABTS radical clearance;
FIG. 5 shows ABTS free radical scavenging IC of a test agent50Comparing the graphs;
fig. 6 is a graph comparing the rate of change of skin elasticity.
Detailed Description
The technical solutions of the present invention are further illustrated by the following specific examples, which do not represent limitations to the scope of the present invention. Insubstantial modifications and adaptations of the present invention by others of the concepts fall within the scope of the invention.
In the examples of the present invention, the Wild strain of Spirulina platensis (Wild type, WT, hereinafter referred to as Wild strain WT) and the mutagenized Spirulina strain H11 (the algal species is designated as SCSIO 44012-H11, the preservation number of the strain is CCTCC M2017772, hereinafter referred to as mutagenized strain H11) were compared for testing. Wherein, for the mutagenic strain H11, the mutagenic strain carries a 'practice ten' return satellite, returns to the ground after operating on orbit for 12 days, and has a preservation number: CCTCC M2017772. The wild strain WT and the mutant strain H11 used in the experiment are preserved by the alga resource and the alga species library of the biotechnological subject group of the Nanhai ocean institute of Chinese academy of sciences.
1. The culture method of the spirulina comprises the following steps:
the culture medium of the spirulina comprises the following components: 5g/L NaHCO3，0.5g/L NaNO3，0.02g/L K2HPO4，0.01g/L FeSO4·7H2O，0.08g/L Na2EDTA, 1mL/L A5 solution, prepared by taking the upper seawater of the south sea area as a matrix. The salinity is controlled to be 28 +/-1 per thousand by supplementing deionized water. The spirulina mutagenic strain H11 and the wild strain WT of the invention are respectively subjected to parallel comparative culture tests outdoors. Inoculating in plastic bucket with 50L volume for over-supplementing CO2Controlling the pH of the culture mediumStirring at a fixed time of 9.0-9.5 days, culturing for 4 weeks at an average temperature of 27 deg.C, collecting Spirulina, freeze drying, and making into powder.
2. The extraction method of the spirulina H11 strain extract comprises the following steps:
20g of mutagenic strain H11 and wild strain WT spirulina powder were weighed out in a weight ratio of 1: 25 of the feed liquid ratio, adding 500ml of absolute ethyl alcohol, shaking the table at 26 ℃, shaking the table at 180rpm overnight and extracting. The next day, the residue was filtered off, the liquid was collected, ethanol was removed by rotary evaporation at 40 ℃, a small amount of deionized water was added several times to dissolve out the active substance, yielding a crude extract of 25 ml.
The crude extract obtained is subjected to a purification operation.
The AB-8 macroporous resin is pretreated, and the method comprises the following specific steps: weighing an appropriate amount of AB-8 macroporous resin, loading the AB-8 macroporous resin on a column by a wet method, filling the column until the volume of the column is about 50mL, adding 95% ethanol which is 10cm higher than the resin layer into the column, soaking the column for 24 hours, and then leaching the column with 95% ethanol until an effluent liquid is diluted by water in a test tube and is not turbid. And finally washing with water until the ethanol content is less than 1% or no obvious ethanol smell exists. And 2-5 mm of liquid is kept on the resin layer surface so as to avoid drying the column. The column was then soaked with 150mL of 5% HCl solution for 2-4 hours and then washed with water until the effluent pH was neutral. The column was soaked with 150mL of 2% NaOH solution for 2-4 hours and then washed with water until the pH of the effluent was neutral.
The extract of the mutagenic strain spirulina and the crude extract of the wild spirulina are passed through an exchange column, the flow rate at the outlet is controlled to be 1 drop/second, and air bubbles cannot exist in the resin layer. The stock solution in the resin layer was replaced with 150mL of deionized water, and the resin layer was washed. The stock solution in the resin layer was replaced with 150mL of 30% ethanol, and the resin layer was washed. The stock solution in the resin layer was replaced with 150mL of 60% ethanol, and the resin layer was washed. The stock solution in the resin layer was replaced with 150mL of 95% ethanol, and the resin layer was washed. The collected 60% ethanol solution is subjected to rotary evaporation at 40 ℃ to remove ethanol and water, and then deionized water is added to dissolve out the active substances, so that the pigment and the smell of the extract can be remarkably reduced. Thus, an antioxidant test was conducted.
3. And (4) active ingredient determination.
In this example, the polyphenol content in the corresponding mutant strain H11 and wild strain WT was characterized by measuring the gallic acid content. Wherein, the determination of the content of the gallic acid is obtained by determining the absorbance of the gallic acid at the wavelength of 760nm and comparing the absorbance with a gallic acid standard curve.
Specifically, the determination method of the content (equivalent) of the gallic acid comprises the following steps:
diluting Spirulina extract by 10 times, adding 1mL into test tubes with plugs (1 mL deionized water as blank control), respectively adding 1mL deionized water into each tube, adding 0.5mL Folin-phenol solution diluted by one time (0.5 mL deionized water as negative control), and adding 3mL Na with mass volume fraction of 13.4%2CO3The solution is added with deionized water to be constant volume to 10mL, and is reacted for 2h in the dark at room temperature, and the absorbance is carried out at the wavelength of 760 nm.
The determination method of the gallic acid standard curve comprises the following steps:
preparing 1mg/mL gallic acid standard solution: accurately weighing 100mg of gallic acid standard, completely dissolving, and adding into a 100mL volumetric flask to constant volume.
Accurately measured 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0mL of 1mg/mL gallic acid standard solution, prepared into 0, 10, 20, 40, 60, 80, 100. mu.g/mL standard series in a 50mL volumetric flask, and subjected to the standard curve measurement according to the gallic acid measurement method.
Further, with reference to fig. 1 and table 1 below, table 1 is a table corresponding to the relationship between gallic acid concentration and absorbance; FIG. 1 is a gallic acid standard curve graph, which is plotted with gallic acid concentration as abscissa and absorbance as ordinate.
The obtained gallic acid has standard curve equation of A-0.0885 x +0.0768 and linear coefficient R2The gallic acid concentration was found to be in a good linear relationship in the range of 1 to 10. mu.g/mL (0.9927).
Table 1: gallic acid concentration as abscissa concentration and absorbance relation table
| Concentration (μ g/mL) | 1 | 2 | 4 | 6 | 8 | 10 |
| Absorbance of the solution | 0.132 | 0.269 | 0.444 | 0.618 | 0.812 | 0.928 |
Calculating gallic acid equivalent of total phenols in wild strain WT Spirulina extract to 47.819 μ g/mL according to standard curve, and yield to 59.774 μ g/g (dry weight); the gallic acid equivalent of total phenols in the extract of the mutant strain H11 spirulina was 70.305. mu.g/mL, and the yield was 87.881. mu.g/g (dry weight).
TABLE 2 Total phenol content in Spirulina
| OD760 | Total phenol (gallic acid equivalent) concentration (. mu.g/ml) | |
| Mutagenic strain H11 Spirulina extract (1/10) | 0.699 | 70.305 |
| Wild strain WT Spirulina extract (1/10) | 0.500 | 47.819 |
Based on the determination result of gallic acid, the screened mutant strain H11 has higher total phenol content than wild spirulina strain, and is more valuable than wild spirulina, so that the production application of spirulina H11 extract as antioxidant is realized.
4. The spirulina was tested for its antioxidant capacity.
Mutagenic strain H11 Spirulina extract was tested for antioxidant properties, including DPPH free radical scavenging ability and ABTS free radical scavenging ability.
DPPH radical scavenging assay.
Preparing a DPPH working solution: 0.0082g of DPPH powder is dissolved in 100mL of ethanol and stored at 4 ℃ in the dark.
Detecting the antioxidant activity of the test substance: adding 200 μ L of test substance into 1mL of DPPH working solution, reacting for 30min in dark, reading A at wavelength of 517nm1. Adding 200 μ L deionized water into 1ml of working solution of LDPPH, reacting for 30min in dark, and reading A at wavelength of 517nm2. Adding 200 μ L of the test substance into 1mL of ethanol, reacting for 30min in the dark, and reading A at a wavelength of 517nm3In this example, ascorbic acid (i.e., vitamin C), D-alpha-tocopherol (i.e., vitamin E) and Trolox were used together as positive controls。
Wherein, the calculation formula of the DPPH free radical clearance rate is as follows: clearance rate [ [ (A)2+A3-A1)/A2]*100％
The DPPH radical scavenging rate curves are plotted based on the DPPH radical scavenging rates obtained from the test substances at different concentrations, as shown in Table 3 below.
Table 3: DPPH radical clearance rate of test substance
FIG. 2 and FIG. 3 show graphs of the removal rate of DPPH free radicals from the test substance, and FIG. 3 shows the removal IC of DPPH free radicals from the test substance50As can be seen, the effect of scavenging DPPH free radicals is significant and dose dependent. The IC of each component can be obtained according to the dose curve50As shown in figure 3, the DPPH free radical scavenging effect of the mutagenic strain H11 spirulina extract is better than that of the wild strain WT spirulina extract, ascorbic acid, D-alpha-tocopherol and Trolox, which shows that the composition of active ingredients of the mutagenic strain H11 spirulina is changed after space breeding, and the DPPH free radical scavenging performance of the spirulina extract is improved.
ABTS free radical scavenging assay.
Preparing ABTS working solution: and (3) preparing ABTS solution from 0.0286g to 5mL of ABTS powder in deionized water, and storing in dark. 5mL of the antioxidant assay was mixed with 88. mu.L of potassium persulfate (140mmol/L), and the mixture was left to stand overnight in a 37 ℃ thermostat water bath in the dark. The overnight standing stock ABTS was diluted to OD with deionized water734The value is kept at 0.7 +/-0.02, and the working solution is used as the working solution (the working solution is prepared for use).
Detecting the antioxidant activity of the test substance: adding 200 μ L of test substance into 1mL of ABTS working solution, reacting for 30min in dark, and reading A at 734nm4. Adding 200 μ L deionized water into 1mL ABTS working solution, reacting for 30min in dark place, and reading A at 734nm5. Adding 200 μ L of the sample into 1mL of deionized water, and keeping out of the sunThe reaction time was 30min and the reading was A at a wavelength of 734nm6Ascorbic acid, D-alpha-tocopherol and Trolox were used as positive controls.
Clearance rate [ [ (A)5+A6-A4)/A5]*100％
The ABTS free radical scavenging rate curves were plotted as shown in Table 4 below, by which the ABTS free radical scavenging rates were obtained for various concentrations of test substances.
Table 4: clearance rate of test substance to ABTS free radical
FIG. 4 and FIG. 5 show graphs of ABTS free radical scavenging rate of the test substance, and FIG. 5 shows ABTS free radical scavenging IC of the test substance50As can be seen, the respective active ingredients have significant scavenging effects on ABTS radicals, all in an increased dose-increasing effect. The IC of each component can be obtained according to the dose curve50As shown in figure 3, the DPPH free radical scavenging effect of the mutagenic strain H11 spirulina extract is superior to that of the wild strain WT spirulina extract, D-alpha-tocopherol and Trolox, and is close to ascorbic acid, which shows that the composition of active ingredients of the mutagenic strain H11 spirulina is changed through space breeding, and the ABTS free radical scavenging performance of the spirulina H11 strain extract is improved.
Based on this, it was found that the mutagenic strain H11 spirulina extract of the present invention is superior to ascorbic acid, D- α -tocopherol and Trolox in DPPH clearance; is close to ascorbic acid in ABTS clearance and is superior to D-alpha-tocopherol and Trolox. The capacity of the extract of the spirulina strain H11 to scavenge DPPH and ABTS free radicals is also significantly changed compared with the wild strain spirulina. Therefore, the extract of the spirulina H11 strain has wide application as an antioxidant, and has great potential as the antioxidant in the preparation of cosmetics.
In another aspect of the present invention, the present invention provides a skin care product, in which the formulation ratios of examples and comparative examples are as shown in the following table. In example 1, example 2 and example 3, the mutant strain H11 spirulina extract of the present invention was added as an antioxidant, and the mutant strain H11 spirulina strain was deposited as follows: CCTCC M2017772, the wild strain WT spirulina extract was used as a comparative example in comparative examples 1-3, and the wild strain WT spirulina extract without addition of mutagenic strain H11 was used as a blank control.
Table 5: the formula of the skin care product is in proportion
The preparation process of the skin care product with the formula in the embodiment comprises the following steps:
1. adding A, B and C phase raw materials into a stirring pot, stirring and heating to 82-85 ℃;
2. cooling to 42 ℃, adding D, E, F phase and stirring evenly;
3. cooling to 37 ℃, discharging after the inspection is qualified, and standing for 24 hours to obtain the skin care product.
Wherein, A, B, C, D, E, F phases in the preparation process are respectively A phase: deionized water;
phase B: butylene glycol, propylene glycol, glycerol, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, ammonium acryloyldimethyl taurate/VP copolymer, sodium hyaluronate, ceramide 2, panthenol, dipropylene glycol, glycerol polyacrylate;
and C phase: methylparaben, allantoin, glyceryl polyether-26, PEG/PPG-17/6 copolymer, citric acid, and sodium citrate;
phase D: PEG-40 hydrogenated castor oil, essence;
phase E: phenoxyethanol, beta-glucan, nicotinamide, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate;
and (3) phase F: spirulina extract, Lactobacillus/fermented soybean product extract.
In this example, the skin care product was prepared as a skin lotion, and the skin elasticity test was performed on the obtained skin lotion.
Method for testing skin elasticity: the test principle is based on the principle of suction and stretching, where a negative pressure is generated on the skin surface to be tested to suck the skin into a specific test probe, and the depth of the skin sucked into the test probe is measured by a non-contact optical test system. The test probe includes a transmitter and receiver of light, the ratio of which (the ratio of transmitted light to received light) is proportional to the depth of skin being absorbed, thus obtaining a time-dependent curve of the length of skin stretched.
Measuring the skin elasticity of the subject by using a probe PVM600 and a skin elasticity measuring instrument MPA580 of German CK company, selecting a parameter R2 as a comparison index (R2: the ratio of the skin rebound quantity without negative pressure to the maximum stretching quantity with negative pressure is closer to 1, the skin elasticity is better) and measuring for 3 times in total, and taking an average value; the improvement of the skin elasticity of the tested area by the product was evaluated by measuring the change in the skin elasticity value R2 before and after use of the product.
The number of the testees is 33, the test period is 8 weeks, the skin care products of the examples 1-3 and the skin care products of the comparative examples 1-3 and the blank control group are selected as test samples in the test, 7 different areas are divided on the forearm of the testees, the skin care products of the blank control group, the comparative examples 1-3 and the examples 1-3 are smeared on the different areas on the inner side of the forearm every morning and evening, and the smearing amount is about 2mg/cm2And the smearing position of each test sample is kept unchanged in the test period, and then the skin elasticity of the tested area before the test and when the test sample is used for 8 weeks is respectively measured, so that the change rate of the skin elasticity is represented.
Referring to fig. 6 and the following table, table 6 shows the skin elasticity change rate and a comparison of the skin elasticity change rate.
TABLE 6 rate of change of skin elasticity
Therefore, the spirulina extract can be added into the skin care product to improve the oxidation resistance of the skin care product, further improve the skin elasticity and play a role in improving the skin. Meanwhile, as the amount of the spirulina extract is increased, the effect thereof is increased. Compared with the spirulina extract added with wild strains, the spirulina H11 strain extract adopted by the invention can obtain better effect under the condition of adding less amount, the effect is increased along with the increase of the adding amount, the cost and the effect are comprehensively compared, and the adding amount of the spirulina H11 strain extract is 2 percent, which is a better embodiment.
The above detailed description of the preferred embodiment of the present invention is the result of many experiments with great expenditure of labor, money and time.
It should be understood that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same, and those skilled in the art can modify the technical solutions described in the above embodiments, or make equivalent substitutions for some technical features; and all such modifications and alterations are intended to fall within the scope of the appended claims.
Claims (12)
1. The application of the spirulina H11 strain extract is characterized in that the spirulina H11 strain is preserved in a number of: CCTCC M2017772, and the application of the spirulina H11 strain extract as an antioxidant.
2. Use of an extract of strain spirulina H11 as claimed in claim 1, wherein the extract of strain spirulina H11 is used as an antioxidant in cosmetics.
3. The use of an extract of strain spirulina H11 as claimed in claim 2, wherein the extract of strain spirulina H11 is prepared by the following method:
culturing and collecting spirulina H11 strain, drying and pulverizing to obtain spirulina powder, adding ethanol into the spirulina powder, shaking by a shaking table, and filtering to obtain extractive solution;
evaporating the extracting solution by using a rotary evaporator, and dissolving the extracting solution by using deionized water to obtain a crude extract;
adsorbing the crude extract with pretreated AB-8 macroporous resin column, sequentially eluting with deionized water, 20-40% ethanol, 50-70% ethanol, and 95% ethanol to obtain ethanol solution, and concentrating the ethanol solution by evaporation to obtain extract.
4. A skin care product comprises, by mass percent,
penetration enhancer: 0.01 to 5 percent;
humectant: 0.01-20%;
thickening agent: 0.02-0.8%;
pH regulators: 0.01-1%;
preservative: 0.01 to 0.15 percent;
skin conditioner: 0.01 to 5 percent;
solubilizer: 0.01 to 0.5 percent;
skin whitening agent: 0.01 to 5 percent;
aromatic agent: 0.005-0.5%;
antioxidant: 0.01-2.5%, wherein the antioxidant is extract of spirulina H11 strain, and the spirulina H11 strain has a preservation number of: CCTCC M2017772;
water: and (4) the balance.
5. The skin care product of claim 3 wherein the penetration enhancer is bis-diethoxydiethylene glycol cyclohexane 1, 4-dicarboxylate.
6. The skin care product of claim 3, wherein the moisturizer comprises one or a combination of two or more of dipropylene glycol, panthenol, butylene glycol, glycerin, PEG/PPG-17/6 copolymer, glycerin polyacrylate, glyceryl polyether-26, and sodium hyaluronate.
7. The skin care product of claim 3 wherein the thickener comprises one or a combination of two or more of acrylic acid/C10-30 alkanol acrylate crosspolymer, hydroxyethylcellulose, xanthan gum, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, carbomer, and ammonium acryloyldimethyl taurate/VP copolymer.
8. The skin care product of claim 3 wherein the solubilizing agent comprises one or a combination of more than two of polysorbate-20, PEG-40 hydrogenated castor oil, PPG-26-Butanew-26, glyceryl ether-25 PCA isostearate.
9. The skin care product of claim 3, wherein the pH regulator comprises one or more of aminomethyl propanol, citric acid, sodium citrate, potassium hydroxide, sodium hydroxide, arginine, etc.
10. The skin care product of claim 3, wherein the skin conditioning agent comprises one or a combination of two or more of allantoin, bisabolol, ulva extract, oat bran extract, phaeodactylum tricornutum extract, macroalgae extract, fucus extract, chlorella fermentation product, hydrolyzed collagen, ceramide 2, beta-glucan, trehalose, brown algae extract, witch hazel water, mistletoe leaf extract of ginkgo, lalang grass rhizome extract, cactus extract, lactobacillus/soybean fermentation product extract.
11. The skin care product according to claim 3, wherein the skin whitening agent comprises one or a combination of two or more of niacinamide, magnolia sieboldii extract, kojic acid and its derivatives, arbutin and its derivatives.
12. The skin care product according to any one of claims 3 to 10, wherein the antioxidant is added in an amount of 1 to 2.5% by mass.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115844742A (en) * | 2022-11-28 | 2023-03-28 | 广州市科能化妆品科研有限公司 | Moisturizing composition, preparation method thereof and cosmetic |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108265014A (en) * | 2017-12-28 | 2018-07-10 | 中国科学院南海海洋研究所 | One plant passes through high-quality seawater spirulina of space breeding acquisition and application thereof |
| CN109394677A (en) * | 2018-12-27 | 2019-03-01 | 杭州宝格丽生物科技有限公司 | It is a kind of with anti-oxidant, anti-aging effects cosmetics |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108265014A (en) * | 2017-12-28 | 2018-07-10 | 中国科学院南海海洋研究所 | One plant passes through high-quality seawater spirulina of space breeding acquisition and application thereof |
| CN109394677A (en) * | 2018-12-27 | 2019-03-01 | 杭州宝格丽生物科技有限公司 | It is a kind of with anti-oxidant, anti-aging effects cosmetics |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115844742A (en) * | 2022-11-28 | 2023-03-28 | 广州市科能化妆品科研有限公司 | Moisturizing composition, preparation method thereof and cosmetic |
| CN115844742B (en) * | 2022-11-28 | 2023-10-20 | 广州市科能化妆品科研有限公司 | Moisturizing composition, preparation method thereof and cosmetic |
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