CN114948789A - SOD composition with high-efficiency blue light resisting effect and preparation method thereof - Google Patents

SOD composition with high-efficiency blue light resisting effect and preparation method thereof Download PDF

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CN114948789A
CN114948789A CN202210503117.6A CN202210503117A CN114948789A CN 114948789 A CN114948789 A CN 114948789A CN 202210503117 A CN202210503117 A CN 202210503117A CN 114948789 A CN114948789 A CN 114948789A
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sod
blue light
composition
blue
sod composition
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CN114948789B (en
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胡程伟
赖红星
姚元锋
夏玉平
郑振强
何子华
陈泽贤
陈小敏
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Zhuhai Jinbaikang Biological Technology Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/66Enzymes
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34Alcohols
    • A61K8/345Alcohols containing more than one hydroxy group
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/35Ketones, e.g. benzophenone
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A61K8/4946Imidazoles or their condensed derivatives, e.g. benzimidazoles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60Sugars; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73Polysaccharides
    • A61K8/735Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73Polysaccharides
    • A61K8/738Cyclodextrins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/004Aftersun preparations
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/49Solubiliser, Solubilising system
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    • A61K2800/52Stabilizers
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    • A61K2800/59Mixtures
    • A61K2800/592Mixtures of compounds complementing their respective functions
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    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to an SOD composition with a high-efficiency blue-light-resistant effect and a preparation method thereof. The SOD composition with the high-efficiency blue light resistance provided by the invention takes SOD as a core raw material, is matched with other optimized polypeptide, stabilizer and synergistic agent, solves the problems of poor stability, low absorption and utilization rate and the like of SOD in application through specific proportion compatibility, can greatly improve the blue light resistance and damage repair effect of SOD, and is an ideal cosmetic composition for resisting blue light and repairing skin damage.

Description

SOD composition with high-efficiency blue light resisting effect and preparation method thereof
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to an SOD composition with a high-efficiency blue-light-resistant effect and a preparation method thereof.
Background
Research shows that all the sunlight wave band light can damage human skin, including infrared ray, visible light and ultraviolet ray. Blue light is an important component of visible light, and is also common in daily life as well as electronic screens. Blue light is the highest energy band in visible light, and the penetrating power is also the strongest, even stronger than ultraviolet light, so that the blue light can cause skin damage similar to that caused by ultraviolet light.
The blue light irradiates keratinocytes to enable the cells to generate oxidative stress, the cells mainly generate superoxide anions when being irradiated by the blue light, and mainly generate singlet oxygen when being irradiated by UVA ultraviolet rays with the wavelength of 320-420 nm, both of the superoxide anions and the singlet oxygen mediate skin injury through a Reactive Oxygen Species (ROS) path, induce the cells to generate free radicals, reduce the oxidation resistance of fibroblasts, inhibit the proliferation of the fibroblasts, greatly reduce the extracellular matrix of the fibroblasts in the dermis of the skin, and further accelerate the aging speed of the skin.
At present, the blue light resistant cosmetics in the market are generally added with components such as plant extracts or plant essential oils to prevent the skin from being damaged by blue light. For example: patent document CN 106727027a discloses a blue light resistant, oxidation resistant, radical scavenging composition. The composition for resisting blue light, resisting oxidation and removing free radicals mainly comprises a compound of marigold flower extract and safflower oil, a tea extract and a leontopodium alpinum extract, wherein the compound of the marigold flower extract and the safflower oil can enhance the blue light filtering capability of human skin, prevent the damage of blue light to the skin, capture free radicals in human skin tissues and delay aging caused by skin oxidation.
Patent document CN 109464308A discloses an essential oil composition with blue light resisting and skin caring effects, which mainly comprises orange flower essential oil, pomegranate essential oil, ginger essential oil and lavender essential oil, wherein the prepared essential oil composition can absorb blue light to avoid the damage of the blue light to the skin, and meanwhile, the essential oil composition also has the functions of promoting blood circulation, whitening, removing freckles, promoting cell growth, promoting blood circulation to remove blood stasis and resisting free radicals, and can eliminate skin damage and skin aging caused by the blue light, so that the purpose of resisting the blue light is achieved.
However, the blue light resistant cosmetics added with components such as plant extracts or plant essential oils have the defects of poor blue light resistant effect, short repair time, easy volatilization of the plant essential oils and quick reduction of the blue light resistant effect in the use process. Superoxide dismutase (SOD) is a metalloprotease containing copper, zinc, iron and manganese, has the functions of catalyzing superoxide anion free radicals to carry out disproportionation reaction and balancing oxygen free radicals in an organism, and is expected to become an important component for resisting the skin aging problem caused by blue light radiation.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides an SOD composition with high-efficiency blue light resistance and a preparation method thereof. The SOD composition with the efficient blue light resisting effect provided by the invention takes SOD as a core raw material, is matched with other optimized polypeptide, stabilizer and synergistic agent, solves the problems of poor stability, low utilization rate and the like of SOD in application through specific proportion compatibility, and can also greatly improve the blue light resisting and damage repairing effects of the SOD.
The invention provides a SOD composition with high-efficiency blue light resistance, which comprises the following components in percentage by mass:
0.01-2% of superoxide dismutase, 0.01-2% of polypeptide, 5-10% of stabilizer, 2-5% of synergist, 10-30% of solubilizer and the balance of purified water to 100%.
Further, the SOD composition with the high-efficiency blue-light resisting effect comprises the following components in percentage by mass:
0.1-2% of superoxide dismutase, 0.1-2% of polypeptide, 5-8% of stabilizer, 2-5% of synergist, 10-28% of solubilizer and purified water added to 100%.
Further, the polypeptide is one or the combination of more than two of carnosine, glutathione, acetyl tetrapeptide-2 and palmitoyl tetrapeptide-10.
Further, the stabilizer is one or two of hydroxyethyl piperazine ethane sulfonic acid, trehalose and sodium hyaluronate.
Further, the synergist is p-hydroxyacetophenone.
Further, the solubilizer is one or two of butanediol and pentanediol.
Further, the SOD composition with the efficient blue light resisting effect also comprises 5-10% of cyclodextrin in percentage by mass.
In addition, the invention also provides a preparation method of the SOD composition with high-efficiency blue light resisting effect, which comprises the following steps:
s1, adding the synergist into purified water, heating to 80-100 ℃, and stirring at 250-350 rpm until the synergist is clear and transparent to obtain a mixed solution;
s2, cooling the mixed liquid prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, a stabilizer and a solubilizer, and stirring at 450-550 rpm until the mixed liquid is clear and transparent, so as to obtain the product.
Further, the preparation method of the SOD composition with high-efficiency anti-blue light effect further comprises step S3, specifically: adding cyclodextrin into purified water to prepare a cyclodextrin saturated solution, adding the mixed solution prepared in the step S2 into the cyclodextrin saturated solution, heating to 60-80 ℃, stirring for 2-3 h, cooling to room temperature, stirring at room temperature for 6-8 h, standing at the temperature of 2-8 ℃ for 12h, taking out, filtering, taking out crystals, adding purified water, and stirring to a clear and transparent state, wherein the material-to-liquid ratio of the crystals to the purified water is 1g:6 mL.
At present, although SOD has been widely used in skin care products, there are still many unsolved problems in the cosmetic industry, such as: SOD has poor stability, and is easily influenced by factors such as dissolved oxygen, illumination, electrolyte, temperature and the like in a water-based system to be degraded actively; and the molecular weight of the natural SOD is about 32000Da, the natural SOD belongs to macromolecular protein, no specific receptor exists on a cell membrane, the natural SOD is difficult to rapidly enter cells to play a role, and the effect of the SOD is greatly limited to play.
In order to solve the above problems, the present inventors have creatively proposed an SOD composition having high stability and remarkable anti-blue light effect and skin damage repairing effect. The SOD composition takes SOD as a core raw material, is matched with polypeptide, stabilizer and synergistic agent, solves the problems of poor stability and low utilization rate of the SOD through specific proportion compatibility, and can also greatly improve the blue light resistance of the SOD.
During the research process, the inventor unexpectedly finds that: the composition of the SOD cosmetic comprises a stabilizer formed by mixing hydroxyethyl piperazine ethane sulfonic acid with one or two of trehalose and sodium hyaluronate, p-hydroxyacetophenone as a synergist, butanediol and pentanediol as solubilizers, and can improve the stability of SOD.
The inventor finds in the research process that although trehalose and sodium hyaluronate are generally used as moisturizers in cosmetics and are really more used as moisturizing ingredients in terms of human body efficacy, the inventor finds that the raw materials are two different levels of understanding concepts in terms of formula structure effect and human body efficacy effect. The stabilizer formed by mixing the hydroxyethyl piperazine ethanesulfonic acid with one or two of trehalose and sodium hyaluronate is mainly used for maintaining the stability of the pH value of a system and weakening the efficiency of Newtonian fluid molecules and Brownian movement in a formula system, and the problem of enzyme activity loss caused by autodigestion reaction of the enzyme activity of SOD in a solution is solved, so that the enzyme activity of the SOD in the system is maintained, and the absorption rate of the SOD can be improved. The p-hydroxyacetophenone serving as the synergist has a synergistic effect on blue light prevention and oxidation resistance, so that the blue light prevention and oxidation resistance of the system is further improved; butanediol and pentanediol are used as solubilizers, so that the solubility of the system can be improved, the system is in a stable, clear and transparent state, and the stability of the product is better facilitated.
In addition, the inventor also provides that the cyclodextrin with a certain proportion is added to carry out closed wrapping on the SOD, so that the problem that the SOD is influenced by factors such as dissolved oxygen, illumination, electrolyte, temperature, microorganisms and the like in a water-based system to influence enzymatic activity degradation can be better solved, the stability of the SOD is further improved, and the effects of resisting blue light and repairing damage of the SOD can be better played.
The SOD composition with the efficient blue light resisting effect is transparent viscous liquid with colorless to light green appearance. Tests show that the prepared SOD composition has long-term higher activity stability, the activity damage of the SOD is lower when the SOD composition is tested at a high temperature of 40 ℃ for 90 days, and the prepared SOD composition has more efficient blue light damage protection effect within the wavelength range of 400-500 nm.
Compared with the prior art, the SOD composition with the efficient blue light resisting effect has the advantages of high stability, good skin absorption rate, remarkable blue light resisting and damage repairing effects, capability of maintaining the blue light resisting and skin damage repairing effects for a long time, simple preparation process, suitability for large-scale production and use, and suitability for ideal blue light resisting and skin damage repairing cosmetic compositions.
Detailed Description
The present invention is further described in the following description of the specific embodiments, which is not intended to limit the invention, but various modifications and improvements can be made by those skilled in the art according to the basic idea of the invention, within the scope of the invention, as long as they do not depart from the basic idea of the invention. The materials and reagents involved in the present invention can be obtained by means of commercial or ordinary techniques in the art. Wherein: SOD and polypeptide were purchased from Biotech GmbH, Nanjing Sibaike, and cyclodextrin was purchased from Dachang Yangtze (Shanghai).
Example 1 SOD composition with high anti-blue light efficacy
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.2% of superoxide dismutase, 0.2% of polypeptide, 6% of stabilizing agent, 5% of p-hydroxyacetophenone, 10% of solubilizer and purified water added to 100%;
the polypeptide is formed by mixing 0.1% of carnosine and 0.1% of glutathione by mass percent.
The stabilizer is formed by mixing 5% of hydroxyethyl piperazine ethane sulfonic acid and 1% of sodium hyaluronate.
The solubilizer is formed by mixing 5 mass percent of butanediol and 5 mass percent of pentanediol.
The preparation process comprises the following steps:
s1, adding p-hydroxyacetophenone into purified water, heating to 80 ℃, and stirring at 350rpm until the mixture is clear and transparent to obtain a mixed solution;
s2 cooling the mixed solution prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, stabilizer and solubilizer, and stirring at 550rpm to be clear and transparent.
Example 2SOD composition with high anti-blue light efficacy
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.5 percent of superoxide dismutase, 0.5 percent of polypeptide, 7 percent of stabilizing agent, 2 percent of p-hydroxyacetophenone, 15 percent of solubilizer and purified water added to 100 percent;
the polypeptide is mixed and combined by 0.25 percent of glutathione and 0.25 percent of acetyl tetrapeptide-2;
the stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 1% by mass of trehalose and 1% by mass of sodium hyaluronate;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The preparation process comprises the following steps:
s1, adding p-hydroxyacetophenone into purified water, heating to 90 ℃, and stirring at 300rpm until the mixture is clear and transparent to obtain a mixed solution;
s2 cooling the mixed solution prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, stabilizer and solubilizer, and stirring at 400rpm until the mixture is clear and transparent.
Example 3 SOD composition with high anti-blue light efficacy
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
1% of superoxide dismutase, 1% of polypeptide, 10% of stabilizing agent, 5% of p-hydroxyacetophenone, 25% of solubilizer and purified water added to 100%;
the polypeptide is formed by mixing 0.25% of carnosine, 0.25% of glutathione and 0.5% of palmitoyl tetrapeptide-10 in percentage by mass.
The stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 4% by mass of trehalose and 1% by mass of sodium hyaluronate.
The solubilizer is formed by mixing and combining 5% of butanediol and 20% of pentanediol.
The preparation process comprises the following steps:
s1, adding p-hydroxyacetophenone into purified water, heating to 100 ℃, and stirring at 250rpm until the mixture is clear and transparent to obtain a mixed solution;
s2 cooling the mixed solution prepared in the step S1 to normal temperature, then adding superoxide dismutase, polypeptide, stabilizer and solubilizer, and stirring at 450rpm until the mixture is clear and transparent.
Comparative example 1 SOD composition with high-efficiency anti-blue light effect
0.5 percent of superoxide dismutase and the purified water is added to 100 percent.
The preparation process comprises the following steps:
stirring superoxide dismutase and purified water at 300rpm until the system is clear and transparent.
Comparative example 2SOD composition with high-efficiency anti-blue light effect
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
1% of superoxide dismutase, 7% of stabilizing agent, 2% of p-hydroxyacetophenone, 15% of solubilizer and purified water added to 100%;
the stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 1% by mass of trehalose and 1% by mass of sodium hyaluronate;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The difference from example 2 is that: lacking the polypeptide.
The preparation process is similar to that of example 2.
Comparative example 3 SOD composition with high-efficiency blue light resistance
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.5% of superoxide dismutase, 0.5% of polypeptide, 7% of stabilizing agent, 15% of solubilizer and purified water added to 100%;
the polypeptide is mixed and combined by 0.25 percent of glutathione and 0.25 percent of acetyl tetrapeptide-2;
the stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 1% by mass of trehalose and 1% by mass of sodium hyaluronate;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The difference from example 2 is that: p-hydroxyacetophenone is lacking.
The preparation process is similar to that of example 2.
Comparative example 4 SOD composition with high-efficiency anti-blue light effect
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.5% of superoxide dismutase, 0.5% of polypeptide, 7% of hydroxyethyl piperazine ethane sulfonic acid, 2% of p-hydroxyacetophenone, 15% of solubilizer and purified water which is added to 100%;
the polypeptide is mixed and combined by 0.25 percent of glutathione and 0.25 percent of acetyl tetrapeptide-2;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The difference from example 2 is that: the stabilizer is hydroxyethyl piperazine ethane sulfonic acid.
The preparation process is similar to that of example 2.
Comparative example 5 SOD composition with high-efficiency anti-blue-light effect
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.5 percent of superoxide dismutase, 0.5 percent of polypeptide, 7 percent of stabilizing agent, 2 percent of p-hydroxyacetophenone, 15 percent of solubilizer and purified water added to 100 percent;
the polypeptide is mixed and combined by 0.25 percent of glutathione and 0.25 percent of acetyl tetrapeptide-2;
the stabilizer is formed by mixing 6% of trehalose and 1% of sodium hyaluronate in percentage by mass;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The difference from example 2 is that: the stabilizer is formed by mixing 3.5 mass percent of trehalose and 3.5 mass percent of sodium hyaluronate.
The preparation process is similar to that of example 2.
Example 4 SOD composition with high anti-blue-light efficacy
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
0.5% of superoxide dismutase, 0.5% of polypeptide, 7% of stabilizing agent, 2% of p-hydroxyacetophenone, 15% of solubilizer, 5% of cyclodextrin and purified water which is added to 100%;
the polypeptide is mixed and combined by 0.25 percent of glutathione and 0.25 percent of acetyl tetrapeptide-2;
the stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 1% by mass of trehalose and 1% by mass of sodium hyaluronate;
the solubilizer is formed by mixing and combining butanediol with the mass percent of 10% and pentanediol with the mass percent of 5%.
The preparation process comprises the following steps:
s1, adding p-hydroxyacetophenone into purified water, heating to 90 ℃, and stirring at 300rpm until the mixture is clear and transparent to obtain a mixed solution;
s2, cooling the mixed solution prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, stabilizer and solubilizer, and stirring at 400rpm until the mixture is clear and transparent;
s3 adding cyclodextrin into purified water to prepare a cyclodextrin saturated solution, adding the mixed solution prepared in the step S2 into the cyclodextrin saturated solution, heating to 70 ℃, stirring for 2 hours, cooling to room temperature, stirring at room temperature for 7 hours, standing at 4 ℃ for 12 hours, taking out, filtering, taking out crystals, adding purified water, and stirring to a clear and transparent state, wherein the material-to-liquid ratio of the crystals to the purified water is 1g:6 mL.
Example 5 SOD composition with high anti-blue light efficacy
The SOD composition with the efficient blue light resisting effect comprises the following components in percentage by mass:
1% of superoxide dismutase, 1% of polypeptide, 10% of stabilizing agent, 8% of p-hydroxyacetophenone, 25% of solubilizer, 8% of cyclodextrin and purified water added to 100%;
the polypeptide is formed by mixing 0.25% of carnosine, 0.25% of glutathione and 0.5% of palmitoyl tetrapeptide-10 in percentage by mass.
The stabilizer is formed by mixing 5% by mass of hydroxyethyl piperazine ethane sulfonic acid, 4% by mass of trehalose and 1% by mass of sodium hyaluronate.
The solubilizer is formed by mixing and combining 5 mass percent of butanediol and 20 mass percent of pentanediol.
The preparation process comprises the following steps:
s1, adding p-hydroxyacetophenone into purified water, heating to 90 ℃, and stirring at 300rpm until the mixture is clear and transparent to obtain a mixed solution;
s2, cooling the mixed solution prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, stabilizer and solubilizer, and stirring at 400rpm until the mixture is clear and transparent;
s3 adding cyclodextrin into purified water to prepare a cyclodextrin saturated solution, adding the mixed solution prepared in the step S2 into the cyclodextrin saturated solution, heating to 60 ℃, stirring for 2 hours, cooling to room temperature, stirring for 8 hours at room temperature, standing at 4 ℃ for 12 hours, taking out, filtering, taking out crystals, adding purified water, stirring to a clear and transparent state, wherein the material-to-liquid ratio of the crystals to the purified water is 1g:6mL, and thus obtaining the cyclodextrin saturated solution. Test example I test for stability of SOD composition having high anti-blue light efficacy
1. Test subjects:
example 1, example 2, example 3, example 4, example 5, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5 prepared SOD compositions having high efficiency of blue light resistance.
2. The test method comprises the following steps:
the superoxide dismutase remaining activity and the activity damage ratio of the test sample are determined by taking 10mL of each of the SOD compositions with high-efficiency blue light resistance prepared in example 1, example 2, example 3, example 4, example 5, comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5, placing the SOD compositions in a sealed penicillin bottle, placing the SOD compositions in a constant-temperature heating box at 40 ℃, and periodically sampling (5 days, 10 days, 30 days, 60 days and 90 days) to perform autoxidation rate determination.
Determination of superoxide dismutase remaining Activity in test samples: by means of the fast autoxidation of pyrogallol under the alkaline condition to release O 2- The intermediate product is produced in a colored form, the reaction mixture turns yellow-brown after the start of the reaction, turns green after a few minutes and turns yellow again after a few hours, as a result of the oxidation of the intermediate product produced. In the presence of superoxide dismutase (SOD), because it can catalyze O 2- And H + Combined to form O 2 And H 2 O 2 Thus, the accumulation of the intermediate product is prevented, and the enzymatic activity of SOD in the test sample can be determined by calculation.
The light absorption value is measured at the wavelength of 325nm by means of a multifunctional micropore detector, the reading is carried out once every 30s, and the change of the light absorption value per minute within 4min is measured.
Drawing by taking the light absorption value as a vertical coordinate and the time as a horizontal coordinate; in the above graph of absorbance versus time, the following equation is calculated:
Figure BDA0003636204060000101
in the formula: Δ a325 is the reaction rate of the autoxidation tube;
Δ A' 325 is the reaction rate of the sample tube;
d is the dilution multiple of the test sample.
Vt is the total volume of the reaction solution and is expressed by ml;
v is the volume of the added test sample to be tested, and the unit is ml;
3. and (3) test results:
the test results are shown in table 1.
TABLE 1 stability test data for SOD compositions with high potency against blue light
Figure BDA0003636204060000102
Figure BDA0003636204060000111
As can be seen from table 1, the SOD compositions with high-efficiency blue light resistance prepared in embodiments 1 to 3 of the present invention have high stability at a high temperature of 40 ℃, and especially, the SOD compositions with high-efficiency blue light resistance prepared in embodiments 4 to 5 in which cyclodextrin is added as a coating agent have better stability. The stability of the SOD compositions with high-efficiency blue-light resisting efficacy prepared in the comparative examples 2-5 is poor, which shows that the stability of the SOD compositions is improved due to the synergistic effect of the components in the formula of the SOD compositions with high-efficiency blue-light resisting efficacy.
Test example two, fibroblast blue light damage protection test
1. Test subjects:
SOD compositions having high anti-blue light effects, prepared in example 2, example 4 and comparative example 1.
2. The test method comprises the following steps:
2.1 establishing blue light damage model based on mouse embryo fibroblast
Cells were plated at 1.5X 10 3 The cells were inoculated in a 96-well plate at a density of one cell per well, and after 24 hours of culture, LED-BL (blue light) at a wavelength of λ 450nm at 5, 15, 25, 45, 85J/cm 2 The blank control group was wrapped with tinfoil paper to protect from light. The medium in the 96-well plate was discarded and PBS was added before irradiation with LED-BL. Immediately after irradiation, complete medium was added again and the mixture was placed in an incubator for cultivation. After 24 hours, cell viability assays were performed using MTT.
2.2 testing of the Pre-protective Effect of SOD compositions on blue light Damage
The test is carried out by setting 5 test samples, setting 4 repeated holes under each test sample, and setting a zero setting hole, a model hole and a control hole (DMEM culture medium containing 10% fetal calf serum) in the test. The test adopts an MTT method to detect the cell activity, and the specific operation is as follows:
2.2.1, seed plate:
digesting the cells in logarithmic growth phase with pancreatin containing EDTA, centrifuging to remove supernatant, and adjusting cell density to 1.5 x 10 4 After/ml, the 96-well plates were seeded at 100uL per well.
2.2.2 design configuration of different concentrations of test substances: SOD composition samples with high-efficiency blue-light-resistant efficacy prepared in example 2, example 4 and comparative example 1 are prepared into test samples with different concentration gradients, and the specific concentrations are as follows: 100ug/mL, 50ug/mL, 25ug/mL, 12.5ug/mL, and 6.2 ug/mL.
2.2.3 administration:
the cells were dosed until they were fully adherent (24 h post inoculation), 100uL per well. After the administration, the mixture was left at 37 ℃ with 5% CO 2 In an incubator.
2.2.4 molding:
cells were incubated for 24h with LED-BL (blue light) at wavelength λ 450nm at 45J/cm 2 The dose of irradiation was applied to the drug administration group and the model group, and the blank control group was wrapped with tinfoil paper to be protected from light. The medium in the 96-well plate was discarded and PBS was added before irradiation with LED-BL. Immediately after irradiation, complete medium was added again and the mixture was placed in an incubator for cultivation.
2.2.5 cell Activity assay:
after 24h of cell culture, the 96-well plate was washed twice with PBS, 100uL of 0.5mg/mL MTT-DMEM basal medium solution was added to each well, and incubated at 37 ℃ for 3 hours in the absence of light. Then 100uL of DMSO was added to each well, shaken for 15min on a shaker, and the OD was read at 570nm in a microplate reader.
2.2.6 calculation formula:
cell viability ═ 100% (administration well OD-zeroed OD)/(control well OD-zeroed OD) ×
3. And (3) test results:
3.1 based on mouse embryosBlue light damage results of fetal fibroblasts: the cells are set from low to high at 5, 15, 25, 45 and 85J/cm 2 The total 5 blue light irradiation doses, the blue light damage result of which is 45J/cm 2 A damage peak occurs.
3.2, the test data of the pre-protection effect of SOD composition on blue light damage are shown in Table 2.
TABLE 2 Pre-protection test data for SOD compositions against blue light damage
Figure BDA0003636204060000121
Figure BDA0003636204060000131
As shown in Table 2, the SOD composition with high-efficiency blue light resistance prepared by the invention has better effects of blue light resistance and skin damage repair.
The foregoing embodiments are merely illustrative of the principles of the present invention and its efficacy, and are not to be construed as limiting the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims (9)

1. The SOD composition with the efficient blue light resisting effect is characterized by comprising the following components in percentage by mass:
0.01-2% of superoxide dismutase, 0.01-2% of polypeptide, 5-10% of stabilizer, 2-5% of synergist, 10-30% of solubilizer and purified water added to 100%.
2. The SOD composition with high-efficiency blue-light resisting effect as claimed in claim 1, which comprises the following components in percentage by mass:
0.1-2% of superoxide dismutase, 0.1-2% of polypeptide, 5-8% of stabilizer, 2-5% of synergist, 10-28% of solubilizer and purified water added to 100%.
3. The SOD composition with high anti-blue light efficacy according to claim 1 or 2, wherein the polypeptide is one or a combination of two or more of carnosine, glutathione, acetyl tetrapeptide-2, and palmitoyl tetrapeptide-10.
4. The SOD composition with high anti-blue light effect according to claim 1 or 2, wherein the stabilizer is hydroxyethylpiperazineethanesulfonic acid in combination with one or both of trehalose and sodium hyaluronate.
5. The SOD composition with high anti-blue-light efficacy according to claim 1 or 2, wherein the synergist is p-hydroxyacetophenone.
6. The SOD composition with high anti-blue light efficacy according to claim 1 or 2, wherein the solubilizer is one or a combination of two of butanediol and pentanediol.
7. The SOD composition with high-efficiency blue-light resisting effect as claimed in claim 1 or 2, wherein the SOD composition with high-efficiency blue-light resisting effect further comprises 5-10% by mass of cyclodextrin.
8. The method for preparing the SOD composition with high-efficiency anti-blue-light effect as defined in any one of claims 1-7, comprising the steps of:
s1, adding the synergist into purified water, heating to 80-100 ℃, and stirring at 250-350 rpm until the synergist is clear and transparent to obtain a mixed solution;
s2, cooling the mixed liquid prepared in the step S1 to normal temperature, adding superoxide dismutase, polypeptide, a stabilizer and a solubilizer, and stirring at 450-550 rpm until the mixed liquid is clear and transparent, so as to obtain the product.
9. The method for preparing the SOD composition with high anti-blue-light effect as claimed in claim 8, further comprising a step S3, which comprises: adding cyclodextrin into purified water to prepare a cyclodextrin saturated solution, adding the mixed solution prepared in the step S2 into the cyclodextrin saturated solution, heating to 60-80 ℃, stirring for 2-3 h, cooling to room temperature, stirring at room temperature for 6-8 h, standing at the temperature of 2-8 ℃ for 12h, taking out, filtering, taking out crystals, adding purified water, and stirring to a clear and transparent state, wherein the material-to-liquid ratio of the crystals to the purified water is 1g:6 mL.
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