WO2023103595A1 - 一种具有清除自由基功效的组合物及其应用 - Google Patents

一种具有清除自由基功效的组合物及其应用 Download PDF

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WO2023103595A1
WO2023103595A1 PCT/CN2022/125931 CN2022125931W WO2023103595A1 WO 2023103595 A1 WO2023103595 A1 WO 2023103595A1 CN 2022125931 W CN2022125931 W CN 2022125931W WO 2023103595 A1 WO2023103595 A1 WO 2023103595A1
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parts
oil
acid
composition
group
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English (en)
French (fr)
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叶睿
胡璠
胡晓芸
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懿奈(上海)生物科技有限公司
德肤科(上海)生物科技有限公司
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Publication of WO2023103595A1 publication Critical patent/WO2023103595A1/zh

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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
    • A61K8/67—Vitamins
    • A61K8/676—Ascorbic acid, i.e. vitamin C
    • 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
    • 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
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • 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
    • A61K8/67—Vitamins
    • 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
    • A61K8/67—Vitamins
    • A61K8/678—Tocopherol, i.e. vitamin E
    • 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
    • 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
    • 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
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • 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
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/592—Mixtures of compounds complementing their respective functions
    • A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the application relates to the technical field of cosmetics, in particular to a composition with the effect of scavenging free radicals and its application.
  • Free radicals are very unstable, have strong oxidative properties, and have a strong ability to absorb electrons. They need to capture electrons from cell molecules in body tissues to pair themselves, and when cell molecules release an electron, they also become free radicals. , but also to snatch the electrons in the cell membrane or nucleus molecules, so that new free radicals are constantly generated. Free radicals can cause protein denaturation and cross-linking, causing many enzymes and hormones in the body to lose their biological activity, reducing the vitality of the body's immunity, nerve reflexes, and exercise capabilities. At the same time, it can also destroy the structure of nucleic acids and cause metabolic disorders in the entire body, etc. , and eventually cause disease in the body.
  • the purpose of this application is to provide a composition with the effect of scavenging free radicals and its application.
  • the first aspect of the present application provides a composition with the effect of scavenging free radicals, in parts by mass, the composition comprises: 3-o-ethyl ascorbic acid: 1 to 3 parts; ascorbic acid tetraiso Palmitate: 2-4 parts; Tocopherol: 0.3-0.7 parts; Acetylcysteine: 0.1-0.3 parts.
  • the composition provided by the present application has the effect of scavenging free radicals, comprising in parts by mass: 3-o-ethyl ascorbic acid: 2 parts; ascorbyl tetraisopalmitate: 3 parts; tocopherol: 0.5 parts; acetyl Cysteine: 0.2 parts.
  • the second aspect of the present application provides the application of the composition having the effect of scavenging free radicals described in the first aspect of the present application in cosmetics and/or skin care products.
  • the cosmetic and/or skin care product is applied to the outer surface of the skin.
  • the cosmetic and/or skin care products are selected from one or more of eye cream, face cream, essence, lotion, and mask essence.
  • the composition having the effect of scavenging free radicals is dispersed in the matrix material as an additive, and the matrix material is selected from one of oily raw materials, powdery raw materials, and colloidal raw materials or several.
  • the cosmetics and/or skin care products also include: surfactants, flavors and fragrances, pigments, moisturizers, preservatives, antioxidants, ultraviolet absorbers, chelating agents, astringents One or more of additives, penetration aids, and nutritional additives.
  • the present application uses 1 to 3 parts of 3-o-ethyl ascorbic acid, 2 to 4 parts of ascorbic acid tetraisopalmitate, 0.3 to 0.7 parts of tocopherol, and 0.1 parts of acetylcysteine ⁇ 0.3 parts, to achieve synergistic effect. Therefore, applying the composition with the effect of scavenging free radicals provided by the present application to cosmetics and/or skin care products can exert obvious skin anti-oxidation and anti-aging effects at a relatively low application amount.
  • Fig. 1 is the comparison chart of DPPH free radical scavenging activity of Et-VC group, VC-IP group, VE group, NAC group, Et-VC/VC-IP/VE/NAC group;
  • Fig. 2 is the comparison chart of DPPH free radical scavenging activity of Et-VC group, VC-IP group, VE group, NAC group, contrast group 1;
  • Fig. 3 is a comparison chart of DPPH free radical scavenging activities of Et-VC group, VC-IP group, VE group, NAC group and comparison group 2.
  • the embodiment of the present application provides a composition with the effect of scavenging free radicals, in terms of parts by mass, the composition comprises: 3-o-ethyl ascorbic acid: 1 to 3 parts; ascorbyl tetraisopalmitate: 2 to 4 parts parts; tocopherol: 0.3 to 0.7 parts; acetylcysteine: 0.1 to 0.3 parts.
  • the provided composition having the effect of scavenging free radicals comprises, in parts by mass: 3-o-ethyl ascorbic acid: 2 parts; ascorbyl tetraisopalmitate: 3 parts; tocopherol: 0.5 parts; Acetylcysteine: 0.2 parts.
  • composition having the effect of scavenging free radicals provided by the embodiments of the present application can be applied in cosmetics and/or skin care products, and the cosmetics and/or skin care products are applied to the outer surface of the skin.
  • the cosmetic and/or skin care products using the composition having the effect of scavenging free radicals are selected from one or more of eye cream, face cream, essence, lotion, and mask essence.
  • the composition having the effect of scavenging free radicals is dispersed in the matrix material as an additive, and the matrix material is selected from oily raw materials, powdery raw materials, colloidal raw materials, oily raw materials, and solvent raw materials. one or several.
  • the oleaginous material can be any one or several combinations of the following base materials: soybean oil, olive oil, almond oil, castor oil, peanut oil, cottonseed oil, tea seed oil, jojoba oil, avocado oil oil, coconut oil, palm oil, cocoa butter, rice bran oil, evening primrose oil, wheat germ oil, corn germ oil, palm wax, wood wax, mink oil, turtle oil, snake oil, egg butter, tallow, horse tallow, Pig fat, deer fat, lecithin, lanolin, beeswax, spermaceti, shellac wax, paraffin wax, petrolatum, microcrystalline wax, ozokerite, squalane, lanolin wax, lanolin alcohol, lanolin alcohol esters, Acetylated lanolin, acetylated lanolin alcohol, lanolin acid, lanolin esters, polyoxyethylene lanolin, polyoxyethylene lanolin ether, polyoxypropylene lanolin ether, hydrogenated lanolin, alkoxy
  • the Cn fatty acids such as: lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, erucic acid, capric acid.
  • the Cn fatty alcohols are such as: lauryl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, coconut oil alcohol.
  • the Cn fatty acid esters, Cn fatty alcohol esters such as: isopropyl myristate, myristyl myristate, myristyl lactate, cetyl lactate, isopropyl palmitate, palm tree isopropyl Capryl, Butyl Stearate, Isooctyl Stearate, Monoglyceryl Stearate, Polyethylene Glycol Stearate, Decyl Oleate, Glyceryl Caprylate, Glyceryl Caprate, Triolein ester.
  • the silty raw material can be any one or several combinations of the following matrix raw materials: mica, magnesium carbonate, titanium dioxide, zinc white powder, calcium hydroxide, calcium carbonate, kaolin, calcium stearate, zinc stearate , Magnesium Stearate, Talc, Silicon Dioxide, Aluminum Hydroxide, Calcium Pyrophosphate, Calcium Bicarbonate, Bentonite Clay.
  • the oily raw material may be any one or more of vegetable oily raw materials, animal oily raw materials, mineral oily raw materials, synthetic oily raw materials, and semi-synthetic oily raw materials.
  • vegetable oily raw materials animal oily raw materials, mineral oily raw materials, synthetic oily raw materials, and semi-synthetic oily raw materials.
  • the solvent raw material can be any one or several combinations of the following matrix raw materials: water, ethanol, propanol, isopropanol, isobutanol, ethylene glycol, 1,2-propylene glycol, cyclohexane , dichlorodifluoromethane, tetrafluorodichloroethane, ethylene glycol monoethyl ether, diethylene glycol monoethyl ether, acetone, methyl isobutyl ketone, ethyl acetate, butyl acetate, amyl acetate, phthalic acid Dibutyl ester.
  • matrix raw materials water, ethanol, propanol, isopropanol, isobutanol, ethylene glycol, 1,2-propylene glycol, cyclohexane , dichlorodifluoromethane, tetrafluorodichloroethane, ethylene glycol monoethyl ether, diethylene glycol
  • the cosmetics and/or skin care products using the composition with the effect of scavenging free radicals also include: surfactants, flavors and fragrances, pigments, moisturizers, preservatives, antioxidants, ultraviolet rays One or more of absorbents, chelating agents, astringents, penetration aids, pH regulators, and nutritional additives.
  • the surfactant can be: 1) anionic surfactant, such as: C12-14 fatty alcohol ammonium sulfate, C12-14 fatty alcohol polyoxyethylene ether carboxylate, lauryl sulfate diethanolamine salt, C12 -14 Fatty Alcohol Polyoxyethylene Ether Ammonium Sulfate, Triethanolamine Lauryl Sulfate, Alcohol Ether Phthalate Monoester Sodium Salt, Acylated Peptide, Sodium Lauric Acid Monoethanolamide Sulfate, n-Octyl Polyoxypropylene Ether Amber Acid monoester sulfonate, cetyl alcohol succinic acid monoester sulfonate, sulfosuccinic acid monoester disodium salt, succinic acid monostearyl disodium salt, alcohol ether sulfosuccinic acid monoester disodium salt, Fatty alcohol polyoxyethylene ether (sodium) sulfosuccinic acid monoester ammonium salt,
  • the essence and spices can be selected from: 1) natural spices, such as absinthe oil, eucalyptus oil, star anise oil, lime oil, white clove extract, clove oil, white orchid oil, white orchid extract , white orchid leaf oil, cypress oil, peppermint oil, atractylodes stearin, sweet rhinoceros extract, orange leaf oil, clove basil oil, big flower jasmine extract, wintergreen oil, angelica oil, angelica extract, lichenic acid, sandalwood oil , Toi Toi Flower Oil, Toi Toi Leaf Oil, Hyacinth Extract, Liquidambar Extract, Cinnamomum Oil, Sassafras Oil, Nardine Oil, Osmanthus Extract, Tangerine Oil, Carrot Seed Oil, Castoreum, Ambrette oil, yellow orchid oil, patchouli oil, artemisia oil, gourami extract, acacia extract, ginger oil, orange oil, peppermint oil, chrysanthemum extract, cumin oil, amber, am
  • the pigment can be one or more selected from the following components: zirconium dioxide, lead acetate, silver nitrate, chlorophyll, copper chlorophyllin, ferric ferrocyanide, guanine, fast red, food red I , Vat Red I, Carmine, Edible Bayberry Red, Edible Cherry Red, Food Red 17, Amaranth, Fast Green, Food Yellow 3, Edible Lemon Yellow, Edible Sudan Yellow, Edible Indigo, Food Blue 2, Fluorescent Pink, ⁇ -Carotene, Henna, Gardenia Red Pigment, Gardenia Green Pigment, Gardenia Blue Pigment, Gardenia Yellow Pigment, Titanium Mica Pearlescent Pigment, Iron Oxide Red, Iron Oxide Yellow, Iron Oxide Black, Chrome Oxide Green, Oxychlorine Bismuth, basic peach red, vermilion R, quinoline yellow, shikonin, lac red pigment, ultramarine blue, acid red 87, acid green 25, acid fluorescent yellow, acid orange 7, golden light red 53, Lisolbao Red
  • the preservative can be one or several combinations selected from the following components: sorbic acid and its salts, propionate, parabens, benzyl alcohol, benzoic acid and its salts, 5- Chloro-2-methyl-4-isothiazolin-3-one, -2-methyl-4-isothiazolin-3-one, carson, imidazolidinyl urea, dehydroacetic acid and its salts, sodium sulfite, Sodium bisulfite, sodium metabisulfite, 2,2-bromo-2-nitro-1,3-propanediol (Bropol), isoascorbic acid and its salts, kojic acid and its esters, azelaic acid, usnic acid , Resorcinol.
  • the antioxidant can be one or several combinations selected from the following components: SOD, butyl hydroxyanisole, gallate, ascorbic acid and its salts, erythorbic acid and its salts, sitosterol, reed Ding, tocopherol.
  • the moisturizing agent can be one or several combinations selected from the following components: 1,2-butanediol, sorbitol, xylitol, glycerin, lactate, polyethylene glycol, DL- Pyrrolidone carboxylate, D-glucose, kojic acid and its esters, uric acid, orotic acid, pectinic acid, laminin, collagen, trehalose, cholesteryl 12-hydroxystearate.
  • the ultraviolet absorber can be one or several combinations selected from the following components: pyridoxine hydrochloride, kojic acid and esters thereof, rutin, aloin, caffeine, hesperetin, 2-Ethylhexyl salicylate, 2-ethylhexyl p-dimethylaminobenzoate, 2-ethylhexyl p-methoxycinnamate, phenyl salicylate, anthranilate , 2-Hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, 2-hydroxy-4-octyloxybenzophenone.
  • the chelating agent can be one or several combinations selected from the following components: EDTA disodium salt, bipyridinethione zinc, and sodium thiosulfate.
  • the astringent can be one or several combinations selected from the following components: zinc phenol p-sulfonate, aluminum hydroxychloride, aluminum sulfate, potassium aluminum sulfate, zinc sulfate, aluminum chloride, zinc chloride , Calamine.
  • the penetration aid may be one or several combinations selected from the following components: dimethyl sulfoxide, 1-n-dodecylazepan-2-one, and 2-aminoethanol.
  • the pH regulator may be one or several combinations selected from the following components: tartaric acid, lactic acid, boric acid, and succinic acid.
  • the nutritional additive can be one or several combinations selected from the following components: chitin, chitosan, silk peptide, silk fibroin, lecithin, epidermal growth factor, trehalose, lactose, pearl Powder, collagen, collagen hydrolyzate, hyaluronic acid and its salts, egg membrane protein, phytosterol, lysozyme chloride, sodium chondroitin sulfate, bee milk, propolis, honey, glycyrrhizic acid, glycyrrhetinic acid, gamma-flax Acid, kojic acid and its esters, gibberellic acid, glycine, L-aspartic acid and its salts, L-cystine and its salts, DL-alanine, L-serine, DL-serine, L-methionine , DL-methionine, L-lysine hydrochloride, L-threonine, inos, in
  • the instruments, kits, cells and reagents used in the experimental examples can be purchased through open commercial channels, and are commonly used instruments, kits, cells and reagents in this field, and the operation steps of the instruments and kits are guided by the instructions conduct.
  • the DPPH free radical scavenging test was used to evaluate the scavenging effect of the experimental group and the control group on free radicals.
  • Raw material 1 name: 3-o-ethyl ascorbic acid; code name: Et-VC; properties: white to off-white crystalline powder.
  • Raw Material 2 Name: Ascorbyl Tetraisopalmitate; Code: VC-IP; Purity: Properties: Colorless to pale yellow transparent oily liquid.
  • Raw material 3 Name: Tocopherol; Code: VE; Properties: Yellow to brown-red viscous oily liquid.
  • Raw material 4 Name: Acetylcysteine; Code: NAC; Properties: White crystalline powder.
  • Reagent 1 DPPH: 100 mg/bottle, purchased from: Sigma.
  • test substance or reference substance was prepared into a mother solution with DMSO or ddH 2 O, and then diluted to an appropriate concentration with serum-free medium or PBS, and the test substances of the groups shown in Table 1 were respectively obtained:
  • test group and contrast group are respectively set up with the following detection systems A, B, C and D (the test substance consumption is 20ul, and the DPPH consumption 180ul):
  • the above detection steps are carried out in a 96-well plate, set at least 3 multiple wells, add the required analytes, DPPH, PBS or absolute ethanol, mix with a row gun, and place at room temperature in the dark for 30 minutes.
  • the DPPH scavenging activity was statistically processed by Excel input and Prism (Version7) software package, expressed as mean ⁇ standard error. One-way analysis of variance among each group was considered to be statistically significant at p ⁇ 0.05.
  • Fig. 1 is the comparison chart of DPPH free radical scavenging activity of Et-VC group, VC-IP group, VE group, NAC group, Et-VC/VC-IP/VE/NAC group;
  • Fig. 2 is the comparison chart of DPPH free radical scavenging activity of Et-VC group, VC-IP group, VE group, NAC group, contrast group 1;
  • Fig. 3 is a comparison chart of DPPH free radical scavenging activities of Et-VC group, VC-IP group, VE group, NAC group and comparison group 2.
  • n 3, Mean ⁇ SEM. "**” means p ⁇ 0.05 compared with Et-VC/VC-IP/VE/NAC group, and "***” means p ⁇ 0.001 compared with Et-VC/VC-IP/VE/NAC group.
  • n 3, Mean ⁇ SEM. "*” means p ⁇ 0.05 compared with the control group, and "***” means p ⁇ 0.001 compared with the control group.

Abstract

本申请涉及化妆品技术领域，公开了一种具有清除自由基功效的组合物及其应用。本申请提供的具有清除自由基功效的组合物以质量份计包含：3-o-乙基抗坏血酸：1～3份；抗坏血酸四异棕榈酸酯：2～4份；生育酚：0.3～0.7份；乙酰半胱氨酸：0.1～0.3份。本申请通过采用3-o-乙基抗坏血酸、抗坏血酸四异棕榈酸酯、生育酚、乙酰半胱氨酸按照一定的质量份配比混合得到组合物，该组合物在清除自由基功效方面具有协同增效作用，能够以较低施用量获得较好的皮肤抗氧化、抗衰老作用。

Description

一种具有清除自由基功效的组合物及其应用
相关申请的交叉引用
本申请基于申请号为“202111506049.0”、申请日为2021年12月10日的中国专利申请提出，并要求该中国专利申请的优先权，该中国专利申请的全部内容在此以引入方式并入本申请。
技术领域
本申请涉及化妆品技术领域，特别涉及一种具有清除自由基功效的组合物及其应用。
背景技术
在机体代谢过程中，氧的一个电子被夺去后，不具有成对电子的氧原子就成为了自由基。自由基很不稳定，具有强氧化性，吸收的电子能力极强，它需要在身体组织的细胞分子中再夺取电子来使自己配对，而当细胞分子推陈出新一个电子后，它也变成自由基，又要去抢夺细胞膜或细胞核分子中的电子，这样不断产生新的自由基。自由基能引起蛋白质变性和交联，使体内的许多酶及激素失去生物活性，机体的免疫能力、神经反射能力、运动能力等系统活力降低，同时还能破坏核酸结构和导致整个机体代谢失常等，最终使机体发生病变。
从空气中吸入人体的氧气在酶的作用下，有相当一部分氧转化成超氧阴离子，用于消灭病毒等外来入侵者，起到保护机体的作用，而过剩的超氧阴离子则成为促使人体细胞衰老的自由基。人体自身可以产生内源性抗氧化物质催化分解超氧阴离子自由基，使人体达到安全平衡状态。但随着年龄的 增长，人体产生的内源性抗氧化水平会逐渐降低，此时过剩的超氧阴离子自由基就会攻击细胞膜上的不饱和磷脂，使细胞过早地老化，产生色素沉积，形成老年斑，导致人体衰老。
发明内容
本申请的目的在于提供一种具有清除自由基功效的组合物及其应用。
为解决上述技术问题，本申请的第一方面提供一种具有清除自由基功效的组合物，以质量份计，该组合物包含：3-o-乙基抗坏血酸：1～3份；抗坏血酸四异棕榈酸酯：2～4份；生育酚：0.3～0.7份；乙酰半胱氨酸：0.1～0.3份。
优选地，本申请提供的具有清除自由基功效的组合物，以质量份计包含：3-o-乙基抗坏血酸：2份；抗坏血酸四异棕榈酸酯：3份；生育酚：0.5份；乙酰半胱氨酸：0.2份。
本申请的第二方面提供了本申请第一方面所述的具有清除自由基功效的组合物在化妆品和/或护肤品中的应用。
本申请所提供的应用中，所述化妆品和/或护肤品施用于皮肤外表面。
优选地，本申请所提供的应用中，所述化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
优选地，本申请所提供的应用中，所述具有清除自由基功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料中的一种或几种。
优选地，本申请所提供的应用中，所述化妆品和/或护肤品还包括：表面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、营养性添加剂中的一种或几种。
相对于现有技术而言，本申请采用3-o-乙基抗坏血酸1～3份、抗坏血酸四异棕榈酸酯：2～4份、生育酚：0.3～0.7份、乙酰半胱氨酸：0.1～0.3份，实现协同增效作用。因而，将本申请提供的具有清除自由基功效的组合物应用于化妆品和/或护肤品中，可在较低施用量的情况下发挥明显的皮肤抗氧化、抗衰老作用。
附图说明
图1为Et-VC组、VC-IP组、VE组、NAC组、Et-VC/VC-IP/VE/NAC组的DPPH自由基清除活性对比图；
图2为Et-VC组、VC-IP组、VE组、NAC组、对比组1的DPPH自由基清除活性对比图；
图3为Et-VC组、VC-IP组、VE组、NAC组、对比组2的DPPH自由基清除活性对比图。
具体实施方式
为使本申请的目的、技术方案和优点更加清楚，下面将对本申请的各实施方式进行详细的阐述。然而，本领域的普通技术人员可以理解，在本申请各实施方式中，为了使读者更好地理解本申请而提出了许多技术细节。但是，即使没有这些技术细节和基于以下各实施方式的种种变化和修改，也可以实现本申请所要求保护的技术方案。
本申请的实施方式提供一种具有清除自由基功效的组合物，以质量份计，该组合物包含：3-o-乙基抗坏血酸：1～3份；抗坏血酸四异棕榈酸酯：2～4份；生育酚：0.3～0.7份；乙酰半胱氨酸：0.1～0.3份。
在本申请部分实施方式中，所提供的具有清除自由基功效的组合物， 以质量份计包含：3-o-乙基抗坏血酸：2份；抗坏血酸四异棕榈酸酯：3份；生育酚：0.5份；乙酰半胱氨酸：0.2份。
本申请的实施方式所提供的具有清除自由基功效的组合物可应用于化妆品和/或护肤品中，所述化妆品和/或护肤品施用于皮肤外表面。
在本申请部分实施方式中，应用所述具有清除自由基功效的组合物的化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
本申请部分实施方式中，所述具有清除自由基功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料、油脂原料、溶剂原料中的一种或几种。
其中，所述油质原料可以是下述基质原料中的任意一种或几种组合：大豆油、橄榄油、杏仁油、蓖麻油、花生油、棉籽油、茶籽油、霍霍巴油、鳄梨油、椰子油、棕榈油、可可脂、米糠油、月见草油、小麦胚芽油、玉米胚芽油、棕榈蜡、木蜡、水貂油、海龟油、蛇油、蛋黄油、牛脂、马脂、猪脂、鹿脂、卵磷脂、羊毛脂、蜂蜡、鲸蜡、虫胶蜡、石蜡、凡士林、微晶蜡、地蜡、角鲨烷、羊毛脂蜡、羊毛脂醇、羊毛脂醇酯化物、乙酰化羊毛脂、乙酰化羊毛脂醇、羊毛脂酸、羊毛脂酸酯化物、聚氧乙烯羊毛脂、聚氧乙烯羊毛醇醚、聚氧丙烯羊毛醇醚、氢化羊毛脂、烷氧基化氢化羊毛脂、硅油(如：二甲基硅油、八甲基硅油、甲基苯基聚硅氧烷)、聚硅氧烷-聚氧烷基嵌段共聚物、甲基含氢硅油、Cn脂肪酸及其酯、Cn脂肪醇及其酯；其中，n≥10，并优选为10-24，更优选为12-20，如14、16、17、18。所述Cn脂肪酸如：月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、油酸、亚油酸、芥酸、癸酸。所述Cn脂肪醇如：月桂醇、鲸蜡醇、硬脂醇、异硬脂醇、油醇、椰油醇。所述Cn脂肪酸酯、Cn脂肪醇酯如：肉豆蔻酸异丙酯、肉豆蔻酸肉豆蔻酯、十四醇乳酸酯、十六醇乳酸酯、棕榈酸异丙酯、棕榈树异辛酯、硬脂酸丁酯、硬 脂酸异辛酯、硬脂酸单甘油脂、硬脂酸聚乙二醇酯、油酸癸酯、辛酸甘油酯、癸酸甘油酯、甘油三油酸酯。
所述粉质原料可以是下述基质原料中的任意一种或几种组合：云母、碳酸镁、钛白粉、锌白粉、氢氧化钙、碳酸钙、高岭土、硬脂酸钙、硬脂酸锌、硬脂酸镁、滑石粉、二氧化硅、氢氧化铝、焦磷酸钙、碳酸氢钙、膨润土。
其中，所述油脂原料可以是植物油质原料、动物油质原料、矿物油质原料、合成油质原料、半合成油质原料中的任意一种或几种。如：淀粉、环糊精、黄原胶、角叉胶、瓜尔豆胶、阿拉伯胶、黄芪胶、琼脂、虫胶、海藻酸钠、明胶、甲基纤维素、乙基纤维素、羧甲基纤维素钠、羟乙基纤维素、羟丙基纤维素、聚纤维素醚季铵盐、聚乙烯醇、聚乙烯吡咯烷酮、聚丙烯酸钠、聚氧乙烯、乙烯基吡咯烷酮/乙酸乙烯酯共聚物、卡波树脂、硅酸铝镁、C12-15醇苯甲酸酯。
其中，所述溶剂原料可以是下述基质原料中的任意一种或几种组合：水、乙醇、丙醇、异丙醇、异丁醇、乙二醇、1,2-丙二醇、环己烷、二氯二氟甲烷、四氟二氯乙烷、乙二醇单乙醚、二甘醇单乙醚、丙酮、甲基异丁基甲酮、醋酸乙酯、醋酸丁酯、乙酸戊酯、邻苯二甲酸二丁酯。
在本申请部分实施方式中，应用所述具有清除自由基功效的组合物的化妆品和/或护肤品还包括：表面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、pH值调节剂、营养性添加剂中的一种或几种。
其中，所述表面活性剂可以是：1)阴离子表面活性剂，如：C12-14脂肪醇硫酸铵、C12-14脂肪醇聚氧乙烯醚羧酸盐、十二烷基硫酸二乙醇胺盐、C12-14脂肪醇聚氧乙烯醚硫酸铵、十二烷基硫酸三乙醇胺、醇醚邻苯二甲酸单酯钠盐、酰化肽、月桂酸单乙醇酰胺硫酸钠、正辛醇聚氧丙烯醚琥珀酸 单酯磺酸盐、十六醇琥珀酸单酯磺酸盐、磺基琥珀酸单酯二钠盐、琥珀酸单十八酯二钠盐、醇醚磺基琥珀酸单酯二钠盐、脂肪醇聚氧乙烯醚(钠)磺基琥珀酸单酯铵盐、油酰胺磺基琥珀酸单酯二钠盐、N-酰基谷氨酸钾盐、吡咯烷酮羧酸钠、聚丙烯酸钠、脂肪醇聚氧乙烯醚磷酸酯、脂肪醇聚氧乙烯醚磷酸单酯及其盐、脂肪醇聚氧乙烯醚磷酸单酯乙醇胺盐、烷基酚聚氧乙烯醚磷酸单酯及其盐、壬基酚醚磷酸单酯乙醇胺盐、烷基磷酸酯盐、邻苯二甲酸单月桂醇酯盐、十一碳烯酸锌、阴离子型氨基酸表面活性剂、N-C12-18酰基谷氨酸钠、N-月桂酰L-丙氨酸钠、N-硬脂酰基谷氨酸单钠盐、N-椰子酰基谷氨酸单钠盐、N-混合脂肪酰基谷氨酸单钠盐、长直链烷基芳醚磺酸钠TH；2)非离子表面活性剂，如：油醇聚氧乙烯醚(如乳化剂VO系列)、烷基酚聚氧乙烯醚(如乳化剂OPE系列)、硬脂酸聚氧乙烯酯、乙二醇单硬脂酸酯、二硬脂酸乙二醇酯、聚乙二醇单硬脂酸酯、聚乙二醇双硬脂酸酯、聚乙二醇双月桂酸酯、双硬脂酸甘油酯、三硬脂酸甘油酯、蔗糖硬脂酸酯、聚氧乙烯甘油醚单硬脂酸酯、N-月桂酰基谷氨酸双烷醇酯、苯甲酸脂肪醇酯、山梨醇单脂肪酸酯(司盘，Span，如司盘-20、司盘60、司盘65、司盘-80、司盘-83、司盘-85)、聚氧乙烯醚山梨醇单脂肪酸酯(吐温，Tween，如吐温-20、吐温-40、吐温-60、吐温-61、吐温-80)、二乙醇胺、三乙醇胺、脂肪酸单乙醇酰胺(如椰子油酸单乙醇酰胺、月桂酸单乙醇酰胺、棕榈酸单乙醇酰胺)、椰子油酸二乙醇酰胺、月桂酸二乙醇酰胺、甲基葡萄糖苷硬脂酸酯、聚氧乙烯甲基葡萄糖苷硬脂酸酯、乙二醇葡萄糖苷硬脂酸酯、丙二醇基葡萄糖苷硬脂酸酯、C8-16烷基葡萄糖苷、甘油葡萄糖苷硬脂酸酯、琥珀酸月桂醇醚单酯、月桂醇甘油酯聚氧乙烯醚、硬脂酸甘露醇酐酯、聚氧乙烯貂油、貂油酸甲酯、貂油酸异丙酯；3)阳离子表面活性剂，如氯化C12-18烷基三烷基铵(如氯化十六烷基二甲基烷基铵、氯化十八烷基三甲基铵、氯化十六烷基三甲基铵)、氯化油酰胺丙基二羟丙基二甲基铵、氯化二羟丙基二甲基十二烷基铵、氯化二甲基二烯丙基铵/丙烯酰胺共聚物、氯化油酰胺丙基-2,3-二羟丙基二甲基 铵、乙基硫酸油酰胺丙基二甲基乙基铵、氯化双C12-18基二甲基铵、聚季铵-11调理剂、M-505聚季铵-7调理剂、貂油酰胺丙基胺-壳聚糖、貂油酰胺丙基胺-水解蛋白、阳离子蛋白肽、瓜尔胶-羟丙基三甲基氯化铵盐、聚纤维素醚季铵盐、单油酸三乙醇胺酯、DNP系列；4)两性离子表面活性剂，如C12-18烷基二羟乙基甜菜碱、N-C12-18烷基-N-(2-羟乙基)-N-(2-甲酰胺基乙基)铵乙酸盐、C12-18烷氧基羟丙基甜菜碱、氧化椰油酰胺基丙基胺、椰油羟乙基磺酸钠、氧化十八烷基二甲基胺、椰油酰二乙醇胺氧化胺、N-烷基-β-氨基丙酰二乙醇胺、氧化月桂酰基丙胺、N-C12-18酰基谷氨酸、羟乙基癸酸咪唑啉甜菜碱、羟乙基肉豆蔻酸咪唑啉甜菜碱、羟乙基棕榈酸咪唑啉甜菜碱。
其中，所述香精与香料可以是选自：1)天然香料，如苦艾油、桉叶油、八角茴香油、白柠檬油、白丁香浸膏、丁香油、白兰花油、白兰花浸膏、白兰叶油、柏木油、薄荷油、苍术硬脂、草木犀浸膏、橙叶油、丁香罗勒油、大花茉莉浸膏、冬青油、当归油、当归浸膏、地衣酸、地檀香油、玳玳花油、玳玳叶油、风信子浸膏、枫香浸膏、芳樟油、黄樟油、甘松油、桂花浸膏、红橘油、胡萝卜籽油、海狸香、黄葵油、黄兰油、藿香油、黄蒿油、九里香浸膏、金合欢浸膏、姜油、橘子油、椒样薄荷油、菊花浸膏、枯茗油、龙涎香、留兰香油、灵猫香、腊梅油、铃兰浸膏、木香根油、米兰花油、玫瑰香叶油、玫瑰油、茉莉浸膏、牡蛎油、墨红浸膏、墨红油、柠檬桉叶油、柠檬油、七里香浸膏、楠叶油、芹菜籽油、肉豆蔻油、肉桂油、山苍子油、水貂香膏酊、水仙浸膏、松针油、树苔浸膏、素馨浸膏、麝香、麝鼠香膏、甜橙油、檀香油、晚香玉油、晚香玉浸膏、万寿菊精油、小茴香油、血桧木油、香茅油、香叶油、康乃馨油、紫苏油、香紫苏油、香附油、香柠檬薄荷油、岩兰草油、香薷油、香薷草浸膏、悬钩子蔷薇花油、悬钩子蔷薇花浸膏、薰衣草油、月桂叶油、依兰依兰油、芫荽籽油、鱼香草油、岩蔷薇浸膏、岩桂油、柚皮油、鸢尾油、烟草花油、香橼叶油、栀子花浸膏、紫穗槐油、紫罗 兰叶浸膏、愈创木酚、4-甲基愈创木酚、4-乙基愈创木酚、天然香兰素、樟脑；2)合成香精，如柠檬烯、长叶烯、乙位石竹烯、异长叶烯、异长叶烯酮、异长叶烷酮、溴代苏合香烯、二苯甲烷、二苯醚、间甲基二苯醚、丁香酚、异丁香酚、乙位萘甲醚、乙位萘乙醚、对甲基茴香醚、异丁香酚苄醚、玫瑰醚、檀香醚、水仙醚(或茉莉醚)、降龙涎香醚、甲基柏木醚、环氧柏木烷、埃林塔尔、埃林塔尔N、3-己烯醇、癸醇、月桂醇、苯甲醇、苯乙醇、对甲氧基苄醇、肉桂醇、二甲基苄基甲醇、香叶醇、橙花醇、芳樟醇、玫瑰醇、松油醇、香茅醇、消旋薄荷脑、薄荷脑、冰片、柏木醇、柏木烷酮、甲基柏木酮、3,4-二氧亚甲基苯甲醇、α-甲基-3,4-二氢亚甲基苯丙醛、2-叔丁基-4-甲基环己醇、香紫苏醇、檀香醇、3-甲基-5-(2’,2’,3’-三甲基环戊烯-1’-基)戊-2-醇、苯甲醛、苯乙酮、苯乙酸、对甲基苯乙酮二苯甲酮、月桂醛、肉桂醛、肉桂酸、α-戊基-β-苯基丙烯醛、α-苯基丙醛、铃兰醛、新铃兰醛、兔耳草醛、3,4-二氧亚甲基苯甲醛、3,7-二甲基-6-辛烯醛、3,7-二甲基-7-羟基辛醛、柠檬醛、乙位环高柠檬醛、艾薇醛、柑青醛、4-羟基-3-甲氧基苯甲醛、α-二氢突厥酮、β-突厥烯酮、紫罗兰酮、6-甲基紫罗兰酮、橙花酮、香芹酮、胡椒基丙酮、覆盆子酮、药草酮、顺式茉莉酮、庚醛乙二醇缩醛、苯乙醛二甲缩醛、辛醛二甲缩醛、α-戊基桂醛二甲缩醛、柠檬醛二甲缩醛、柠檬醛二乙缩醛、茴香醛二甲缩醛、蘑菇醛、叶青素、风信子素、苹果酯、吉普缩酮、甲酸香叶酯、甲酸香茅酯、阿弗曼酯、乙酸叶醇酯、乙酸苄酯、乙酸苯乙酯、乙酸肉桂酯、乙酸松油酯、乙酸龙脑酯、乙酸芳樟酯、乙酸香叶酯、乙酸对叔丁基环己酯、乙酸邻叔丁基环己酯、乙酸香根酯、乙酸琥珀酯、乙酸阿弗曼酯、乙酸十氢萘酯、乙酸柏木酯、丙酸香叶酯、乙酸三氯甲基苯甲酯、丁酸苄酯、丁酸叶醇酯、丁酸香叶酯、2-甲基戊酸苯乙酯、异丁酸对甲酚酯、2-壬烯酸甲酯、苯甲酸乙酯、苯甲酸甲酯、苯甲酸叶醇酯、苯甲酸苄酯、苯乙酸乙酯、苯乙酸对甲酚酯、肉桂酸乙酯、水杨酸丁酯、水杨酸异丁酯、水杨酸异戊酯、水杨酸叶醇酯、水杨酸己酯、水杨酸苄酯、水杨酸苯乙酯、水 杨酸环己酯、茉莉素、茉莉酯、2-戊基环戊酮醋酸甲酯、橙花素、γ-壬内酯、γ-十一内酯、香豆素、吲哚、3-甲基吲哚、3,7-二甲基-2,6-辛二烯腈、对甲氧基苯甲腈、二甲苯麝香、酮麝香、2,6-二硝基-3-甲氧基-4-叔丁基甲苯、1,1-二甲基-4-乙酰基-6-叔丁基茚满、六甲基三环异色满麝香、1,1,2,3,3,6-六甲基-5-乙酰基茚满、麝香-T、麝香-M、麝香-L、麝香-F、11-氧杂环十六内酯、吐纳麝香；3)香精，玫瑰香精、茉莉香精、檀香香精、白兰香精。
其中，所述色素可以是选自如下组分中的一种或几种：二氧化锆、醋酸铅、硝酸银、叶绿素、叶绿素铜、亚铁氰化铁、鸟嘌呤、坚牢红、食品红I、还原红I、胭脂红、食用杨梅红、食用樱桃红、食品红17、苋菜红、坚牢绿、食品黄3、食用柠檬黄、食用苏丹黄、食用靛蓝、食品蓝2、荧光桃红、β-胡萝卜素、指甲花、栀子红色素、栀子绿色素、栀子蓝色素、栀子黄色素、钛云母珠光颜料、氧化铁红、氧化铁黄、氧化铁黑、氧化铬绿、氧氯化铋、碱性桃红、银朱R、喹啉黄、紫草宁、紫胶红色素、群青、酸性红87、酸性绿25、酸性荧光黄、酸性橙7、金光红53、立索尔宝红BK、立索尔宝红2R、永固橙。
其中，所述防腐剂可以是选自如下组分中的一种或几种组合：山梨酸及其盐、丙酸盐、对羟基苯甲酸酯、苯甲醇、苯甲酸及其盐、5-氯-2-甲基-4-异噻唑啉-3-酮、-2-甲基-4-异噻唑啉-3-酮、卡松、咪唑烷基脲、脱氢醋酸及其盐、亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠、2,2-溴-2-硝基-1,3-丙二醇(布罗波尔)、异抗坏血酸及其盐、曲酸及其酯、壬二酸、松萝酸、间苯二酚。
其中，所述抗氧剂可以是选自如下组分中的一种或几种组合：SOD、丁基羟基茴香醚、没食子酸酯、抗坏血酸及其盐、异抗坏血酸及其盐、谷甾醇、芦丁、生育酚。
其中，所述保湿剂可以是选自如下组分中的一种或几种组合：1,2-丁二醇、山梨醇、木糖醇、甘油、乳酸盐、聚乙二醇、DL-吡咯烷酮羧酸盐、 D-葡萄糖、曲酸及其酯、尿酸、乳清酸、果胶酸、昆布氨酸、胶原蛋白、海藻糖、12-羟基硬脂酸胆甾醇酯。
其中，所述紫外线吸收剂可以是选自如下组分中的一种或几种组合：吡哆醇盐酸盐、曲酸及其酯、芦丁、芦荟苷、咖啡氨酸、橙皮素、2-乙基己基水杨酸酯、2-乙基己基对二甲基氨基苯酸酯、2-乙基己基对甲氧基肉桂酸酯、水杨酸苯酯、基邻氨基苯甲酸酯、2-羟基-4-甲氧基二苯甲酮、2-羟基-4-甲氧基二苯甲酮-5-磺酸、2-羟基-4-辛氧基二苯甲酮。
其中，所述螯合剂可以是选自如下组分中的一种或几种组合：EDTA二钠盐、双吡啶硫酮锌、硫代硫酸钠。
其中，所述收敛剂可以是选自如下组分中的一种或几种组合：苯酚对磺酸锌、羟基氯化铝、硫酸铝、硫酸铝钾、硫酸锌、氯化铝、氯化锌、卡拉明。
其中，所述助渗剂可以是选自如下组分中的一种或几种组合：二甲基亚砜、1-正十二烷基氮杂环庚-2-酮、2-氨基乙醇。
其中，所述pH值调节剂可以是选自如下组分中的一种或几种组合：酒石酸、乳酸、硼酸、琥珀酸。
其中，所述营养性添加剂可以是选自如下组分中的一种或几种组合：甲壳素、壳聚糖、丝肽、丝素、卵磷脂、表皮细胞生长因子、海藻糖、乳糖、珍珠粉、胶原蛋白、胶原水解物、透明质酸及其盐、蛋膜素、植物甾醇、氯化溶菌酶、硫酸软骨素钠、蜂乳、蜂胶、蜂蜜、甘草酸、甘草次酸、γ-亚麻酸、曲酸及其酯、赤霉酸、甘氨酸、L-天冬氨酸及其盐、L-胱氨酸及其盐、DL-丙氨酸、L-丝氨酸、DL-丝氨酸、L-蛋氨酸、DL-蛋氨酸、L赖氨酸盐酸盐、L苏氨酸、肌醇、麦芽醇、泛酸盐、D-泛醇、视黄醇醋酸酯、尿囊素、吡哆醇盐酸盐、可的松、烟酰胺、维生素、雌激素、苏木素、尿酸、乳清酸、芦荟苷、壬二酸、松香酸、松萝酸、昆布氨酸、胸腺素、茜草素、柠檬烯、 黄芩黄素、脱氧胆酸、愈疮木萸、橙皮素、鞣酸、二硫化硒、茶皂苷、柠檬酸三乙酯、丁基羟基甲苯、植酸、聚氯化二甲基二烯丙基铵。
实验例
实验例中用到的仪器、试剂盒、细胞和试剂均可通过公开的商业渠道购买，为本领域常用的仪器、试剂盒、细胞和试剂，仪器和试剂盒的使用操作步骤按照其说明书的指导进行。
实验目的
使用DPPH自由基清除试验，评价实验组和对照组对自由基的清除作用。
主要实验原料和试剂
原料1：名称：3-o-乙基抗坏血酸；代号：Et-VC；性状：白色至类白色结晶粉末。
原料2：名称：抗坏血酸四异棕榈酸酯；代号：VC-IP；纯度：性状：无色至淡黄色透明油状液体。
原料3：名称：生育酚；代号：VE；性状：黄色至棕红色粘性油状液体。
原料4：名称：乙酰半胱氨酸；代号：NAC；性状：白色结晶性粉末。
试剂1：DPPH：100mg/瓶，购自：Sigma。
实验方法
(1)DPPH工作液配制：取一管10mM的DPPH溶液(500ul)，用无水乙醇稀释至25ml(0.2mM)。
(2)待测物或对照品用DMSO或者ddH 2O配制成母液，再用无血清培养基或者PBS稀释至合适浓度，分别获得如表1所示组别的待测物：
表1
Figure PCTCN2022125931-appb-000001
(3)使用DPPH试剂检测各实验组、对比组的待测物的自由基清除能力，各实验组、对比组分别设如下检测体系A、B、C、D(待测物用量20ul，DPPH用量180ul)：
A：不加待测物，只加DPPH，待测物体积用PBS代替；
B：待测物和DPPH都不加，待测物体积用PBS代替，DPPH体积用无水乙醇代替；
C：待测物和DPPH都加；
D：不加DPPH，只加待测物，DPPH体积用无水乙醇代替。
上述检测步骤在96孔板中进行，设置至少3复孔，加入所需的待测物、DPPH、PBS或无水乙醇后用排枪混匀，室温避光放置30min.
(4)对检测体系A、B、C、D于540nm下测定吸光度值(本实验以0.01mg/ml EGCG为阳性对照)，分别得到吸光度A、吸光度B、吸光度C、吸光度D，并计算各个实验组、对比组的DPPH清除活性(％)。
DPPH清除活性计算：
Figure PCTCN2022125931-appb-000002
(5)数据录入和统计分析：
采用Excel录入和Prism(Version7)软件包统计处理DPPH清除活性，以均数±标准误表示。各组间做单因素方差分析,认为p<0.05有统计学差异。
实验结果
图1为Et-VC组、VC-IP组、VE组、NAC组、Et-VC/VC-IP/VE/NAC组的DPPH自由基清除活性对比图；
图2为Et-VC组、VC-IP组、VE组、NAC组、对比组1的DPPH自由基清除活性对比图；
图3为Et-VC组、VC-IP组、VE组、NAC组、对比组2的DPPH自由基清除活性对比图。
图1中：n＝3，Mean±SEM。“**”表示与Et-VC/VC-IP/VE/NAC组相比p＜0.05，“***”表示与Et-VC/VC-IP/VE/NAC组相比p＜0.001。
图2、图3中：n＝3，Mean±SEM。“*”表示与对比组相比p＜0.05，“***”表示与对比组相比p＜0.001。
根据图1，采用Et-VC/VC-IP/VE/NAC组(其中，按质量份计：Et-VC：VC-IP：VE：NAC＝2:3:0.5:0.2)给药，与单独给药的Et-VC组、VC-IP组、VE组、NAC组相比，Et-VC/VC-IP/VE/NAC组的自由基清除活性显著上升，说明Et-VC/VC-IP/VE/NAC组在自由基清除活性方面具有协同增效作用。
根据图2和图3，采用对比组1(其中，按质量份计：Et-VC：VC-IP：VE：NAC＝2:3:0.5:0.01)给药，或对比组2(其中，按质量份计：Et-VC：VC-IP：VE：NAC＝2:3:0.5:0.02)给药，与单独给药的Et-VC组、VC-IP组、VE组、NAC组相比，对比组1和对比组2在自由基清除活性方面并不具有协同增效作用。
本领域的普通技术人员可以理解，上述各实施方式是实现本申请的具体实施例，而在实际应用中，可以在形式上和细节上对其作各种改变，而不偏离本申请的精神和范围。

Claims (7)

  1. 一种具有清除自由基功效的组合物，以质量份计包含：
    3-o-乙基抗坏血酸：1～3份；
    抗坏血酸四异棕榈酸酯：2～4份；
    生育酚：0.3～0.7份；
    乙酰半胱氨酸：0.1～0.3份。
  2. 根据权利要求1所述的具有清除自由基功效的组合物，以质量份计包含：
    3-o-乙基抗坏血酸：2份；
    抗坏血酸四异棕榈酸酯：3份；
    生育酚：0.5份；
    乙酰半胱氨酸：0.2份。
  3. 权利要求1至2中任一项所述的具有清除自由基功效的组合物在化妆品和/或护肤品中的应用。
  4. 根据权利要求3所述的应用，其中，所述化妆品和/或护肤品施用于皮肤外表面。
  5. 根据权利要求3所述的应用，其中，所述化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
  6. 根据权利要求3所述的应用，其中，所述具有清除自由基功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料中的一种或几种。
  7. 根据权利要求3所述的应用，其中，所述化妆品和/或护肤品还包括： 表面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、营养性添加剂中的一种或几种。
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WO2015046389A1 (ja) * 2013-09-27 2015-04-02 ロート製薬株式会社 新規ベンジリデンアゾリジン誘導体またはその塩を含む皮膚外用組成物
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WO2017069157A1 (ja) * 2015-10-20 2017-04-27 ロート製薬株式会社 皮膚外用組成物
CN107334649A (zh) * 2016-04-28 2017-11-10 日本乐敦制药株式会社 外用组合物
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WO2015046389A1 (ja) * 2013-09-27 2015-04-02 ロート製薬株式会社 新規ベンジリデンアゾリジン誘導体またはその塩を含む皮膚外用組成物
WO2017069157A1 (ja) * 2015-10-20 2017-04-27 ロート製薬株式会社 皮膚外用組成物
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