CN107296769B - Application of praecox extract - Google Patents
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- CN107296769B CN107296769B CN201710629720.8A CN201710629720A CN107296769B CN 107296769 B CN107296769 B CN 107296769B CN 201710629720 A CN201710629720 A CN 201710629720A CN 107296769 B CN107296769 B CN 107296769B
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Images
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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
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Dermatology (AREA)
- Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a praecox extract and application thereof in cosmetics, wherein the praecox extract is a praecox water extract, a praecox alcohol extract or any mixture of the praecox water extract and the praecox alcohol extract. The preparation method comprises ultrasonic extracting or extracting under heat preservation the stem marrow of the praecox in an extraction solvent, filtering, concentrating, and drying to obtain the extract. The praecox extract has the functions of removing free radicals and inhibiting tyrosinase activity, has the functions of resisting aging and whitening, and can be used for preparing cosmetics with the functions of resisting aging and whitening.
Description
Technical Field
The invention relates to the field of plant extracts and cosmetics, in particular to a Stachys extract and application thereof in cosmetics.
Technical Field
Recent studies have demonstrated that pigmentation is caused by ultraviolet activation of melanin-producing enzymes in melanocytes in the epidermis to produce pigments. Melanocytes located in the basal layer of the human epidermis contact and associate with surrounding keratinocytes to form an "epidermal melanin unit", in which melanin is a nitrogen-containing complex that is synthesized in the melanosome. Generally, melanin is an important factor in determining skin color in terms of quantity and quality of melanin. During melanin production, tyrosinase converts tyrosine into dopa and dopaquinone, which are then polymerized by non-enzymatic oxidation to produce macromolecular melanin. Therefore, the melanin generation can be limited and the skin pigmentation can be improved by inhibiting the activity of tyrosinase.
Praecox is a family specific to east asia and belongs to the family of praecox (Stachyuraceae), genus Stachyuraceae. This family is about 15, 6 varieties. The Chinese is made of 10 varieties and 5 varieties, and is produced in regions of south provinces of Qinling mountain. The rambutan variety is various, including rambutan (i.e. western domain rambutan hook. f. et Thoms ex Benth.), Chinese rambutan (S.chinensis Franch.), kombu (S.obova (Red.) hand. and-Mazz.), Yuye (S.retussus Yang.) hand. and Yunan (S.yunnanensis Franch.) etc., which are collectively called as "medulla junci", stem marrow entered drug, and has the efficacy of diuresis, lactation promotion and heat clearing.
Himalayan praecox (Stachyurus himalaicus hook. f. et Thoms ex Benth.) is also called as Stachyuratus praecox, rambutan kola, rambutan ramosissimum and the like, is a shrub, is distributed in provinces such as Shaanxi, Zhejiang, Hunan, Hubei, Sichuan, Guizhou, Taiwan, Guangdong, Guangxi, Yunnan, Xizang and the like, is grown under mountain slope broad-leaved forests with the elevation of 400-3000 meters or in shrubs, and is also distributed in North India, Nipol, Plumbum and Burmese. Its stem marrow is white, and can be used as Chinese medicine "Tongcao", and has the functions of promoting diuresis, promoting lactation and clearing damp-heat, and can be used for curing the diseases of febrile disease, polydipsia, scanty urine or anuresis, acute cystitis, nephritis, edema, dysuresia and galactostasis, etc., and its tender stem and leaf also can be used for curing venomous snake bite and fracture.
The current research on praecox is mainly focused on the pharmaceutical aspect, and the research on the extraction process, chemical components, pharmacological action and the like of praecox is limited; the pharmacological effects of the praecox reported in the literature are that the total polysaccharide active site improves the activity of whole blood SOD by reducing the content of aging-related metabolites (lipid peroxide products LPO and lipofuscin LF) and finally improves the antioxidation effect; the extraction of effective components of the praecox brevicaulis mainly to obtain total polysaccharide by adopting a water reflux extraction process. At present, no report is found on the research on the praecox and its chemical components for improving skin function, delaying skin aging, achieving anti-aging and whitening effects by scavenging free radicals and inhibiting tyrosinase activity, and applying the praecox and its chemical components to skin external preparations such as skin care products or cosmetics.
Disclosure of Invention
The present invention aims to provide a extract of praecox.
The present invention also uses the aforementioned extract of praecox as an external preparation for skin.
The technical scheme of the invention is as follows: a praecox extract is praecox water extract, praecox alcohol extract or their mixture.
The praecox is at least one of himalaya praecox, Chinese praecox, fallen egg leaf praecox, concave leaf praecox, Sichuan praecox or Yunnan praecox.
The alcohol extract of praecox extract is prepared by method I, comprising the steps of:
(1) ultrasonic extracting the stem pith of the praecox in alcohol solution;
(2) concentrating the filtrate and drying.
Preferably, in the step (1), the stem pith of the praecox is ultrasonically extracted with an alcohol solution under 300-1000W for 1-3 times, each time for 0.5-2 hours, and the filtrates are combined. The ultrasonic extraction can be carried out at 10-40 ℃, and is usually room temperature.
Preferably, the ratio of the alcohol solution to the stem pith of the praecox is 30-60L/kg per ultrasonic extraction.
Preferably, the alcohol solution is 60-85% ethanol solution.
Preferably, the concentration drying method in the step (2) is rotary evaporation drying or freeze drying, and the temperature of the rotary evaporation drying is 35-80 ℃, and more preferably 40-60 ℃.
The extract of praecox is prepared by method II, comprising the steps of:
(A) putting the stem pith of the praecox into water for heat preservation and extraction;
(B) concentrating the filtrate and drying.
Preferably, the heat-preservation leaching in the step (A) is as follows: the stem pith of the praecox is extracted for 1-3 times by water at 90-98 ℃, each time lasts for 0.5-2 hours, and the filtrates are combined. The preferable leaching temperature is 94-97 ℃.
The ratio of water to the stem pith of the praecox in each extraction is 30-80L/kg, preferably 35-75L/kg.
Preferably, the concentration drying method in step (B) is rotary evaporation drying or freeze drying, especially freeze drying.
The water content of the stem pith of the praecox in the methods I and II is not more than 10 percent, and the grain diameter is 25-800 meshes. More preferably, the water content does not exceed 5%.
The concentrating and drying method is rotary evaporation or freeze drying. Preferably, the temperature of the rotary evaporation is 40-60 ℃.
The praecox extract has the effects of removing free radicals, inhibiting tyrosinase activity and inhibiting melanin generation, and can be used as an active ingredient for preparing skin external preparations, such as skin care products or cosmetics. At a content of 0.05%, the efficiency of scavenging free radicals exceeds 50%; when the content is 0.001%, the tyrosinase activity can be obviously inhibited, the melanin content is obviously reduced, and the whitening function is realized.
Therefore, the extract of the praecox of the present invention can be used as an active ingredient for whitening or anti-aging function, and can be used for preparing skin external preparations.
In the external preparation for skin, the addition amount of the extract of Stachys praecox is 0.0001-5 wt%, preferably 0.005-2 wt%.
The skin external preparation can be cosmetic water, essence, lotion, cream, etc.
The invention has the advantages that the provided extract of the praecox has good functions of removing free radicals, inhibiting tyrosinase activity, inhibiting melanin generation, improving pigmentation and the like, and has good whitening and anti-aging effects; and is natural, safe and effective, does not irritate skin, and can be used as functional additive to be added into cosmetics or skin care products.
The preparation method of the praecox extract is simple and easy to implement, has low cost, and the obtained extract has high activity and can exert the effect under the condition of extremely low content.
Drawings
FIG. 1 is a diagram of co-culture of primary melanoma cells and keratinocytes in example 4
FIG. 2 is a graph of the mean optical density of the co-culture plots of the cells of example 4.
FIG. 3 is a graph showing the effect of the extract of Stachys praecox of example 4 on the inhibition of tyrosinase activity.
Detailed Description
The invention is further illustrated with reference to specific examples. The examples are provided to aid in the understanding of the invention, and are not to be construed as limiting the invention. The experimental method not indicating the specific operation method and conditions is selected according to the conventional operation method and experimental conditions or the commercial instructions.
Example 1 alcohol extract of Stachys praecox
Drying the stem pith of the Himalayan praecox until the water content is less than or equal to 5%, taking 100g of the dried stem pith, crushing the dried stem pith into coarse powder with the particle size of 25-800 meshes, putting the coarse powder into 4L of 70% (v/v) ethanol aqueous solution, and performing ultrasonic extraction at room temperature for 1 hour at 500W;
filtering to obtain solid, and repeating the above extraction steps once.
The filtrates are combined and rotated to dryness at 40-60 ℃ to obtain about 2g of ethanol extract of himalayan praecox.
Example 2 aqueous extract of Stachys praecox
Drying the stem pith of the Himalayan praecox until the water content is less than or equal to 5%, taking 100g of the dried stem pith, crushing the dried stem pith into coarse powder with the particle size of 25-800, putting the coarse powder into 6L of deionized water, heating the coarse powder in a water bath at 96 ℃ for 1 hour, and filtering and separating solid and filtrate;
the above extraction steps were repeated once for the solid.
Mixing filtrates, concentrating, and freeze drying to obtain about 2g of Himalayan Stachys water extract.
EXAMPLE 3DPPH scavenging Activity test
1, 1-Diphenyl-2-trinitrophenylhydrazine (1, 1-Diphenyl-2-piperidinylhydrazyl, DPPH) is a stable nitrogen center organic free radical, and an ethanol solution of the stable nitrogen center organic free radical is dark purple and has strong absorption at about 517 nm; in the presence of the free radical scavenger, DPPH absorption is diminished by its single electron pairing, and gradually disappears. The degree of color fading of the DPPH ethanol solution is in a quantitative relation with the number of electrons received by the DPPH ethanol solution, so that a spectrophotometer can be used for carrying out rapid quantitative analysis, detecting the condition of scavenging free radicals and evaluating the capacity of the antioxidant for scavenging free radicals, the antioxidant capacity and the anti-aging capacity.
Preparing a DPPH ethanol solution: 6mg of DPPH was weighed out accurately, dissolved in 95% ethanol and placed in a 50mL volumetric flask, and dissolved with the aid of ultrasound. When in use, the mixture is diluted by four times by 95% ethanol to obtain a DPPH ethanol solution with the concentration of 3mg/100mL (76 mu M), and the DPPH ethanol solution is stored in a dark place (0-4 ℃).
Pretreatment of a sample to be detected: the product of example 1 or 2 was dissolved in distilled water and diluted with distilled water to a suitable concentration for use.
The experimental steps are as follows: adding 2mL of sample solution to be detected into a test tube, then adding 2mL of ethanol solution of LDPPH, and uniformly mixing; the content of Himalayan praecox extract in the mixed system is 0.5 mg/mL. Reacting in dark place at room temperature for 30min in the dark place in a dark place, and measuring absorbance Abs at 525 nm; in addition to the sample group, a sample solvent control group, a blank control group and a solvent control group (solvent for DPPH) were provided, respectively.
Clearance I (%) ═ 1- (T-T)0)/(C-C0)]×100％
The clearance rate is calculated according to the formula, the larger the clearance rate is, the stronger the oxidation resistance is, and the clearance rate is more than or equal to 10 percent, the oxidation resistance is considered to be realized.
In the formula: t is0: adding the absorbance of equal volume of 95% ethanol into the sample solution to be detected (sample solvent control group);
t: adding the absorbance of the sample solution to be detected and an isopyknic DPPH solution (sample group);
C0: absorbance of distilled water plus equal volume of 95% ethanol (solvent control);
c: absorbance of distilled water plus an equal volume of DPPH solution (blank control).
TABLE 1 radical scavenging effect of extract of praecox in western region
Example 4 whitening Activity test
The keratinocyte and melanoma cell co-culture model is a recognized model for simulating 'epidermal melanin unit' in vitro at present and is closer to the physiological characteristics of skin. The method comprises the steps of mixing and inoculating cultured primary melanoma cells and keratinocyte cells according to a certain proportion, and co-culturing to form an in vitro model of an epidermal melanin unit; and then adding a sample for treatment, and detecting the melanin content and the activity of tyrosinase by a dopa staining method.
The "epidermal melanin unit" in vitro model (primary melanoma cells and keratinocytes were mixed and inoculated at a ratio of 1: 4) was divided into a blank group (NT) without any addition, a positive control group to which Arbutin (Arbutin, 500 μ g/mL) was added, and a sample group to which the alcohol extract of Stachys japonicus (10 μ g/mL) of example 1 was added. And (3) respectively culturing the in vitro models of each group for 6-7 days, and detecting the melanin content and the activity of tyrosinase by using a dopa staining method.
The photograph of the cell co-culture is shown in FIG. 1, and the cell growth state of the sample group treated with the himalayan praecox extract is good, which indicates that the extract has no cytotoxicity; and the melanocytes of the sample group became lighter in color compared to NT, similar to that of the positive control group (arbutin 500. mu.g/mL).
The average optical density (white) analysis of the cell co-culture pictures by the software showed in fig. 2, the average optical density of the sample group was higher than that of the control group (NT) and the arbutin positive control group, which proved that the melanin content in the cells was decreased after the alcohol extract of the Stachys (10 ug/mL) treatment.
The tyrosinase activity inhibition rate is shown in figure 3. The results show that the tyrosinase activity is obviously inhibited in the sample group treated by the alcohol extract of himalayan praecox, which is far higher than that of the arbutin positive control group.
The results show that the alcohol extract of himalayan praecox has effects of inhibiting melanin generation, improving skin pigmentation, and whitening skin.
The results of the whitening activity test of the himalayan praecox extract of example 2 were the same as the results of the himalayan praecox alcohol extract, and the melanin content of the in vitro model of "epidermal melanin unit" was significantly reduced and the tyrosinase activity was inhibited after the himalayan praecox extract was added.
The alcohol extracts and water extracts of himalayan praecox of examples 1 and 2 were extracted with water in a ratio of 1: 1, the obtained compound also has the whitening effect, can obviously reduce the melanin content and inhibit the activity of tyrosinase.
The extract of Stachys praecox has effects of scavenging free radicals, preventing melanin formation, inhibiting tyrosinase activity, whitening skin and resisting aging.
Claims (2)
1. Use of a praecox extract for the preparation of a skin external preparation having whitening and anti-aging effects, characterized in that the praecox extract is a water extract, an alcohol extract or any mixture thereof; the alcohol extract of praecox is prepared by the method I, and comprises the following steps:
(1) putting the stem pith of the praecox into an alcohol solution for ultrasonic extraction; the alcoholic solution is 60-85% of ethanol solution;
(2) concentrating the filtrate and drying;
the praecox water extract is prepared by the method II, and comprises the following steps:
(A) putting the stem pith of praecox into water, preserving heat and leaching;
(B) concentrating the filtrate and drying;
the water content of the stem pith of the praecox in the methods I and II is not more than 10 percent, and the grain diameter is 25-800 meshes;
in the step (1) of the method I, the stem pith of the praecox is ultrasonically extracted for 1 to 3 times by using an alcoholic solution under the condition of 300 to 1000W, each time lasts for 0.5 to 2 hours, and the filtrates are combined; the dosage ratio of the alcoholic solution to the stem pith of the praecox is 30-60L/kg during each ultrasonic extraction;
in the method II, the heat-preservation leaching in the step (A) comprises the following steps: extracting the stem pith of praecox with water at 90-98 ℃ for 1-3 times, each time for 0.5-2 hours, and combining the filtrates; the ratio of water to the stem pith of the praecox in each extraction is 30-80L/kg.
2. The use according to claim 1, wherein said praecox is at least one of himalayan praecox, chinese praecox, peridok, praecox, surname or yunnan praecox.
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Address after: 201403 Shanghai City, Fengxian District Lifenglu No. 12 Patentee after: Shanghai Natural Hall Group Co.,Ltd. Country or region after: China Address before: 201403 Shanghai City, Fengxian District Lifenglu No. 12 Patentee before: JALA Group Co. Country or region before: China |