CN109363956B - Application of ganoderma lucidum extract, whitening water lotion and preparation method thereof - Google Patents
Application of ganoderma lucidum extract, whitening water lotion and preparation method thereof Download PDFInfo
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- CN109363956B CN109363956B CN201811639703.3A CN201811639703A CN109363956B CN 109363956 B CN109363956 B CN 109363956B CN 201811639703 A CN201811639703 A CN 201811639703A CN 109363956 B CN109363956 B CN 109363956B
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
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- 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
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
The invention relates to application of a Lei-Lai-Zhi extract, whitening water lotion and a preparation method thereof. The ganoderma lucidum extract of the present invention has an excellent effect of inhibiting tyrosinase activity. Furthermore, the whitening water lotion comprises the ganoderma lucidum extract, and the addition amount of the ganoderma lucidum extract is 0.01-12%, preferably 0.1-8% based on the total mass of the whitening water lotion. The whitening water lotion can reduce melanin generation and has excellent whitening effect. In addition, the whitening water lotion also has excellent moisturizing effect and no side effect on human body.
Description
Technical Field
The invention relates to application of a Lei-Lai-Zhi extract, whitening water lotion and a preparation method thereof, belonging to the field of cosmetics.
Background
Melanin (Melanin) is mainly produced by melanocytes (melanocytes) in the basal layer of the epidermis of human skin, and it reduces the damage of ultraviolet rays to the skin. The content and distribution of melanin determine the color of the skin. However, abnormal accumulation of melanin in the basal layer causes hyperpigmentation to cause melasma, freckles, age spots, etc., affecting the quality of life of people. With the progress of the research on melanin biosynthesis, researchers found that tyrosinase plays an important role in melanin biosynthesis, which is a major rate-limiting enzyme in the conversion of tyrosine and dopa to melanin, and that its overexpression is a major cause of hyperpigmentation. Therefore, inhibiting the activity of tyrosinase can block the biosynthesis reaction chain of melanin, reduce the generation of melanin and realize the whitening effect.
The main routes of skin whitening and skin care include the following for the pathways and influencing factors of melanin production: (1) reducing the number of melanocytes produced by inhibiting the proliferation of melanocytes of the skin; (2) the biosynthesis of melanin is reduced by inhibiting the catalytic activity of tyrosinase, a key enzyme for skin melanin generation, so that the purpose of whitening is achieved; (3) through sun protection, the damage of ultraviolet rays in sunlight to the skin (sunburn and sunburn) is prevented, and skin aging is slowed down; (4) is prepared by removing oxygen free radicals from skin, and relieving melanogenesis and skin aging by using antioxidant.
In the prior art, in order to prevent skin from becoming black, spots, freckles, etc., and to maintain the original whiteness of the skin, whitening lotions containing various whitening agents such as hydroquinone, kojic acid, arbutin, etc. have been proposed. However, if these substances are mixed in a large amount, problems may occur in the sense of use and safety. Such as hydroquinone, is very irritating to the skin; kojic acid is easy to discolor and has certain irritation to skin; arbutin has single action and poor effect, etc.
With the increase of attention of people to skin health, the development of a natural whitening agent with safety, stability, obvious effect and high cost performance has become one of the main research directions of the current pharmaceutical and cosmetic industries, and has a very good development prospect.
Disclosure of Invention
Problems to be solved by the invention
In view of the prior art, hydroquinone is very irritating to the skin; kojic acid is easy to discolor and has certain irritation to skin; the invention provides the application of the ganoderma extract for inhibiting the activity of tyrosinase.
The invention also provides a whitening water lotion which can reduce melanin generation and has excellent whitening effect. In addition, the whitening water lotion also has excellent moisturizing effect and no side effect on human body.
Further, the invention also provides a preparation method of the whitening water lotion, which is simple to operate and easy to obtain raw materials.
Means for solving the problems
The invention provides an application of a ganoderma lucidum extract in inhibiting tyrosinase activity.
According to the application of the invention, the application of the ganoderma lucidum extract in preparing cosmetics and/or skin care products for inhibiting tyrosinase activity is provided.
According to the use of the invention, the preparation method of the ganoderma lucidum extract comprises the following steps: pulverizing the raw material of the ganoderma, and extracting by using water and alcohol; preferably, the first and second electrodes are formed of a metal,
the mass ratio of the ganoderma loddigesii raw material, water and alcohol is 1: 0.05-5: 0.05 to 5; and/or
The extraction temperature is 80-100 ℃; and/or
The extraction times are 1-5 times; and/or
The extraction time is 1-5 hours.
The invention also provides whitening moisture lotion which comprises the ganoderma lucidum extract, wherein the addition amount of the ganoderma lucidum extract is 0.01-12%, preferably 0.1-8% by mass of the total mass of the whitening moisture lotion.
The whitening water lotion further comprises one or the combination of more than two of a humectant, a thickening agent, a pH regulator, an emulsifier, grease, a skin conditioner, an antioxidant, a chelating agent, a preservative and an aromatic; wherein the content of the first and second substances,
the addition amount of the humectant is 0.01-20% based on the total mass of the whitening water lotion; the addition amount of the thickening agent is 0.02-1.8%; the addition amount of the pH regulator is 0.01-1%; the addition amount of the emulsifier is 0.01-2%; the addition amount of the grease is 1-8%; the addition amount of the skin conditioner is 0.01-10%; the addition amount of the antioxidant is 0.01-2%; the addition amount of the chelating agent is 0.01-1%; the addition amount of the preservative is 0.01-1.5%; the addition amount of the aromatic is 0.005-0.5%.
The whitening water lotion comprises one or more of glycerin, dipropylene glycol, glyceryl polyether-26, sodium hyaluronate, panthenol, PEG/PPG-17/6 copolymer, butanediol, xylitol, betaine, glycerin polyacrylate, propylene glycol, mannose and trehalose; and/or
The grease comprises one or the combination of more than two of cyclopentadimethyl siloxane, cyclodimethyl siloxane, a compound of dimethyl siloxane and dimethiconol, oleyl erucate, shea butter, ethylhexyl palmitate, hydrogenated polydecene, cyclohexasiloxane, hydrogenated polyisobutene, C20-24 alkyl dimethyl siloxane, C13-14 isoparaffin and C12-15 alcohol benzoate.
The whitening water lotion comprises one or more than two of carbomer, xanthan gum, sclerotium rolfsii, behenyl alcohol, acryloyl dimethyl taurate/VP copolymer, hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer, acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer and a compound of isohexadecane and polysorbate-60; and/or
The pH regulator comprises one or more of aminomethyl propanol, arginine, citric acid, sodium citrate and sodium hydroxide.
The whitening water lotion comprises one or more of PEG-20 methyl glucose sesquistearate, sorbitan isostearate, polyglycerol-3 methyl glucose distearate, laureth-7 and isosteareth-20; and/or
The antioxidant comprises one or more of vitamin E, tocopherol acetate, butylated hydroxytoluene, lycopene, and ascorbic acid ethyl ether.
The whitening water lotion comprises one or more of hydrolyzed collagen, oat peptide, ceramide 3, fucus extract, chlorella fermentation product, chlorella extract, brown algae extract, witch hazel water, bisabolol, allantoin, ginkgo mistletoe leaf extract, cogongrass rhizome extract, serine, kelp extract, beta-glucan and cactus extract.
The invention also provides a preparation method of the whitening water lotion, which comprises the step of mixing the components of the whitening water lotion.
ADVANTAGEOUS EFFECTS OF INVENTION
The ganoderma lucidum extract has excellent effect of inhibiting tyrosinase activity;
further, the whitening water lotion of the present invention can reduce melanin production and has excellent whitening effects. In addition, the whitening water lotion also has excellent moisturizing effect and no side effect on human body.
In addition, the preparation method of the whitening water lotion is simple to operate, and raw materials are easy to obtain.
Drawings
FIG. 1 is a graph showing tyrosinase inhibition as a function of sample concentration of a Nephelus extract, wherein the sample concentration is the mass concentration of the Nephelus extract;
FIG. 2 is a graph showing the results of a mechanism for inhibition of tyrosinase activity by Leptospira extract;
FIG. 3 is a graph showing the results of a type of inhibition assay for tyrosinase activity by Leptospira extract;
FIG. 4 is a graph showing a comparison of the melanin change rates of application examples 1 to 5 and application comparative examples 1 to 5 of the present invention.
Detailed Description
Various exemplary embodiments, features and aspects of the invention will be described in detail below. The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.
Furthermore, in the following detailed description, numerous specific details are set forth in order to provide a better understanding of the present invention. It will be understood by those skilled in the art that the present invention may be practiced without some of these specific details. In other instances, methods, means, devices and steps which are well known to those skilled in the art have not been described in detail so as not to obscure the invention.
First embodiment
In a first embodiment of the invention, the use of an extract of Leptospira for inhibiting tyrosinase activity is provided. The extract of Lei Zhi has excellent inhibitory activity on tyrosinase, and has excellent whitening effect when used in cosmetics, especially in whitening water lotion.
The Himalayan mushroom is a natural substance formed by decomposing a specific plant by microorganisms for a long time, and mainly exists in Himalayan mountain areas. It is an effective and safe dietary supplement, restores energy balance and can prevent certain diseases. The Xilaizhi mainly comprises humic substances, including fulvic acid, which accounts for 60-80% of trace elements including total nutrient compounds and selenium with anti-aging property. Considering that Lei Zhi is a unique plant complex and is rich in fulvic acid, it is well known for its properties such as antioxidant, anti-inflammatory and memory enhancement.
The inventor of the invention finds that the ganoderma lucidum extract has excellent inhibitory effect on the activity of tyrosinase. The tyrosinase inhibition rate gradually increases with the increase of the mass concentration of the ganoderma extract, and tends to balance when reaching a certain degree.
In the present invention, the preparation method of the ganoderma lucidum extract may comprise: pulverizing the Lei-Lai-Zhi raw material, and extracting with water and alcohol to obtain Lei-Lai-Zhi extract; preferably, the first and second electrodes are formed of a metal,
the mass ratio of the ganoderma loddigesii raw material, water and alcohol is 1: 0.05-5: 0.05 to 5, preferably 1: 0.1-4: 0.1 to 4, more preferably 1: 0.2-2: 0.2-2. When the ratio of the ganoderma lucidum raw material, water and alcohol is 1: 0.05-5: when the content is within 0.05-5, the obtained Ganoderma extract has high content of effective components, and can further effectively inhibit tyrosinase activity.
In the present invention, the temperature of the extraction is 80 to 100 ℃, preferably 90 to 100 ℃. The number of times of extraction is 1-5 times, preferably 2-4 times. The extraction time is 1 to 5 hours, preferably 2 to 4 hours. In general, the alcohol may be methanol, ethanol, butanediol, and the like.
Further, filtering the obtained extract, storing in a storage tank, slowly flowing the filtrate in the storage tank into an external circulation tank and a settling tank for concentrating to obtain liquid components and precipitates, spray drying the liquid components, drying the precipitates, and sieving with a 80-100 mesh sieve to obtain the extract with proper particle size.
By adopting the preparation method, the prepared ganoderma lucidum extract is brown powder, and the ash content, the moisture content, the heavy metal content and the arsenic content are respectively below 10% and 5% by mass, 10ppm by mass, 2ppm by mass and 3ppm by mass, and the total number of bacteria is below 1000 CFU/g; the total amount of mould and yeast is below 100CFU/g, and no bacteria such as salmonella and Escherichia coli are existed.
The ganoderma extract prepared by the method can effectively inhibit the activity of tyrosinase.
Second embodiment
A second embodiment of the present invention provides a whitening moisture lotion comprising the ganoderma lucidum extract of the first embodiment. The addition amount of the ganoderma lucidum extract is 0.01-12%, preferably 0.1-8% based on the total mass of the whitening water lotion, for example: the content of the extract of Lei-Ganoderma can be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 9%, etc. When the addition amount of the Ganoderma extract is 0.01-12%, the melanin content in skin is reduced by using the whitening water lotion. When the addition amount of the ganoderma extract is less than 0.01%, the content of melanin is reduced a little, and the whitening effect cannot be achieved; when the addition amount of the ganoderma lucidum extract is more than 12%, the content of the ganoderma lucidum extract is too high, the cost is too high, and the whitening effect cannot be further increased.
In the invention, the whitening water lotion further comprises one or the combination of more than two of a humectant, a thickening agent, a pH regulator, an emulsifier, grease, a skin conditioner, an antioxidant, a chelating agent, a preservative and an aromatic; the whitening water lotion has mild formula composition, and can fully exert the efficacy of the ganoderma lucidum extract. Specifically, the method comprises the following steps:
the addition amount of the humectant is 0.01-20% by the total mass of the whitening moisture lotion, and the whitening moisture lotion can play a role in moisturizing. In order to further exert the efficacy of the humectant, the humectant can be added in an amount of 1-18%, 2-16%, 3-14%, 4-15%, 5-14%, 6-13%, or 7-12%. When the content of the humectant is less than 0.01%, the moisturizing effect is not obvious; when the content of the humectant is more than 20%, the whitening moisture lotion has a sticky feeling.
In the invention, the humectant comprises one or more of glycerin, dipropylene glycol, glyceryl polyether-26, sodium hyaluronate, panthenol, PEG/PPG-17/6 copolymer, butanediol, xylitol, betaine, glycerin polyacrylate, propylene glycol, mannose and trehalose. The whitening water lotion disclosed by the invention has more excellent moisturizing and hydrating effects by using the combination of various humectants.
The thickener is added in an amount of 0.02 to 1.8% based on the total mass of the whitening water lotion, and for example, the thickener may be added in an amount of 0.1%, 0.2%, 0.4%, 0.5%, 0.6%, 0.8%, 1.0%, 1.2%, 1.5%, or the like. When the addition amount of the thickener is between 0.02 and 1.8 percent, the low-viscosity feeling and excellent use feeling are achieved, the dispersibility is good, and the absorption is fast. When the addition amount of the thickener is less than 0.02%, the whitening water lotion has thin texture and no sticky feeling; when the thickener is added in an amount of more than 1.8%, the whitening lotion is too thick and heavy, which may increase the burden on the skin.
The thickening agent comprises one or the combination of more than two of carbomer, xanthan gum, sclerotium rolfsii, behenyl alcohol, acryloyl dimethyl taurate/VP copolymer, hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer, acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, hydroxyethyl acrylate/acryloyl dimethyl taurate copolymer and a compound of isohexadecane and polysorbate-60.
According to the invention, the stability of the whitening water lotion can be improved by using the complex thickening agent such as hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, a complex of isohexadecane and polysorbate-60 and the like.
The addition amount of the pH regulator is 0.01-1% of the total mass of the whitening water lotion. The pH value of the whitening water lotion is more suitable for human skin by adding the pH regulator. Preferably, the amount of the pH adjuster of the present invention added may be 0.03 to 0.8%, 0.06 to 0.5%, 0.1 to 0.3%, or the like. When the amount of the pH adjuster added is more than 1% or less than 0.01%, whitening water lotion with an appropriate pH value cannot be obtained. The pH regulator comprises one or more of aminomethyl propanol, arginine, citric acid, sodium citrate and sodium hydroxide.
In the present invention, the emulsifier is added in an amount of 0.01 to 2% based on the total mass of the whitening water lotion, for example: may be 0.1 to 1.5%, may be 0.5 to 1.2%, etc. When the dosage of the emulsifier is less than 0.01%, the emulsification is insufficient, so that the system is unstable; when the dosage of the emulsifier is more than 2 percent, certain influence can be caused on the stability of the product.
In the present invention, the emulsifier includes one or a combination of two or more of PEG-20 methyl glucose sesquistearate, sorbitan isostearate, polyglyceryl-3 methyl glucose distearate, laureth-7, isosteareth-20.
In the invention, the addition amount of the grease is 1-8% of the total mass of the whitening water lotion. For example, the amount of the oil or fat added may be 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, or the like. The skin whitening lotion can reduce the evaporation of water on the skin surface and prevent skin chap by adding oil and fat. In addition, by adding oil or fat, a hydrophobic film can be formed on the skin surface to prevent the invasion of harmful substances from the outside. When the content of the oil is less than 1%, evaporation of moisture on the skin surface cannot be reduced, and intrusion of harmful substances cannot be effectively prevented; when the content of the oil is more than 8%, the whitening water is too greasy, and the use feeling is reduced.
In the present invention, the oil or fat includes one or a combination of two or more of cyclopentadimethylsiloxane, cyclomethicone, a compound of dimethicone and dimethiconol, oleyl erucate, shea butter, ethylhexyl palmitate, hydrogenated polydecene, cyclohexasiloxane, hydrogenated polyisobutene, C20-24 alkyl dimethicone, C13-14 isoparaffin, and C12-15 alcohol benzoate.
In order to further improve the efficacy of the whitening water lotion, the whitening water lotion further comprises the skin conditioner. The skin conditioner is added to calm the skin, so that the facial skin can be relieved from injury and redness and swelling, and wrinkles can be reduced. In addition, the whitening moisture lotion can improve the oil yielding condition of skin, is not greasy in texture and can adjust the water-oil balance.
The addition amount of the skin conditioner is 0.01-10% of the total mass of the whitening water lotion. Preferably, the skin conditioning agent may be added in an amount of 0.1 to 8%, may be 0.3 to 5%, may be 0.5 to 4%, and the like. When the amount of the skin conditioner added is less than 0.01%, the corresponding effect cannot be achieved.
In the present invention, the skin conditioner includes one or a combination of two or more of hydrolyzed collagen, avenin, ceramide 3, fucus extract, chlorella fermentation product, chlorella extract, brown algae extract, hamamelis water, bisabolol, allantoin, ginkgo mistletoe leaf extract, cogongrass rhizome extract, serine, macroalgae extract, beta-glucan, and cactus extract.
Wherein the avenin is a fragment of avenin and a component of avenin. Oat protein has unique function, and the amino acid species balance of oat protein is far superior to other grains. After the oat protein is decomposed, small molecular peptides with high quality can be generated, and the small molecular peptides are easy to be absorbed by skin. The oat peptide has the effects of resisting oxidation, increasing skin activity, delaying skin aging, moisturizing, reducing wrinkles, resisting allergy and the like.
Among them, ceramide 3 is a lipid substance, which has a structure similar to that of a substance constituting the horny layer of the skin, can rapidly permeate into the skin, helps the renewal of the natural protective layer of the skin, promotes the balance of natural hydration, and combines with water in the horny layer to form a network structure to lock water.
Wherein, the beta-glucan is natural plant mucilage glycan, and the component also has certain anti-allergy and anti-inflammatory effects and the like. The beta-glucan has good compatibility with most raw materials, has no anaphylactic reaction, and can improve the skin moisturizing capability. In addition, the beta-glucan can also play a role in resisting oxidation and protecting the skin from being damaged by ultraviolet rays, can reduce skin wrinkles and improve the capability of the skin in resisting external damage.
Among them, allantoin can reduce the adhesion of stratum corneum cells, accelerate epidermal cell renewal, enhance skin repair ability, and has high safety.
Wherein the kelp extract is derived from kelp, and the kelp is proliferated in a large amount by using kelp cells, so that the kelp extract contains high content of natural plant protein. Therefore, the kelp extract has the effects of supplementing cell nutrition and effectively preserving moisture.
The addition amount of the antioxidant is 0.01-2% of the total mass of the whitening water lotion. The invention can prevent the oily components of oil, wax, hydrocarbon and the like of the cosmetics from contacting with oxygen in the air to generate oxidation, generate peroxide, aldehyde, acid and the like, and lead the cosmetics to discolor, rancidity, quality reduction and the like by using the antioxidant. The antioxidant comprises one or more of vitamin E, tocopherol acetate, butylated hydroxytoluene, lycopene, and ascorbic acid ethyl ether.
The whitening water lotion of the invention can be added with a chelating agent properly. The addition amount of the chelating agent is 0.01-1% of the total mass of the whitening water lotion. The chelating agent may be disodium EDTA and/or tetrasodium EDTA and the like.
In addition, the whitening water lotion also contains a preservative and an aromatic. The addition amount of the preservative in the whitening water lotion is 0.01-1.5% and the addition amount of the aromatic is 0.005-0.5% based on the total mass of the whitening water lotion. The preservative can comprise one or the combination of more than two of phenoxyethanol, methyl hydroxybenzoate, propyl hydroxybenzoate, benzoic acid and salts thereof. The aromatic may be a perfume, etc.
The whitening moisture lotion disclosed by the invention not only can play a whitening role, but also has the effects of moisturizing and locking water, can prevent moisture loss and creates a moisturizing barrier for skin. Meanwhile, the whitening moisture lotion provided by the invention has a repairing effect on skin and activates the function of the skin.
Third embodiment
A third embodiment of the present invention provides a method for preparing the whitening water lotion of the second embodiment, including a step of mixing the components of the whitening water lotion.
Specifically, the preparation method of the whitening water lotion comprises the following steps:
1. adding water, chelating agent, partial skin conditioner, humectant, partial thickener and emulsifier into a stirring pot, stirring, and heating to 80-85 deg.C;
2. cooling to 55-65 deg.C, adding oil, pH regulator, the rest thickener, antioxidant, and part of antiseptic, and stirring;
3. cooling to 40-50 deg.C, adding the above extract, rest skin conditioner, rest antiseptic, and aromatic, and stirring;
4. cooling to 30-40 deg.C, discharging after qualified inspection, and standing for 12-24 hr;
5. and (5) after the inspection is qualified, subpackaging, packaging, inspecting again, and warehousing the finished product.
Examples
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
In the examples, the preparation method of the ganoderma lucidum extract comprises the following steps:
pulverizing Lei-Lai-Zhi raw material, adding into an extraction tank, adding water and ethanol at 100 deg.C, wherein the mass ratio of Lei-Lai-Zhi raw material to water to ethanol is 4:3:3, decocting and extracting for 3 times, each time for 2.5 hr, to obtain Lei-Lai-Zhi extractive solution. Filtering the obtained extract, storing in a storage tank, allowing the filtrate in the storage tank to slowly flow into an external circulation tank and a settling tank for concentration to obtain liquid components and precipitate, spray drying the liquid components, drying the precipitate, sieving with 80-100 mesh sieve to obtain extract, and detecting the extract to obtain extract with ash content of 7% and water content of 4%.
Examples 1 to 5
The extract of Leptospira japonica was dissolved in 5 different volumes of phosphate buffer pH 6.8 to give 5 test solutions of different concentrations. The mass concentrations of the extract of Leptospira japonica when tested for tyrosinase inhibitory activity are shown in Table 1 below.
Inhibition of tyrosinase Activity assay
Tyrosinase activity is measured as the tyrosinase activity that catalyzes the production of dopaquinone from L-dopa. The method specifically comprises the following steps:
taking 1 test tube, adding 0.4mL (1.0mg/mL) of substrate L-dopa solution, adding 2.4mL of phosphate buffer solution with pH of 6.8, carrying out water bath at 30 ℃ for 10min, and then adding 0.2mL (250U/mL) of tyrosinase to obtain reaction solution 1.
After taking 1 tube, 0.2mL (250U/mL) of tyrosinase was added, and 2.8mL of a phosphate buffer solution having a pH of 6.8 was added to the tube, thereby obtaining a reaction solution 2.
0.4mL (1.0mg/mL) of the substrate L-dopa solution was added to each of 5 tubes, then 2.4mL of the test solutions of examples 1 to 5 was added, and after incubation in a water bath at 30 ℃ for 10min, the amount of tyrosinase was then added in an amount of 0.2mL (250U/mL) to obtain a reaction solution 3.
2.4mL of the test solutions of examples 1 to 5 were added to 5 tubes, 0.4mL of a phosphate buffer solution having a pH of 6.8 was added thereto, and after incubation in a 30 ℃ water bath for 10 minutes, 0.2mL (250U/mL) of tyrosinase was added to obtain a reaction solution 4.
The enzymatic reaction converts L-dopa to the red product dopaquinone, with maximum absorption at 475 nm. Read the absorbance A at 475nm 1min from the start of the mixing475The rate of the enzymatic reaction is defined as A per minute475Is indicated by an increased value of (a). The absorbance A at 475nm of the above reaction solution 1min after the start of the reaction was measured, and the inhibition rate of tyrosinase activity was calculated according to the following formula, and the results are shown in Table 1 and FIG. 1. Wherein, the mass concentration of the ganoderma lucidum extract in the table is the final mass concentration of the ganoderma lucidum extract in the test system.
Inhibition rate ═ 1- (A)3-A4)/(A1-A2)]×100％
In the formula: a. the1Absorbance when substrate alone is contained without adding the extract of Leisha Crassifolia;
A2absorbance when no Ganoderma extract is added and no substrate is added;
A3is absorbance of the mixture containing the above extract and substrate;
A4the absorbance of the extract containing Ganoderma extract without adding substrate.
TABLE 1
As can be seen from Table 1 and FIG. 1, the extract of Leptospira has excellent inhibitory effect on tyrosinase activity. The inhibition rate of tyrosinase is gradually increased along with the increase of the mass concentration of the ganoderma extract, and gradually approaches to balance when the mass concentration of the ganoderma extract is more than 2 mg/ml.
Comparative example 1
5 different volumes of phosphate buffer at pH 6.8 were taken as test solutions for comparative example 1. Wherein, when the inhibition mechanism for inhibiting tyrosinase activity is determined, the mass concentration of the ganoderma lucidum extract is 0mg/L (shown in Table 2).
Examples 6 to 9
4 parts of the extract of Leptospira japonica are dissolved in 5 volumes of phosphate buffer solutions of pH 6.8 corresponding to comparative example 1, respectively, to obtain 4 groups of test solutions of different concentrations corresponding to comparative example 1. The mass concentration of the extract of Lei-Ganoderma when measured for the inhibition mechanism of tyrosinase activity is shown in Table 2 below.
Determination of inhibition mechanism of ganoderma lucidum extract for inhibiting tyrosinase activity
0.4mL (1.0mg/mL) of the substrate L-dopa solution was added to each of 25 tubes, then 2.4mL of the test solutions of comparative example 1 and examples 6 to 9 was added, and after incubation in a water bath at 30 ℃ for 10 minutes, 0.2mL, 0.16mL, 0.12mL, 0.08mL, and 0.04mL of tyrosinase (250U/mL) were added, and a phosphate buffer solution having a pH of 6.8 was added to a final volume of 3.0mL to obtain a reaction solution M.
2.4mL of the test solutions of comparative example 1 and examples 6 to 9 were added to 25 tubes, and after incubation in a 30 ℃ water bath for 10min, 0.2mL, 0.16mL, 0.12mL, 0.08mL, and 0.04mL of tyrosinase (250U/mL) were added, and a phosphate buffer solution having a pH of 6.8 was added to a final volume of 3.0mL to obtain reaction solution N. Wherein, the mass concentration of the ganoderma lucidum extract and the concentration of tyrosinase in the table are the final mass concentration of the ganoderma lucidum extract in a test system.
Measuring absorbance A at 475nm of the reaction mixture M and the reaction mixture N at 1min after the start of the reaction, and calculating to obtain Δ1A475The specific results are shown in table 2 below:
Δ1A475＝AM-AN
wherein A isMIs absorbance of the mixture containing the above extract and substrate;
ANthe absorbance of the extract containing Ganoderma extract without adding substrate.
Then, the concentration of tyrosinase (U/mL) was plotted on the abscissa, and the initial reaction rate of the enzyme (the rate of change of absorbance. DELTA.) was measured1A475) As an ordinate, the concentration-absorbance change rate curves of tyrosinase of comparative example 1 and examples 6 to 9 were prepared, and the results are shown in FIG. 2.
Specifically, if the concentration-absorbance change rate curves of tyrosinase of examples 6-9 and the concentration-absorbance change rate curve of tyrosinase of comparative example 1 intersect at the origin, reversibility inhibition is determined; irreversible inhibition was determined if the concentration-absorbance change rate curves for the tyrosinases of examples 6-9 were parallel to the concentration-absorbance change rate curve for the tyrosinase of comparative example 1 and did not pass through the origin.
TABLE 2
As can be seen from Table 2 and FIG. 2, the extract of Leptospira japonica was reversible in inhibiting tyrosinase activity.
Comparative example 2
5 different volumes of phosphate buffer at pH 6.8 were taken as test solutions for comparative example 2. Wherein, the mass concentration of the ganoderma lucidum extract is 0mg/L (shown in table 3) when the inhibition type of tyrosinase is determined.
Examples 10 to 13
4 parts of the extract of Leptospira japonica are dissolved in 5 volumes of phosphate buffer solutions of pH 6.8 corresponding to comparative example 2, respectively, to obtain 4 groups of test solutions of different concentrations corresponding to comparative example 2. The mass concentration of the extract of Lei's Ganoderma when determining the type of tyrosinase inhibition is shown in Table 3 below.
Determination of tyrosinase inhibition type of Lei-Zhi extract
To 25 tubes were added 0.4mL, 0.32mL, 0.24mL, 0.16mL, and 0.08mL of the substrate L-dopa solution (1.0mg/mL), respectively, then 2.4mL of the test solutions of comparative example 2 and examples 10 to 13, respectively, and a phosphate buffer solution having a pH of 6.8 was added to a final volume of 2.8mL, and after incubation in a water bath at 30 ℃ for 10min, then an amount of tyrosinase of 0.2mL (250U/mL) was added to obtain a reaction solution m.
2.4mL of the test solutions of comparative example 2 and examples 10 to 13 were added to 25 tubes, 0.4mL of a phosphate buffer solution having a pH of 6.8 was added, and after incubation in a 30 ℃ water bath for 10 minutes, an amount of tyrosinase of 0.2mL (250U/mL) was added to obtain a reaction solution n. Wherein, the mass concentration of the Lei-Ganoderma extract and the molar concentration of the substrate L-dopa are the final mass concentration of the Lei-Ganoderma extract in the test system.
Measuring the absorbance A at 475nm of the reaction mixture m and the reaction mixture n at 1min from the start of the reactionmAnd AnThe value of Δ is calculated by the following formula2A475The specific results are shown in table 3 below:
Δ2A475＝Am-An
wherein A ismIs absorbance of the mixture containing the above extract and substrate;
Anthe absorbance of the extract containing Ganoderma extract without adding substrate.
The reciprocal of the substrate concentration was plotted as the abscissa, and the initial reaction rate of the enzyme (rate of change of absorbance. DELTA.)2A475) The reciprocal of (d) is a double reciprocal curve with the ordinate representing the reciprocal of the substrate concentration-the reciprocal of the rate of change of absorbance.
Specifically, if the reciprocal double curves of examples 10 to 13 and comparative example 1 intersect on the ordinate axis and the intercept of the abscissa becomes smaller as the concentration of the extract of Achilles japonica increases, the inhibition is demonstrated and the maximum rate of the enzymatic reaction is not affected, and it can be judged as competitive inhibition.
Non-competitive inhibition can be judged if the reciprocal curves of examples 10-13 intersect the reciprocal curve of comparative example 1 on the abscissa and the intercept of the ordinate becomes larger as the concentration of the extract of Leptospira increases.
The mixed inhibition can be judged if the reciprocal curves of examples 10-13 intersect the reciprocal curve of comparative example 1 at the second quadrant (on the non-coordinate axis) and the intercept with the ordinate becomes larger as the concentration of the extract of Lepidium meyenii increases.
TABLE 3
As can be seen from Table 3 and FIG. 3, the Himalayan Ganoderma extract has competitive and noncompetitive inhibition of tyrosinase activity, and is a mixed type of inhibition.
Application examples 1 to 5
The whitening water lotion was prepared according to the contents (mass percentages) of the components in the formulations of the whitening water lotions of application examples 1 to 5 in the following tables 4 to 5, and according to the following production process steps. The production process comprises the following steps:
1. adding the phase A into a stirring pot, stirring and heating to 82-85 ℃;
2. cooling to 60 ℃, adding B, C phase and stirring evenly;
3. cooling to 42 deg.C, adding phase D, and stirring;
4. cooling to 37 ℃, discharging after the inspection is qualified, and standing for 24 hours;
5. and (5) after the inspection is qualified, subpackaging, packaging, inspecting again, and warehousing the finished product.
Note: the A, B, C, D phases in the process are respectively
Phase A: water, disodium EDTA, allantoin, mannose, butanediol, glycerol, sodium hyaluronate, xanthan gum, acrylic acid/C10-30 alkanol acrylate crosspolymer, panthenol, betaine, PEG-20 methyl glucose sesquistearate;
phase B: a compound of polydimethylsiloxane and dimethiconol, hydrogenated polyisobutene, cyclomethicone, cyclohexasiloxane, tocopheryl acetate, a complex of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and isohexadecane and polysorbate-60, methylparaben;
and C phase: aminomethyl propanol;
phase D: the composition comprises ganoderma extract, oat peptide, serine, ceramide 3, beta-glucan, kelp extract, phenoxyethanol and essence.
In the formula, mannose, butanediol, glycerol, sodium hyaluronate, panthenol and betaine are humectants;
xanthan gum, acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer and a complex of isohexadecane and polysorbate-60 are thickeners;
the compound of polydimethylsiloxane and dimethiconol, hydrogenated polyisobutene, cyclomethicone and cyclohexasiloxane are grease;
PEG-20 methyl glucose sesquistearate is an emulsifier; aminomethyl propanol is a pH adjusting agent.
Oat peptide, serine, ceramide 3, beta-glucan, macroalgae extract and allantoin are skin conditioners;
disodium EDTA is a chelating agent; tocopheryl acetate is an antioxidant;
phenoxyethanol and methyl hydroxybenzoate as antiseptic; the essence is an aromatic.
In the invention, the compound of hydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer, isohexadecane and polysorbate-60 is prepared by a manufacturer: the company, Saybolt, trade name: simugel FL.
Composite of polydimethylsiloxane and dimethiconol, manufacturer: dao corning, usa, trade mark: PMX-1403 silicone oil.
Table 4 application examples 1-5
Application of comparative examples 1 to 5
Moisture lotions were prepared in accordance with the contents (mass percentages) of the respective components in the formulations of the moisture lotions of comparative application examples 1 to 5 in the following table 5 in the same manner as in application examples 1 to 5.
TABLE 5 application comparative examples 1-5
Whitening efficacy test
Method for testing skin melanin change rate: the content of melanin in the skin is determined by measuring the reflection of light of a specific wavelength on the skin of a human body. The emitter of the probe emits light with three wavelengths of 568nm, 660nm and 880nm to the skin surface, the receiver measures the light reflected by the skin and measures the amount of light absorbed by the skin, so as to obtain the change rate of skin melanin.
The test of the skin melanin change rate is carried out by using a pigment detector of CK company in Germany, wherein a test probe MX 18 of the skin melanin consists of a light source emitter and a light source receiver, and a spring is additionally arranged to keep the pressure on the skin constant during the detection. The measuring range of the probe is 0-999, and the higher the measuring value is, the higher the content of melanin in the skin can be indicated.
The number of the subjects is 33, the test period is 4 weeks, the whitening water lotion of application examples 1-5 and the water lotion of application comparative examples 1-5 are selected in the test, the whitening water lotion is smeared on different areas of the inner side of the forearm every morning and evening, the reflection quantity of the skin of the tested area before the test (Day1) and when the test area is used for 28 days (Day28) after the illumination of specific wavelength is respectively measured to determine the content of melanin in the skin, and then the change rate of the melanin is characterized, and the result of the specific melanin change rate is shown in fig. 4.
As can be seen from fig. 4, the change rate of melanin is large, i.e., the melanin content is reduced, in the application examples 1 to 5 of the present application, and thus, the use of the ganoderma lucidum extract can effectively whiten the skin.
In the application comparative examples 1 to 5, when the ganoderma lucidum extract is not used, the change rate of the melanin content is small, the melanin content is reduced little, even increased, and therefore, the whitening effect of the application comparative examples 1 to 5 is poor.
The above examples of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (8)
1. The whitening water lotion is characterized by comprising a ganoderma lucidum extract, a humectant, a thickener, a pH regulator, an emulsifier, grease, a skin conditioner and an antioxidant, wherein the addition amount of the ganoderma lucidum extract is 0.01-12% by mass of the total whitening water lotion; the addition amount of the humectant is 0.01-20%; the addition amount of the thickening agent is 0.02-1.8%; the addition amount of the pH regulator is 0.01-1%; the addition amount of the emulsifier is 0.01-2%; the addition amount of the grease is 1-8%; the addition amount of the skin conditioner is 0.01-10%; the addition amount of the antioxidant is 0.01-2%; wherein the content of the first and second substances,
the ganoderma lucidum extract can inhibit the activity of tyrosinase, and the ash content of the ganoderma lucidum extract is less than 10% and the water content of the ganoderma lucidum extract is less than 5% by mass;
the preparation method of the ganoderma loddigesii extract comprises the following steps: pulverizing the Lei-Lai-Zhi raw material, and extracting with water and ethanol to obtain Lei-Lai-Zhi extract;
the mass ratio of the ganoderma loddigesii raw material to the water to the ethanol is 4:3: 3;
the extraction temperature is 80-100 ℃;
the extraction times are 2-4 times;
the extraction time is 2-4 hours;
filtering the obtained extract, storing in a storage tank, allowing the filtrate in the storage tank to flow into an external circulation tank and a settling tank for concentrating to obtain liquid components and precipitate, spray drying the liquid components, oven drying the precipitate, and sieving with 80-100 mesh sieve to obtain the extract.
2. The whitening water lotion according to claim 1, wherein the addition amount of the ganoderma lucidum extract is 0.1-8% by mass of the total whitening water lotion.
3. The whitening water lotion according to claim 1 or 2, further comprising one or a combination of two or more of a chelating agent, a preservative, and an aromatic; wherein the content of the first and second substances,
the addition amount of the chelating agent is 0.01-1% of the total mass of the whitening water lotion; the addition amount of the preservative is 0.01-1.5%; the addition amount of the aromatic is 0.005-0.5%.
4. The whitening water lotion of claim 1 or 2, wherein the moisturizer comprises one or a combination of more than two of glycerin, dipropylene glycol, glyceryl polyether-26, sodium hyaluronate, panthenol, PEG/PPG-17/6 copolymer, butylene glycol, xylitol, betaine, glyceryl polyacrylate, propylene glycol, mannose, and trehalose; and/or
The grease comprises one or the combination of more than two of cyclopentadimethyl siloxane, cyclodimethyl siloxane, a compound of dimethyl siloxane and dimethiconol, oleyl erucate, shea butter, ethylhexyl palmitate, hydrogenated polydecene, cyclohexasiloxane, hydrogenated polyisobutene, C20-24 alkyl dimethyl siloxane, C13-14 isoparaffin and C12-15 alcohol benzoate.
5. The whitening water lotion according to claim 1 or 2, wherein the thickener comprises one or a combination of two or more of carbomer, xanthan gum, sclerotium rolfsii, behenyl alcohol, ammonium acryloyldimethyltaurate/VP copolymer, hydroxyethyl acrylate/sodium acryloyldimethyltaurate copolymer, acrylic acid (ester)/C10-30 alkanol acrylate crosspolymer, hydroxyethyl acrylate/sodium acryloyldimethyltaurate copolymer, and a complex of isohexadecane and polysorbate-60; and/or
The pH regulator comprises one or more of aminomethyl propanol, arginine, citric acid, sodium citrate and sodium hydroxide.
6. The whitening water lotion according to claim 1 or 2, wherein the emulsifier comprises one or a combination of two or more of PEG-20 methyl glucose sesquistearate, sorbitan isostearate, polyglyceryl-3 methyl glucose distearate, laureth-7, isosteareth-20; and/or
The antioxidant comprises one or more of vitamin E, tocopherol acetate, butylated hydroxytoluene, lycopene, and ascorbic acid ethyl ether.
7. The whitening water lotion according to claim 1 or 2, wherein the skin conditioner comprises one or a combination of two or more of hydrolyzed collagen, oat peptide, ceramide 3, fucus extract, chlorella fermentation product, chlorella extract, brown algae extract, witch hazel water, bisabolol, allantoin, ginkgo mistletoe leaf extract, cogongrass rhizome extract, serine, kelp extract, beta-glucan, and cactus extract.
8. A method for preparing the whitening water lotion according to any one of claims 1 to 7, which comprises the step of mixing the components of the whitening water lotion.
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