Preparation method of constant-water-coagulation moisturizing component and cosmetic application of constant-water-coagulation moisturizing component

Abstract

The invention discloses a preparation method of a constant-water-coagulation moisturizing component and cosmetic application of the constant-water-coagulation moisturizing component, and relates to the technical field of cosmetics. According to the preparation method of the constant-water-coagulation moisturizing component, glycosylglycerol is prepared by adopting taklamaya desert algae in a mode of combining salt stress and enzyme catalysis, the content of the prepared glycosylglycerol is high, and the glycosylglycerol is more beneficial to activation and promotion of cell regeneration; the cell activity is enhanced and the cell metabolism is accelerated; the cell oxidation resistance is enhanced; and collagen synthesis is promoted. The composition is matched with carbohydrate isomeride, sodium hyaluronate, ceramide, squalane and other components to form a constant-water-coagulation moisturizing component, and the constant-water-coagulation moisturizing component is applied to cosmetics and can improve the water content of the cuticle of the cheek skin and reduce the transepidermal water loss rate of the cheek skin.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a preparation method of a constant water gel moisturizing component and cosmetic applications thereof. Background Art

[0002] Skin is the largest organ in the human body, and water is an important medium for skin cells to survive. It plays an irreplaceable role in maintaining skin moisture and promoting cell function. Normal and healthy skin is smooth, soft and elastic, while dry skin has a rough surface, split layers, loses elasticity, and even cracks. The flexibility of the skin is related to the water content and fat content. There is a natural moisturizing system in the human skin, which is composed of three substances: water, natural moisturizing factor (NMF), and lipids. It keeps the stratum corneum at a certain water content and maintains the moisture and luster of the skin. Especially in the stratum corneum, the natural moisturizing factor relies on sufficient water to keep the skin soft and elastic. In the case of dry environment or aggravated water loss, the proper use of hydrating and moisturizing products can effectively help the skin stay healthy.

[0003] Glyceryl glucoside can enhance the expression of aquaporin in epidermal cells, fundamentally enhancing the ability of cells to actively absorb water. Moreover, the molecular size of glyceryl glucoside is only 1 / 3 of hyaluronic acid, which is easily absorbed by cells. It can significantly increase the transcription of mRNA (messenger RNA) encoding AQP3 in cells, thereby allowing cells to synthesize AQP3 in large quantities, restore the normal level of water channels on the cell membrane, and maintain skin hydration.

[0004] However, the current algae extraction has a low content of glycerol glucoside, resulting in poor moisturizing effect of the products prepared therefrom.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide a preparation method of a constant water gel moisturizing component and its cosmetic application.

[0007] The present invention is achieved in that:

[0008] In a first aspect, the preparation method of the constant water gel moisturizing component of the present invention comprises, by mass percentage, uniformly mixing 50-60% of glycerol glucoside, 20-45% of saccharide isomers, 0.01-0.5% of sodium hyaluronate, 0.001-0.05% of ceramide and 0.1-5% of squalane;

[0009] Wherein, the preparation method of the glycerol glucoside comprises:

[0010] Purify the Taklimakan Desert Algae by solid culture, then pick a single colony of the Taklimakan Desert Algae and inoculate it into a liquid culture medium as a primary seed liquid, and culture it in a constant temperature and light shaking incubator for 4-6 days;

[0011] The seed solution activated to the logarithmic growth phase was inoculated into the culture dish and the algae cells were grown to OD 730 When the value is 2-3, sterilized sodium chloride is added, and CO2 is introduced as a carbon source for salt stress growth. The growth is measured every day at 20-40℃, and the volume of the culture solution that evaporates is supplemented when sampling;

[0012] When the algae cells grow to OD 730 When the value is 2-3, the Taklimakan Desert Algae is separated and purified by a plate line method, and the purified strain is cultured in a BG11 liquid medium at a light temperature of 30-40°C;

[0013] Add 0.3-0.6 mol / L sucrose, 0.8-1.2 mol / L glycerol, 40-60 mmol / L MES buffer and 40-60 μg sucrose phosphorylase to the purified strain, react at 30-40° C. for 40-60 min, boil for 10-15 min to terminate the reaction, and obtain algae liquid;

[0014] The algae liquid is separated after centrifugation, ddH2O is added to the algae cell pellet for resuspending, and then anhydrous ethanol is added and shaken for 5-10 minutes. After mixing, it is water-bathed at 60-70°C for 3-5 hours. After the water bath, it is centrifuged again and separated, and the supernatant is dried to obtain a crude sample. ddH2O is added to the crude sample for dissolution, and then filtered. The filtered liquid is stored at -80°C as glycerol glucoside with a stereoconfiguration of 2-aGG.

[0015] In an optional embodiment, the concentration of sodium chloride in the culture dish is 0.6M-3.0M.

[0016] In an optional embodiment, the volume ratio of the algae liquid, ddH2O for resuspension and the anhydrous ethanol is 1 ml: 100-200 μL: 400-500 μL.

[0017] In an optional embodiment, the centrifugation comprises centrifugation at 10000-15000 rpm for 5-10 min;

[0018] And / or, the dissolving includes adding ddH2O and then placing in an ultrasonic cleaner for dissolving for 5-10 minutes;

[0019] And / or, the filtering comprises filtering with a 0.22 μm filter.

[0020] In an optional embodiment, the pH of the MES buffer is 6-8.

[0021] In an optional embodiment, the components of the constant water gel moisturizing ingredient include, by mass percentage: 55-60% of the glyceryl glucoside, 35-45% of the carbohydrate isomers, 0.05-0.15% of the sodium hyaluronate, 0.02-0.05% of the ceramide and 2.5-3% of the squalane.

[0022] In a second aspect, the present invention provides a constant water gel moisturizing component, which is prepared by the preparation method of the constant water gel moisturizing component as described in any of the above-mentioned embodiments.

[0023] In a third aspect, the present invention provides use of the constant water gel moisturizing ingredient as described in the above embodiment in the preparation of cosmetics.

[0024] In an optional embodiment, the amount of the constant water gel moisturizing ingredient added to the cosmetic is 1-3%.

[0025] In an optional embodiment, the cosmetics include at least one of skin care water, skin care lotion, skin care essence, skin care cream, whitening cream, anti-wrinkle cream, anti-aging cream, sunscreen, BB cream, isolation cream and facial mask.

[0026] The present invention has the following beneficial effects:

[0027] The preparation method of the constant water gel moisturizing component provided by the present invention is to prepare glycerol glucoside by combining salt stress and enzyme catalysis with Taklimakan desert algae. The prepared glycerol glucoside has a high content, which is more conducive to activating and promoting cell regeneration; enhancing cell vitality, accelerating cell metabolism; enhancing cell antioxidant capacity; and promoting collagen synthesis. It is combined with ingredients such as carbohydrate isomers, sodium hyaluronate, ceramide and squalane to form a constant water gel moisturizing component, wherein sodium hyaluronate has a strong water absorption capacity and can absorb a large amount of external water and lock it. Glycerol glucoside can increase the expression of skin water channel protein and promote water transport. The combination of the two can increase the moisture content of the skin from both inside and outside. The carbohydrate isomers bind water on the skin surface and maintain the moist state of the skin surface together with sodium hyaluronate. Ceramide repairs the lipid structure between the cells of the stratum corneum of the skin, improves the skin barrier function, and prevents water from being lost through the epidermis. Squalane forms a protective film on the surface of the skin, which can reduce the stimulation of external factors on the skin, and work synergistically with ceramide to further consolidate the skin barrier and enable the skin to better retain moisture. The protective film formed by squalane can reduce the evaporation of moisture from the skin, and work together with the hydration film formed by sodium hyaluronate and the moisture bound to the sugar isomers to prolong the moisturizing time of the skin. Glyceryl glucoside and ceramide enhance the durability of the entire moisturizing system by improving the internal water transport and barrier function of the skin, so that these ingredients play a synergistic role in moisturizing and maintain the hydration of the skin for a long time. This constant water gel moisturizing ingredient is used in cosmetics to increase the water content of the stratum corneum of the cheek skin and reduce the transepidermal water loss rate of the cheek skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0029] Figure 1 This is a schematic diagram of the effect of temperature on enzyme activity in Experimental Example 1 of the present invention;

[0030] Figure 2 This is a schematic diagram of the effect of pH on enzyme activity in Experimental Example 1 of the present invention;

[0031] Figure 3 This is a schematic diagram of the difference in the content of glycerol glucoside synthesized by different algae strains in Experimental Example 2 of the present invention;

[0032] Figure 4 This is a schematic diagram of the difference in the content of glycerol glucoside synthesized by different processes in Experimental Example 3 of the present invention;

[0033] Figure 5 Schematic diagram of the moisturizing effect of the constant water gel moisturizing ingredient in Experimental Example 4 of the present invention;

[0034] Figure 6 This is a statistical chart of the water content of the stratum corneum of the skin of the constant water gel emulsion in Experimental Example 5 of the present invention;

[0035] Figure 7 This is a statistical chart of the transepidermal water loss rate of the constant water gel emulsion in Experimental Example 5 of the present invention. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0037] The present invention provides a method for preparing a constant water gel moisturizing component, which comprises, by mass percentage, uniformly mixing 50-60% of glycerol glucoside, 20-45% of saccharide isomers, 0.01-0.5% of sodium hyaluronate, 0.001-0.05% of ceramide and 0.1-5% of squalane;

[0038] Wherein, the preparation method of glycerol glucoside comprises:

[0039] The Taklimakan Desert Algae was purified by solid culture, and then a single Taklimakan Desert Algae colony was selected and inoculated into a liquid culture medium as a primary seed liquid, and then cultured in a constant temperature and light shaking incubator for 4-6 days;

[0040] The seed solution activated to the logarithmic growth phase was inoculated into the culture dish and the algae cells were grown to OD 730 When the value is 2-3, sterilized sodium chloride is added, and CO2 is introduced as a carbon source for salt stress growth. The growth is measured every day at 20-40℃, and the volume of the culture solution that evaporates is supplemented when sampling;

[0041] When the algae cells grow to OD 730 When the value was 2-3, the Taklimakan Desert Algae was isolated and purified by the plate line method, and the purified strains were cultured in BG11 liquid medium at a light temperature of 30-40°C;

[0042] Add 0.3-0.6 mol / L sucrose, 0.8-1.2 mol / L glycerol, 40-60 mmol / L MES buffer and 40-60 μg sucrose phosphorylase to the purified strain, react at 30-40°C for 40-60 min, boil for 10-15 min to terminate the reaction, and obtain algae liquid;

[0043] The algae liquid was separated after centrifugation, ddH2O was added to the algae cell pellet for resuspending, and then anhydrous ethanol was added for oscillation for 5-10 minutes. After mixing, it was water bathed at 60-70°C for 3-5 hours. After the water bath, it was centrifuged again and separated. The supernatant was dried to obtain a crude sample. ddH2O was added to the crude sample for dissolution, and then filtered. The filtered liquid was stored at -80°C as glycerol glucoside with a stereo configuration of 2-aGG.

[0044] In the present invention, the Taklimakan Desert Algae is used for purification and extraction, wherein the Taklimakan Desert Algae can produce a higher content of glycerol glucoside under the action of salt stress and enzyme catalysis.

[0045] Specifically, under salt stress, the Taklimakan Desert algae can rapidly synthesize small molecule compatible substances, such as glycerol glucoside, followed by enzyme catalysis, in which sucrose is used to provide glucose molecules and glycerol is used to provide glycerol molecules. Under the action of enzymes, glucose molecules and glycerol molecules are connected by glycosidic bonds to form natural active small molecule glycerol glucoside.

[0046] The concentration of sodium chloride during salt stress is 0.6M-3M. The enzyme used in enzyme catalysis is sucrose phosphorylase. Through the above process, the Taklimakan Desert Algae can produce more glycerol glucoside and increase the content of glycerol glucoside.

[0047] Furthermore, the volume ratio of algae liquid, ddH2O for resuspension and anhydrous ethanol is 1ml: 100-200μL: 400-500μL. The algae cell precipitate can be evenly dispersed in ddH2O by resuspension, and then anhydrous ethanol is added to fully extract the algae cells in ddH2O. A water bath of 60-70°C can ensure complete extraction. After the water bath, centrifugation is performed again for separation, and the centrifugation includes centrifugation at 10000-15000rpm for 5-10min; impurities insoluble in anhydrous ethanol can be removed, thereby improving the purity of the sample. Then the supernatant is dried to obtain a crude sample, ddH2O is added to the crude sample and placed in an ultrasonic cleaning instrument for dissolution for 5-10min. The dissolution effect of the crude sample is better with ultrasound, and then it is filtered with a 0.22μm filter to filter out fine insoluble matter and impurities in the solution. The filtered liquid is stored at -80°C as glycerol glucoside with a stereo configuration of 2-aGG.

[0048] The glyceryl glucoside prepared by the above preparation method has a 2-aGG stereo configuration, and the obtained product has a high content of glyceryl glucoside. Glyceryl glucoside has excellent moisturizing ability. Glyceryl glucoside with a 2-aGG configuration can significantly increase the expression of aquaporin AQP3, and AQP3 protein plays an important role in the water retention function of skin epithelial cells. At the same time, glyceryl glucoside has good skin permeability and can effectively reduce water loss. In addition, 2-αGG has a variety of physiological activities, such as activating and promoting cell regeneration; enhancing cell vitality, accelerating cell metabolism; enhancing cell antioxidant capacity; promoting collagen synthesis, etc. The above characteristics make this substance useful as a cosmetic raw material.

[0049] The present invention prepares a constant water gel moisturizing component by mixing 50-60% of the above-mentioned glycerol glucoside, 20-45% of sugar isomers, 0.01-0.5% of sodium hyaluronate, 0.001-0.05% of ceramide and 0.1-5% of squalane according to mass percentage. The component has good moisturizing effect, can increase the water content of the stratum corneum of the cheek skin, and reduce the transepidermal water loss rate of the cheek skin.

[0050] Specifically, the glycerol glucoside and its derivatives found in nature can be divided into α configuration and β configuration according to the different glycosidic bond conformations. The configuration of the glycerol glucoside in the present invention is the 2-aGG stereo configuration, which is more suitable for use in cosmetics.

[0051] Sugar isomers are mainly used as moisturizers and repair ingredients in skin care products. They have multiple functions such as locking in moisture, repairing the skin barrier, promoting skin metabolism, repairing skin damage, enhancing skin resistance and anti-aging, and are highly safe.

[0052] The molecular weight distribution range of sodium hyaluronate is very wide, ranging from 1kDa to 3000kDa. The molecular weight of high molecular weight sodium hyaluronate is higher than 2000KDa, the molecular weight of medium molecular weight sodium hyaluronate is between 500KDa and 2000KDa, and the molecular weight of low molecular weight sodium hyaluronate is between 10KDa and 500KDa. The present invention preferably uses low molecular weight sodium hyaluronate, which can penetrate into the dermis of the skin, play a role inside the skin, lock moisture, promote skin metabolism, and make the skin moist, smooth, delicate, tender and elastic.

[0053] The present invention preferably uses high-purity ceramide of natural origin. Ceramide (Ceramide) exists naturally in the skin. It is a class of amide compounds formed by dehydration of long-chain fatty acids and the amino group of sphingosine. It mainly contains ceramide phosphorylcholine and ceramide phosphorylethanolamine. Phospholipids are the main components of cell membranes. There are natural moisturizing factors in the stratum corneum of the skin, and ceramide NP is an important component thereof. Its molecular structure contains hydrophilic hydroxyl groups and hydrophobic fatty acid chains, which can be combined with the moisture in the stratum corneum to form a mesh structure, which firmly locks the moisture and prevents the evaporation of moisture, thereby playing a moisturizing effect. The skin barrier is mainly composed of keratinocytes and intercellular lipids, and ceramide NP is one of the main components of intercellular lipids. When the skin barrier is damaged, the content of ceramide NP will decrease, causing the connection between cells to become loose, the permeability of the skin to increase, and it is easily damaged by external stimuli. Supplementing ceramide NP can restore the normal structure of intercellular lipids, enhance the adhesion between keratinocytes, and repair damaged skin barriers.

[0054] Squalane is a natural moisturizing oil with excellent antioxidant and thermal stability. Squalane is the closest lipid to human sebum, with strong affinity, and can integrate with the human body's own sebum membrane to form a natural barrier on the skin surface. It can also inhibit the peroxidation of skin lipids, effectively penetrate the skin, and promote the proliferation of skin basal cells. It has obvious physiological effects on delaying skin aging, improving and eliminating chloasma; squalane can also open skin pores, promote blood circulation, enhance cell metabolism, and help repair damaged cells.

[0055] Preferably, the components of the constant water gel moisturizing ingredient include, by mass percentage, 55-60% glyceryl glucoside, 35-45% saccharide isomers, 0.05-0.15% sodium hyaluronate, 0.02-0.05% ceramide and 2.5-3% squalane.

[0056] Furthermore, the present invention also provides the use of the above-mentioned Hengshui gel moisturizing component in the preparation of cosmetics, wherein the amount of the Hengshui gel moisturizing component added to the cosmetics is 1-3%. The cosmetics include but are not limited to at least one of skin care water, skin care lotion, skin care essence, skin care cream, whitening cream, anti-wrinkle cream, anti-aging cream, sunscreen, BB cream, isolation cream and facial mask.

[0057] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0058] Example 1

[0059] This embodiment provides a method for synthesizing glycerol glucoside, and its preparation method is as follows:

[0060] S1. Use a sterile inoculation loop to pick a single colony of Taklimakan Desert Algae (commercially purchased from Xinjiang Yeli Biotechnology Co., Ltd.) on the BG11 solid plate into a 50 mL conical flask containing 20 mL of BG11 liquid culture medium as the primary seed liquid, and place it in a constant temperature and light shaking incubator for about 5 days.

[0061] S2. The seed solution activated to the logarithmic growth phase was inoculated into a culture dish for evaluation. The inoculation volume was 60 mL. 730 When the value is about 2-3, add 0.9M sterilized sodium chloride, and use a gas valve to pass 3% CO2 as a carbon source. Measure its growth every day at 30°C. When sampling, replenish the volume of the culture medium that has evaporated. Sterile water can be used to supplement the culture medium.

[0062] S3, wait for the algae cells to grow to OD 730 When the value is about 2-3, the Taklimakan Desert Algae is separated and purified by the plate line method, and the purified strain is cultured in BG11 liquid culture medium at a light temperature of 30-40°C.

[0063] S4. Add 0.4 mol / L sucrose, 1 mol / L glycerol, 50 mmol / L MES buffer (pH 7.0), and 50 μg sucrose phosphorylase (commercially available from Merck) to the purified strain, react at 35°C for 40 min, and boil for 10 min to terminate the reaction.

[0064] S5. Take 1 mL of algae solution into a 1.5 mL EP tube, centrifuge at 12000 rpm for 5 min, separate the supernatant (extracellular compatible substances) from the precipitate, add 100 μL ddH2O to the algae cell precipitate to resuspend the cells, then add 400 μL of anhydrous ethanol and oscillate with a vortexer for 5 min. After mixing, place in a water bath at 65°C for 4 h. After the water bath ends, centrifuge the sample at 13000 rpm for 5 min, carefully take the supernatant into a 10-15 mL centrifuge tube (be careful not to suck up the precipitate), blow dry it completely with a nitrogen blower at 55°C, add 1 mL ddH2O to the dried sample to dissolve it. If the sample on the tube wall is difficult to dissolve, place it in an ultrasonic cleaner for 5 min. Filter the dissolved sample with a 0.22 μm filter, and store the treated sample at -80°C.

[0065] Experimental Example 1: Enzyme Catalysis Parameter Selection

[0066] (1) Effect of temperature on enzyme activity

[0067] The temperature of step S4 in Example 1 was adjusted to 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, and 60°C, respectively, and the effects of different temperatures on enzyme activity were tested. The test results are shown in FIG. Figure 1As shown, it can be seen that with the increase of temperature, the enzyme activity first increases and then decreases. Therefore, the preferred enzyme catalysis temperature in the present invention is 30-40°C.

[0068] (2) Effect of pH on enzyme activity

[0069] The pH of step S4 in Example 1 was adjusted to pH=4, pH=5, pH=6, pH=7, pH=8, pH=9, and pH=10, respectively, and the pH was adjusted by selecting different buffers, including citric acid-sodium citrate buffer (pH4.0-6.0), MES buffer (pH 6.0-8.0), and glycine-sodium hydroxide buffer (pH 8.0-10.0), to detect the effect of different pH values ​​on enzyme activity. The test results are as follows: Figure 2 As shown, it can be seen that with the increase of pH, the enzyme activity first increases and then decreases. Therefore, the preferred pH for enzyme catalysis in the present invention is 6-8.

[0070] Experimental Example 2: Selection of different algae strains

[0071] This experimental example investigates the difference in the content of glycerol glucoside synthesized by different algae strains. In this experimental example, Spirulina, Nostoc, Anabaena and Microcystis were selected as control examples to compare with the Taklimakan Desert algae in Example 1. Spirulina, Nostoc, Anabaena and Microcystis also synthesized glycerol glucoside in the same manner as in Example 1 to obtain different samples. The filtered liquid was placed in a chromatographic bottle and measured by ion chromatography. The data results are shown in Figure 3 .

[0072] It can be seen that the content of glycerol glucoside in different algae strains is quite different. The Taklimakan Desert algae selected in the present invention to synthesize glycerol glucoside has the highest content.

[0073] Experimental Example 3: Selection of different synthesis processes

[0074] This example examines different routes in the synthesis process and provides Comparative Examples 1 to 3.

[0075] Comparative Example 1

[0076] This comparative example is basically the same as Example 1, except that the salt stress in step S2 of Example 1 is omitted in Comparative Example 1. Specifically, the following steps are included:

[0077] S1 is the same as Example 1.

[0078] S2. The seed solution activated to the logarithmic growth phase was inoculated into a culture dish for evaluation. The inoculation volume was 60 mL. 730When the value is about 2-3, 3% CO2 is passed through a gas valve as a carbon source, and its growth is measured every day at 30°C. When sampling, the volume of the culture medium that has evaporated is supplemented, and sterile water can be used to supplement the culture medium.

[0079] S3-S5 are the same as in Example 1.

[0080] Comparative Example 2

[0081] This comparative example is basically the same as Example 1, except that the enzyme catalysis in step S4 of Example 1 is omitted in Comparative Example 2. Specifically, the following steps are included:

[0082] S1-S3 are the same as in Example 1.

[0083] S4, omitted.

[0084] S5. Take 1 mL of algae solution into a 1.5 mL EP tube, centrifuge at 12000 rpm for 5 min, separate the supernatant (extracellular compatible substances) from the precipitate, add 100 μL ddH2O to the algae cell precipitate to resuspend the cells, then add 400 μL of anhydrous ethanol and oscillate with a vortexer for 5 min. After mixing, place in a water bath at 65°C for 4 h. After the water bath ends, centrifuge the sample at 13000 rpm for 5 min, carefully take the supernatant into a 10-15 mL centrifuge tube (be careful not to suck up the precipitate), blow dry it completely with a nitrogen blower at 55°C, add 1 mL ddH2O to the dried sample to dissolve it. If the sample on the tube wall is difficult to dissolve, place it in an ultrasonic cleaner for 5 min. Filter the dissolved sample with a 0.22 μm filter, and store the treated sample at -80°C.

[0085] Comparative Example 3

[0086] This comparative example is substantially the same as Example 1, except that, in Comparative Example 3, the concentration of sodium chloride used for salt stress in Example 1 is 5 M.

[0087] The samples obtained in Example 1 and Comparative Examples 1-3 were placed in chromatographic bottles and measured by ion chromatography. The data results are shown in Figure 4 .

[0088] It can be seen that the content of glycerol glucoside in the sample obtained in Example 1 of the present invention is significantly greater than that in Comparative Examples 1-3, wherein the step of salt stress is omitted in Comparative Example 1, which cannot promote the production of glycerol glucoside by Taklimakan Desert Algae under salt stress conditions, so that the content of glycerol glucoside is significantly lower than that in Example 1. The step of enzyme catalysis is omitted in Comparative Example 2, which also results in a significantly lower content of glycerol glucoside than that in Example 1, and it can be seen from the data of Comparative Examples 1 and 2 that the combination of salt stress and enzyme catalysis can play a synergistic role, significantly increasing the amount of glycerol glucoside produced by Taklimakan Desert Algae. In Comparative Example 3, the concentration of sodium chloride during salt stress is increased, and excessive concentration will inhibit the production of glycerol glucoside by Taklimakan Desert Algae, thereby resulting in a significantly lower content of glycerol glucoside than that in Example 1.

[0089] Composition Examples and Comparative Examples

[0090] The glycerol glucoside involved in the following composition examples and composition comparative examples all adopts the glycerol glucoside provided in Example 1.

[0091] Composition Example 1: Prepare a constant water gel moisturizing ingredient according to the following components and ratios: 40% glyceryl glucoside, 55% carbohydrate isomers, 0.1% sodium hyaluronate, 0.1% ceramide, and 4.8% squalane, and stir and mix them evenly at room temperature to obtain the product.

[0092] Composition Example 2: Prepare a constant water gel moisturizing ingredient according to the following components and proportions: 55% glyceryl glucoside, 40% carbohydrate isomers, 0.5% sodium hyaluronate, 0.01% ceramide, and 4.49% squalane, and stir and mix evenly at room temperature to obtain the obtained ingredient.

[0093] Composition Example 3: Prepare a constant water gel moisturizing ingredient according to the following components and proportions: 53% glyceryl glucoside, 45% carbohydrate isomers, 0.2% sodium hyaluronate, 0.01% ceramide, and 1.79% squalane, and stir and mix evenly at room temperature to obtain the obtained ingredient.

[0094] Composition Example 4: A constant water gel moisturizing ingredient is prepared according to the following components and proportions: 45% glyceryl glucoside, 50% carbohydrate isomers, 0.1% sodium hyaluronate, 0.05% ceramide, and 4.85% squalane, and stirred and mixed evenly at room temperature to obtain the obtained ingredient.

[0095] Composition Example 5: Prepare a constant water gel moisturizing ingredient according to the following components and proportions: 57% glyceryl glucoside, 40% carbohydrate isomers, 0.1% sodium hyaluronate, 0.05% ceramide, and 2.85% squalane, and stir and mix evenly at room temperature to obtain the obtained ingredient.

[0096] Composition Example 6: Prepare a constant water gel moisturizing ingredient according to the following components and proportions: 60% glyceryl glucoside, 35% carbohydrate isomers, 0.5% sodium hyaluronate, 0.1% ceramide, and 1.4% squalane, and stir and mix evenly at room temperature to obtain the obtained ingredient.

[0097] The comparative example 1 of the composition is prepared according to the following components and proportions: 40% of carbohydrate isomers, 0.1% of sodium hyaluronate, 0.05% of ceramide, 2.85% of squalane, 57% of purified water are added, and the mixture is stirred and mixed uniformly at room temperature to obtain the composition.

[0098] The comparative example 2 of the composition is prepared according to the following components and proportions: 57% glyceryl glucoside, 0.1% sodium hyaluronate, 0.05% ceramide, 2.85% squalane, 40% purified water is added, and the mixture is stirred and mixed uniformly at room temperature to obtain the composition.

[0099] The comparative composition example 3 is prepared according to the following components and proportions: 57% glyceryl glucoside, 40% carbohydrate isomers, 0.05% ceramide, 2.85% squalane, 0.1% purified water is added, and the mixture is stirred and mixed uniformly at room temperature to obtain the product.

[0100] The comparative composition example 4 is prepared according to the following components and proportions: 57% glyceryl glucoside, 40% carbohydrate isomers, 0.1% sodium hyaluronate, 2.85% squalane, 0.05% purified water is added, and the mixture is stirred and mixed uniformly at room temperature to obtain the product.

[0101] The comparative composition example 5 is prepared according to the following components and proportions: 57% glyceryl glucoside, 40% carbohydrate isomers, 0.05% ceramide, 0.1% sodium hyaluronate, 2.85% purified water is added, and the mixture is stirred and mixed uniformly at room temperature to obtain the product.

[0102] The composition comparative example 6 is prepared according to the following components and proportions: 57% of glycerol glucoside is added with 43% of purified water, and the mixture is stirred and mixed uniformly at room temperature to obtain the product.

[0103] The proportions of active ingredients in the above examples and comparative examples are shown in the following table:

[0104]

[0105] Experimental Example 4: Determination of moisturizing effect by in vitro weighing method

[0106] (1) Place the prepared saturated potassium acetate solution and saturated ammonium sulfate solution into a sealed box respectively and wait for the sealed box to reach the required humidity.

[0107] (2) Use an analytical balance to weigh the mass of the glass plate with 3 cm of tape on it (M0).

[0108] (3) Use a glass rod to evenly spread the samples of the examples and comparative examples on a glass plate with a 3 cm adhesive tape, and weigh the mass (M1) using an analytical balance. Then place the samples in a desiccator at a required constant humidity.

[0109] (4) After 4 hours and 8 hours, take out the glass plates respectively, weigh the mass (M2), test its moisture retention rate, and immediately put them into a desiccator with constant humidity after weighing.

[0110] The formula for calculating the moisturizing rate is as follows:

[0111] Moisture retention rate = (M2-M0) / (M1-M0)×100%; where: M0 is the mass of the empty plate (g); M1 is the mass of the glass plate after adding the sample (g); M2 is the mass after being placed in the desiccator for several hours (g).

[0112] The results are shown in Figure 5 ,from Figure 5 It can be seen that the 4h and 8h moisturizing rates of composition examples 1-6 of the present application are significantly better than those of composition examples 1-6, especially significantly better than the effect of composition comparative example 1 lacking glyceryl glucoside, while composition comparative example 6 only contains glyceryl glucoside. Combining the effects of composition comparative example 1 and composition comparative example 6, it can be seen that there is a synergistic effect between glyceryl glucoside and other components of saccharide isomers, sodium hyaluronate, ceramide and squalane. Further, combined with composition comparative example 2, it can be seen that the effect of omitting saccharide isomers is also significantly worse than that of the embodiment, and the sum of the effects of composition comparative example 1 and composition comparative example 2 is also significantly lower than that of the embodiment.

[0113] The mechanism of action of the composition:

[0114] Sodium hyaluronate has a strong water absorption capacity and can absorb a large amount of external water and lock it. Glyceryl glucoside can increase the expression of skin water channel protein and promote water transport. The combination of the two can increase the moisture content of the skin from both inside and outside. The carbohydrate isomers bind water on the skin surface and work with sodium hyaluronate to maintain the hydration of the skin surface. Ceramide repairs the lipid structure between the cells of the stratum corneum of the skin, improves the skin barrier function, and prevents water from being lost through the epidermis. Squalane forms a protective film on the skin surface, which can reduce the stimulation of external factors on the skin, and synergizes with ceramide to further consolidate the skin barrier and enable the skin to better retain moisture. The protective film formed by squalane can reduce the evaporation of skin moisture, and work together with the hydration film formed by sodium hyaluronate and the water bound by carbohydrate isomers to prolong the moisturizing time of the skin. Glyceryl glucoside and ceramide enhance the durability of the entire moisturizing system from the aspects of improving the internal water transport and barrier function of the skin, so that these ingredients play a synergistic role in moisturizing and maintain the hydration of the skin for a long time.

[0115] The constant water gel moisturizing component provided in the above composition Example 5 is used as a raw material to prepare a constant water gel moisturizing emulsion.

[0116] The formula ingredients of Hengshui Gel Lotion are as follows:

[0117]

[0118] The preparation steps of the constant water gel emulsion are as follows:

[0119] Add behenyl alcohol, dimethicone, caprylic / capric triglyceride, trilaureth-4 phosphate and glyceryl stearate in phase B into the clean oil phase pot in turn, heat to 80°C, stir to dissolve evenly, and keep warm for 10 minutes; add purified water, ammonium acryloyldimethyltaurate / VP copolymer and pentanediol in phase A into the clean water phase pot in turn, homogenize at high speed for 3 minutes, heat to 80°C, keep warm for 15 minutes, and stir thoroughly until completely dissolved; pump phase A and phase B into the emulsifier pot, homogenize at high speed for 5 minutes, and continue stirring for 8 minutes; cool the water to 45°C, add phenoxyethanol, hexylene glycol and ethylhexylglycerin in phase C in turn; finally add the constant water gel moisturizing ingredients in phase D; stir thoroughly and evenly, cool to 38°C, filter the material, and obtain an emulsion sample after passing the inspection.

[0120] Experimental Example 5: Evaluation of the Moisturizing Efficacy of Lotion

[0121] According to the prepared lotion samples, the moisturizing efficacy of the product was evaluated before and after use.

[0122] Volunteers: healthy Chinese female subjects aged between 30 and 55 years old;

[0123] The facial skin is red; the basic value of transepidermal water loss (TEWL) of the cheek area should be >10g / m when measured by the transepidermal water loss tester during the inclusion and exclusion screening and baseline value (D0) measurement. 2 / h; Willing not to use cosmetics, health products, medicines or other types of products with the same effects as the test product, such as moisturizing effects, in addition to the test product during the trial period;

[0124] Application area: The entire face area

[0125] How to use: Take an amount the size of a pearl and apply evenly on five places on the face (forehead, bridge of nose, sides of cheeks, chin). Use your fingertips to apply evenly from the sides of the nose to the base of the ears, from the inside to the outside, lifting and tightening the contours in turn. Preheat your hands and press gently to allow it to be fully absorbed.

[0126] Test cycle: Before product use (week 0) and 4 weeks after continuous use, facial skin parameters were collected using a non-invasive skin testing instrument and full-face photos were taken.

[0127] Test parameters: water content of skin stratum corneum, transepidermal water loss (TEWL).

[0128] SPSS 21.0 statistical software was used for data analysis. Repeated measures analysis of variance was used to analyze the differences between the baseline values ​​(D0) and the follow-up values ​​(D28) of each parameter. At the D28 follow-up, the binomial test was used to analyze the differences in the self-assessment questionnaire of the subjects. Statistically significant differences were marked with p<0.05*, p<0.01**, and p<0.001***.

[0129] The moisture content of the skin stratum corneum is measured by Corneometer, CM825. The larger the value, the higher the moisture content of the skin stratum corneum. The more the moisture content of the skin stratum corneum increases compared with the baseline value (D0) at different follow-up time points, the better the moisturizing effect of the product.

[0130] After using the test product, the test results of the moisture content of the stratum corneum of the cheek skin are as follows Figure 6 As shown: Compared with the baseline value (D0), the water content of the stratum corneum of the cheek skin increased by 43.88% and 52.52% at the D0T15min and D28 revisits, respectively. After repeated measures analysis of variance, compared with D0, the water content of the stratum corneum of the cheek skin at each revisit time point showed significant differences (p<0.001).

[0131] Trans-epidermal water loss (TEWL) is measured by Tewameter, TM Hex and is an important parameter that reflects skin water loss and barrier function. The lower the value, the better the skin barrier function.

[0132] After using the test product, the test results of the transepidermal water loss rate of cheek skin are as follows Figure 7 As shown: Compared with the baseline value (D0), the transepidermal water loss rate of cheek skin decreased by 12.13% and 25.40% at D0T15min and D28 respectively. After repeated measures analysis of variance, compared with D0, the transepidermal water loss rate of cheek skin at each visit time point showed significant differences (p<0.001).

[0133] It can be seen from the above experiments that the constant water gel moisturizing component provided by the present invention is added to the emulsion, which can increase the water content of the stratum corneum of the cheek skin and reduce the transepidermal water loss rate of the cheek skin.

[0134] In summary, the preparation method of glycerol glucoside provided by the present invention is prepared by combining Taklimakan Desert algae with salt stress and enzyme catalysis. The content of the prepared glycerol glucoside is high, which is more conducive to activating and promoting cell regeneration; enhancing cell vitality, accelerating cell metabolism; enhancing cell antioxidant capacity; and promoting collagen synthesis. It is combined with ingredients such as carbohydrate isomers, sodium hyaluronate, ceramide and squalane to form a constant water gel moisturizing ingredient, among which sodium hyaluronate has a strong water absorption capacity and can absorb a large amount of external water and lock it. Glycerol glucoside can increase the expression of skin water channel protein and promote water transport. The combination of the two can increase the moisture content of the skin from both inside and outside. The carbohydrate isomers bind water on the skin surface and maintain the moist state of the skin surface together with sodium hyaluronate. Ceramide repairs the lipid structure between the cells of the stratum corneum of the skin, improves the skin barrier function, and prevents water from being lost through the epidermis. Squalane forms a protective film on the surface of the skin, which can reduce the stimulation of external factors on the skin, and work synergistically with ceramide to further consolidate the skin barrier and enable the skin to better retain moisture. The protective film formed by squalane can reduce the evaporation of moisture from the skin, and work together with the hydration film formed by sodium hyaluronate and the moisture bound to the sugar isomers to prolong the moisturizing time of the skin. Glyceryl glucoside and ceramide enhance the durability of the entire moisturizing system by improving the internal water transport and barrier function of the skin, so that these ingredients play a synergistic role in moisturizing and maintain the hydration of the skin for a long time. This constant water gel moisturizing ingredient is used in cosmetics to increase the water content of the stratum corneum of the cheek skin and reduce the transepidermal water loss rate of the cheek skin.

[0135] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a constant water gel moisturizing ingredient, characterized in that: Calculated by mass percentage, it comprises uniformly mixing 50-60% of glyceryl glucoside, 20-45% of saccharide isomers, 0.01-0.5% of sodium hyaluronate, 0.001-0.05% of ceramide and 0.1-5% of squalane; The preparation method of the glycerol glucoside comprises: solid culture and purification of Taklimakan Desert Algae, then picking a single colony of the Taklimakan Desert Algae and inoculating it into a liquid culture medium as a primary seed liquid, and culturing it in a constant temperature and light shaking incubator for 4-6 days; The seed solution activated to the logarithmic growth phase was inoculated into the culture dish and the algae cells were grown to OD 730 When the value is 2-3, sterilized sodium chloride is added, and CO2 is introduced as a carbon source for salt stress growth. The growth is measured every day at 20-40℃, and the volume of the culture solution that evaporates is supplemented when sampling; When the algae cells grow to OD 730 When the value is 2-3, the Taklimakan Desert Algae is separated and purified by a plate line method, and the purified strain is cultured in a BG11 liquid medium at a light temperature of 30-40°C; Add 0.3-0.6 mol / L sucrose, 0.8-1.2 mol / L glycerol, 40-60 mmol / L MES buffer and 40-60 μg sucrose phosphorylase to the purified strain, react at 30-40° C. for 40-60 min, boil for 10-15 min to terminate the reaction, and obtain algae liquid; The algae liquid is separated after centrifugation, ddH2O is added to the algae cell pellet for resuspending, and then anhydrous ethanol is added and shaken for 5-10 minutes. After mixing, it is water-bathed at 60-70°C for 3-5 hours. After the water bath, it is centrifuged again and separated, and the supernatant is dried to obtain a crude sample. ddH2O is added to the crude sample for dissolution, and then filtered. The filtered liquid is stored at -80°C as glycerol glucoside with a stereoconfiguration of 2-aGG.

2. The method for preparing the constant water gel moisturizing component according to claim 1, characterized in that: The concentration of sodium chloride in the culture dish is 0.6M-3.0M.

3. The method for preparing the constant water gel moisturizing component according to claim 1, characterized in that: The volume ratio of the algae solution, ddH2O for resuspension and the anhydrous ethanol is 1 ml: 100-200 μL: 400-500 μL.

4. The method for preparing the constant water gel moisturizing component according to claim 1, characterized in that: The centrifugation includes centrifugation at 10000-15000 rpm for 5-10 min; And / or, the dissolving includes adding ddH2O and then placing in an ultrasonic cleaner for dissolving for 5-10 minutes; And / or, the filtering comprises filtering with a 0.22 μm filter.

5. The method for preparing the constant water gel moisturizing component according to claim 1, characterized in that: The pH of the MES buffer is 6-8.

6. The method for preparing the constant water gel moisturizing component according to claim 1, characterized in that: The components of the constant water gel moisturizing ingredient include, by mass percentage: 55-60% of the glyceryl glucoside, 35-45% of the saccharide isomers, 0.05-0.15% of the sodium hyaluronate, 0.02-0.05% of the ceramide and 2.5-3% of the squalane.

7. A constant water gel moisturizing ingredient, characterized in that: The invention is prepared by the method for preparing the constant water gel moisturizing component according to any one of claims 1 to 6.

8. Use of the constant water gel moisturizing ingredient as claimed in claim 7 in the preparation of cosmetics.

9. The use according to claim 8, characterized in that: The amount of the constant water gel moisturizing component added to the cosmetic is 1-3%.

10. The use according to claim 8, characterized in that: The cosmetics include at least one of skin care water, skin care lotion, skin care essence, skin care cream, whitening cream, anti-wrinkle cream, anti-aging cream, sunscreen, BB cream, isolation cream and facial mask.

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