Tea flower composition with antioxidant and whitening effects and application thereof

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a tea flower composition with antioxidant and whitening effects and application thereof. The tea flower composition is prepared from the following components in parts by weight: 0.1 to 5 parts of tea flower extract, 0.05 to 1 part of alfalfa seed extract and 0.5 to 4 parts of black willow bark extract. The tea flower composition disclosed by the invention combines the tea flower extract, the alfalfa seed extract and the black willow bark extract, and the three extracts are compounded in a preferable range of the tea flower composition and can fully play a synergistic effect, so that mild and efficient care is brought to skin, anti-oxidation protection can be provided, a whitening effect can be realized while moisturizing is realized, and the tea flower composition has a good application prospect. And the composition is natural and safe, is suitable for various skin types, and is especially suitable for consumers who pursue mild and efficient skin care experience.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and in particular relates to a tea flower composition with antioxidant and whitening effects and application thereof. Background Art

[0002] As people pay more attention to health and beauty, antioxidant and whitening functional products have gradually become popular in the market. Existing antioxidant and whitening products mostly use traditional plant extracts or chemical ingredients. Although some ingredients can effectively inhibit free radicals, slow down aging, and brighten the skin, they still have the following drawbacks:

[0003] First, existing antioxidants and whitening ingredients often pose skin irritation issues. Many chemical whitening ingredients can cause allergies and irritation, and long-term use can damage the skin barrier and cause skin sensitivity. Furthermore, while some natural plant extracts have a milder effect, their antioxidant capacity and whitening effects are often limited, failing to meet the demands of modern consumers for effective and long-lasting results.

[0004] Secondly, existing technologies for antioxidant and whitening products often use single ingredients or simple formula combinations, lacking comprehensive, synergistic, and multifunctional formulas. This results in limited product effectiveness, particularly in terms of long-term antioxidant and whitening effects and sustained efficacy.

[0005] Therefore, there is an urgent need for a composite natural plant composition that can not only effectively resist oxidation and protect the skin from free radical damage, but also gently whiten the skin and inhibit melanin formation, so as to meet the market demand for efficient and safe skin care products. Summary of the Invention

[0006] The purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a tea flower composition with antioxidant and whitening effects, which significantly enhances the product's multiple effects such as antioxidant and whitening through the synergistic effect between the components.

[0007] To achieve the above objectives, the technical solutions adopted by the present invention include:

[0008] In a first aspect, the present invention provides a tea flower composition with antioxidant and whitening effects, wherein the tea flower composition comprises the following components in parts by weight: 0.1-5 parts of tea flower extract, 0.05-1 parts of alfalfa seed extract, and 0.5-4 parts of black willow bark extract.

[0009] Tea tree flowers contain tea polyphenols, tea polysaccharides, tea saponins, proteins, amino acids, vitamins, and other ingredients beneficial to the human body. Modern medical research indicates that tea tree flowers have detoxifying, lipid-lowering, blood sugar-lowering, anti-aging, anti-cancer, nourishing, and beauty-enhancing properties. The active ingredients in tea tree flowers exhibit excellent antioxidant, antibacterial, and anti-inflammatory properties, offering potential for the prevention and treatment of several chronic diseases.

[0010] Alfalfa seed extract is rich in protein, vitamins, minerals, and a variety of bioactive ingredients, such as alfalfa polysaccharides and flavonoids. Alfalfa polysaccharides moisturize and promote skin cell metabolism, while flavonoids have antioxidant and anti-inflammatory properties, protecting the skin from free radical damage, reducing inflammation, helping to prevent skin aging, and improving skin radiance and elasticity.

[0011] The main active ingredient of black willow bark extract is salicin, which is converted into salicylic acid in the human body. It has anti-inflammatory, antibacterial and skin-soothing effects. It can reduce skin inflammation, inhibit acne formation, and promote the metabolism of the skin's stratum corneum, making the skin smoother and more delicate. It is helpful to improve skin roughness and fade fine lines.

[0012] The tea flower composition of the present invention combines tea flower extract, alfalfa seed extract and black willow bark extract. The three ingredients are compounded within the preferred range of the present invention to fully exert a synergistic effect, thereby providing gentle and efficient care for the skin. It can not only provide antioxidant protection, but also achieve whitening effects while moisturizing. The composition is natural and safe, suitable for all skin types, and is especially suitable for consumers who pursue a gentle and efficient skin care experience.

[0013] Preferably, the tea tree flower composition comprises the following components in parts by weight: 0.5-1 part of tea tree flower extract, 0.1-0.5 part of alfalfa seed extract and 1-2 parts of black willow bark extract.

[0014] More preferably, the tea tree flower composition comprises the following components in parts by weight: 1 part of tea tree flower extract, 0.5 parts of alfalfa seed extract and 2 parts of black willow bark extract.

[0015] Through experimental research, it was found that when the three components are combined using the optimal dosage ratio specified above, their respective functions can be maximized, thereby ensuring that the prepared composition has excellent effects in anti-oxidation, moisturizing and whitening, with the best cost-effectiveness and the most stable properties.

[0016] Preferably, the preparation method of the tea tree flower extract comprises the following steps:

[0017] S1, dissolving tea tree pollen in water, stirring and soaking, and then performing ultrasonic extraction to obtain an extract;

[0018] S2. The extract is subjected to coarse filtration and fine filtration to obtain the tea tree flower extract.

[0019] The present invention adopts the above-mentioned purification process to prepare the tea tree flower extract, which can effectively remove impurities in the extract and prevent the impurity components from reacting with the other two plant components, thereby ensuring the stability of the raw material combination and formula; and after extraction by the above-mentioned purification process, the active ingredients of the tea tree flower extract are concentrated, thereby further enhancing its antioxidant and other effects.

[0020] Preferably, in step S1, the specific method of stirring and soaking is: soaking at 20-30°C for 8-12 minutes, and the stirring rate is 300-500rpm; the specific method of ultrasonic extraction is: ultrasonic extraction at 25-30°C for 10-20 minutes, and the ultrasonic power is 100-300W; the mass ratio of the tea tree pollen to water is 10:(25-35).

[0021] More preferably, in step S1, the specific method of stirring and soaking is: soaking at 25°C for 10 minutes, with a stirring rate of 400 rpm; the specific method of ultrasonic extraction is: ultrasonic extraction at 28°C for 15 minutes, with an ultrasonic power of 200 W; and the mass ratio of tea tree pollen to water is 10:30.

[0022] Preferably, in step S2, the specific method of the coarse filtration is: using 80-120 mesh filter cloth for coarse filtration; the specific method of the fine filtration is: using a filter membrane with a pore size of 0.22-0.55 μm for fine filtration.

[0023] More preferably, in step S2, the specific method of the coarse filtration is: using a 100-mesh filter cloth for coarse filtration; the specific method of the fine filtration is: using a filter membrane with a pore size of 0.45 μm for fine filtration.

[0024] Through experimental research, it was found that when tea flowers are extracted purely using the above-mentioned limited parameter conditions, the impurities in the extract can be minimized to the greatest extent, thereby effectively ensuring the long-term stability of the composition, avoiding precipitation and precipitation problems, and further enhancing the product's efficacy in whitening, anti-oxidation, moisturizing and other aspects.

[0025] In a second aspect, the present invention provides the use of the tea flower composition with antioxidant and whitening effects in cosmetics.

[0026] Preferably, the cosmetics include essences, creams, sunscreens, facial masks and makeup.

[0027] In a third aspect, the present invention provides an essence with antioxidant and whitening effects, wherein the essence comprises the tea flower composition.

[0028] Preferably, the essence further comprises a moisturizer, an oil phase, a thickener, a preservative and a pH adjuster;

[0029] The moisturizer includes at least one of glycerin, propylene glycol, butylene glycol, dipropylene glycol and glyceryl polyether-26; and / or the oil phase includes at least one of octyl cocoate, polydimethylsiloxane and glyceryl monostearate; and / or the thickener includes carbomer; and / or the preservative includes at least one of p-hydroxyacetophenone, phenoxyethanol, methylparaben and chlorphenesin; and / or the pH adjuster includes sodium citrate or sodium hydroxide.

[0030] In a fourth aspect, the present invention provides a method for preparing the essence having antioxidant and whitening effects, comprising the following steps:

[0031] S1. Dissolve the alfalfa seed extract in water, set the speed to 400-500 r / min, and stir for 10-20 min to obtain an alfalfa seed extract solution;

[0032] S2. stirring and mixing the moisturizing agent and water at a speed of 250-350 r / min to obtain an aqueous phase;

[0033] S3. Heat the oil phase to 70-80° C. and stir at 200-300 r / min for 5-10 min to obtain an oil phase;

[0034] S4. Heat the water phase to 65-75°C, add the oil phase and stir to mix. Control the stirring speed to 750-850 r / min and stir for 3-7 minutes to form a microemulsion.

[0035] S5. Cool the microemulsion to 40-50° C., add a thickener and a pH adjuster, adjust the pH value to 5.5-6.5, then add the tea tree flower extract, alfalfa seed extract solution, black willow bark extract and a preservative, and stir evenly to obtain the essence.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1) The tea flower composition of the present invention combines tea flower extract, alfalfa seed extract and black willow bark extract. The three ingredients are compounded within the preferred range of the present invention to fully exert a synergistic effect, thereby providing gentle and efficient care for the skin, providing antioxidant protection, and achieving a whitening effect while moisturizing. The composition is natural, safe, and suitable for all skin types, especially for consumers who pursue a gentle and efficient skin care experience.

[0038] (2) The specific purification process of the present invention is used to prepare the tea tree flower extract, which can effectively remove impurities in the extract and prevent the impurity components from reacting with the other two plant components, thereby ensuring the stability of the raw material combination and formula; and after extraction by the above-mentioned purification process, the active ingredients of the tea tree flower extract are concentrated, thereby further enhancing its antioxidant and other effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the extraction process diagram of tea tree flower extract;

[0040] Figure 2 This is a comparative diagram of the moisturizing rates of the samples described in Effect Example 2. DETAILED DESCRIPTION

[0041] To better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0042] Unless otherwise specified, all raw materials involved in the following examples and comparative examples are commercially available general materials.

[0043] The tea flower described below is a powder purchased from Qimen Xie Yongzhong Black Tea Co., Ltd.; the alfalfa seed extract is a powder purchased from Fufeng Snow Biotechnology Co., Ltd.; the black willow bark extract is a liquid purchased from Guangzhou Huayang Biotechnology Co., Ltd.

[0044] Examples 1-4

[0045] Examples 1-4 provide an essence with antioxidant and whitening effects, the formula of which is shown in Table 1 (total weight parts is 100 parts). The preparation method of the essence comprises the following steps:

[0046] S1. Pretreatment: First, dissolve the alfalfa seed extract in water, set the speed to 400-500 r / min, and stir for 15 minutes to obtain an alfalfa seed extract solution;

[0047] S2. Preparation of aqueous phase: Add deionized water to a stirring pot, turn on the stirrer and set the speed to 300 r / min. While stirring, add glycerol and propylene glycol in sequence and stir evenly to ensure complete dissolution;

[0048] S3. Preparation of oil phase: In another stirring pot, mix octyl cocoate, polydimethylsiloxane, and glyceryl monostearate, heat to 75°C, and stir at 250 rpm for 8 minutes to fully dissolve and mix the ingredients to form a uniform oil phase;

[0049] S4. Heat the water phase to 70° C., keep stirring, and slowly add the oil phase to the water phase, controlling the addition rate to avoid generating excessive bubbles; during the addition of the oil phase, gradually increase the stirring speed to 800 r / min and continue stirring for 5 minutes to fully emulsify the oil phase and the water phase to form a microemulsion;

[0050] S5. Slowly cool the microemulsion to 45° C., add carbomer while stirring, stir evenly to fully disperse the carbomer, continue cooling to room temperature, slowly add sodium hydroxide solution for neutralization, and adjust the pH value to 5.5-6.5;

[0051] S6. While stirring, add the tea tree flower extract, the alfalfa seed extract solution, the black willow bark extract and the phenoxyethanol preservative in sequence and stir evenly to obtain the essence.

[0052] The preparation process of the tea tree flower extract is as follows:

[0053] (1) Extraction: Dissolve 10 g of powdered tea tree flowers in 30 g of water, stir at 400 rpm at 25°C, soak for 10 minutes, and then extract in an ultrasonic device at 200 W of ultrasonic power, 15 minutes of extraction time, and 28°C of temperature.

[0054] (2) Coarse filtration: After the extraction is completed, preliminary filtration is performed through a 100-mesh filter cloth to remove larger particles of impurities, such as incompletely crushed plant residues;

[0055] (3) Fine filtration: To further remove fine impurities and suspended particles, microporous membrane filtration can be used. A filter membrane with a pore size of 0.45 μm is used and a vacuum filtration device is used to obtain a clear, slightly yellowish tea flower extract.

[0056] Table 1 Formula of the essence described in Examples 1-4 (parts by weight)

[0057] Components Example 1 Example 2 Example 3 Example 4 Tea Tree Flower Extract 0.1 5 0.5 1 Alfalfa seed extract 1 0.05 0.1 0.5 Black Willow Bark Extract 4 0.5 1 2 glycerin 10 10 10 10 Propylene glycol 5 5 5 5 Octyl Cocoate 5 5 5 5 Polydimethylsiloxane 3 3 3 3 Glyceryl Monostearate 2 2 2 2 Carbomer 0.5 0.5 0.5 0.5 Sodium hydroxide solution 0.1 0.1 0.1 0.1 Phenoxyethanol 0.5 0.5 0.5 0.5 water margin margin margin margin

[0058] Example 5

[0059] This embodiment provides an essence with antioxidant and whitening effects, which differs from Example 4 only in that the preparation process of the tea tree flower extract is different.

[0060] The preparation process of the tea tree flower extract described in this embodiment is as follows:

[0061] (1) Extraction: Dissolve 10 g of powdered tea tree flowers in 25 g of water, stir at 500 rpm at 20°C, soak for 8 minutes, and then place in an ultrasonic device for extraction. The conditions are ultrasonic power 100 W, extraction time 20 minutes, and temperature 30°C.

[0062] (2) Coarse filtration: After the extraction is completed, preliminary filtration is performed through a 100-mesh filter cloth to remove larger particles of impurities, such as incompletely crushed plant residues;

[0063] (3) Fine filtration: To further remove fine impurities and suspended particles, microporous membrane filtration can be used. A filter membrane with a pore size of 0.45 μm is used and a vacuum filtration device is used to obtain a clear, slightly yellowish tea flower extract.

[0064] Example 6

[0065] This embodiment provides an essence with antioxidant and whitening effects, which differs from Example 4 only in that the preparation process of the tea tree flower extract is different.

[0066] The preparation process of the tea tree flower extract described in this embodiment is as follows:

[0067] (1) Extraction: Dissolve 10 g of powdered tea tree flowers in 35 g of water, stir at 300 rpm at 30°C, soak for 12 minutes, and then extract in an ultrasonic device at 300 W of ultrasonic power, 10 minutes of extraction, and 25°C of temperature.

[0068] (2) Coarse filtration: After the extraction is completed, preliminary filtration is performed through a 100-mesh filter cloth to remove larger particles of impurities, such as incompletely crushed plant residues;

[0069] (3) Fine filtration: To further remove fine impurities and suspended particles, microporous membrane filtration can be used. A filter membrane with a pore size of 0.45 μm is used and a vacuum filtration device is used to obtain a clear, slightly yellowish tea flower extract.

[0070] Comparative Examples 1-8

[0071] This comparative example provides an essence with antioxidant and whitening effects, the formula of which is shown in Table 2 (total weight parts is 100 parts). The preparation method of the essence is the same as that of Example 4, the only difference being the formula.

[0072] Table 2 Formula of the essence described in Comparative Examples 1-8 (parts by weight)

[0073]

[0074]

[0075] Comparative Example 9

[0076] This comparative example provides an essence with antioxidant and whitening effects, which differs from Example 4 only in that an equal amount of grape seed extract is used to replace alfalfa seed extract, and the remaining components and amounts are the same as those in Example 4.

[0077] Comparative Example 10

[0078] This comparative example provides an essence with antioxidant and whitening effects, which differs from Example 4 only in that an equal amount of witch hazel extract is used to replace the black willow bark extract, and the remaining components and amounts are the same as those in Example 4.

[0079] Comparative Example 11

[0080] This comparative example provides an essence with antioxidant and whitening effects. The only difference between this essence and Example 4 is that the preparation process of the tea tree flower extract is different from that of Example 4. The preparation process of the tea tree flower extract in this comparative example is as follows:

[0081] (1) Extraction: Dissolve 10 g of powdered tea tree flowers in 30 g of water, stir at 400 rpm at 25°C, soak for 25 minutes, and then extract in an ultrasonic device at 200 W of ultrasonic power, 15 minutes of extraction, and 28°C of temperature.

[0082] (2) Coarse filtration: After the extraction is completed, preliminary filtration is performed through a 100-mesh filter cloth to remove larger particle impurities, such as incompletely crushed plant residues, to obtain the tea tree flower extract.

[0083] Effect Example 1

[0084] This effect example uses the essence prepared in Examples 1-6 and Comparative Examples 1-11 as samples, and adopts ABTS + The free radical scavenging effect was used to test its antioxidant activity. The specific method is as follows:

[0085] Dilute the test sample with water to a concentration of 0.1%, and use PBS buffer as a solvent to prepare the sample solution for testing. Set up a sample tube (As), a sample background (Ab), and a sample blank tube (A0). For each sample tube (As) of each tested concentration, three parallel tubes must be set up, and three parallel sample blank tubes (A0) must also be set up. Add 0.2 mL of sample solution of the same concentration to each sample tube (As) and sample background (Ab), add 0.2 mL of PBS buffer to the sample blank tube (A0), and add 0.8 mL of ABTS to each of As and A0. + For the working solution, add 0.8 mL of PBS buffer to Ab and react in the dark for 6 minutes. Transfer the solution in each reaction tube into a 1 cm cuvette and measure the absorbance at 734 nm. Calculate the half-clearance concentration IC50 using the following formula:

[0086]

[0087] The specific test results are shown in Table 3.

[0088] Table 3 Antioxidant test results

[0089] Sample source Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 <![CDATA[IC 50 (ABTS + )]]> 0.0097％ 0.0082％ 0.0078％ 0.0064％ 0.0073％ 0.0081％ Sample source Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 <![CDATA[IC 50 (ABTS + )]]> 0.0696％ 0.0532％ 0.0548％ 0.1104％ 0.1792％ 0.1346％ Sample source Comparative Example 7 Comparative Example 8 Comparative Example 9 Comparative Example 10 Comparative Example 11 <![CDATA[IC 50 (ABTS + )]]> 0.0219％ 0.0163％ 0.0398％ 0.0478％ 0.0114％

[0090] The results in Table 3 show that the essence containing the tea flower composition prepared by the technical solution of the present invention has significantly improved scavenging ability of ABTS free radicals, thereby exerting a better antioxidant effect, especially the essence containing the composition in Example 4 has the best antioxidant effect.

[0091] When any one or two of the tea tree flower extract, alfalfa seed extract, and black willow bark extract in the composition are missing in Comparative Examples 1-6, the prepared essence's ability to scavenge ABTS free radicals is significantly weakened, that is, its antioxidant activity decreases. When the dosage ratio of the three active ingredients in Comparative Examples 7-8 exceeds the specified range of the present invention, the antioxidant activity of the prepared essence also decreases. The above results show that the tea tree flower extract, alfalfa seed extract, and black willow bark extract components selected by the present invention can better exert a synergistic effect when combined within a specific amount ratio range, thereby significantly increasing the antioxidant activity of the product. Once the amount ratio exceeds the specified range, the synergistic effect between the components will be insignificant or even antagonistic, thereby reducing the antioxidant efficacy of the product and reducing the cost-effectiveness. In Comparative Examples 8-9, when other components with similar efficacy were used to replace the alfalfa seed extract and the black willow bark extract, the antioxidant effect of the prepared essence also decreased to a certain extent, indicating that the components in the composition of the present invention cannot be conventionally replaced, otherwise the synergy between the components will be affected. When the tea flowers prepared by other purification processes are used to prepare products in Comparative Example 11, the antioxidant properties of the products will also decrease, indicating that the purification process described in the present invention can efficiently utilize the tea flowers, thereby helping to further enhance the antioxidant and other effects of the products.

[0092] Effect Example 2

[0093] This effect example uses the essences prepared in Examples 1-6 and Comparative Examples 1-11 as samples to test their moisturizing effects. The specific method is as follows:

[0094] (1) Test purpose and principle:

[0095] Skin hydration is important for maintaining the epidermal barrier function, various enzyme activities, and normal cell activities such as differentiation and desquamation of skin tissue. The water content of the skin determines the softness of the skin and the exchange of substances inside and outside the cells. It is related to the health of the skin. The local use of moisturizing products is the main method to maintain skin moisture and improve adverse symptoms.

[0096] An ideal moisturizing product should have sustained moisturizing properties. This test method continuously monitors the weight change of the test sample and plots a curve of the sample's moisture retention rate versus time. Based on the trend of the moisture retention rate curve, the moisturizing capacity of the test sample is observed and the moisturizing efficacy of the sample on the skin is evaluated. This test method is an in vitro method and is suitable for water-based cosmetics.

[0097] (2) Test index: Moisturizing rate. The higher the moisturizing rate, the stronger the moisturizing effect of the sample.

[0098] (3) Experimental Method: Samples were weighed, and then pretreated with the sample and positive control (5% glycerol). The sample and positive control were placed in a constant temperature and humidity chamber at 20°C and 40% humidity. Finally, the sample was weighed regularly, and the moisture retention rate was calculated: Moisture retention rate (%) = [(initial sample weight - final sample weight) / initial sample weight] × 100%. The specific test results are shown in Table 4.

[0099] Table 4 Moisturizing rate (%) test results

[0100]

[0101]

[0102] The results in Table 4 show that the essence containing the tea flower composition prepared according to the technical solution of the present invention has good moisturizing properties and can effectively inhibit the loss of skin moisture. However, the essences prepared in Comparative Examples 1-11 have significantly lower moisturizing effects than those in the examples.

[0103] Effect Example 3

[0104] This effect example uses the essences prepared in Examples 1-6 and Comparative Examples 1-11 as samples, and tests their whitening efficacy by inhibiting melanocyte tyrosinase activity in vitro. The specific method is as follows:

[0105] (1) Principle: Tyrosinase is a copper-containing metalloenzyme belonging to the polyphenol oxidase class. It is an important rate-limiting enzyme in a series of chemical reactions during the production of melanin. Tyrosinase catalyzes L-dopa to produce dopachrome, which has a characteristic absorption peak at 492nm, and the activity of tyrosinase is then measured. B16 mouse melanoma cells can be used as a cell model to study the inhibition rate of tyrosinase activity in cosmetics. On the basis of meeting the effectiveness verification, the inhibition rate of tyrosinase activity in the experimental group was significantly increased compared with the blank control group, and there was a significant difference (P<0.05), indicating that the experimental sample has the ability to inhibit tyrosinase activity at the tested concentration, which can be used as one of the evidences to support the name of whitening cosmetic raw materials.

[0106] (2) Determination method:

[0107] 1. Cell plating: B16 mouse melanoma cells are normally cultured using RPMI 1640 complete medium (RPMI 1640 complete medium does not produce melanin). Remove the cells from the carbon dioxide incubator, observe the cell growth under a microscope, and randomly select two fields of view to take pictures using a low-power microscope (4X) and a high-power microscope (10X). When the cell density reaches 80%-90%, it is confirmed that it can be harvested. Wipe the outer surface of the culture flask with a dust-free cloth containing alcohol and transfer it to the biosafety cabinet. Gently shake the culture flask back and forth, remove the old culture medium with a pipette, wash the cell surface once with an appropriate amount of PBS, digest with 0.25% trypsin for 2 minutes, and observe under a microscope. When most cells become round and in suspension, add about 2-3 times the volume of RPMI containing serum. 1640 complete medium culture to terminate digestion, and collect into centrifuge tube, 1500rpm, centrifuge for 5min. After the centrifugation is completed. Discard the supernatant, add a certain volume of DMEM high glucose complete medium to the centrifuge tube (at this time, resuspend the cells and use DMEM high glucose medium in the next experiment. B16 cells will produce melanin in DMEM medium), resuspend evenly and count using a cell counter. Take 20μL for cell counting. Inoculate into 24-well plates at a density of 100,000 cells / well. The blank control group is not inoculated with cells and only DMEM high glucose medium is added for 24h.

[0108] 2. Sample treatment: Prepare the treated samples, 2 mL for each group, blank control group: DMEM high-glucose complete medium, (nicotinamide) positive control experimental group: 200 μL nicotinamide (1 mg / mL stock solution) + 2 mL DMEM high-glucose complete medium, test sample experimental group: 200 μL essence sample (1 mg / mL stock solution) + 2 mL DMEM high-glucose complete medium; discard the cell culture supernatant, add 500 μL blank control reagent, positive control group reagent, and experimental group reagent, respectively, and culture for 72 hours. During this period, observe the cell status and culture medium every day and replace with fresh blank control reagent, positive control group reagent, and experimental group reagent.

[0109] 3. Cell Harvesting: After removing the culture supernatant, wash once with an appropriate amount of PBS. Add 200 μL of 0.25% trypsin to each well to digest the cells at 37°C for 2 minutes. Add 400 μL of PBS containing 10% FBS to terminate digestion and pipette the cells. Collect the cells into a 1.5 mL centrifuge tube and centrifuge at 4000 rpm for 5 minutes. Discard the supernatant.

[0110] 4. Detection: Add 100 μL of 1% Triton x 100 to each tube to lyse cells. Freeze at 80°C for 30 minutes, then place at room temperature for 10 minutes. Centrifuge at 12,000 rpm at 4°C for 20 minutes to remove cell debris. Collect 50 μL of the supernatant and add 150 μL of 2 mg / mL LDOPA solution. Incubate at 37°C for 1 hour. Measure absorbance at 492 nm using a microplate reader.

[0111] 5. Data analysis: Enzyme activity inhibition rate = [1-(As-Ab) / (Ac-Ab)] × 100%, As: absorbance of the experimental group or positive control group, Ac: absorbance of the negative control well, Ab: absorbance of the blank control group. The results are shown in Table 5.

[0112] Table 5 In vitro melanocyte tyrosinase activity inhibition assay results

[0113]

[0114] The results in Table 5 show that the essence containing the tea flower composition prepared according to the technical solution of the present invention can effectively inhibit tyrosinase activity, thereby achieving a good whitening effect. In particular, the essence containing the composition in Example 4 has the best whitening effect. However, the lack of active ingredients in Comparative Examples 1-6 and the fact that the active ingredient ratios in Comparative Examples 7-8 exceed the preferred range of the present invention will lead to a decrease in the tyrosinase inhibitory effect of the products. Similarly, the replacement of the active ingredients in Comparative Examples 9-10 will also lead to a decrease in the tyrosinase inhibitory effect of the products.

[0115] Effect Example 4

[0116] This effect example uses the essences prepared in Examples 4-6 and Comparative Example 11 as samples to test their stability. The specific method is as follows:

[0117] After the physical and chemical indicators and appearance indicators of the test samples meet the standards, the samples are tested under different conditions:

[0118] (1) Normal temperature test group, placed at 25°C for 1 day, 7 days, and 45 days;

[0119] (2) Heat resistance test group, placed at 45°C for 1 day, 7 days, and 45 days;

[0120] The test samples were observed on the first, seventh, and forty-fifth days after treatment, with the observation items being the appearance, color, and skin feel of the test samples. The specific test results are shown in Table 6.

[0121] Table 6 Stability test results

[0122]

[0123] The results in Table 6 show that the essence containing the composition prepared using the technical solution of the present invention has good stability, with no obvious abnormalities under different test environments. However, when the tea tree flower extract prepared using other purification processes in Comparative Example 11 was used to formulate the essence, the product was prone to precipitation and precipitation after long-term storage and high temperature conditions, resulting in poor stability.

[0124] Finally, it should be noted that the above embodiments are intended to illustrate the technical solutions of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A tea flower composition with antioxidant and whitening effects, characterized in that: The tea tree flower composition comprises the following components in parts by weight: 0.1-5 parts of tea tree flower extract, 0.05-1 parts of alfalfa seed extract and 0.5-4 parts of black willow bark extract.

2. The tea flower composition with antioxidant and whitening effects according to claim 1, characterized in that: The tea tree flower composition comprises the following components in parts by weight: 0.5-1 part of tea tree flower extract, 0.1-0.5 part of alfalfa seed extract and 1-2 parts of black willow bark extract.

3. The tea flower composition with antioxidant and whitening effects according to claim 1 or 2, characterized in that: The preparation method of the tea tree flower extract comprises the following steps: S1, dissolving tea tree pollen in water, stirring and soaking, and then performing ultrasonic extraction to obtain an extract; S2. The extract is subjected to coarse filtration and fine filtration to obtain the tea tree flower extract.

4. The tea flower composition with antioxidant and whitening effects according to claim 3, characterized in that: In step S1, the specific method of stirring and soaking is: soaking at 20-30°C for 8-12 minutes, and the stirring rate is 300-500rpm; the specific method of ultrasonic extraction is: ultrasonic extraction at 25-30°C for 10-20 minutes, and the ultrasonic power is 100-300W; the mass ratio of the tea tree pollen to water is 10:(25-35).

5. The tea flower composition with antioxidant and whitening effects according to claim 3, characterized in that: In the step S2, the specific method of the coarse filtration is: using 80-120 mesh filter cloth for coarse filtration; the specific method of the fine filtration is: using a filter membrane with a pore size of 0.22-0.55 μm for fine filtration.

6. Use of the tea flower composition with antioxidant and whitening effects as claimed in any one of claims 1 to 5 in cosmetics.

7. An essence with antioxidant and whitening effects, characterized in that: The essence comprises the tea tree flower composition according to any one of claims 1 to 5.

8. The essence with antioxidant and whitening effects according to claim 7, characterized in that: The essence also includes a moisturizer, an oil phase, a thickener, a preservative and a pH adjuster; The moisturizer includes at least one of glycerin, propylene glycol, butylene glycol, dipropylene glycol and glyceryl polyether-26; and / or the oil phase includes at least one of octyl cocoate, polydimethylsiloxane and glyceryl monostearate; and / or the thickener includes carbomer; and / or the preservative includes at least one of p-hydroxyacetophenone, phenoxyethanol, methylparaben and chlorphenesin; and / or the pH adjuster includes sodium citrate or sodium hydroxide.

9. The method for preparing the essence with antioxidant and whitening effects according to claim 8, wherein: The following steps are involved: S1. Dissolve the alfalfa seed extract in water, set the speed to 400-500 r / min, and stir for 10-20 min to obtain an alfalfa seed extract solution; S2. stirring and mixing the moisturizing agent and water at a speed of 250-350 r / min to obtain an aqueous phase; S3. Heat the oil phase to 70-80° C. and stir at 200-300 r / min for 5-10 min to obtain an oil phase; S4. Heat the water phase to 65-75°C, add the oil phase and stir to mix. Control the stirring speed to 750-850 r / min and stir for 3-7 minutes to form a microemulsion. S5. Cool the microemulsion to 40-50° C., add a thickener and a pH adjuster, adjust the pH value to 5.5-6.5, then add the tea tree flower extract, alfalfa seed extract solution, black willow bark extract and a preservative, and stir evenly to obtain the essence.

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