Soothing composition containing white tea extract, essence and preparation method

Abstract

The invention discloses a soothing composition containing a white tea extract. The soothing composition comprises the white tea extract, a eucommia ulmoides bark extract, a citrus fruit extract, an artemia extract and a gardenia fruit extract. The extract of Fuding white tea picked at the beginning of April is adopted, so that the anti-inflammatory effect is more excellent, and irritation is avoided under the condition of a relatively high addition amount in actual product application. The white tea extract, the citrus extract, the artemia extract, the eucommia bark extract and the gardenia fruit extract are combined, so that the reaction threshold value of the skin can be remarkably increased, the thermal pressure of the oily sensitive skin can be relieved in multiple dimensions, and the high-temperature irritation resistance of the skin can be optimized. The composition can provide an auxiliary treatment scheme for face care of people after sun drying and after medical art when side reactions such as dryness, redness and dryness-heat occur, the active matter raw materials in the composition are safe in source and easy to obtain, and the application prospect is good.

Description

Technical Field

[0001] The invention relates to the field of makeup preparations, and in particular to a soothing composition containing white tea extract, an essence and a preparation method thereof. Background Art

[0002] As life pressure increases and external environmental pollution intensifies, people's skin problems emerge in an endless stream, and the number of people with sensitive skin increases, especially oily sensitive skin, which not only has the problem of oil secretion in the T-zone, but also has local redness, low tolerance and easy irritation and inflammation. Traditional skin care products for oily skin are mostly believed to seriously clog pores due to oil secretion and severe skin keratinization. Therefore, the irritation of oily skin skin care products is relatively high, but they are not suitable for oily sensitive skin. Oily sensitive skin has a low tolerance threshold for cold and heat, and it will cause heat stress problems when alternating between high and low temperatures. Therefore, it is very important to develop a composition suitable for oily sensitive skin to relieve the heat stress of oily sensitive skin.

[0003] Chinese invention patent CN114588078B discloses a self-emulsifying functional oil phase and its preparation method and application. It adopts a self-emulsifying functional plant oil phase to fill the gap in the market demand for functional oil phase. Oil is an oil-in-water emulsifier that can moisturize the skin and repair keratin, but it cannot alleviate the problem of heat stress of oily sensitive skin. Chinese invention patent CN116617134A discloses a skin soothing composition and its preparation method and products. The nerve soothing component is a combination of extracts such as citrus fruit extract, and the skin barrier repairing component is a combination of extracts such as black tea seed oil. The outer layer is wrapped with lecithin and / or hydrogenated lecithin. The effect of soothing sensitive skin is achieved by repairing damaged skin barrier function, inhibiting inflammatory response and reducing nerve response. But it is not friendly to oily skin. Summary of the invention

[0004] In order to develop a composition suitable for oily sensitive skin and relieving heat stress of oily sensitive skin, the first aspect of the present invention provides a soothing composition containing white tea extract, wherein the soothing composition includes white tea (Hippophaerhamnoides Linn.) extract, Eucommia ulmoides oliv (Eucommia ulmoides oliv) bark extract, citrus fruit (Citrus reticulata Blanco) extract, Artemia (Brine Shrimp) extract, and Gardeniae Fructus extract.

[0005] As a preferred embodiment, the white tea extract is extracted from Fuding white tea, and the Fuding white tea is harvested in April-May in the northern hemisphere.

[0006] As a preferred embodiment, the Fuding white tea is harvested in early April in the northern hemisphere.

[0007] As a preferred embodiment, the content of tea polyphenols in the white tea extract is 6.0-6.5 mg / mL.

[0008] During the experiment, the inventors found that by extracting white tea grown in Fuding, Fujian and picked in early April, a better anti-inflammatory effect can be obtained. The possible reasons are speculated to be: the white tea grown in Fuding, Fujian and picked in early April contains a higher content of tea polyphenols, and the white tea is a slightly fermented tea. During the fermentation process, the content of tea polyphenols decreases less, and the oxidation reaction of tea polyphenols in the micro-fermentation process is weaker, and theanine produced in the micro-fermentation process has higher anti-inflammatory and antioxidant effects. The tea polyphenol content in the white tea extract used in the present invention reaches 6-6.5 mg / mL, which has a better anti-inflammatory effect.

[0009] The inventors further discovered that the composition of the present invention is non-irritating at a relatively high concentration. The reason may be that the tea polyphenols and theanine in the white tea extract have a good synergistic effect, rather than an enrichment effect of an added effect. Therefore, the combined effect is more coordinated and stable, and is less likely to cause skin irritation.

[0010] As a preferred embodiment, the soothing composition includes, by weight, 0.1-2 parts of white tea extract, 0.05-1 parts of eucommia bark extract, 0.1-2 parts of citrus fruit extract, 0.1-3 parts of artemia extract, and 0.1-2 parts of gardenia fruit extract.

[0011] As a preferred embodiment, the soothing composition includes, by weight, 0.1-1 part of white tea extract, 0.05-0.5 part of eucommia bark extract, 0.1-1.5 parts of citrus fruit extract, 0.1-1 part of artemia extract, and 0.1-1 part of gardenia fruit extract.

[0012] As a preferred embodiment, the soothing composition includes, by weight, 0.1 parts of white tea extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0013] As a preferred embodiment, the weight ratio of the white tea extract to the eucommia bark extract is (1-3):1; the weight ratio of the white tea extract to the citrus fruit extract is (0.5-1.5):1.

[0014] As a preferred embodiment, the weight ratio of the white tea extract to the Artemia extract is (0.5-1.5):1; the weight ratio of the white tea extract to the Gardenia fruit extract is (0.5-3):1.

[0015] The inventors found in the experimental process that the combination of white tea extract, citrus extract, Artemia extract, Eucommia bark extract, and Gardenia fruit extract can significantly increase the skin's reaction threshold, relieve the thermal stress of oily sensitive skin in multiple dimensions, and optimize the skin's resistance to high temperature irritation. The possible reason is that the combination of white tea extract, citrus extract, Artemia extract, Eucommia bark extract, and Gardenia fruit extract can increase the expression of HSP70 gene, promote the upregulation of heat shock protein expression when external high temperature stimulation occurs, stabilize protein function, improve skin stability, and avoid the skin's heat stress response when alternating between cold and hot.

[0016] In addition to excessive sebum overflow, oily and sensitive skin also has non-immune mechanisms of facial skin that overreact to various factors (including external invasions such as pollution), and the tolerance threshold is reduced, resulting in a fragile and reddening epidermal barrier. Especially under the influence of high temperature, sebaceous gland secretion increases by 10% for every degree increase in temperature; thermal stimulation and excessive temperature difference will also accelerate the loss of moisture from the skin, and symptoms of redness and itching are prone to occur. In the present invention, the Eucommia bark extract can reduce skin temperature, calm the skin, inhibit the expression of heat stress genes, and help the skin resist and absorb infrared and ultraviolet radiation that cause temperature changes and thermal aging with Artemia extract. Because the barrier is damaged and vascular reactivity increases under the stimulation of inflammation, the increase in the sensitivity of skin nerve afferents will also extend to vascular reactions. Citrus fruit extract can reduce the influx of calcium ions, inhibit the activation of TRPV1 receptors, relieve local tingling and burning sensations of the skin, and synergize with white tea extract to relieve redness, relieving the heat pressure damage of oily sensitive skin in multiple dimensions.

[0017] The second aspect of the present invention provides an application of a soothing composition containing white tea extract, which is applied to soothing essence, skin care water, skin care lotion, skin care ointment, skin care cream, and shower gel.

[0018] As a preferred embodiment, based on the weight percentage of the soothing essence, the preparation raw materials include 2-18% of the oil phase, 4-8% of the soothing composition containing white tea extract, and the water phase is supplemented to 100%.

[0019] As a preferred embodiment, based on the weight percentage of the soothing essence, the oil phase includes 2-11% moisturizer, 0.5-2% thickener, and 0.2-5% skin conditioner.

[0020] As a preferred embodiment, the moisturizing agent is selected from at least one of glycerin, allantoin, panthenol, sodium hyaluronate, butylene glycol, 1,2-pentanediol, and glyceryl polyether-26.

[0021] As a preferred embodiment, the moisturizing agent includes a combination of glycerin, panthenol and sodium hyaluronate.

[0022] As a preferred embodiment, the weight ratio of glycerol, panthenol and sodium hyaluronate is (1-10): (1-5): (0.1-1).

[0023] As a preferred embodiment, the weight ratio of glycerol, panthenol and sodium hyaluronate is (1-5): (1-3): (0.1-0.5).

[0024] As a preferred embodiment, the weight ratio of glycerin, panthenol and sodium hyaluronate is 3:2.7:0.2.

[0025] As a preferred embodiment, the thickener is selected from at least one of PEG-150 distearate, acrylic acid (ester) copolymer, carbomer, xanthan gum, and sodium acrylic acid (ester) copolymer.

[0026] As a preferred embodiment, the thickener is xanthan gum.

[0027] As a preferred embodiment, the skin conditioning agent includes but is not limited to p-hydroxyacetophenone, dipotassium glycyrrhizinate, lotus extract, olive fruit extract, niacinamide, allantoin, flaxseed extract, Dendrobium nobile extract, Capparis spinosa extract, carboxymethyl de-chitosan, and Ophiopogon japonicus root extract.

[0028] The soothing composition containing white tea extract of the present invention is applied to essence, which can make the essence added with olive fruit appear clear and transparent. It is well known to those skilled in the art that olive fruit is prone to precipitation and discoloration problems, but within the addition ratio range of the composition of the present invention, the essence will not have precipitation of raw materials or flocculent precipitation.

[0029] As a preferred embodiment, the skin conditioning agent is a combination of p-hydroxyacetophenone and dipotassium glycyrrhizinate.

[0030] As a preferred embodiment, the aqueous phase includes deionized water and panthenol. The weight ratio of the panthenol is 1-3% based on the weight percentage of the soothing essence. Preferably, the weight ratio of the panthenol is 2% based on the weight percentage of the soothing essence.

[0031] The third aspect of the present invention provides a method for preparing a soothing essence, comprising the following steps:

[0032] S1 premixes the moisturizer and the thickener to obtain a premixed liquid A;

[0033] Heat the S2 reactor in a water bath, start stirring, add premix A, and then add other raw materials of the oil phase, heat to 70-90°C, keep stirring for 8-15 minutes, and homogenize at 5000-7000rpm for 4-6 minutes;

[0034] S3 cools down to 40-50℃, adds water phase and stirs evenly;

[0035] S4 cools down to 30-50°C, adds the soothing composition containing white tea extract, stirs the functional phase evenly, and passes the test.

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

[0037] (1) The soothing composition containing white tea extract of the present invention uses the extract of Fuding white tea picked in early April, has a more excellent anti-inflammatory effect, and is non-irritating at a higher addition amount in actual product applications.

[0038] (2) The soothing composition containing white tea extract of the present invention adopts a combination of white tea extract, citrus extract, Artemia extract, Eucommia bark extract, and Gardenia fruit extract, which can significantly improve the skin's reaction threshold, relieve the thermal stress of oily sensitive skin in multiple dimensions, and optimize the skin's resistance to high temperature irritation.

[0039] (3) The soothing composition containing white tea extract of the present invention, wherein the weight ratio of white tea extract to eucommia bark extract is (1-3):1, and the weight ratio of white tea extract to citrus fruit extract is (0.5-1.5):1, can reduce skin temperature, calm the skin, and inhibit the expression of heat stress genes.

[0040] (4) The soothing composition containing white tea extract of the present invention, wherein the weight ratio of white tea extract to Artemia extract is (0.5-1.5):1; the weight ratio of white tea extract to Gardenia fruit extract is (0.5-3:1), can relieve local stinging and burning sensation of the skin, synergistically soothe and reduce redness with white tea extract, and relieve heat-pressure damage to oily sensitive skin in multiple dimensions.

[0041] (5) After the soothing composition containing white tea extract of the present invention is applied to the preparation, it can provide an auxiliary treatment plan for the facial care of people after sun exposure and medical aesthetics when side effects such as dryness, redness, and heat occur. In addition, the raw materials of the active ingredients in the functional composition of the present invention are safe and easily available, and have good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 The essences prepared in Application Example 1 and Application Comparative Example 1 are shown on the left and Application Example 1 on the right.

[0043] Figure 2 The photos are of the essence prepared in Comparative Example 1 after being stored in the environments of 25°C, 42°C, 5°C and -15°C for 3 months, from left to right: 25°C, 42°C, 5°C and -15°C.

[0044] Figure 3 The photos are of the essence prepared in Example 1 after being stored in the environments of 25°C, 42°C, 5°C and -15°C for 3 months, from left to right: 25°C, 42°C, 5°C and -15°C. DETAILED DESCRIPTION

[0045] Example 1

[0046] A soothing composition containing white tea extract, comprising, by weight, 0.1 part of white tea extract, 0.05 part of eucommia bark extract, 0.1 part of citrus fruit extract, 0.2 part of artemia extract, and 0.1 part of gardenia fruit extract.

[0047] The white tea extract is extracted from Fuding white tea picked in early April in the northern hemisphere and is purchased from Shanhai Innovation Xuanjia, with the trade name: White Tea Extract Pro.

[0048] The Eucommia bark extract was purchased from Biolande with the trade name: EUCOOLAN.

[0049] The citrus fruit extract was purchased from Shanghai Jiakai, with the trade name: Calmnerv CR.

[0050] The Artemia extract was purchased from Ashland, trade name: gp4g TM pb biofunctional.

[0051] The gardenia fruit extract was purchased from Shanghai Jinzhi, trade name: GarPepair.

[0052] The white tea extract, the eucommia bark extract, the citrus fruit extract, the Artemia extract and the gardenia fruit extract were uniformly mixed to obtain a soothing composition containing the white tea extract, and then a performance test was performed.

[0053] Example 2

[0054] A soothing composition containing white tea extract, comprising, by weight, 0.05 parts of white tea extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0055] The rest of the implementation is the same as Example 1.

[0056] Example 3

[0057] A soothing composition containing white tea extract, comprising, by weight, 0.15 parts of white tea extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0058] The rest of the implementation is the same as Example 1.

[0059] Example 4

[0060] A soothing composition containing white tea extract, which comprises, by weight, 0 parts of white tea extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0061] The rest of the implementation is the same as Example 1.

[0062] Example 5

[0063] A soothing composition containing white tea extract, comprising, by weight, 0.025 parts of olive leaf extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0064] The olive leaf extract was purchased from Draco, with the trade name: olive polyol extract.

[0065] The rest of the implementation is the same as Example 1.

[0066] Example 6

[0067] A soothing composition containing white tea extract, comprising, by weight, 0.05 parts of olive leaf extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0068] The white tea extract was purchased from Luojie, trade name: White Tea Extract CI.

[0069] Example 7

[0070] A soothing composition containing white tea extract, comprising, by weight, 0.05 parts of olive leaf extract, 0.05 parts of eucommia bark extract, 0.1 parts of citrus fruit extract, 0.2 parts of artemia extract, and 0.1 parts of gardenia fruit extract.

[0071] The white tea extract was purchased from Croda with the trade name: CRODAROM WHITE TEA EC.

[0072] Application Example 1

[0073] The invention discloses an application of a soothing composition containing white tea extract, which is applied to a soothing essence.

[0074] The raw materials for preparing the soothing essence include, by weight percentage, 7.3% of an oil phase, 0.2% of an efficacy phase, 3.69% of a soothing composition containing white tea extract, and the water phase is supplemented to 100%.

[0075] The oil phase includes humectant 5.9%, thickener 0.8%, and skin conditioner 0.6%.

[0076] The moisturizer includes a combination of glycerin, panthenol and sodium hyaluronate in a weight ratio of 3:2.7:0.2.

[0077] The thickener is xanthan gum.

[0078] The skin conditioning agent is a combination of p-hydroxyacetophenone and dipotassium glycyrrhizinate in a weight ratio of 0.2:0.4.

[0079] The water phase includes deionized water and panthenol. Based on the weight percentage of the soothing essence, the weight ratio of the panthenol is 2%.

[0080] The functional phase is olive fruit extract.

[0081] The soothing composition containing white tea extract comprises a combination of white tea extract, citrus extract, eucommia bark extract, gardenia fruit extract and Artemia extract, with a weight ratio of 0.1:0.05:0.1:0.2:0.1.

[0082] A method for preparing a soothing essence comprises the following steps:

[0083] S1 premixes the moisturizer and the thickener to obtain a premixed liquid A;

[0084] The S2 reactor was heated in a water bath, stirred, and premixed liquid A was added, and then other raw materials of the oil phase were added, heated to 80°C, stirred at this temperature for 10 minutes, and homogenized at 6000 rpm for 5 minutes;

[0085] S3 cools down to 45°C, adds water phase and stirs evenly;

[0086] S4 cools down to 40°C, adds the soothing composition containing white tea extract, stirs the functional phase evenly, and passes the test.

[0087] Application Comparative Example 1

[0088] The invention discloses an application of a soothing composition containing white tea extract, which is applied to a soothing essence.

[0089] The raw materials for preparing the soothing essence include, by weight percentage, 7.3% of the oil phase, 0.2% of the soothing composition, and the water phase is supplemented to 100%.

[0090] The oil phase includes humectant 5.9%, thickener 0.8%, and skin conditioner 0.6%.

[0091] The moisturizer includes a combination of glycerin, panthenol and sodium hyaluronate in a weight ratio of 3:2.7:0.2.

[0092] The thickener is xanthan gum.

[0093] The skin conditioning agent is a combination of p-hydroxyacetophenone and dipotassium glycyrrhizinate in a weight ratio of 0.2:0.4.

[0094] The water phase includes deionized water and panthenol. Based on the weight percentage of the soothing essence, the weight ratio of the panthenol is 2%.

[0095] The soothing composition is an olive leaf extract purchased from Draco with the trade name: olive polyol extract.

[0096] A method for preparing a soothing essence comprises the following steps:

[0097] S1 premixes the moisturizer and the thickener to obtain a premixed liquid A;

[0098] The S2 reactor was heated in a water bath, stirred, and premixed liquid A was added, and then other raw materials of the oil phase were added, heated to 80°C, stirred at this temperature for 10 minutes, and homogenized at 6000 rpm for 5 minutes;

[0099] S3 cools down to 45°C, adds water phase and stirs evenly;

[0100] S4 cools down to 40°C, adds the soothing composition, stirs evenly, and passes the test.

[0101] Performance Testing

[0102] The soothing composition containing white tea extract prepared in the example is diluted with deionized water to a corresponding concentration and set aside.

[0103] 1. Cytotoxicity test: The cell viability of mouse mononuclear macrophages (RAW264.7) was tested using the cytotoxicity test method (MTT method) with reference to the Standard Operating Procedure for Cosmetic Cytotoxicity Assay.

[0104] The mitochondria of living cells contain succinate dehydrogenase, which can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan and deposit it in cells, while dead cells do not have this function. Dimethyl sulfoxide solution (DMSO) can dissolve the formazan in cells, and the absorbance value at a wavelength of 490nm is measured using a multifunctional microplate reader. Since the amount of crystal formation is proportional to the number of living cells, the absorbance value can indirectly reflect the number of living cells.

[0105] 1. Inoculate HaCaT and RAW264.7 cells (from Wuhan Pronocell Life Science Co., Ltd.) in 96-well plates (5×10 3 Each experimental group of cells: 5×10 3 Incubate at 37°C, 5% CO2 for 24 hours.

[0106] 2. Administration: fresh culture medium (DMEM+10% FBS) containing corresponding concentrations of the examples was added to the sample group, fresh culture medium (DMEM+10% FBS) was added to the normal control group, and blank culture medium (DMEM) was added to the blank group. The cells were incubated at 37° C. and 5% CO 2 for 24 h.

[0107] 3. After the incubation, add 200 μL of 0.05 wt% MTT solution to each well and continue culturing for 4 h.

[0108] 4. Remove the culture medium, add 200 μL DMSO solution, shake to mix, and measure the absorbance at 490 nm.

[0109]

[0110] OD 490 : Absorbance value at 490nm. The test results are shown in Table 1.

[0111] Table 1

[0112]

[0113]

[0114]

[0115] The composition of Example 1 has no cytotoxicity at a relatively high concentration and is highly safe to use.

[0116] 2. Anti-inflammatory effect test: refer to the "Standard operating procedures for the determination of inflammatory factors in mouse macrophages in vitro" standard to test the relative expression of inflammatory factors in mouse mononuclear macrophages. Human immortalized keratinocytes (HaCaT), mouse mononuclear macrophages (RAW264.7).

[0117] Inflammatory response is a series of local cell and vascular defense responses that occur after tissue damage. It is a protective mechanism for the body to remove damaging factors through various immune cells (such as macrophages, etc.). Macrophages are important inflammatory cells in the body. They participate in the initiation of inflammatory responses and secrete a large number of proinflammatory mediators and proinflammatory factors, playing an important role in the pathogenesis of inflammatory diseases. Cyclooxygenase (COX), also known as prostaglandin endoxidase reductase, is a bifunctional enzyme with cyclooxygenase and catalase activities. It is a key enzyme that catalyzes the conversion of arachidonic acid into prostaglandins. Among them, COX-2 is an inducible enzyme. The activity of COX-2 in normal tissue cells is extremely low. When cells are stimulated by inflammation, its expression level in inflammatory cells can increase to 10-80 times the normal level, causing an increase in the content of PEG2, PGI2 and PGE1 in the inflammatory site, leading to inflammatory response and tissue damage. In this experiment, LPS was used to induce macrophages, and the relative expression of COX-2 gene was detected to evaluate whether the sample had anti-inflammatory effects.

[0118] 1. Inoculate RAW264.7 (from Wuhan Pronocell Life Science Co., Ltd.) in a 6-well plate (6×10 5 Each experimental group of cells: 6×10 5 The cells were incubated at 37°C, 5% CO2 for 24 h.

[0119] 2. After the incubation, the culture medium was removed and washed 1-2 times with D-Hanks (biological buffer). The normal control group was filled with fresh culture medium, the model control group was filled with fresh culture medium containing the modeling agent (0.5 μg / mL LPS), and the sample group was filled with fresh culture medium containing the example and the modeling agent (0.5 μg / mL LPS). The cells were incubated at 37°C and 5% CO2 for 24 h.

[0120] 3. Extract total RNA from each experimental group, synthesize cDNA, and use q-PCR to detect the gene expression of β-actin and target genes.

[0121] The specific steps are as follows: Use RNA rapid extraction kit to extract total RNA from each experimental group

[0122] 3.1 Lyse cells with Lysis Buffer; add an equal volume of anhydrous ethanol and mix thoroughly.

[0123] 3.2 Add the above sample lysate to the centrifuge column and centrifuge at 4000g for 1 min at room temperature.

[0124] 3.3 Add 500 μL Wash Buffer to the centrifuge tube and centrifuge at 12000g for 1 min at room temperature.

[0125] 3.4 Transfer the centrifuge column to a new EP tube, open the cover and let it dry for 2 minutes; add 20 μL Elution Buffer to the centrifuge column.

[0126] 3.5 Place at room temperature for 1-2 minutes, centrifuge at 12000g for 1 minute at room temperature, discard the centrifuge column, and the centrifuge product is RNA.

[0127] 3.6 Synthesize cDNA using cDNA reverse transcription kit

[0128] 3.6.1 Reference The remaining genomic DNA was removed according to the instructions of Ⅲ1st Strand cDNA Synthesis SuperMix for qPCR (gDNAdigester plus) and incubated at 42°C for 2 min.

[0129] 3.6.2 Standard reverse transcription procedure: incubate at 25°C for 5 min, 55°C for 15 min, and 85°C for 5 min.

[0130] 3.7. Detect the gene expression of β-actin and target genes using q-PCR.

[0131] 3.7.1 Refer to Hieff Universal Blue qPCR SYBR Green Master Mix says

[0132] Follow the instructions to configure the q-PCR reaction system.

[0133] 3.7.2 The standard RT-QPCR procedure is as follows:

[0134]

[0135] 4. Use β-actin as an internal reference for gene expression and calculate the relative RNA expression of the target gene.

[0136] Relative RNA expression = 2 ΔΔC(t)

[0137]

[0138] ΔC(t)=C(t) 目的基因 -C(t) β-actin

[0139] The test results are shown in Table 2.

[0140] Table 2

[0141] Detection concentration (wt%) Relative expression of COX-2 gene Example 1 1.25 0.516 Example 2 1.25 0.576 Example 3 0.156 0.600 Example 4 0.312 0.643 Example 5 0.312 0.647 Example 6 0.312 0.639 Example 7 1.25 0.630

[0142] The expression of COX-2 was significantly downregulated, indicating that the expression of inflammatory factors can be reduced.

[0143] 3. High temperature stimulation test

[0144] In addition to natural aging caused by age, skin aging also includes common photoaging and temperature aging. Among them, skin aging caused by light stimulation such as ultraviolet rays or blue light has been fully explained and evaluated. However, there are fewer studies on the aging effects caused by temperature stimulation. The two most common types of proteins: heat shock proteins and cold shock proteins play an important regulatory role in the body's resistance to external temperature changes. For example, when the external temperature rises, the heat shock proteins in the cells will be upregulated to stabilize functional proteins, maintain cell homeostasis, and reduce cell damage. HSP70 plays an important role in the body's resistance to adverse stress, especially temperature stress. This experiment uses high temperature to induce human immortalized keratinocytes, and detects the relative expression of the HSP70 gene to evaluate whether the sample has resistance to high temperature stimulation.

[0145] 1. Inoculate HaCaT (from Wuhan Pronocell Life Science Co., Ltd.) in a 6-well plate (6×10 5 Each experimental group of cells: 6×10 5 The cells were incubated at 37°C, 5% CO2 for 24 h.

[0146] 2. Pre-sampling: The normal control group and the model control group were given fresh culture medium (DMEM+10% FBS), and the sample group was given fresh culture medium containing samples, and the cells were incubated at 37°C and 5% CO2 for 24 hours.

[0147] 3. After the incubation, remove the culture medium, wash with D-Hanks for 1-2 times, and incubate the model control group and sample group in a 44°C incubator for 30 minutes.

[0148] 4. Remove D-Hanks, replace the normal control group and model control group with fresh culture medium, and replace the sample group with fresh culture medium containing samples, and continue to incubate at 37°C, 5% CO2 for 24 hours.

[0149] 5. Extract total RNA from each experimental group, synthesize cDNA, and use q-PCR to detect the gene expression of β-actin and target genes.

[0150] The specific steps are as follows: Use RNA rapid extraction kit to extract total RNA from each experimental group

[0151] 5.1 Lyse cells with Lysis Buffer; add an equal volume of anhydrous ethanol and mix thoroughly.

[0152] 5.2 Add the above sample lysate to the centrifuge column and centrifuge at 4000g for 1 min at room temperature.

[0153] 5.3 Add 500 μL Wash Buffer to the centrifuge tube and centrifuge at 12000g for 1 min at room temperature.

[0154] 5.4 Transfer the centrifuge column to a new EP tube, open the cover and let it dry for 2 minutes; add 20 μL Elution Buffer to the centrifuge column.

[0155] 5.5 Place at room temperature for 1-2 minutes, centrifuge at 12000g for 1 minute at room temperature, discard the centrifuge column, and the centrifuge product is RNA.

[0156] 5.6 Synthesize cDNA using cDNA reverse transcription kit

[0157] 5.6.1 Reference The operation was performed according to the instructions of Ⅲ1st Strand cDNA Synthesis SuperMix for qPCR (gDNAdigester plus), residual genomic DNA was removed, and the mixture was incubated at 42°C for 2 min.

[0158] 5.6.2 Standard reverse transcription procedure: incubate at 25°C for 5 min, 55°C for 15 min, and 85°C for 5 min.

[0159] 5.7. Detect the gene expression of β-actin and target gene using q-PCR.

[0160] 5.7.1 Refer to Hieff Follow the instructions of Universal Blue qPCR SYBR Green Master Mix to configure the q-PCR reaction system.

[0161] 5.7.2 The standard RT-QPCR procedure is as follows:

[0162]

[0163] 6. Use β-actin as an internal reference for gene expression and calculate the relative RNA expression of the target gene.

[0164] Relative RNA expression = 2 ΔΔC(t)

[0165]

[0166] ΔC(t)=C(t) 目的基因 -C(t) β-actin

[0167] The test results are shown in Table 3.

[0168] Table 3

[0169] Detection concentration (wt%) Relative expression of COX-2 gene Example 1 5 0.645 Example 2 5 0.701 Example 3 0.156 0.724 Example 4 0.156 0.747 Example 5 0.312 0.736 Example 6 0.312 0.754 Example 7 5 0.727

[0170] The composition of Example 1 can significantly down-regulate the transcription levels of HSP70 and CIRBP caused by high temperature stimulation, which preliminarily proves that the sample has a protective effect on the aging reaction caused by temperature stimulation.

[0171] 4. Stability test

[0172] The essences of Application Example 1 and Application Comparative Example 1 were placed in transparent bottles and the state of the essences was observed at room temperature. Figure 1 , Left: Application Comparative Example 1; Right: Application Example 1. Figure 1 It can be seen that the state of Application Example 1 is clearer.

[0173] The essences of Application Example 1 and Application Comparative Example 1 were placed in transparent bottles and stored at 25°C, 42°C, 5°C, and -15°C for 3 months. Figure 2 , see Application Example 1 Figure 3 . From left to right: 25℃, 42℃, 5℃, -15℃.

[0174] The essence of Comparative Example 1 changes color significantly at high temperature and is turbid, with poor stability.

[0175] The essence of Application Example 1 has less obvious color change at high temperature and is in a clear state with high stability.

Claims

1. A soothing composition containing white tea extract, characterized in that: The soothing composition comprises white tea extract, eucommia bark extract, citrus fruit extract, artemia extract, and gardenia fruit extract.

2. The soothing composition containing white tea extract according to claim 1, characterized in that: The white tea extract is extracted from Fuding white tea, and the harvest time of Fuding white tea is April to May in the northern hemisphere.

3. The soothing composition containing white tea extract according to claim 1, characterized in that: The content of tea polyphenols in the white tea extract is 6.0-6.5 mg / mL.

4. The soothing composition containing white tea extract according to claim 1, characterized in that: In parts by weight, the soothing composition comprises 0.1-2 parts of white tea extract, 0.05-1 parts of eucommia bark extract, 0.1-2 parts of citrus fruit extract, 0.1-3 parts of artemia extract, and 0.1-2 parts of gardenia fruit extract.

5. The soothing composition containing white tea extract according to claim 4, characterized in that: The weight ratio of the white tea extract to the eucommia bark extract is (1-3):1; the weight ratio of the white tea extract to the citrus fruit extract is (0.5-1.5):

1.

6. The soothing composition containing white tea extract according to claim 4, characterized in that: The weight ratio of the white tea extract to the Artemia extract is (0.5-1.5):1; the weight ratio of the white tea extract to the Gardenia fruit extract is (0.5-3):

1.

7. An application of the soothing composition containing white tea extract according to any one of claims 1 to 6, characterized in that: Used in soothing essence, skin care water, skin care lotion, skin care ointment and skin care cream.

8. The use of the soothing composition containing white tea extract according to claim 7, characterized in that: The soothing essence comprises, by weight percentage, 2-18% of an oil phase, 0.1-3% of an efficacy phase, 4-8% of a soothing composition containing white tea extract, and the water phase is supplemented to 100%.

9. The use of the soothing composition containing white tea extract according to claim 8, characterized in that: Calculated by weight percentage of the soothing essence, the oil phase includes 2-11% moisturizer, 0.5-2% thickener, and 0.2-5% skin conditioner.

10. A method for preparing the soothing essence according to claim 9, characterized in that: The following steps are involved: S1 premixes the moisturizer and the thickener to obtain a premixed liquid A; Heat the S2 reactor in a water bath, start stirring, add premix A, and then add other raw materials of the oil phase, heat to 70-90°C, keep stirring for 8-15 minutes, and homogenize at 5000-7000rpm for 4-6 minutes; S3 cools down to 40-50℃, adds water phase and stirs evenly; S4 cools down to 30-50℃, adds the soothing composition containing white tea extract, stirs the functional phase evenly, and passes the test.

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