Whitening and soothing eurotium cristatum tea fermentation composition as well as preparation method and application thereof

Abstract

The invention discloses a whitening and soothing eurotium cristatum tea fermentation composition as well as a preparation method and application thereof, and relates to the technical field of skin products. The invention provides a whitening and soothing eurotium cristatum tea fermentation composition. The whitening and soothing eurotium cristatum tea fermentation composition is prepared from the following components in parts by weight: 0.5 to 2 parts of eurotium cristatum tea fermentation filtrate, 0.1 to 0.8 part of alpha-arbutin and 0.01 to 1.5 parts of rhizoma gastrodiae root extract. When the eurotium cristatum tea fermentation filtrate, the gastrodia elata root extract and the alpha-arbutin are cooperatively used, the limitation of a single effect can be broken through, and the effects of safely whitening, quickly relieving and persistently shrinking pores are considered.

Description

Technical Field

[0001] The invention relates to the technical field of skin products, in particular to a whitening and soothing golden flower fungus tea fermentation composition and a preparation method and application thereof. Background Art

[0002] As consumers' comprehensive demands for skin health and appearance continue to increase, skin products with whitening, soothing and pore care functions have become a hot spot in the market. However, the existing products with the above multiple functions generally have problems such as single mechanism of action, limited effect and insufficient safety, which makes it difficult to meet consumers' demand for efficient and gentle skin care.

[0003] In the field of whitening, traditional products rely on highly toxic ingredients (such as hydroquinone) or inefficient and mild ingredients (such as arbutin). The former has the risk of sensitization and pigment abnormalities, and the latter is limited by transdermal absorption rate and formula stability, resulting in slow and insignificant whitening effects. At the same time, modern environmental pressures (such as ultraviolet rays, pollution) and improper skin care behaviors can easily cause skin sensitivity and inflammation. Existing soothing products are difficult to take into account both immediate soothing and long-term barrier protection while whitening care due to the potential irritation of chemical synthetic ingredients or the unstable penetration efficiency and content of plant active ingredients. In addition, the improvement of the problem of enlarged pores still remains at the surface convergence level. Although ingredients such as alcohol and witch hazel can temporarily tighten the stratum corneum, they will destroy the water and oil balance of the skin, aggravate the discomfort of sensitive skin, and lack a deep mechanism for regulating sebaceous gland secretion and collagen regeneration, resulting in repeated pore problems.

[0004] In summary, the realization of whitening, soothing and pore care effects in the existing technology has bottlenecks in ingredient safety, depth of action and multi-effect synergy. It is urgent to develop an innovative formula that can break through the limitation of a single effect and take into account safe whitening, rapid soothing and lasting pore shrinking to meet consumers' comprehensive skin care needs for "safe whitening, rapid soothing and lasting pore shrinking". Summary of the invention

[0005] Based on this, the purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a whitening and soothing golden flower fungus tea ferment composition that is safe, whitening, quickly soothing, and lastingly shrinks pores, as well as a preparation method and application thereof.

[0006] To achieve the above-mentioned purpose, in the first aspect of the present application, the technical solution adopted by the present invention is: a whitening and soothing golden flower fungus tea fermentation composition, comprising the following components in parts by weight: 0.5-2 parts of golden flower fungus tea fermentation filtrate, 0.1-0.8 parts of α-arbutin, and 0.01-1.5 parts of Gastrodia elata root extract.

[0007] Preferably, the whitening and soothing Flos Caryophyllata tea fermentation composition comprises the following components in parts by weight: 1-1.5 parts of Flos Caryophyllata tea fermentation filtrate, 0.3-0.5 parts of α-arbutin, and 0.5-1 part of Gastrodia elata root extract.

[0008] Preferably, in the whitening and soothing Flos Caryophyllata tea fermentation composition, based on the total weight of the whitening and soothing Flos Caryophyllata tea fermentation composition, the weight percentage of the Flos Caryophyllata tea fermentation filtrate is 60-70%.

[0009] The Flos Caryophyllata tea fermentation filtrate is an active liquid extracted from tea as a matrix through aerobic fermentation by artificially inoculating Eurotium cristatum (also known as Flos Caryophyllata). Since Flos Caryophyllata itself originates from tea, it is more suitable for fermentation with tea. The resulting filtrate is rich in metabolites of Flos Caryophyllata, such as small molecule polypeptides, polysaccharides, organic acids, etc., combining the natural components of tea and the advantages of microbial fermentation. It can scavenge free radicals, inhibit lipid peroxidation, and delay skin photoaging and collagen loss. The Gastrodia elata root extract is a natural active ingredient extracted from the dried tuber of the orchid plant Gastrodia elata, containing gastrodin, polysaccharides, phenolic compounds, p-hydroxybenzyl alcohol, vanillin and other components. It is widely used in pharmacy and has sedative, anti-inflammatory and neuroprotective effects, participating in the regulation of cardiovascular function and antioxidant reactions. Due to the presence of components such as chlorogenic acid, it has the effects of enhancing immunity and anti-aging. α-Arbutin has become a core ingredient of high-end skin care products due to its high-efficiency whitening, low irritation and multi-path anti-aging characteristics. Its fermentation production process and strict quality control ensure safety and efficacy, and it is applicable to multiple fields such as skin care products, medicine and agriculture. It is an epimer of β-arbutin with a different glycosidic bond direction, having stronger tyrosinase inhibitory ability and stability.

[0010] When the fermented filtrate of Flos Lonicerae and Camellia sinensis, Gastrodia elata root extract, and α-arbutin are used in combination, they can break through the limitations of single efficacy and achieve three effects: safe whitening, rapid soothing, and long-lasting pore shrinking. In terms of whitening, α-arbutin inhibits the activity of tyrosinase, blocks the melanin production pathway, and reduces the formation of age spots. The fermented filtrate of Flos Lonicerae and Camellia sinensis contains organic acids that gently exfoliate the cutin, accelerating the metabolism of old cutin containing melanin. Its polypeptide components may promote epidermal renewal and indirectly brighten the skin tone. Gastrodia elata root extract is rich in antioxidants (such as phenols), reducing the excessive synthesis of melanin caused by oxidative stress. At the same time, its anti-inflammatory effect prevents post-inflammatory hyperpigmentation. The three work together from three aspects: inhibiting production, accelerating metabolism, and preventing the cause, which may achieve more efficient and long-lasting whitening effects. In terms of soothing, the prebiotic components in the fermented filtrate of Flos Lonicerae and Camellia sinensis strengthen the skin barrier, and the organic acids regulate the pH value, alleviating external stimuli. The anti-inflammatory active components (such as gastrodin) in Gastrodia elata root extract reduce redness and sensitivity, soothing skin discomfort caused by ultraviolet rays or irritation. α-arbutin complements other soothing components. The three work together from the aspects of anti-inflammation and barrier repair, which is suitable for sensitive skin. In terms of pore shrinking, the organic acids (such as lactic acid and acetic acid) in the fermented filtrate of Flos Lonicerae and Camellia sinensis have astringency, and at the same time, their good oil control effect reduces sebum secretion and prevents pore blockage. Gastrodia elata root extract can prevent pore thickening caused by inflammation around the hair follicles; α-arbutin can reduce pigment deposition, improve uneven skin tone around the pores, and make the pores visually more concealed. The three improve the pore appearance and enhance the skin fineness through multiple ways such as oil control, anti-inflammation, firming, and skin tone homogenization.

[0011] Preferably, the fermented filtrate of Flos Lonicerae and Camellia sinensis can be a commercially available product or can be prepared at home.

[0012] In a specific embodiment, the fermented filtrate of Flos Lonicerae and Camellia sinensis includes at least one of the fermented filtrate of Flos Lonicerae and Camellia sinensis of Mingqi Biology, the fermented filtrate of black tea of Guangzhou Xiyuan, and the compound fermented filtrate of Eurotium cristatum tea jointly developed by Guangzhou Zhongzhuang Beauty Industry and Hangzhou Tea Research Institute of All China Federation of Supply and Marketing Cooperatives.

[0013] Preferably, the preparation method of the fermented filtrate of Flos Lonicerae and Camellia sinensis refers to the patent application number: 202411759723.X, and the patent name: A method for improving the anti-aging effect of compound fermented liquid of Eurotium cristatum tea.

[0014] In a specific embodiment, the preparation method of the fermented filtrate of Flos Lonicerae and Camellia sinensis includes the following steps:

[0015] (1) Inoculate the activated seed liquid of Flos Lonicerae and Camellia sinensis into the tea-sea buckthorn leaf fermentation medium for primary fermentation, and culture it on a shaker at 28±2°C for 24-72h to obtain the tea-sea buckthorn leaf fermentation culture solution;

[0016] (2) Inoculate the yeast seed solution into the tea - seabuckthorn leaf fermentation culture medium, and culture it on a shaker at 28 ± 2 °C for co - fermentation for 24 - 36 h;

[0017] (3) After the fermentation is completed, centrifuge the co - fermentation culture medium to obtain the supernatant;

[0018] (4) Pass the obtained supernatant through a sterilizing filter to obtain a sterile tea - seabuckthorn leaf composite fermentation broth.

[0019] Preferably, the Jinhuajun is Eurotium cristatum K8824, which was deposited at the China Center for Type Culture Collection on April 7, 2024. The deposit number of Eurotium cristatum K8824 is: CCTCC M2024643, and the Latin name is Eurotium cristatum.

[0020] Preferably, the preparation method of the activated seed solution of Eurotium cristatum K8824 is as follows: Streak the deposited Eurotium cristatum K8824 on a PDA plate medium, activate it at 28 ± 1 °C for 48 - 72 h, scrape the spores on the plate, after thorough washing with sterile water, fully shake and mix the spore solution into a spore suspension, and adjust the concentration of the spore suspension to 1.0×10 7 CFU / mL to obtain the activated seed solution of Eurotium cristatum K8824.

[0021] Preferably, the preparation method of the fermentation medium is as follows:

[0022] (a) Grind tea and seabuckthorn leaves into fine powders of 40 - 100 mesh respectively, and then mix the tea and seabuckthorn leaves evenly according to the mass ratio of 7:3 - 5:4 to obtain a tea - seabuckthorn leaf fermentation substrate;

[0023] (b) Mix the tea - seabuckthorn leaf fermentation substrate with pure water according to the ratio of 1:30 - 1:9 and stir evenly to obtain a fermentation medium, and sterilize it at 121 °C for 15 min.

[0024] Among them, the yeast includes at least one of Angel yeast and Saccharomyces cerevisiae.

[0025] Among them, Saccharomyces cerevisiae is deposited at the China Center for Type Culture Collection, and the strain deposit number is CCTCC KY2008617.

[0026] Among them, the tea selected is black tea, which is one of black teas such as Keemun black tea, Yingde black tea, and Lapsang Souchong.

[0027] The preparation method of the Saccharomyces cerevisiae seed liquid is as follows: inoculate yeast into YPD liquid medium and culture for 24 - 36 h to obtain yeast seed liquid, and then adjust the spore suspension to 1.0×10 8 CFU / mL to obtain the Saccharomyces cerevisiae seed liquid.

[0028] The preparation method of the Angel yeast seed liquid is as follows: open the bagged Angel high - activity dry yeast beside the alcohol lamp flame, take 2 g of yeast powder and add it to the sterilized 5% glucose solution, place it on a shaker at 28 ± 1 °C and shake - culture and activate for 3 - 6 h to obtain the Angel yeast seed liquid, and then adjust the concentration to 1.0×10 8 CFU / mL to obtain the Angel yeast seed liquid.

[0029] Among them, the inoculation amount of the Eurotium cristatum K8824 seed liquid is 3 - 8%.

[0030] Among them, the inoculation amount of the yeast is 2 - 5%.

[0031] Among them, the shaker speed is 120 - 200 r / min.

[0032] Among them, the centrifugation speed is 2800 - 3500 r / min.

[0033] Among them, the sterilizing filter is a sterilizing filter with a pore size of 0.15 - 0.30 μm.

[0034] In the second aspect of the present application, the present application provides the application of the whitening and soothing Eurotium cristatum tea - yeast composition in the preparation of skin care products.

[0035] Preferably, the skin care products include color cosmetics and skin care products; the skin care products include any one of lotion, emulsion, cream, facial mask, essence, essence oil, spray, sunscreen; the color cosmetics include any one of liquid foundation, lip balm.

[0036] In the third aspect of the present application, the present application provides a cream, and the cream includes the following components in mass percentage: the whitening and soothing Eurotium cristatum tea - yeast composition 0.5 - 3%, the skin care product matrix 2 - 40%, and the balance is water.

[0037] Preferably, the present application provides a cream, including the following components in mass percentage: the whitening and soothing Eurotium cristatum tea - yeast composition 0.5 - 3%, humectant 10 - 20%, emulsifier 1 - 5%, thickener 0.3 - 1%, skin conditioner 5 - 10%, preservative 0.2 - 0.8%, pH regulator 0.1 - 0.5%, essence 0.001 - 0.1%, and the balance is water.

[0038] Preferably, the skin care product matrix includes at least one of a thickener, a humectant, an emulsifier, a skin conditioner, a preservative, a fragrance, and a pH regulator.

[0039] Preferably, the thickener includes at least one of xanthan gum, carbomer, polyvinyl alcohol, sodium polyacrylate, hydroxyethyl cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.; the humectant includes at least one of glycerol, hyaluronic acid, trehalose, glycerol, butanediol, propylene glycol, etc.; the emulsifier includes at least one of fatty acid glycerides, polysorbates, sodium lauryl sulfate, potassium cetyl phosphate, etc.; the skin conditioner includes at least one of squalane, sweet almond oil, jojoba oil, shea butter, allantoin, vitamin E, panthenol, etc.; the preservative includes at least one of phenoxyethanol, potassium sorbate, sodium benzoate, etc.; the pH regulator includes at least one of citric acid, sodium citrate, sodium hydroxide, triethanolamine, lactic acid, etc.

[0040] Compared with the prior art, the beneficial effects of the present invention are as follows: when the fermented filtrate of Flos Chrysanthemi Indici tea, the extract of Gastrodia elata root, and α-arbutin are used in combination, the limitation of single efficacy can be broken through, and the effects of safe whitening, rapid soothing, and long-lasting pore shrinking can be taken into account. Description of the Drawings

[0041] Figure 1 It is a diagram of pore shrinking on the human body for Application Example 1. Detailed Embodiments

[0042] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with the drawings and specific embodiments. The purpose is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention. The experimental reagents and instruments designed in the embodiments and comparative examples of the present invention are all common ordinary reagents and instruments unless otherwise specified, and can be obtained through commercial channels. In the embodiments and comparative examples, the experimental methods used are all conventional methods unless otherwise specified; and unless otherwise specified, the raw materials used in parallel experiments are of the same batch.

[0043] Now, the raw materials used in the embodiments of the present application will be further described, but are not limited to the following raw materials:

[0044] α-Arbutin: Shenzhen Shanhai Innovation Technology Co., Ltd.;

[0045] Extract of Gastrodia elata root: Guangzhou Baihaobo Co., Ltd.;

[0046] Fermented filtrate of probiotic tea: Hangzhou Jiajiale Biotechnology Co., Ltd.;

[0047] β-arbutin: Shenzhen Shanhai Innovation Technology Co., Ltd.;

[0048] Gastrodia elata extract: Shanyang Lianfeng Biotechnology Co., Ltd.

[0049] The preparation method of the activated seed liquid of Eurotium cristatum K8824 is as follows: Streak the preserved Eurotium cristatum K8824 on a PDA plate medium, activate it at 28 ± 1 °C for 72 h, scrape the spores on the plate, and after thoroughly washing with sterile water, fully shake and mix the spore liquid into a spore suspension, and adjust the concentration of the spore suspension to 1.0×10 7 CFU / mL to obtain the activated seed liquid of Eurotium cristatum K8824.

[0050] The preparation method of the Saccharomyces cerevisiae seed liquid is as follows: Inoculate the yeast into a YPD liquid medium and culture for 36 h to obtain a yeast seed liquid, and then adjust the spore suspension to 1.0×10 8 CFU / mL to obtain the Saccharomyces cerevisiae seed liquid.

[0051] The preparation method of the Angel yeast seed liquid is as follows: Open the bagged Angel high-activity dry yeast beside the alcohol lamp flame, take 2 g of yeast powder and add it to the sterilized 5% glucose solution, place it on a shaker at 28 ± 1 °C and shake and culture for 6 h to obtain the Angel yeast seed liquid, and then adjust the concentration to 1.0×10 8 CFU / mL to obtain the Angel yeast seed liquid.

[0052] The golden flower fungus is Eurotium cristatum K8824, which was deposited at the China Center for Type Culture Collection on April 7, 2024. The deposit number of this Eurotium cristatum K8824 is: CCTCC M 2024643; the Saccharomyces cerevisiae is deposited at the China Center for Type Culture Collection, and the strain deposit number is CCTCC KY2008617.

[0053] Golden flower fungus tea fermentation filtrate-1: Self-made, the preparation method is as follows:

[0054] (1) Grind tea and seabuckthorn leaves into fine powders of 40 meshes respectively, and mix them evenly at a mass ratio of tea to seabuckthorn leaves of 6:4 to obtain a tea-seabuckthorn leaf fermentation substrate; the tea selected is Keemun black tea;

[0055] (2) Mix the tea-seabuckthorn leaf fermentation substrate with pure water at a ratio of 1:30 and stir evenly to obtain a fermentation medium, sterilize it at 121 °C for 15 min;

[0056] (3) Inoculate the Eurotium cristatum seed liquid into the fermentation medium for primary fermentation. The inoculation amount of Eurotium cristatum is 5%, and culture it on a shaker at 28 ± 2 °C for 48 h with a shaker speed of 150 r / min;

[0057] (4) After 48 h of fermentation of Eurotium cristatum K8824, inoculate the Angel yeast seed liquid into the tea-seabuckthorn leaf fermentation liquid for co-fermentation. The inoculation amount is 3%, and co-culture it on a shaker at 28 ± 2 °C for 24 h with a shaker speed of 150 r / min.

[0058] (5) After the fermentation is completed, centrifuge the fermentation medium to obtain the supernatant at a centrifugation speed of 3000 r / min.

[0059] (6) Pass the obtained supernatant through a 0.22 μm sterilizing filter to obtain a sterile tea-seabuckthorn leaf composite fermentation liquid, namely Jinhuajun tea fermentation filtrate-1.

[0060] Jinhuajun tea fermentation filtrate-2: Self-made. The preparation method is different from that of Jinhuajun tea fermentation filtrate-1 only in step (4). Specifically, after 48 h of fermentation of Eurotium cristatum K8824, inoculate the Saccharomyces cerevisiae seed liquid into the tea-seabuckthorn leaf fermentation liquid for co-fermentation. The inoculation amount is 3%, and co-culture it on a shaker at 28 ± 2 °C for 24 h with a shaker speed of 150 r / min.

[0061] Jinhuajun tea fermentation filtrate-3: Self-made. The preparation method is different from that of Jinhuajun tea fermentation filtrate-1 only in steps (4)-(5). Ferment Eurotium cristatum alone without adding yeast. After 48 h of fermentation of Eurotium cristatum K8824, centrifuge the fermentation medium to obtain the supernatant at a centrifugation speed of 3000 r / min.

[0062] Jinhuajun tea fermentation filtrate-4: Self-made. The preparation method is different from that of Jinhuajun tea fermentation filtrate-1 only in step (1). Do not add seabuckthorn leaves, and keep the total mass of tea the same as the total mass of tea and seabuckthorn leaves in Jinhuajun tea fermentation filtrate-1.

[0063] Jinhuajun tea fermentation filtrate-5: Jinhuahong tea fermentation filtrate, Mingqi Biotechnology Co., Ltd.;

[0064] Jinhuajun tea fermentation filtrate-6: Jinhuhei tea fermentation liquid, Mingqi Biotechnology Co., Ltd.;

[0065] Examples and comparative examples

[0066] The present invention provides a whitening and soothing Jinhuajun tea fermentation composition. The components and weight parts of the whitening and soothing Jinhuajun tea fermentation composition are selected as shown in Table 1-2. The preparation method of the whitening and soothing Jinhuajun tea fermentation composition includes the following steps: Mix the components evenly to obtain the whitening and soothing Jinhuajun tea fermentation composition.

[0067] Table 1

[0068]

[0069]

[0070] Table 2

[0071]

[0072]

[0073] Application Examples

[0074] The application examples, comparative application examples and blank application examples of the present invention provide a facial cream, the components (mass percentage) of the facial cream are shown in Table 3; wherein the whitening and soothing golden flower fungus tea ferment compositions used in application examples 1-10 are respectively the whitening and soothing golden flower fungus tea ferment compositions prepared in embodiments 1-10, for example, the whitening and soothing golden flower fungus tea ferment compositions in embodiment 1 are used in application example 1, the whitening and soothing golden flower fungus tea ferment compositions in embodiment 2 are used in application example 2, and so on; the whitening and soothing golden flower fungus tea ferment compositions used in comparative application examples 1-9 are respectively the whitening and soothing golden flower fungus tea ferment compositions prepared in comparative examples 1-9; the whitening and soothing golden flower fungus tea ferment compositions used in application example 11 are the whitening and soothing golden flower fungus tea ferment compositions prepared in embodiment 1;

[0075] Table 3

[0076]

[0077]

[0078] The preparation method of the facial cream in Application Example 1 comprises the following steps. The preparation methods of the other application examples are similar to those in Application Example 1, except that no relevant components are added:

[0079] Preparation: Accurately weigh the various raw materials in the above formula, prepare clean stirring equipment, heating equipment (such as a constant temperature water bath), emulsifying equipment (such as a homogenizer) and storage containers. Ensure that the operating environment is clean and hygienic to avoid impurities and microbial contamination.

[0080] Preparation of oil phase: Add squalane, sweet almond oil, jojoba oil, shea butter, polyglyceryl-6 stearate and sorbitan oleate into a clean container, place in a constant temperature water bath, heat to 75-80°C, stir to completely melt and mix evenly to form an oil phase.

[0081] Aqueous phase preparation: In another clean container, add panthenol, trehalose, glycerin, tea fermentation filtrate, Gastrodia elata root extract, α-arbutin, xanthan gum and deionized water, stir evenly, heat to 75 - 80 °C to ensure that all components are completely dissolved to form an aqueous phase.

[0082] Emulsification process: Under stirring, slowly add the aqueous phase to the oil phase while stirring, and then use a homogenizer for homogenization and emulsification to fully mix the oil phase and the aqueous phase to form a uniform and delicate cream. The emulsification time is generally 10 - 15 minutes, and the specific time can be adjusted according to the actual situation.

[0083] Adjust the pH value: When the temperature of the cream drops to 50 - 55 °C, slowly add triethanolamine while stirring, and at the same time monitor the pH value of the cream with pH test paper or a pH meter, and adjust the pH value to between 5.5 - 7.0 to make it suitable for skin use.

[0084] Add fragrance and preservative: When the temperature of the cream drops below 40 °C, add phenoxyethanol and fragrance in sequence and stir evenly to make them evenly dispersed in the cream.

[0085] Packaging and storage: After cooling the prepared cream to room temperature, pack it into a clean container and seal it for storage. During storage, avoid direct sunlight and high-temperature environment to ensure the quality and stability of the cream.

[0086] Performance test - 1 Soothing experiment - Macrophage experiment

[0087] Test samples: Use the samples from Application Examples 1 - 11, Comparative Application Examples 1 - 9, and Blank Application Example.

[0088] Experimental principle: Inflammatory response is a basic lesion of skin aging and is closely related to the integrity of the skin barrier. As an important part of the immune system, macrophages play a crucial role in the occurrence of inflammation. When the body is stimulated by external factors, macrophages will secrete a large number of inflammatory mediators to play an immunomodulatory role and maintain the immune homeostasis of the body. Inflammatory response is mediated by multiple molecular mechanisms. Among them, NO and PGE2 are two important inflammatory mediators involved in inflammatory response. A large amount of inflammatory mediators NO and PGE2 will be released by cells during inflammation. PTGS-2 is the key upstream enzyme for the synthesis of NO and PEG2. NO is synthesized by NOS, and NOS is divided into type I, type II and type III. Among them, type III NOS is mainly highly expressed in macrophages and various white blood cells under the stimulation of LPS, etc., and is the inducible NOS, that is, iNOS. The expression of iNOS is induced to increase by pro-inflammatory substances and inflammatory mediators, producing a large amount of NO, which promotes the inflammatory response. Therefore, regulating the release of NO mediated by iNOS is an important target for treating inflammatory response. PGE2 is generated by PTGS-2 catalyzing arachidonic acid. PTGS-2 starts to be highly expressed after being stimulated and plays an important role in the inflammatory response, mainly catalyzing the production of the inflammatory mediator PEG2. LPS is one of the components of the outer membrane of the cell wall of Gram-negative bacteria and has high activity. Its induction of inflammatory response mainly occurs through two pathways, NF-kB and MAPKs. Among them, the NF-kB signaling pathway is particularly crucial. LPS induces the expression of iNOS and PTGS-2 by activating NF-kB, and the phosphorylation of NF-kB is an important step in its activation.

[0089] Experimental method:

[0090] (1) Detection of NO and PGE2 content: Take cells in the logarithmic growth phase, collect the supernatant, and add 2 mL per well, about 2×10 5 cells to each well of a 6-well plate and culture until 80% of the bottom of the plate is covered. Except for the normal control group without drug treatment, the LPS model group cells are only added with LPS (1 μg / mL), and each experimental group is added with LPS (1 μg / mL) and the test sample. After continuing to culture for 24 h, use a NO detection kit and an ELISA kit to detect the contents of NO and PGE2 in the cell supernatant respectively.

[0091] (2) Determination of the expression of cellular iNOS and PTSG2 mRNA: When the cells were cultured in a 6-well plate until 80% of the bottom of the plate was covered, except for the normal control group without drug treatment, the cells in the LPS model group were only added with LPS (1 μg / mL), and the cells in each experimental group were added with LPS (1 μg / mL) and the test sample. After continuous culture for 24 h, the total RNA of the cells in the 6-well plate was extracted, and reverse transcription was performed to synthesize First-strand cDNA according to the RevertAidTM FirstStrand cDNA Synthesis Kit. The primer information of 18S, iNOS, and PTSG2 genes is shown in Table 4 below. Mix 9.5 μL of sterilized ultrapure water, 1 μL of cDNA, 1 μL each of the specific primers F forward and R reverse (final concentration is 1 μmol / L), and 12.5 μL of MaaximaTM SYBR Green, Fluorescein qPCR MasterMix (2×), and place it in an IQTM5 type fluorescence quantitative PCR instrument for amplification. The amplification steps are as follows: pre-denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 50 s, 45 cycles; extension at 72 °C for 7 min. Each amplification was set with 18S as an internal reference, and the normal control group was used as the unit "1", and the 2-△△Ct method was used for calculation and analysis.

[0092] Table 4 Primer sequences of amplified genes 18S, iNOS, and PTSG2

[0093]

[0094]

[0095] The test results are shown in Table 5.

[0096] Table 5

[0097]

[0098]

[0099] As can be seen from the above table, the gene down-regulation change rate of the cream prepared by the present invention reaches more than 50%. When the fermented filtrate of Flos Lonicerae fermented by Flos Lonicerae, the extract of Gastrodia elata roots, and α-arbutin are used in combination, a good soothing effect can be achieved.

[0100] It can be seen from the comparison of Application Example 1 and Application Examples 7-10 that the weight parts of the components in the composition will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the soothing effect of the obtained composition is better. It can be seen from the comparison of Application Examples 1-6 that the source of the fermented filtrate of Flos Lonicerae fermented by Flos Lonicerae will affect the performance of the product, and the fermented filtrate of Flos Lonicerae fermented by Flos Lonicerae obtained by a specific preparation method has a better soothing effect.

[0101] As can be seen from the comparison between Application Example 1 and Comparative Application Examples 1-9, when one or two of the components are not added, or when other component parts are used to make up for the non-addition, the effects of the present invention cannot be achieved; when replaced with other similar components, the corresponding effects of the present invention cannot be obtained either. When the weight parts of the components in the composition are not within the scope of this application, good synergistic effects cannot be exerted.

[0102] Performance Test - 2 Test Method for Human Skin Whitening Effect - Skin ITA°

[0103] Test Process:

[0104] 1. Test products: Essence prepared from Application Examples 1-11, Comparative Application Examples 1-9 and Blank Application Example;

[0105] 2. Negative control: Blank control in the blackened area.

[0106] 3. Positive control: 7% ascorbic acid (vitamin C) product, prepared according to the formula in Appendix I of the Notice No. 17 of 2021 of the National Medical Products Administration in the Technical Specifications for the Safety of Skin Care Products (2015 Edition).

[0107] 4. Subjects: 5 people in each test group, a total of 23 groups, and 115 effective subjects completed the test, aged 19 to 59 years old, meeting the volunteer inclusion and exclusion criteria for subjects.

[0108] 5. Instrument and equipment

[0109] 5.1 Solar simulator: All performance indicators meet the requirements of the measurement specification according to the Technical Specifications for the Safety of Skin Care Products.

[0110] 5.2 Skin colorimeter: An instrument capable of measuring the data of the CIE L*a*b color space formulated by the International Commission on Illumination (CIE).

[0111] 6. Test method: Conduct the test according to the specific requirements of the Technical Specifications for the Safety of Skin Care Products (2015 Edition).

[0112] Select the non-exposed area of the subject's thigh as the test site, and the area of each blackened test area should be no less than 0.5 cm 2, and it should be located within each painted sample area. Determine the MED of the test site of the subject. Use a daylight simulator to irradiate once a day at the same irradiation point with a dose of 0.75 times the MED for 4 consecutive days. The 4 days after the irradiation is the skin darkening period, and no treatment is given. On the 5th day after the irradiation, visually evaluate and detect the skin color of each test area with a skin color instrument, and eliminate the test areas with poor consistency. Starting from the 5th day, apply the corresponding test substance to each darkened test area according to the random table, and continuously apply the test substance for at least 4 weeks. The skin color should be visually evaluated and detected at 1 week, 2 weeks, 3 weeks, and 4 weeks after application, and the records should be made.

[0113] 7. Calculation method: 1). Use independent samples t-test (or rank sum test) for statistical analysis. Compare with the negative control, *p < 0.05. 2). Difference = skin parameter after product use - skin parameter before product use.

[0114] Test results: As shown in Table 6.

[0115] Table 6 Skin ITA° difference results (mean ± standard deviation) before and after using the sample

[0116]

[0117]

[0118]

[0119] As can be seen from the above table, after using the cream prepared by the present invention for 4 weeks, the difference in skin ITA° before and after use is more than 5.5. When the fermented filtrate of Flos Chrysanthemi Indici, the extract of Gastrodia elata root, and α-arbutin are used in combination, a good whitening effect can be achieved.

[0120] From the comparison of Application Example 1 and Application Examples 7 - 10, it can be seen that the weight parts of the components in the composition will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the whitening effect of the obtained composition is better. From the comparison of Application Examples 1 - 6, it can be seen that the source of the fermented filtrate of Flos Chrysanthemi Indici will affect the performance of the product, and the fermented filtrate of Flos Chrysanthemi Indici obtained by a specific preparation method has a better whitening effect.

[0121] From the comparison of Application Example 1 and Comparative Application Examples 1 - 9, it can be seen that when one or two components are not added, or when other component parts are used to make up for it while not adding, the effects of the present invention cannot be achieved; when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained. When the weight parts of the components in the composition are not within the scope of the present application, a good synergistic effect cannot be exerted.

[0122] Performance test - 3 Pore - shrinking human experiment

[0123] Randomly select 105 volunteers and divide them into 21 groups, with 5 people in each group. They use the facial creams prepared in the above Application Examples 1-11, Comparative Application Examples 1-9, and Blank Application Example for 4 weeks, and conduct efficacy evaluations through laboratory instrument tests. The specific efficacy evaluation methods are as follows:

[0124] Evaluation items: The number and area of pores before and 4 weeks after using the sample were tested with a VisioFace 1000D facial image analyzer;

[0125] Test basis: HAB / R-WI-242 "Evaluation of Oil Control Efficacy of Facial Care Skin Products - Human Test Method";

[0126] Judgment principle: If the number and area of skin pores in the test area show a significant decrease before and after using the product, it indicates that the tested sample has the effect of shrinking pores; otherwise, it does not have the effect of shrinking pores.

[0127] Calculation formula:

[0128] (The number value of pores at 4 weeks - the number value of pores before using the product) / the number value of pores before using the product = the change rate of pore number %; (The area value of pores at 4 weeks - the area value of pores before using the product) / the area value of pores before using the product = the change rate of pore area %.

[0129] The test results are shown in Table 7.

[0130] Table 7

[0131]

[0132]

[0133]

[0134] As can be seen from the above table, after using the facial cream prepared by the present invention for 4 weeks, both the number and area of pores have been significantly improved. When the fermented filtrate of Flos Chrysanthemi Indici, the extract of Gastrodia elata root, and α-arbutin are used in combination, a good pore shrinking effect can be achieved. Figure 1 It is the human pore shrinking diagram of Application Example 1. It can be seen that a good pore shrinking effect can be achieved after using it for 4 weeks.

[0135] By comparing Application Example 1 with Application Examples 7-10, it can be known that the weight parts of the components in the composition will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the pore shrinking effect of the obtained composition is better. By comparing Application Examples 1-6, it can be known that the source of the fermented filtrate of Flos Chrysanthemi Indici will affect the performance of the product, and the fermented filtrate of Flos Chrysanthemi Indici obtained by a specific preparation method has a better pore shrinking effect.

[0136] From the comparison between Application Example 1 and Comparative Application Examples 1-9, it can be seen that when one or two of the components are not added, or when the amounts of other components are used to make up for the non-added components, the effects of the present invention cannot be achieved; when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained either. When the weight parts of the components in the composition are not within the scope of the present application, good synergistic effects cannot be exerted.

[0137] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently substituted without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A whitening and soothing chrysanthemum fermented tea composition, characterized in that, Comprising the following components in parts by weight: 0.5 - 2 parts of fermented filtrate of Flos Lonicerae and Camellia sinensis, 0.1 - 0.8 part of α-arbutin, 0.01 - 1.5 parts of Gastrodia elata root extract.

2. The whitening and soothing chrysanthemum flower mushroom tea fermentation composition according to claim 1, wherein, Comprising the following components in parts by weight: 1 - 1.5 parts of fermented filtrate of Flos Lonicerae and Camellia sinensis, 0.3 - 0.5 part of α-arbutin, 0.5 - 1 part of Gastrodia elata root extract.

3. The whitening and soothing chrysanthemum fermented tea composition according to claim 1, characterized in that Based on the total weight of the whitening and soothing fermented composition of Flos Lonicerae and Camellia sinensis, the weight percentage content of the fermented filtrate of Flos Lonicerae and Camellia sinensis is 60 - 70%.

4. The whitening and soothing chrysanthemum flower mushroom tea fermentation composition according to claim 1, wherein The preparation method of the fermented filtrate of Flos Lonicerae and Camellia sinensis comprises the following steps: (1) Inoculate the activated seed liquid of Flos Lonicerae and Camellia sinensis into a tea - seabuckthorn leaf fermentation medium for primary fermentation, and culture it on a shaker at 28 ± 2°C for 24 - 72 h to obtain a tea - seabuckthorn leaf fermentation culture solution; (2) Inoculate the yeast seed liquid into the tea - seabuckthorn leaf fermentation culture solution, and culture it on a shaker at 28 ± 2°C for 24 - 36 h for co - fermentation; (3) After fermentation, centrifuge the co - fermentation culture solution to obtain a supernatant; (4) Pass the obtained supernatant through a sterilizing filter to obtain a sterile tea - seabuckthorn leaf composite fermentation liquid.

5. Use of the whitening and soothing fermented composition of Flos Lonicerae and Camellia sinensis according to any one of claims 1 - 4 in the preparation of skin care products.

6. A skin care product, characterized in that, Comprising the whitening and soothing fermented composition of Flos Lonicerae and Camellia sinensis according to any one of claims 1 - 4.

7. The skin care product according to claim 6, wherein, The skin care products include color cosmetics and skin care products; the skin care products include any one of lotion, emulsion, cream, facial mask, essence, essential oil, spray, sunscreen; the color cosmetics include any one of liquid foundation, lipstick.

8. The skin care product according to claim 6, wherein, The skin care product is a cream.

9. A cream, characterized in that, The cream comprises the following components in mass percentage: 0.5 - 3% of the whitening and soothing fermented composition of Flos Lonicerae and Camellia sinensis according to any one of claims 1 - 4, 2 - 40% of the skin care product matrix, and the balance is water.

10. The facial cream according to claim 9, characterized in that, The skin care product matrix comprises at least one of a thickener, a humectant, an emulsifier, a skin conditioner, a preservative, a fragrance, a pH regulator.

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