Fermentation product with moisturizing and soothing effects, and preparation method and application thereof

Abstract

This invention relates to a fermented product with moisturizing and soothing effects, its preparation method, and its application. The preparation method of the fermented product with moisturizing and soothing effects includes the following steps: mixing kiwifruit and blueberry with a carbon source, water, and fermentation bacteria; fermenting; centrifuging to separate the fermentation broth; and filtering the fermentation broth to obtain a fermented product filtrate. This invention creatively uses the fermented product of kiwifruit and blueberry as an active ingredient in cosmetics. Kiwifruit and blueberry are rich in antioxidants, which can help the skin resist free radical damage and slow down the skin aging process. The fermented product of kiwifruit and blueberry contains moisturizing ingredients, which help retain skin moisture, reduce dryness, and make the skin smooth and soft.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a fermentation product with moisturizing and soothing effects, its preparation method, and its application. Background Technology

[0002] Kiwifruit (Latin name: *Actinidia chinensis* Planch.), also known as Chinese kiwifruit, is a plant belonging to the genus *Actinidia* in the family Actinidiaceae, order Ericales. There are 66 species in the genus *Actinidia*, 62 of which are naturally distributed in China. The two main species cultivated globally are *Actinidia deliciosa* and *Actinidia chinensis*. Major cultivated varieties in my country include Hayward, Qinmei, Cuixiang, Xuxiang, Guichang, Hongyang, Donghong, Jintao, Jinyan, and Huayou. Fruit cores can be green, red, or yellow, and fruit shapes include oval, short oval, ovate, cylindrical, and oblong-cylindrical. In 2021, China's kiwifruit planting area was 199,140 hectares, accounting for 69.4% of the global total; China's kiwifruit production was 2.3807 million tons, accounting for 53.3% of the global total.

[0003] Currently, the main market application of kiwifruit is in food, such as direct consumption as fruit, as well as kiwifruit juice, kiwifruit beverages, and kiwifruit wine. In the cosmetics field, kiwifruit is used in products including kiwifruit fruit, kiwifruit water, kiwifruit extract, kiwifruit juice, kiwifruit seeds, kiwifruit seed oil, and kiwifruit seed extract. No kiwifruit fermentation products have yet been found to be used in cosmetics.

[0004] Blueberry (Vaccinium vitis-idaea L.) is a small shrub belonging to the genus Vaccinium in the family Ericaceae. It has slender, creeping rhizomes underground. The stems are thin, erect or prostrate at the base, and the branches and young branches are covered with grayish-white short soft hairs. The leaves are densely arranged, leathery, and elliptical or obovate. The inflorescences are short racemes, growing at the tips of last year's branches. The berries are spherical and purplish-red. Flowering occurs from June to July, and fruiting from August to September.

[0005] Bilberry is rich in a variety of active ingredients. Studies have shown that these active ingredients can fight skin oxidation and delay aging. However, there are few reports on the application of bilberry fermentation products in cosmetics. Summary of the Invention

[0006] In view of the shortcomings of the prior art, the purpose of this invention is to provide a fermentation product with moisturizing and soothing effects, its preparation method and application.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a method for preparing a fermented product with moisturizing and soothing effects. The method for preparing the fermented product with moisturizing and soothing effects includes the following steps: mixing kiwifruit and blueberry with a carbon source, water and fermentation bacteria, fermenting, centrifuging to separate the fermentation broth, and filtering the fermentation broth to obtain a fermentation product filtrate.

[0009] This invention creatively uses fermented products of kiwifruit and blueberry as active cosmetic ingredients. Kiwifruit and blueberry are rich in antioxidants, which can help the skin resist free radical damage and slow down the skin aging process. The fermented products of kiwifruit and blueberry contain moisturizing ingredients, which help retain skin moisture, reduce dryness, and make the skin smooth and soft.

[0010] Preferably, the amount of carbon source added in step (1) is 5%-30% (w / w).

[0011] The amount of carbon source added can be, for example, 5%, 10%, 15%, 20%, 25%, 30%, etc. Specific values ​​within the above range can be selected, and will not be elaborated further here.

[0012] Preferably, the carbon source includes any one or a combination of at least two of glucose, maltose, sucrose, lactose, or dextrin.

[0013] Preferably, the amount of water added in step (1) is 5-15 times the total weight of the kiwifruit, blueberries and carbon source.

[0014] The amount of water added can be, for example, 5 times, 8 times, 10 times, 13 times, 15 times, etc.

[0015] Preferably, the mass ratio of kiwifruit to blueberries in step (1) is 1:3-3:1.

[0016] Both kiwifruit and blueberry contain a high amount of active skincare ingredients. When used together for fermentation, the resulting fermented product contains more active substances than that obtained by fermenting a single component, resulting in superior skincare, whitening, anti-allergy, and repair effects.

[0017] In step (1), the mass ratio of kiwifruit to blueberries can be 1:1, 1:2, 1:3, 2:1, 3:1, 3:2, 2:3, etc. Specific values ​​within the above range can be selected, and will not be elaborated further here.

[0018] Preferably, the fermentation bacteria in step (1) include lactic acid bacteria and / or yeast; preferably, a combination of lactic acid bacteria and yeast.

[0019] This invention creatively develops a combination of lactic acid bacteria and yeast. In the fermentation process of kiwifruit and blueberry, the use of these two types of fermentation bacteria makes the components in the fermentation products of kiwifruit and blueberry more active, thus resulting in better effects when made into cosmetics.

[0020] Preferably, the lactic acid bacteria include any one or a combination of at least two of Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus acidophilus, Lactococcus lactis, or Lactococcus gasseri; more preferably, a combination of Lactobacillus plantarum, Lactococcus lactis, and Lactococcus gasseri.

[0021] The synergistic use of the aforementioned lactic acid bacteria can make the fermentation products with moisturizing and soothing effects in this application more bioactive, thereby having better skin care, moisturizing, whitening and anti-allergy effects.

[0022] Preferably, the yeast includes any one or a combination of at least two of Kluyveromyces lactis, Saccharomyces cerevisiae, or Pichia pastoris.

[0023] Preferably, the ratio of the amount of fermenting lactic acid bacteria to yeast inoculated in step (1) is (3-5):1, based on the number of live bacteria.

[0024] The ratios mentioned include, for example, 3:1, 7:2, 11:3, 4:1, 9:2, and 5:1. Specific point values ​​within the above range can be selected, and will not be elaborated upon further here.

[0025] Preferably, the fermentation in step (2) includes fermentation at 10-35°C for 5-60 days.

[0026] The range of 10-35 includes values ​​such as 10, 13, 15, 18, 20, 25, 28, 30, and 35; the range of 5-60 includes values ​​such as 5, 10, 15, 20, 30, 35, 40, 45, 55, and 60. Specific values ​​within the above ranges can be selected, and will not be elaborated upon further here.

[0027] Secondly, this application provides a fermentation product with moisturizing and soothing effects prepared by a method for preparing a fermentation product with moisturizing and soothing effects as described in the first aspect.

[0028] The fermentation product, fermented with selected fermentation bacteria and process parameters, contains a variety of active skin-care substances, resulting in good effects and gentle, non-irritating properties.

[0029] Thirdly, this application provides the application of the fermentation product with moisturizing and soothing effects described in the second aspect in the preparation of daily chemical products.

[0030] The aforementioned fermented products, known for their moisturizing and soothing effects, contain a variety of active ingredients that are safe, gentle, and non-irritating to the skin. These active ingredients help the skin resist free radical damage and slow down the skin aging process. The fermented products of kiwi and blueberry contain moisturizing components that help retain skin moisture, reduce dryness, and leave the skin smooth and soft.

[0031] Fourthly, the present invention provides a skin care product. The skin care product comprises: the fermented product with moisturizing and soothing effects described in the second aspect, a skin conditioning agent, a surfactant, a thickener, a pH adjuster, an ion regulator, and an antibacterial agent.

[0032] Preferably, the skin conditioning agent comprises any one or a combination of at least two of mannitol, glycerin, caprylyl glycol, ethylhexylglycerin, trehalose, β-glucan, sodium hyaluronate, carnosine, oligopeptide-1, acetyl hexapeptide-8, decapeptide-4, ascorbic acid, tranexamic acid, azelaic acid, retinoic acid, or hyaluronic acid.

[0033] Preferably, the skincare products include essence creams, lotions, serums, toners, eye creams, or face masks.

[0034] This invention creatively uses fermented products of kiwifruit and blueberry as active cosmetic ingredients. Kiwifruit and blueberry are rich in antioxidants, which can help the skin resist free radical damage and slow down the skin aging process. The fermented products of kiwifruit and blueberry contain moisturizing ingredients, which help retain skin moisture, reduce dryness, and make the skin smooth and soft. Detailed Implementation

[0035] To facilitate understanding of the present invention, the following embodiments are provided. Those skilled in the art should understand that these embodiments are merely illustrative and should not be construed as limiting the scope of the invention.

[0036] The sources of some components in the examples, comparative examples, application examples, and comparative application examples are as follows:

[0037] Lactobacillus plantarum (ATCC 8014), Lactococcus garvieae (ATCC 43921), Lactococcus lactis (CICC 20408), Lactobacillus paracasei (ATCC 334), Pichia farinosa (CICC 1688), Kluyveromyces lactis (ATCC 8585), Saccharomyces cerevisiae (ATCC 26785), and Lactobacillus rhamnosus (CICC 20408) were all sourced from the Guangdong Provincial Center for Microbial Culture Collection. 20255) was derived from Guangdong Provincial Microbial Culture Collection Center; Lactobacillus helveticus (CICC 24071) was derived from Guangdong Provincial Microbial Culture Collection Center; β-glucan was derived from product G304913 of Shanghai Aladdin Biochemical Technology Co., Ltd.; recombinant human collagen (SFH) was derived from Zhejiang Zhuji Juyuan Biotechnology Co., Ltd. (10g / bag); glycoprotein was derived from product 100g of Guangzhou Jinbaolai Biotechnology Co., Ltd.; and decapeptide-4 was derived from product ST-4 of Shanxi Jinnian Biotechnology Co., Ltd.

[0038] Example 1

[0039] This embodiment provides a fermented product with moisturizing and soothing effects.

[0040] 10 kg of domestic Guichang variety green-fleshed kiwifruit and 10 kg of blueberries were selected. The fruit peels were rinsed with water and removed using an automatic peeling machine. The pulp was then crushed, and 1 kg of sucrose and 80 kg of water were added, and stirred well. 1×10⁻⁶ Lactobacillus plantarum was added. 5 CFU / mL, Lactococcus gasseri 1×10 5 CFU / mL, Lactococcus lactis 1×10 5 CFU / mL, Kluyveromycin 1×10 5 CFU / mL. Seal the container with a breathable membrane and let it ferment at 35°C for 14 days. Centrifuge the fermentation broth at 3500 rpm, and then filter the supernatant through a 200-mesh filter cloth to obtain the fermentation filtrate.

[0041] Example 2

[0042] This embodiment provides a fermented product with moisturizing and soothing effects.

[0043] Select 5 kg of domestically grown red-fleshed kiwifruit (Golden Peach variety) and 15 kg of blueberries. Rinse the fruit peels with water and remove them simultaneously using an automatic peeling machine. Crush the pulp, add 0.4 kg of sucrose and 32 kg of water, and stir well. Add 1×10⁻⁶ Lactobacillus rhamnosus. 5 CFU / mL, Lactobacillus helveticus 1×10 5 CFU / mL, Lactobacillus paracasei 8×10 4 CFU / mL, Pichia pastoris 6×10 4 CFU / mL. Seal the container with a breathable membrane and let it ferment at 30°C for 18 days. Centrifuge the fermentation broth at 3500 rpm, and then filter the supernatant through a 300-mesh filter cloth to obtain the fermentation filtrate.

[0044] Example 3

[0045] This embodiment provides a fermented product with moisturizing and soothing effects.

[0046] Six kilograms of domestically grown Hongyang variety red-fleshed kiwifruit and 12 kilograms of blueberries were selected. The fruit peels were rinsed with water and removed using an automatic peeling machine. The pulp was crushed, and 0.8 kilograms of sucrose and 65 kilograms of water were added and stirred well. 1×10⁻⁶ Lactobacillus plantarum was added. 5 CFU / mL, Lactobacillus helveticus 1×10 5 CFU / mL, Lactococcus lactis 1.2×10 5 CFU / mL, Kluyveromycin 1×10 5 CFU / mL, Saccharomyces cerevisiae 5×10 4 CFU / mL. Seal the container with a breathable membrane and let it ferment at 25°C for 16 days. Centrifuge the fermentation broth at 4000 rpm, and then filter the supernatant through a 250-mesh filter cloth to obtain the fermentation filtrate.

[0047] Example 4

[0048] This embodiment provides a fermented product with moisturizing and soothing effects.

[0049] 12 kg of domestic Hayward green-fleshed kiwifruit and 4 kg of blueberries were selected. The fruit peels were rinsed with water and removed using an automatic peeling machine. The pulp was then crushed, and 1.2 kg of sucrose and 100 kg of water were added, followed by thorough mixing. 6 × 10⁶ Lactobacillus plantarum was added. 4 CFU / mL, Lactobacillus helveticus 5×10 4 CFU / mL, Lactobacillus paracasei 3×10 4CFU / mL, Lactococcus lactis 1.2×10 4 CFU / mL, Kluyveromycin 4×10 4 CFU / mL, Saccharomyces cerevisiae 3×10 4 CFU / mL. Seal the container with a breathable membrane and allow it to ferment at 18°C ​​for 28 days. Centrifuge the fermentation broth at 4000 rpm, and then filter the supernatant through a 250-mesh filter cloth to obtain the fermentation filtrate.

[0050] Example 5

[0051] This embodiment provides a fermented product with moisturizing and soothing effects. The only difference between this product and Example 1 is that it does not contain *Lactobacillus plantarum*. The reduced number of viable bacteria is proportionally distributed among *Lactobacillus gasseri* and *Lactococcus lactis*, while the number of viable lactic acid bacteria remains unchanged. All other conditions remain unchanged, and the preparation method is the same as in Example 1.

[0052] Example 6

[0053] This embodiment provides a fermented product with moisturizing and soothing effects. The only difference between this embodiment and Example 1 is that it does not contain Lactobacillus gasseri. The reduced number of viable bacteria is proportionally distributed among Lactobacillus plantarum and Lactococcus lactis, while the number of viable lactic acid bacteria remains unchanged. All other conditions remain unchanged, and the preparation method is the same as in Example 1.

[0054] Example 7

[0055] This embodiment provides a fermented product with moisturizing and soothing effects. The only difference between this product and Example 1 is that it does not contain Lactococcus lactis. The reduced number of live bacteria is proportionally distributed among Lactobacillus gasseri and Lactobacillus plantarum, while the number of live lactic acid bacteria remains unchanged. All other conditions remain unchanged, and the preparation method is the same as in Example 1.

[0056] Example 8

[0057] This embodiment provides a fermented product with moisturizing and soothing effects. The only difference between the fermented product with moisturizing and soothing effects and that of Example 1 is that it does not contain Kluyveromyces lactis. The reduced portion is distributed proportionally to Lactococcus lactis, Lactobacillus gasseri, and Lactobacillus plantarum. The number of viable bacteria in the fermentation remains unchanged, and all other conditions remain unchanged. The preparation method is the same as that in Example 1.

[0058] Example 9

[0059] This embodiment provides a fermentation product that differs from that in Example 1 only in that it does not contain Lactobacillus plantarum, Lactococcus gasseri, and Lactococcus lactis. The reduced portion is proportionally allocated to Kluyveromyces lactis, the viable count of the fermentation bacteria remains unchanged, and all other conditions remain unchanged. The preparation method is the same as in Example 1.

[0060] Comparative Example 1

[0061] This embodiment provides a kiwifruit fermentation product. The only difference between the kiwifruit fermentation product and that in Embodiment 1 is that the raw materials do not contain blueberries. The reduced portion of blueberries is allocated to the kiwifruit. Other conditions remain unchanged, and the preparation method is the same as in Embodiment 1.

[0062] Comparative Example 2

[0063] This embodiment provides a blueberry fermentation product. The only difference between the blueberry fermentation product and that in Example 1 is that the raw materials do not contain kiwifruit. The reduced portion of kiwifruit is allocated to the blueberries. Other conditions remain unchanged, and the preparation method is the same as in Example 1.

[0064] Application Example 1

[0065] This application example provides a skincare product, which includes:

[0066] Example 1 yielded a fermentation product filtrate with moisturizing and soothing effects containing 8% trehalose, 0.55% β-glucan, 0.45% recombinant human collagen, 0.5% carnosine, 0.15% oligopeptide-1, 0.1% sodium hyaluronate, 0.2% glycerol, 0.03% caprylyl glycol, 0.04% acetyl hexapeptide-8, 0.02% sodium chloride, 0.05% ethylhexylglycerin, 0.002% disodium hydrogen phosphate, 0.05% glycoprotein, 0.02% decapeptide-4, with the remainder being water.

[0067] The preparation method is to mix the above components evenly.

[0068] Application Example 2

[0069] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in an equal amount with the fermentation product filtrate prepared in Example 2, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0070] Application Example 3

[0071] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Example 3, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0072] Application Example 4

[0073] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Example 4, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0074] Application Example 5

[0075] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Example 5, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0076] Application Example 6

[0077] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in an equal amount with the fermentation product filtrate prepared in Example 6, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0078] Application Example 7

[0079] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in an equal amount with the fermentation product filtrate prepared in Example 7, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0080] Application Example 8

[0081] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in an equal amount with the fermentation product filtrate prepared in Example 8, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0082] Application Example 9

[0083] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Example 9, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0084] Comparative Application Example 1

[0085] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Comparative Example 1, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0086] Comparative Application Example 2

[0087] This application example provides a skin care product, which differs from Application Example 1 only in that the fermentation product filtrate with moisturizing and soothing effects prepared in Example 1 is replaced in equal volume with the fermentation product filtrate prepared in Comparative Example 2, while all other conditions remain unchanged, and the preparation method is the same as in Application Example 1.

[0088] Comparative Application Example 3

[0089] This comparative application example provides a composition containing kiwi fruit fermentation product and blueberry fermentation product. The only difference between this composition and application example 1 is that it does not contain the fermentation product filtrate with moisturizing and soothing effects. The reduced mass portion of the filtrate is proportionally allocated to the other components. The preparation method is the same as that in application example 1.

[0090] Test Example 1

[0091] Skin whitening - zebrafish experiment

[0092] Referring to T / ZHCA 012-2021 "Test Method for Whitening Efficacy of Cosmetics - Test Method for Melanin Inhibition Efficacy of Zebrafish Embryos", a zebrafish model was used to evaluate the whitening efficacy of the samples. Zebrafish have three types of pigment cells in their neural crest: melanocytes, xanthocyanins, and iridocytic cells, which differentiate from intermediate progenitor cells. Therefore, zebrafish have pigment on their body surface and can be used as an animal model for evaluating whitening efficacy.

[0093] Wild zebrafish of the AB strain were cultured and bred according to the Westerfield method. The system water pH was set to 7.2-7.4 and conductivity to 450-550 μS / m. The system used purified water, and the pH and conductivity were adjusted to the set parameters by automatically adding sodium chloride and sodium bicarbonate. The fish room temperature was 26-28℃, with a photoperiod of 14 hours light / 10 hours dark. Spawning was stimulated by light, and embryos were collected and rinsed clean with water. Normally developing embryos were selected under a microscope and placed in an incubator for further culture. 24-well plates were used as experimental containers, and zebrafish at 24 hpf embryo stage were randomly placed in each well. Each experimental group (Application Examples 1-9, Control Examples 1-3) used 10 embryos per group. 100 μL of sample was added to each well. The wells were incubated for 48 hours, and the development of melanin on the embryonic surface was observed using a fluorescence microscope. Images were taken and the area of ​​melanin patches was statistically analyzed using ImageJ software (US NIH), and the melanin inhibition rate of the experimental groups was calculated. Statistical analysis was performed using SPSS 26 software.

[0094] Table 1

[0095]

[0096]

[0097] As shown in the table above, the experimental group with added filtrate of kiwifruit and blueberry fermentation products had a higher melanin inhibition rate, indicating that the former had a better whitening effect. The melanin inhibition rate of Application Examples 1-4 was better than that of Application Examples 5-9, indicating that their process and parameters were a better process, and the fermentation products obtained could better reflect the invention's effects.

[0098] Test Example 2

[0099] Moisturizing - Zebrafish Experiment

[0100] Zebrafish skin structure is similar to human skin. By immersing zebrafish in a hypertonic solution to induce skin dehydration, adding samples, and observing the effects of the samples on preventing dehydration and shrinkage and promoting repair in the tail fin and trunk of zebrafish, the moisturizing effect of the samples can be evaluated.

[0101] Wild-type zebrafish with normal development (72 hpf) were selected and divided into three groups: a control group (pure water), a model group (hypertonic 2% sodium chloride solution), and experimental groups (Application Examples 1-9 and Control Application Examples 1-3), with 10 zebrafish in each group. The zebrafish were placed on glass slides and photographed under a stereomicroscope. The zebrafish were then transferred to 96-well plates, one per well, and the water in the wells was aspirated. 200 μL of the corresponding solution was added to each group. The plates were incubated at 28°C for 20 min, and the zebrafish were photographed again under a stereomicroscope. The tail length of the zebrafish was measured and calculated using ImageJ software (NIH, USA), and the tail length contraction rate was calculated compared to the length before treatment. SPSS 26 software was used for statistical analysis of the data.

[0102] Table 2

[0103]

[0104]

[0105] As shown in the table above, the model groups exhibited significant shrinkage rates. Both the experimental groups with added kiwi and blueberry fermentation product filtrate and the unadded groups showed reduced shrinkage rates in the zebrafish tails, but the added groups showed better results, indicating a superior moisturizing effect. The shrinkage rates of Application Examples 1-4 were superior to those of Application Examples 5-9, suggesting that their process is a more efficient one, and the resulting fermentation products better reflect the invention's effects.

[0106] Test Example 3

[0107] Antioxidant - Zebrafish

[0108] When the indicator CM-H2DCFDA is added to the zebrafish juvenile culture system, it can produce relatively stable high-intensity fluorescence under the action of intracellular reactive oxygen species. By detecting the fluorescence intensity, the production of free radicals in zebrafish can be understood, which is suitable for evaluating the antioxidant effect of test samples.

[0109] Ten 3-day-fed zebrafish juveniles of normal development and consistent developmental stage were randomly selected and transferred to 24-well plates. Twelve experimental groups were set up (Examples 1-9 and Control Examples 1-3); the experimental groups were dilutions of the application and control examples. 1 mL of sample was added to each well, and CM-H2DCFDA was added to a concentration of 0.5 μg / mL. The mixture was thoroughly mixed, and the zebrafish were transferred to 96-well plates, one fish per well, with 100 μL of liquid per well. The 96-well plates were wrapped in aluminum foil to protect them from light and incubated at 28°C for 20 h. After incubation, the fluorescence value (S) was detected using a multi-mode microplate reader (Thermo Fisher Varioskan LUX, USA) (excitation wavelength 485 nm, emission wavelength 535 nm). The relative content of reactive oxygen species (ROS) in the zebrafish of the test group was calculated by subtracting the fluorescence value of the untreated group from the fluorescence value of the untreated group. SPSS 26 software was used for statistical analysis of the experimental data.

[0110] Calculate the ROS removal rate using the following formula:

[0111]

[0112] Table 3

[0113]

[0114]

[0115] As can be seen from the table above, the sample examples with added fermentation products that have moisturizing and soothing effects have a higher ROS scavenging rate compared with the control group without added products. Even at a lower concentration, they still have a certain removal effect, indicating that the examples have good antioxidant effects.

[0116] Test Example 4

[0117] Soothing - Human Experimentation

[0118] The soothing effects of the examples and comparative samples were evaluated and compared in accordance with T / CAB 0152-2022 "Test Methods for Seven Efficacy Items of Cosmetics: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing". The soothing effects of the products were verified and compared by assessing the improvement in skin redness, itching, stinging, and other irritation.

[0119] In addition to meeting the standard screening criteria, participants must also undergo a lactic acid stinging test: At room temperature, 50 μL of 10% lactic acid solution is added to the nasolabial fold on either side. Participants are asked about their subjective symptoms at 2.5 min and 5 min, and are scored on a 0-3 scale (0 for no stinging, 1 for mild stinging, 2 for moderate stinging, and 3 for severe stinging). The scores at the two time points are added together; a total score ≥3 is considered positive and eligible for consideration.

[0120] One hundred and thirty healthy subjects, aged 32.4–51.5 years (mean 43.5 years), of unrestricted gender, were selected. They were randomly divided into a control group and an experimental group (Examples 1–9, control examples 1–3), with ten subjects in each group. Before the test, subjects cleaned their faces and patted them dry with lint-free absorbent paper towels. Hemoglobin levels in the facial test areas were measured using a skin melanin and hemoglobin analyzer (Courage+Khazaka electronic GmbH, Mexameter MX18, Germany). The control group received saline solution. Subjects used one tablet every two days, and facial hemoglobin levels were measured at 8, 14, and 28 days. A lactic acid stinging test and subjective assessment were also conducted to compare differences between different time points and groups. SPSS 26 software was used for statistical analysis. Hemoglobin results are shown in Table 4, and stinging scores are shown in Table 5.

[0121] Table 4

[0122] Group / Days 0 days 14 days 28 days 56 days Example 1 432±41 368±28 278±30 155±18 Example 2 440±38 371±34 285±30 139±16 Example 3 428±30 360±35 273±28 128±17 Example 4 435±32 365±37 268±28 130±22 Example 5 426±34 390±40 328±27 231±25 Example 6 434±37 389±34 320±25 234±20 Example 7 435±36 395±36 331±30 209±21 Example 8 427±30 380±29 317±32 217±20 Example 9 431±33 381±31 310±34 238±23 Comparative Example 1 428±32 411±37 360±40 291±27 Comparative Example 2 433±36 409±37 364±38 305±28 Comparative Example 3 430±36 415±35 375±38 324±27 Blank group 438±34 430±35 406±40 387±32

[0123] Table 5

[0124]

[0125]

[0126] According to the table above, the heme content in the control group decreased slightly, but the stinging score was not relieved and remained greater than or equal to 3. Both the application group and the control group showed a significant decrease in heme content and stinging score. Due to the addition of kiwi and blueberry fermentation product filtrate, the application group showed significant advantages in both data compared to the control group after 28 and 56 days, especially in application groups 1-4, indicating that the addition of fermentation products with moisturizing and soothing effects had a significant soothing effect.

[0127] Test Example 5

[0128] Repair - Human Experimentation

[0129] The repair efficacy of the examples and comparative examples was evaluated and compared in accordance with T / CAB 0152-2022 "Test Methods for Seven Efficacy Items of Cosmetics: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing". Changes in the transepidermal water loss rate of the skin were measured to reflect changes in the skin barrier function, thus verifying the product's repair efficacy.

[0130] Subjects, like test case 4, will undergo a transepidermal water loss test using a Tewameter TM 300 (Courage+Khazaka electronic GmbH, Germany) on the face. A value >10 g / m² is required.2 Subjects who meet the h-hour time requirement can be enrolled. Subjects remain relaxed and balanced in the testing environment for 30 minutes. The transepidermal water loss rate in the subject's cheek testing area is measured and recorded as the baseline value.

[0131] The subjects were divided into 13 groups, with 10 people in each group for each sample. The sample of the example and the sample of the comparative example were distributed respectively. The transepidermal water loss rate (TEWL) was measured after using the product for 14 days, 28 days and 56 days. The data are summarized as follows. The experimental data were statistically analyzed using SPSS 26 software: The transepidermal water loss rate is shown in Table 6.

[0132] Table 6

[0133]

[0134]

[0135] As can be seen from the table above, the subjects using the sample containing kiwi and blueberry fermentation product filtrate had a significantly lower transepidermal water loss rate than the corresponding control sample without filtrate, especially the samples using Examples 1-4, which showed the most significant reduction. This indicates that the sample containing the kiwi and blueberry fermentation product filtrate has a better skin repair effect.

[0136] The applicant declares that the detailed process equipment and process flow of this invention are illustrated through the above embodiments, but this invention is not limited to the above detailed process equipment and process flow, that is, it does not mean that this invention must rely on the above detailed process equipment and process flow to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, additions of auxiliary components, and selection of specific methods, all fall within the protection scope and disclosure scope of this invention.

Claims

1. A method for preparing a fermentation product with moisturizing and soothing effects, characterized in that, The preparation method of the fermented product with moisturizing and soothing effects consists of the following steps: Kiwifruit and blueberries are mixed with carbon source, water and fermentation bacteria, fermented, and the fermentation liquid is separated by centrifugation. The fermentation liquid is then filtered to obtain a fermentation product with moisturizing and soothing effects. The fermentation bacteria are a combination of lactic acid bacteria and yeast; The lactic acid bacteria is *Lactobacillus plantarum* (… Lactiplantibacillus plantarum The accession number is ATCC 8014, Lactococcus lactis ( Lactococcus lactis The accession number is CICC 20408 and Lactococcus gasseri ( Lactococcus garvieae The combination with accession number ATCC 43921; The yeast is Kluyveromyces lactis (Lactobacillus) Kluyveromyces lactis The accession number is ATCC 8585; The lactic acid bacteria and yeast strains mentioned were both sourced from the Guangdong Provincial Microbial Culture Collection Center. The mass ratio of kiwifruit to blueberries is 1:3-3:1; The ratio of the inoculation amount of lactic acid bacteria to yeast, based on the number of live bacteria, is (3-5):

1.

2. The method for preparing the fermentation product with moisturizing and soothing effects according to claim 1, characterized in that, The amount of carbon source added is 5%-30% (w / w).

3. The method for preparing the fermentation product with moisturizing and soothing effects according to claim 1, characterized in that, The carbon source includes any one or a combination of at least two of glucose, maltose, sucrose, lactose, or dextrin.

4. The method for preparing the fermentation product with moisturizing and soothing effects according to claim 1, characterized in that, The amount of water added is 5-15 times the total weight of the kiwifruit, blueberries, and carbon source.

5. The method for preparing the fermentation product with moisturizing and soothing effects according to claim 1, characterized in that, The fermentation process involves fermenting at 10-35°C for 5-60 days.

6. A fermentation product with moisturizing and soothing effects prepared by the method of preparing a fermentation product with moisturizing and soothing effects according to any one of claims 1-5.

7. The application of the fermentation product with moisturizing and soothing effects according to claim 6 in the preparation of daily chemical products.

8. A skincare product, characterized in that, The skin care product includes: the fermentation product with moisturizing and soothing effects as described in claim 6, skin conditioning agent, surfactant, thickener, pH adjuster, ion adjuster and antibacterial agent.

9. The skincare product according to claim 8, characterized in that, The skin conditioning agent includes any one or a combination of at least two of the following: mannitol, glycerin, caprylyl glycol, ethylhexylglycerin, trehalose, beta-glucan, sodium hyaluronate, carnosine, oligopeptide-1, acetyl hexapeptide-8, decapeptide-4, ascorbic acid, tranexamic acid, azelaic acid, retinoic acid, or hyaluronic acid.

10. The skincare product according to claim 8 or 9, characterized in that, The skincare products include serums, lotions, or masks.

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