A method for preparing a white tea fermentation product with moisturizing, anti-aging, and soothing effects, as well as its products and applications.

Abstract

This invention relates to a method for preparing a white tea fermentation product with moisturizing, anti-aging, and soothing effects, as well as the product and its applications. The method includes mixing dried white tea leaves with water, soaking them, and then performing a hot extraction followed by solid-liquid separation to obtain a white tea extract. A compound lactobacillus is inoculated into a sterilized culture medium containing the white tea extract for primary fermentation to obtain a primary fermentation product. The primary fermentation product, sorghum rice extract, and wolfberry extract are mixed, sterilized, and then inoculated with a compound yeast for secondary fermentation to obtain a secondary fermentation product. The secondary fermentation product is sterilized, cooled, and then subjected to solid-liquid separation to obtain the white tea fermentation product. The preparation method of the white tea fermentation product involved in this invention is simple, easily scalable for industrial production, and the resulting fermentation product is mild and non-irritating, highly safe, and possesses excellent moisturizing, anti-aging, and soothing effects. It can be used as a raw material in skincare and cosmetic products.

Description

Technical Field

[0001] This invention belongs to the field of plant fermentation technology, and relates to a method for preparing white tea fermentation products with moisturizing, anti-aging and soothing effects, as well as the products and applications thereof. Background Technology

[0002] In recent years, tea and its products have become increasingly popular in the daily chemical industry. The emergence of these products and the rise of tea's application in this sector are inseparable from in-depth research into tea, its components, and its effects. Based on different processing methods and degrees of fermentation, tea is divided into six major categories: green tea (unfermented), white tea (lightly fermented), yellow tea (lightly fermented), oolong tea (semi-fermented), black tea (post-fermented), and red tea (fully fermented). The production of white tea requires very strict control over the variety of tea leaves.

[0003] White tea, a special and precious variety among the six major tea categories, is also the earliest type of tea in history. Named for its unique processing method—neither roasting nor rolling—white tea has a silvery-white to grayish-green appearance, covered in white down, giving it a white appearance. It is mainly produced in Fuding, Zhenghe, Songxi, and Jianyang in Fujian province. Its unique processing techniques result in a fresh and delicate aroma, prominent downy fragrance, and a mellow and refreshing taste, making it widely popular. The unique quality and flavor of white tea are highly appreciated, and its main active ingredients include tea polyphenols, amino acids, caffeine, and soluble sugars.

[0004] Currently, white tea has not fully penetrated domestic and international markets, and there is a need to inject new vitality into the white tea industry and expand its application areas. In particular, there is a lack of necessary experimentation and innovation in the fermentation of white tea. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for preparing white tea fermentation products with moisturizing, anti-aging and soothing effects, as well as the products and applications thereof.

[0006] To achieve this objective, the present invention employs the following technical solution:

[0007] In a first aspect, the present invention provides a method for preparing a white tea fermentation product with moisturizing, anti-aging, and soothing effects, the preparation method comprising the following steps:

[0008] (1) Mix dried white tea leaves with water, soak them, and then perform hot extraction, followed by solid-liquid separation to obtain white tea extract;

[0009] (2) Inoculate the compound lactobacillus into a sterilized culture medium containing white tea extract and carry out a primary fermentation to obtain the primary fermentation product;

[0010] The compound lactobacillus is a combination of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus salivarius.

[0011] (3) Mix the primary fermentation product, sorghum rice extract and wolfberry extract and sterilize them. Inoculate with compound yeast for secondary fermentation to obtain secondary fermentation product.

[0012] The compound yeast is a combination of Bayer conjugated yeast and brewer's yeast;

[0013] (4) After sterilizing the secondary fermentation product, cool it and separate the solid and liquid to obtain the white tea fermentation product.

[0014] This invention develops a novel product using white tea as the main raw material. The preparation method involves using white tea aqueous extract as the primary fermentation substrate, supplemented with sorghum rice extract and wolfberry extract as secondary fermentation substrates. Fermentation is carried out sequentially using lactobacillus and yeast. The complementary and synergistic effects of lactobacillus and yeast are utilized to ferment the white tea aqueous extract, maximizing the preservation of the active components of white tea and giving the fermented product excellent moisturizing, anti-aging, and soothing effects. The addition of sorghum rice extract and wolfberry extract during the secondary fermentation, along with the primary fermentation metabolites of white tea, further enhances the moisturizing, anti-aging, and soothing effects of the product. Furthermore, the microorganisms used in both fermentations have a specific composition, with each species working synergistically to improve the efficacy of the fermented product.

[0015] The preparation method of white tea fermentation products involved in this invention is simple and easy to industrialize. The fermentation products obtained are mild and non-irritating, with high safety, and can be added to skin care products and cosmetics as raw materials.

[0016] Preferably, the ratio of viable counts of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus salivarius is (1-2):(2-4):(1-2):(1-2):(1-2).

[0017] Preferably, the ratio of viable counts of Bayer conjugated yeast and brewer's yeast is (1-3):(1-3).

[0018] Based on the potential synergistic relationship between various fermentation strains, when they meet specific quality ratios, the fermentation products obtained have better effects in moisturizing, anti-aging, and soothing.

[0019] The specific point values ​​in (1-2) above can be 1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 2, etc.; the specific point values ​​in (2-4) can be 2, 2.3, 2.5, 2.7, 3, 3.2, 3.5, 3.8, 4, etc.; the specific point values ​​in (1-3) can be 1, 1.3, 1.5, 1.7, 2, 2.2, 2.5, 2.8, 3, etc.

[0020] Preferably, the ratio of white tea leaves to water in step (1) is 1:(8-12) g / mL, such as 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, 1:11 g / mL, 1:12 g / mL, etc.

[0021] Preferably, the soaking in step (1) is carried out at 40-80℃ (e.g., 40℃, 50℃, 60℃, 70℃, 80℃, etc.) for 1-4 h (e.g., 1 h, 2 h, 3 h, 4 h, etc.).

[0022] Preferably, the thermal extraction in step (1) is carried out at 90-100℃ (e.g., 90℃, 93℃, 95℃, 97℃, 100℃, etc.) for 1-3 h (e.g., 1 h, 1.5 h, 2 h, 3 h, etc.).

[0023] Preferably, the inoculation amount of the compound lactobacillus in step (2) is 2-4%, such as 2%, 2.5%, 3%, 3.5%, 4%, etc.

[0024] Preferably, the temperature of the primary fermentation in step (2) is 35-38℃, such as 35℃, 36℃, 37℃, 38℃, etc.; and the time is 30-48 h, such as 30 h, 32 h, 36 h, 40 h, 42 h, 48 h, etc.

[0025] Preferably, the culture medium in step (2) further contains an external nitrogen source, an external carbon source, and an external inorganic salt.

[0026] The added nitrogen source includes any one or a combination of at least two of yeast powder, tryptone, soy peptone, or beef extract.

[0027] The added carbon source includes any one or a combination of at least two of fructose, glucose, trehalose, lactose, or galactose.

[0028] The added inorganic salts include NaCl and / or K2HPO4.

[0029] Preferably, the culture medium in step (2) comprises, by mass parts, 90-110 parts of white tea extract (e.g., 90, 95, 100, 105, 110, etc.), 1-3 parts of added nitrogen source (e.g., 1, 1.5, 2, 2.5, 3, etc.), 1-3 parts of added carbon source (e.g., 1, 1.5, 2, 2.5, 3, etc.), and 0.1-0.2 parts of added inorganic salt (e.g., 0.1, 0.14, 0.16, 0.2, etc.).

[0030] Preferably, the mass ratio of the primary fermentation product, sorghum rice extract, and wolfberry extract in step (3) is 100:(1-5):(1-5); wherein the specific point values ​​in (1-5) can be selected from 1, 2, 2.5, 3, 3.5, 4, 4.5, 5, etc.

[0031] Preferably, the inoculation amount of the compound lactobacillus in step (3) is 3-5%, such as 3%, 3.5%, 4%, 4.5%, 5%, etc.

[0032] Preferably, the temperature of the secondary fermentation in step (3) is 28-35℃, such as 28℃, 30℃, 32℃, 33℃, 34℃, 35℃, etc.; and the time is 30-48 h, such as 30 h, 36 h, 42 h, 44 h, 48 h, etc.

[0033] Preferably, after the solid-liquid separation in step (4), a drying process is also performed.

[0034] The sorghum rice extract and wolfberry extract used in this invention are preferably prepared by the following method, which has a better effect when combined with white tea for fermentation.

[0035] Preferably, the preparation method of the sorghum rice extract includes the following steps:

[0036] (S1) After pulverizing sorghum rice, it is mixed with an ethanol aqueous solution and then subjected to microwave-ultrasound combined treatment to obtain a treatment solution and sorghum rice residue.

[0037] (S2) The treatment liquid and sorghum residue are refluxed for extraction, and solid-liquid separation is performed to obtain the extract;

[0038] (S3) The extract is concentrated and dried to obtain the sorghum rice extract.

[0039] Before reflux extraction, the sorghum rice extract is pretreated with a combination of microwave and ultrasound. Microwaves can increase the temperature inside plant cells and accelerate cell wall rupture, while ultrasound can produce cavitation effects. Compared with pretreatment methods using microwaves or ultrasound alone, the combined pretreatment method yields better extraction results.

[0040] Preferably, the volume fraction of the ethanol aqueous solution in step (S1) is 70%-90%, such as 70%, 73%, 75%, 78%, 80%, 82%, 85%, 88%, 90%, etc.

[0041] Preferably, the mass ratio of the ethanol aqueous solution to sorghum rice in step (S1) is (5-12):1, for example, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, etc.

[0042] Preferably, the microwave-ultrasound combined treatment in step (S1) is performed at 40-55℃ (e.g., 40℃, 42℃, 45℃, 48℃, 50℃, 52℃, 54℃, 55℃, etc.) for 10-30 min (e.g., 10 min, 15 min, 20 min, 25 min, 30 min, etc.).

[0043] Preferably, the processing power of the microwave in step (S1) is 100-300 W, such as 100 W, 150 W, 200 W, 250 W, 300 W, etc.; the processing power of the ultrasonic wave is 200-400 W, such as 200 W, 250 W, 300 W, 350 W, 400 W, etc.

[0044] Preferably, the reflux extraction in step (S2) is performed 1-3 times, for example, 1 time, 2 times or 3 times; the extraction time for each extraction is independently 0.5-3 h, for example 0.5 h, 0.8 h, 1 h, 1.5 h, 2 h, 2.2 h, 2.5 h, 3 h, etc.

[0045] Preferably, the preparation method of the wolfberry extract includes the following steps:

[0046] (T1) Mix wolfberry with water and extract under vacuum reflux, then separate the solid and liquid to obtain the extract;

[0047] (T2) The extract is decolorized and the solid and liquid are separated to obtain the decolorized solution;

[0048] (T3) The decolorized liquid is subjected to flocculation treatment, solid-liquid separation, filtrate concentration and drying to obtain the wolfberry extract.

[0049] The wolfberries were subjected to decolorization and flocculation treatments after reflux extraction. Compared with the extract without these treatments, the wolfberries exhibited superior efficacy after fermentation.

[0050] Preferably, the mass ratio of water to wolfberry in step (T1) is (5-12):1, for example, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, etc.

[0051] Preferably, the vacuum decompression reflux extraction in step (T1) is performed 1-2 times, and the extraction time for each extraction is independently 0.5-2 h, for example 0.5 h, 1 h, 1.5 h, 2 h, etc.

[0052] Preferably, the decolorization treatment in step (T2) uses 1-5% (e.g., 1%, 2%, 3%, 4%, 5%, etc.) of activated carbon.

[0053] Preferably, the flocculation treatment in step (T3) uses a flocculant with an addition amount of 1-5% (e.g., 1%, 2%, 3%, 4%, 5%, etc.), wherein the flocculant is selected from any one or a combination of at least two of water-soluble polysaccharides, cationic chitosan, guar gum, protein colloids, gelatin, animal glue, cationic starch or tannins.

[0054] In a second aspect, the present invention provides a white tea fermentation product prepared according to the preparation method described in the first aspect.

[0055] Thirdly, the present invention provides the use of the white tea fermentation product according to the second aspect in the preparation of a product having any one or at least two of the following effects;

[0056] 1) Moisturizing; 2) Anti-aging; 3) Antioxidant; 4) Soothing;

[0057] Preferably, the product includes skin care products, cosmetics, pharmaceuticals, or medical materials.

[0058] Other unlisted point values ​​within the numerical ranges involved in this invention can be selected and are all within the protection scope of this invention. Considering the length and brevity of the description, they will not be elaborated here.

[0059] Compared with the prior art, the present invention has the following beneficial effects:

[0060] This invention develops a novel product using white tea as the main raw material. The preparation method involves using white tea aqueous extract as the primary fermentation substrate, supplemented with sorghum rice extract and wolfberry extract as secondary fermentation substrates. Fermentation is carried out sequentially using lactobacillus and yeast. The complementary and synergistic effects of lactobacillus and yeast are utilized to ferment the white tea aqueous extract, maximizing the preservation of the active components of white tea and giving the fermented product excellent moisturizing, anti-aging, and soothing effects. The addition of sorghum rice extract and wolfberry extract during the secondary fermentation, along with the primary fermentation metabolites of white tea, further enhances the moisturizing, anti-aging, and soothing effects of the product. Furthermore, the microorganisms used in both fermentations have a specific composition, with each species working synergistically to improve the efficacy of the fermented product.

[0061] The preparation method of white tea fermentation products involved in this invention is simple and easy to industrialize. The fermentation products obtained are mild and non-irritating, with high safety, and can be added to skin care products and cosmetics as raw materials. Detailed Implementation

[0062] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0063] The following strains of *Lactobacillus plantarum* are ATCC8014; *Lactobacillus acidophilus* are ATCC4356; *Lactobacillus rhamnosus* are ATCC39595; *Lactobacillus casei* are ATCC27139; *Lactobacillus salivarius* are ATCC11741; *Bayer zygosacchariformis* are ATCC MYA-4549; and *Saccharomyces cerevisiae* are ATCC26603.

[0064] Preparation Example 1

[0065] This preparation example provides a sorghum rice extract, prepared by the following method:

[0066] (S1) After pulverizing sorghum rice, mix it with 8 times the mass of 80% ethanol aqueous solution and then perform microwave-ultrasound combined treatment at 50°C for 30 min. The microwave treatment power is 200 W and the ultrasonic treatment power is 300 W, to obtain the treatment liquid and sorghum rice residue.

[0067] (S2) The treatment liquid and sorghum residue were refluxed and extracted twice, each time for 1.5 h, and the solid and liquid were separated. The two extracts were combined.

[0068] (S3) The extract is concentrated under reduced pressure and freeze-dried to obtain the sorghum rice extract.

[0069] This preparation example also provides a wolfberry extract, prepared by the following method:

[0070] (T1) Mix wolfberry with 10 times its weight of water and extract twice under vacuum at 0.05 MPa and 40°C for 1.5 h each time. Combine the two extracts.

[0071] (T2) Add 3% activated carbon to the extract for decolorization, filter, and obtain the decolorized solution;

[0072] (T3) The decolorizing solution was mixed with 1.5% flocculant (made by mixing cationic chitosan and gelatin in a mass ratio of 1:1 with 5 times the mass of 1% acetic acid aqueous solution until a paste is formed, and then mixing with 50 times the mass of water to swell for 12 h) for flocculation treatment, centrifuged, filtered, concentrated under reduced pressure and freeze-dried to obtain the wolfberry extract.

[0073] Preparation Example 2

[0074] This preparation example provides a sorghum rice extract, prepared by the following method:

[0075] (S1) After pulverizing sorghum rice, mix it with 12 times the mass of 70% ethanol aqueous solution and then perform microwave-ultrasound combined treatment at 45°C for 30 min. The microwave treatment power is 150 W and the ultrasonic treatment power is 400 W, to obtain the treatment liquid and sorghum rice residue.

[0076] (S2) The treatment liquid and sorghum residue were refluxed and extracted twice, each time for 2 hours. The solid and liquid were separated and the two extracts were combined.

[0077] (S3) The extract is concentrated under reduced pressure and freeze-dried to obtain the sorghum rice extract.

[0078] This preparation example also provides a wolfberry extract, prepared by the following method:

[0079] (T1) Mix wolfberry with 12 times its weight of water and extract twice under vacuum at 0.05 MPa and 40°C for 2 hours each time. Combine the two extracts.

[0080] (T2) Add 5% activated carbon to the extract for decolorization, filter, and obtain the decolorized solution;

[0081] (T3) The decolorizing solution was mixed with 2% flocculant (cationic chitosan was first mixed with 5 times the mass of 1% acetic acid aqueous solution and stirred until a paste was formed, and then mixed with 50 times the mass of water and swollen for 12 h) for flocculation treatment, centrifuged, filtered, the filtrate was concentrated under reduced pressure and freeze-dried to obtain the wolfberry extract.

[0082] Preparation Example 3

[0083] This preparation example provides a sorghum rice extract, prepared by the following method:

[0084] (S1) After pulverizing sorghum rice, mix it with 5 times the mass of 90% ethanol aqueous solution and then perform microwave-ultrasound combined treatment at 55℃ for 20 min. The microwave treatment power is 250 W and the ultrasonic treatment power is 250 W, to obtain the treatment liquid and sorghum rice residue.

[0085] (S2) The treatment liquid and sorghum residue were refluxed and extracted twice, each time for 1 hour. The solid and liquid were separated and the two extracts were combined.

[0086] (S3) The extract is concentrated under reduced pressure and freeze-dried to obtain the sorghum rice extract.

[0087] This preparation example also provides a wolfberry extract, prepared by the following method:

[0088] (T1) Mix wolfberry with 5 times its weight of water and extract twice under vacuum at 0.05 MPa and 40°C for 1 h each time. Combine the two extracts.

[0089] (T2) Add 3% activated carbon to the extract for decolorization, filter, and obtain the decolorized solution;

[0090] (T3) The decolorizing solution was mixed with 2% flocculant (gelatin was first mixed with 5 times the mass of 1% acetic acid aqueous solution and stirred until it became a paste, and then mixed with 50 times the mass of water and swollen for 12 h) for flocculation treatment, centrifuged, filtered, the filtrate was concentrated under reduced pressure and freeze-dried to obtain the wolfberry extract.

[0091] Example 1

[0092] This embodiment provides a white tea fermentation product, prepared by the following method:

[0093] (1) Mix dried white tea leaves with water at a ratio of 1:10 g / mL, soak at 70℃ for 3 h, then perform hot extraction at 90℃ for 2 h, filter, and obtain white tea extract;

[0094] (2) Prepare seed cultures of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mix them with a viable count of 1:2:1:1:1 to obtain a mixed seed culture;

[0095] A fermentation medium was prepared by mixing 100 parts white tea extract, 2 parts tryptone, 2 parts glucose, 0.1 parts NaCl, and 0.1 parts K2HPO4, and then sterilized under high temperature and high pressure.

[0096] The mixed seed liquid was inoculated at 3% into the sterilized fermentation medium and fermented at 37°C for 48 hours to obtain the primary fermentation product.

[0097] (3) Prepare seed cultures of Bayer conjugated yeast and brewer's yeast respectively, and mix them with a live cell count of 2:1 to obtain a mixed seed culture;

[0098] 100 parts of the primary fermentation product, 2 parts of sorghum rice extract from Preparation Example 1, and 2 parts of wolfberry extract from Preparation Example 1 were mixed and sterilized. The pH was adjusted to 6.8, and the mixed seed liquid was inoculated at 3% and fermented at 32°C for 36 hours to obtain the secondary fermentation product.

[0099] (4) After sterilizing the secondary fermentation product, cool it and filter it through a 10 μm filter plate to obtain the white tea fermentation product. Freeze-dry the filtrate to obtain the white tea fermentation product.

[0100] Example 2

[0101] This embodiment provides a white tea fermentation product, prepared by the following method:

[0102] (1) Mix dried white tea leaves with water at a ratio of 1:12 g / mL, soak at 60℃ for 4 h, then perform hot extraction at 95℃ for 1.5 h, filter, and obtain white tea extract;

[0103] (2) Prepare seed cultures of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mix them with a viable count ratio of 2:3:2:1:1 to obtain a mixed seed culture;

[0104] A fermentation medium was prepared by mixing 100 parts of white tea extract, 1 part of tryptone, 1 part of glucose, 0.1 parts of NaCl, and 0.1 parts of K2HPO4, and then sterilized by high temperature and high pressure.

[0105] The mixed seed liquid was inoculated at 4% into the sterilized fermentation medium and fermented at 35°C for 36 h to obtain the primary fermentation product.

[0106] (3) Prepare seed cultures of Bayer conjugated yeast and brewer's yeast respectively, and mix them with a live cell count of 1:1 to obtain a mixed seed culture;

[0107] 100 parts of the primary fermentation product, 3 parts of the sorghum rice extract from Preparation Example 2, and 1 part of the wolfberry extract from Preparation Example 2 were mixed and sterilized. The pH was adjusted to 6.8, and the mixed seed liquid was inoculated at 4% and fermented at 30°C for 42 hours to obtain the secondary fermentation product.

[0108] (4) After sterilizing the secondary fermentation product, cool it and filter it through a 10 μm filter plate to obtain the white tea fermentation product. Freeze-dry the filtrate to obtain the white tea fermentation product.

[0109] Example 3

[0110] This embodiment provides a white tea fermentation product, prepared by the following method:

[0111] (1) Mix dried white tea leaves with water at a ratio of 1:8 g / mL, soak at 80℃ for 2 h, then perform hot extraction at 95℃ for 1.5 h, filter, and obtain white tea extract;

[0112] (2) Prepare seed cultures of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mix them with a viable count of 1:2:1:2:2 to obtain a mixed seed culture;

[0113] A fermentation medium was prepared by mixing 100 parts of white tea extract, 3 parts of tryptone, 3 parts of glucose, 0.05 parts of NaCl, and 0.05 parts of K2HPO4, and then sterilized by high temperature and high pressure.

[0114] The mixed seed liquid was inoculated at 4% into the sterilized fermentation medium and fermented at 36℃ for 42 h to obtain the primary fermentation product.

[0115] (3) Prepare seed cultures of Bayer conjugated yeast and brewer's yeast respectively, and mix them with a live cell count of 1:1 to obtain a mixed seed culture;

[0116] 100 parts of the primary fermentation product, 1 part of the sorghum rice extract of Preparation Example 3, and 3 parts of the wolfberry extract of Preparation Example 3 were mixed and sterilized. The pH was adjusted to 6.8, and the mixed seed liquid was inoculated at 4% and fermented at 28°C for 48 hours to obtain the secondary fermentation product.

[0117] (4) After sterilizing the secondary fermentation product, cool it and filter it through a 10 μm filter plate to obtain the white tea fermentation product. Freeze-dry the filtrate to obtain the white tea fermentation product.

[0118] Example 4

[0119] This embodiment provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in step (2): seed liquids of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius are prepared separately and mixed with a viable count ratio of 3:1:3:3:1 to obtain a mixed seed liquid. Other conditions remain unchanged.

[0120] Example 5

[0121] This embodiment provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in step (3): seed liquids of Bayer conjugating yeast and brewer's yeast are prepared separately and mixed at a live cell ratio of 5:1 to obtain a mixed seed liquid. Other conditions remain unchanged.

[0122] Comparative Example 1

[0123] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in step (2): seed liquids of Lactobacillus plantarum and Lactobacillus acidophilus are prepared separately and mixed with a live bacteria ratio of 1:2 to obtain a mixed seed liquid. Other conditions remain unchanged.

[0124] Comparative Example 2

[0125] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in step (2): seed liquids of Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius are prepared separately and mixed with a viable count of 1:1:1 to obtain a mixed seed liquid. Other conditions remain unchanged.

[0126] Comparative Example 3

[0127] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in that a single Bayer zygosaccharidosis seed liquid is inoculated in step (3), while the total inoculation amount remains unchanged. Other conditions remain unchanged.

[0128] Comparative Example 4

[0129] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in that a single brewer's yeast seed liquid is inoculated in step (3), while the total inoculation amount remains unchanged. Other conditions remain unchanged.

[0130] Comparative Example 5

[0131] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in that the sorghum rice extract of Example 1 is omitted in step (3) and the wolfberry extract of Example 1 is 4 parts. Other conditions remain unchanged.

[0132] Comparative Example 6

[0133] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in that the wolfberry extract of Example 1 is omitted in step (3) and the sorghum rice extract of Example 1 is 4 parts. Other conditions remain unchanged.

[0134] Comparative Example 7

[0135] This comparative example provides a white tea fermentation product, the preparation method of which differs from that of Example 1 only in that step (3) lacks the wolfberry extract and sorghum rice extract of Example 1, the reduced amount of which is made up by the primary fermentation product. Other conditions remain unchanged.

[0136] Test Example 1

[0137] Safety performance assessment:

[0138] Safety performance was evaluated for the products in Examples 1-5 and Comparative Examples 1-7. Each product group was diluted with PBS to a 10% test sample solution. Thirty eligible volunteers, aged 20-45 years, were randomly divided into three groups, each group trying one of the four different products. The test samples were placed in the small chamber of the plaster applicator; the control well was left untreated. The plaster applicator containing the test substance was applied to the inner forearm of the subject using hypoallergenic adhesive tape, gently pressing it with the palm to ensure even application to the skin, for 24 hours. Skin reactions were observed according to the standards in Table 1 at 30 min, 24 h, and 48 h after removing the plaster applicator.

[0139] Table 1

[0140]

[0141] The results showed that the products of Examples 1-5 and Comparative Examples 1-7 of the present invention all had negative reactions after trial use, indicating that the white tea fermentation products involved in the present invention are safe and mild and will not cause adverse reactions such as skin irritation or sensitization.

[0142] Test Example 2

[0143] Antioxidant effect evaluation test:

[0144] (1) Preparation of test samples: Prepare test samples with a concentration of 2% using the products obtained in Examples 1-5 and Comparative Examples 1-7.

[0145] (2) Experimental procedure: Add 1.0 mL of DPPH test solution with a mass concentration of 40 mg / L and 1.0 mL of the test sample with a concentration of 5% to the test tube. The total volume is made up to 3 mL with anhydrous ethanol. Shake well and react in the dark for 30 min. The absorbance is measured at 517 nm as Ai. Add 1.0 mL of DPPH test solution with a mass concentration of 40 mg / L and 2 mL of anhydrous ethanol to the test tube. Shake well and react in the dark for 30 min. The absorbance is measured as Ac. Add 2 mL of anhydrous ethanol and 1.0 mL of the test sample with a concentration of 5% to the test tube. Shake well and react in the dark for 30 min. The absorbance is measured as Aj.

[0146] Calculate its free radical scavenging rate using the following formula.

[0147] Clearance rate (%) = [1 - (Ai - Aj) / Ac] × 100%

[0148] Where Aj represents the contribution of the sample itself to the absorbance; Ai represents the absorbance of the sample after the DPPH treatment; and Ac represents the absorption of DPPH itself at the measurement wavelength.

[0149] The higher the DPPH scavenging rate, the stronger the antioxidant capacity for scavenging free radicals. The results are shown in Table 2 (each group of samples was measured in parallel 5 times, and the data are presented as average values).

[0150] Test Example 3

[0151] elastase inhibition assay:

[0152] (1) Preparation of test samples: Prepare test samples with a concentration of 2% using the products obtained in Examples 1-5 and Comparative Examples 1-7.

[0153] (2) Experimental procedure: 10 μL of the test sample solution was added to each well of a 96-well plate, and 10 μL of diluent was added to the negative control group. Then, 20 μL of elastase solution with a concentration of 0.1 U / mL was added to the 96-well plate. The 96-well plate was incubated in a constant temperature incubator at 25℃ for 15 min. Then, 50 μL of substrate with a concentration of 1 mg / mL (98% N-succinyl-L-alanyl-L-alanine) was added. The absorbance of the sample at 410 nm was measured using a microplate reader, and the inhibition rate of elastase activity of the test sample was calculated.

[0154] The elastase activity inhibition rate is calculated using the following formula:

[0155] Elastase activity inhibition rate (%) = [(AB) / A] × 100%, where A represents the absorbance value of the negative control group without the test sample, and B represents the absorbance value of different test sample solutions.

[0156] A higher elastase inhibition rate indicates a better anti-aging effect of the product. The results are shown in Table 2 (each group of samples was measured in parallel 5 times, and the data are presented as average values).

[0157] Table 2

[0158]

[0159] As shown in Table 2, compared with the comparative product, the white tea fermentation product involved in this invention, through the selection of specific strains for primary and secondary fermentation, and the supplementation of sorghum rice extract and wolfberry extract as fermentation substrates, exhibits excellent DPPH free radical scavenging activity and elastase inhibitory activity, thus demonstrating superior skin antioxidant and anti-aging potential. Furthermore, the ratio of fermentation strains also influences the antioxidant and anti-aging efficacy of the product to a certain extent.

[0160] Test Example 4

[0161] elastase inhibition assay:

[0162] (1) Preparation of test samples: Prepare test samples with a concentration of 2% using the products obtained in Examples 1-5 and Comparative Examples 1-7.

[0163] (2) Experimental procedure: 0.1 mL of 2.5 mmol / L CaCl2 was added to 0.5 mL of hyaluronidase solution (600 U / mL) and kept at 37°C for 20 min; 0.5 mL of the sample solution to be tested was added and kept at 37°C for 20 min; 2 mL of 0.4 mg / mL sodium hyaluronate was added and kept at 37°C for 40 min; 1 mL of acetylacetone solution was added and heated in a boiling water bath for 15 min, and then immediately cooled with ice water for 5 min; finally, 1.0 mL of Ehrlich reagent was added and the mixture was left at room temperature for 60 min to develop color, and the absorbance was measured at 530 nm.

[0164] The formula for calculating the hyaluronidase activity inhibition rate is:

[0165]

[0166] In the formula, A represents the absorbance of the control solution (using acetate buffer instead of the sample solution); B represents the absorbance of the control blank solution (using acetate buffer instead of the sample solution and enzyme solution); C represents the absorbance of the test sample solution; and D represents the absorbance of the sample blank solution (using acetate buffer instead of the enzyme solution).

[0167] A higher hyaluronidase activity inhibition rate indicates a better soothing effect of the product. The results are shown in Table 3 (each group of samples was measured in triplicate, and the data are presented as averages).

[0168] Table 3

[0169]

[0170] As shown in Table 3, compared with the comparative product, the white tea fermentation product involved in this invention, through the selection of specific strains for primary and secondary fermentation, and the supplementation of sorghum rice extract and wolfberry extract as fermentation substrates, exhibits excellent hyaluronidase activity inhibition, thus demonstrating superior skin-soothing potential. Furthermore, the ratio of fermentation strains also influences the product's soothing efficacy to a certain extent.

[0171] Test Example 5

[0172] Moisturizing evaluation test:

[0173] (1) Preparation of the sample to be tested:

[0174] Essence products containing 5% of the white tea fermentation products of Examples 1-5 and Comparative Examples 1-7 were prepared. The formulation consisted of the following components: 5% white tea fermentation product, 5% squalane, 3% dipotassium glycyrrhizate, 2% 1,2-pentanediol, 1% cetyl ester, 1% caprylyl glycol, and the balance being water. All components were mixed and dissolved evenly at 45°C to obtain the test essence. A blank control product containing the same formulation but without white tea fermentation product was used as the white tea fermentation product.

[0175] (2) 120 women were selected as subjects and divided into 12 groups of 10 people each. A half-face control experiment was conducted. The test serum was applied to the entire half of the face according to the usage instructions, and the blank control serum was applied to the other half of the face. The serum was applied twice a day for 28 consecutive days. Data was collected from both sides of the face on day 0 (before use) and after 28 consecutive days of use.

[0176] (3) Before the test, the subject cleaned their face, allowed it to air dry, and marked the measurement area. Before the test, the subject sat quietly in a standard room for at least 30 minutes, without drinking water, and remained relaxed. The skin moisture content of the test and control areas was measured using a Corneometer CM825 probe. Each area was measured three times in parallel, and the average value was taken. The blank value M0 of each test area was measured first, and the test was continued for 28 days. On the 28th day, the skin moisture content M1 was measured. The test for the same subject was performed by the same person. The rate of increase in skin moisture content (%) was calculated. The test results are shown in Table 4.

[0177] Table 4

[0178]

[0179] As shown in Table 4, compared with the comparative product, the white tea fermentation product involved in this invention, through the selection of specific strains for primary and secondary fermentation, and the supplementation of sorghum rice extract and wolfberry extract as fermentation substrates, exhibits superior skin moisturizing effects. Furthermore, the ratio of fermentation strains also influences the product's moisturizing efficacy to a certain extent.

[0180] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments 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, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0181] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0182] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for preparing a white tea fermentation product having moisturizing, anti-aging, and soothing effects, characterized by, The preparation method comprises the following steps: (1) mixing dry white tea leaves with water, hot extracting at 90-100℃ for 1-3 h after soaking, and then separating the solid and liquid to obtain a white tea extract; (2) inoculating the compound lactobacillus into a sterilized culture medium containing the white tea extract, and then performing primary fermentation at 35-38℃ for 30-48 h to obtain a primary fermentation product; The compound lactobacillus is a combination of lactobacillus plantarum, lactobacillus acidophilus, lactobacillus rhamnosus, lactobacillus casei and lactobacillus salivarius with a ratio of viable bacterial count of (1-2):(2-4):(1-2):(1-2):(1-2); (3) mixing the primary fermentation product, the sorghum extract and the wolfberry extract at a mass ratio of 100:(1-5):(1-5), sterilizing the mixture, inoculating compound yeast, and then performing secondary fermentation at 28-35℃ for 30-48 h to obtain a secondary fermentation product; The compound yeast is a combination of bayer conjugated yeast and saccharomyces cerevisiae with a ratio of viable bacterial count of (1-3):(1-3); The preparation method of the sorghum extract comprises the following steps: (S1) mixing pulverized sorghum powder with an ethanol aqueous solution, and then performing microwave-ultrasonic combined treatment to obtain a treatment liquid and sorghum residue; (S2) performing reflux extraction on the treatment liquid and the sorghum residue, and then separating the solid and liquid to obtain an extract; (S3) concentrating and drying the extract to obtain the sorghum extract; The preparation method of the wolfberry extract comprises the following steps: (T1) mixing wolfberry with water, and then performing vacuum reduced pressure reflux extraction to obtain an extract; (T2) performing decolorization treatment on the extract, and then separating the solid and liquid to obtain a decolorized liquid; (T3) performing flocculation treatment on the decolorized liquid, and then separating the solid and liquid, concentrating and drying the filtrate to obtain the wolfberry extract; (4) sterilizing the secondary fermentation product, cooling, and then separating the solid and liquid to obtain the white tea fermentation product.

2. The production method according to claim 1, characterized by, The soaking in step (1) is performed at 40-80℃ for 1-4 h; The culture medium in step (2) comprises, in mass fractions, 90-110 parts of white tea extract, 1-3 parts of an external nitrogen source, 1-3 parts of an external carbon source, and 0.1-0.2 parts of an external inorganic salt.

3. The preparation method according to claim 1, characterized in that, The volume fraction of the ethanol aqueous solution in step (S1) is 70%-90%; The microwave-ultrasonic combined treatment in step (S1) is performed at 40-55℃ for 10-30 min; The microwave treatment power in step (S1) is 100-300 W, and the ultrasonic treatment power is 200-400 W; The reflux extraction in step (S2) is performed for 1-3 times, and each extraction time is independently 0.5-3 h.

4. The production method according to claim 1, characterized by, The vacuum reduced pressure reflux extraction in step (T1) is performed for 1-2 times, and each extraction time is independently 0.5-2 h; The decolorization treatment in step (T2) adopts activated carbon with an addition amount of 1-5%; The flocculation treatment in step (T3) adopts a flocculant with an addition amount of 1-5%, and the flocculant is selected from any one or a combination of at least two of cationic chitosan, guar gum, gelatin, cationic starch or tannin.

5. A white tea fermentation product prepared according to the method of any one of claims 1-4.

6. Use of the white tea fermentation product of claim 5 in the preparation of a cosmetic product having any one or at least two of the following effects; 1) moisturizing; 2) anti-aging; 3) antioxidant; 4) soothing.

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