Preparation method of white tea fermentation product with moisturizing, anti-aging and soothing effects as well as product and application of white tea fermentation product

Abstract

The invention relates to a preparation method of a white tea fermentation product with moisturizing, anti-aging and soothing effects as well as a product and application thereof. The preparation method comprises the following steps: mixing dry white tea leaves with water, soaking, and then carrying out heat extraction and solid-liquid separation to obtain a white tea extracting solution; inoculating the sterilized culture medium containing the white tea extracting solution with compound lactobacillus, and performing primary fermentation to obtain a primary fermentation product; mixing the primary fermentation product, a sorghum rice extract and a Chinese wolfberry fruit extract, performing sterilization treatment, inoculating compound yeast, and performing secondary fermentation to obtain a secondary fermentation product; and sterilizing the secondary fermentation product, cooling, and carrying out solid-liquid separation to obtain the white tea fermentation product. The preparation method of the white tea fermentation product is simple in process, industrial production is easy to realize, and the prepared fermentation product is mild and non-irritant in component and high in safety, has excellent moisturizing, anti-aging and soothing effects, and can be used as a preparation raw material to be added into skin care products and cosmetics.

Description

Technical Field

[0001] The invention belongs to the technical field of plant fermentation, and relates to a preparation method of a white tea fermentation product with moisturizing, anti-aging and soothing effects, and a product and application thereof. Background Art

[0002] In recent years, tea and its products have been favored by the daily chemical industry. The production of these products and the rise of tea applications in the daily chemical industry are inseparable from the in-depth research on tea and its ingredients and effects. Tea is divided into six categories according to the production method and fermentation degree: green tea (unfermented), white tea (slightly fermented), yellow tea (lightly fermented), oolong tea (semi-fermented), dark tea (post-fermented), and black tea (fully fermented). The production of white tea has very strict requirements on varieties.

[0003] White tea is a special and precious variety among the six major teas, and is also the earliest tea in history. White tea is named for its unique production method, which does not involve frying or rolling. Its appearance is silvery white and grayish green, and its white hairs are all white. It is mainly produced in Fuding, Zhenghe, Songxi, Jianyang and other places in Fujian. Due to its unique processing technology, it has formed the quality characteristics of fresh aroma, prominent hair fragrance, and fresh taste, and is deeply loved by the public. The unique quality and taste of white tea are deeply loved by everyone. The main active ingredients contained in white tea include tea polyphenols, amino acids, caffeine, soluble sugars, etc.

[0004] At present, white tea has not been fully opened up in the domestic and foreign markets. It is necessary to inject new vitality into the white tea industry and expand the application areas of white tea. In particular, there is a lack of necessary attempts and innovations in the field of white tea fermentation. Summary of the invention

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for preparing a white tea fermentation product having moisturizing, anti-aging and soothing effects, as well as a product and application thereof.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

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

[0008] (1) Mixing dried white tea leaves with water, soaking them, performing heat extraction, and separating the solid from the liquid to obtain a white tea extract;

[0009] (2) inoculating the composite lactobacillus into a sterilized culture medium containing white tea extract, performing a primary fermentation, and obtaining a primary fermentation product;

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

[0011] (3) mixing the primary fermentation product, sorghum rice extract, and wolfberry extract, sterilizing the mixture, and inoculating the mixture with a composite yeast to perform a secondary fermentation to obtain a secondary fermentation product;

[0012] The composite yeast is a combination of Bayer's yeast and Saccharomyces cerevisiae;

[0013] (4) sterilizing the secondary fermentation product, cooling it, and separating the solid from the liquid to obtain the white tea fermentation product.

[0014] The invention develops a new product with white tea as the main raw material. The preparation method of the product comprises the following steps: using white tea water extract as a primary fermentation substrate, supplemented with sorghum rice extract and wolfberry fruit extract as secondary fermentation substrates, sequentially performing lactobacillus fermentation and yeast fermentation, wherein the white tea water extract is fermented by utilizing the complementary synergistic effect of lactobacillus and yeast, thereby retaining the active components of white tea to the greatest extent and enabling the fermentation product to have excellent moisturizing, anti-aging and soothing effects; the sorghum rice extract and wolfberry fruit extract added in the secondary fermentation are fermented together with the primary fermentation metabolites of white tea, thereby further improving the moisturizing, anti-aging and soothing effects of the product; and the microorganisms used in the two fermentations have a specific bacterial species composition mode, and the bacterial species cooperate with each other in improving the efficacy of the fermentation product, thereby synergizing and enhancing the effect.

[0015] The preparation method of the white tea fermentation product involved in the present invention has simple process and is easy to realize industrial production. The obtained fermentation product has mild ingredients and is non-irritating and has high safety, and can be added to skin care products and cosmetics as a preparation raw material.

[0016] Preferably, the ratio of the viable cell 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 the viable counts of the Bayer yeast and the brewer's yeast is (1-3):(1-3).

[0018] Based on the potential synergistic relationship between the various fermentation strains, when they meet a specific mass ratio, the obtained fermentation product has better effects in moisturizing, anti-aging and soothing.

[0019] The specific point values ​​in the above (1-2) can be selected from 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 selected from 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 selected from 1, 1.3, 1.5, 1.7, 2, 2.2, 2.5, 2.8, 3, etc.

[0020] Preferably, the solid-liquid ratio of the white tea leaves to water in step (1) is 1:(8-12) g / mL, for example 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°C (e.g. 40°C, 50°C, 60°C, 70°C, 80°C, etc.) for 1-4 h (e.g. 1 h, 2 h, 3 h, 4 h, etc.).

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

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

[0024] Preferably, the temperature of the primary fermentation in step (2) is 35-38°C, for example, 35°C, 36°C, 37°C, 38°C, etc.; the time is 30-48 h, for example, 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 external nitrogen source includes any one of yeast powder, tryptone, soy peptone or beef extract, or a combination of at least two of them.

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

[0028] The added inorganic salt includes NaCl and / or K2HPO4.

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

[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); 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%, for example, 3%, 3.5%, 4%, 4.5%, 5%, etc.

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

[0033] Preferably, the solid-liquid separation in step (4) is followed by drying.

[0034] The sorghum rice extract and wolfberry fruit extract used in the present invention are preferably prepared by the following method, and the effects are better when they are fermented with white tea.

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

[0036] (S1) crushing sorghum rice powder, mixing it with an ethanol aqueous solution, and then subjecting it to a microwave-ultrasonic combined treatment to obtain a treatment liquid and sorghum rice residue;

[0037] (S2) subjecting the treated liquid and sorghum rice residue to reflux extraction and solid-liquid separation to obtain an extract;

[0038] (S3) concentrating and drying the extract to obtain the sorghum rice extract.

[0039] The sorghum rice extract is first pretreated with a microwave-ultrasound combination before reflux extraction. Microwaves can increase the temperature inside plant cells and accelerate the rupture of cell walls, and ultrasound can produce a cavitation effect. Compared with a single microwave or single ultrasound pretreatment method, the combined pretreatment has a better extraction effect.

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

[0041] Preferably, in step (S1), the mass ratio of the ethanol aqueous solution to sorghum rice 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-ultrasonic combined treatment in step (S1) is carried out at 40-55°C (e.g., 40°C, 42°C, 45°C, 48°C, 50°C, 52°C, 54°C, 55°C, 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, for example, 100 W, 150 W, 200 W, 250 W, 300 W, etc.; the processing power of the ultrasonic wave is 200-400 W, for example, 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 time of 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 comprises the following steps:

[0046] (T1) mixing wolfberry with water and subjecting the mixture to vacuum decompression reflux extraction, and performing solid-liquid separation to obtain an extract;

[0047] (T2) decolorizing the extract and separating the solid from the liquid to obtain a decolorized solution;

[0048] (T3) flocculating the decolorized liquid, separating the solid from the liquid, concentrating and drying the filtrate to obtain the wolfberry extract.

[0049] The wolfberry is subjected to decolorization and flocculation treatments in sequence after reflux extraction, and has better efficacy after fermentation than the extracted product without the above treatments.

[0050] Preferably, in step (T1), the mass ratio of water to wolfberry 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 time of each extraction is independently 0.5-2 h, such as 0.5 h, 1 h, 1.5 h, 2 h, etc.

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

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

[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] In a third aspect, the present invention provides a use of the white tea fermentation product according to the second aspect in preparing a product having any one or at least two of the following effects:

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

[0057] Preferably, the product comprises skin care products, cosmetics, medicines or medical materials.

[0058] Other unlisted specific point values ​​within the numerical ranges involved in the present invention can be selected and are all within the protection scope of the present invention. Considering the length of the article and the brevity of the expression, they will not be described one by one here.

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

[0060] The invention develops a new product with white tea as the main raw material. The preparation method of the product comprises the following steps: using white tea water extract as a primary fermentation substrate, supplemented with sorghum rice extract and wolfberry fruit extract as secondary fermentation substrates, sequentially performing lactobacillus fermentation and yeast fermentation, wherein the white tea water extract is fermented by utilizing the complementary synergistic effect of lactobacillus and yeast, thereby retaining the active components of white tea to the greatest extent and enabling the fermentation product to have excellent moisturizing, anti-aging and soothing effects; the sorghum rice extract and wolfberry fruit extract added in the secondary fermentation are fermented together with the primary fermentation metabolites of white tea, thereby further improving the moisturizing, anti-aging and soothing effects of the product; and the microorganisms used in the two fermentations have a specific bacterial species composition mode, and the bacterial species cooperate with each other in improving the efficacy of the fermentation product, thereby synergizing and enhancing the effect.

[0061] The preparation method of the white tea fermentation product involved in the present invention has simple process and is easy to realize industrial production. The obtained fermentation product has mild ingredients and is non-irritating and has high safety, and can be added to skin care products and cosmetics as a preparation raw material. DETAILED DESCRIPTION

[0062] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0063] The plant lactobacillus involved in the following is the ATCC8014 strain; the acidophilus Lactobacillus is the ATCC4356 strain; the rhamnosus Lactobacillus is the ATCC39595 strain; the casei Lactobacillus is the ATCC27139 strain; the salivarius Lactobacillus is the ATCC11741 strain; the bayley yeast is the ATCC MYA-4549 strain; and the cerevisiae yeast is the ATCC26603 strain.

[0064] Preparation Example 1

[0065] This preparation example provides a sorghum rice extract, and the preparation method is as follows:

[0066] (S1) crushing sorghum rice flour and mixing it with 8 times the mass of 80% ethanol aqueous solution, and then subjecting it to microwave-ultrasonic combined treatment at 50° C. for 30 min, with the microwave processing power of 200 W and the ultrasonic processing power of 300 W, to obtain a processing liquid and sorghum rice residue;

[0067] (S2) subjecting the treated liquid and sorghum rice residue to reflux extraction twice, each time for 1.5 h, solid-liquid separation, and combining the two extracts;

[0068] (S3) Concentrating the extract under reduced pressure and freeze-drying to obtain the kaoliang rice extract.

[0069] This preparation example also provides a wolfberry extract, and the preparation method is as follows:

[0070] (T1) Mix wolfberry with 10 times the mass of water, perform vacuum decompression reflux extraction twice at a vacuum degree of 0.05 MPa and a temperature of 40°C, each time for 1.5 h, and combine the two extracts;

[0071] (T2) adding 3% activated carbon to the extract to perform decolorization, filtering, and obtaining a decolorized solution;

[0072] (T3) The decolorized liquid is mixed with 1.5% of a flocculant (cationic chitosan and gelatin in a mass ratio of 1:1 are first mixed with 5 times the mass of a 1% acetic acid aqueous solution and stirred until a paste is formed, and then mixed with 50 times the mass of water and swelled for 12 hours) for flocculation treatment, centrifuged, filtered, and the filtrate is 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, and the preparation method is as follows:

[0075] (S1) crushing sorghum rice flour and mixing it with 12 times the mass of 70% ethanol aqueous solution, and then subjecting it to microwave-ultrasonic combined treatment at 45° C. for 30 min, with the microwave processing power of 150 W and the ultrasonic processing power of 400 W, to obtain a processing liquid and sorghum rice residue;

[0076] (S2) subjecting the treated liquid and sorghum rice residue to reflux extraction twice, each time for 2 h, solid-liquid separation, and combining the two extracts;

[0077] (S3) Concentrating the extract under reduced pressure and freeze-drying to obtain the kaoliang rice extract.

[0078] This preparation example also provides a wolfberry extract, and the preparation method is as follows:

[0079] (T1) Mix wolfberry with 12 times the mass of water, perform vacuum decompression reflux extraction twice at a vacuum degree of 0.05 MPa and a temperature of 40°C, each time for 2 h, and combine the two extracts;

[0080] (T2) adding 5% activated carbon to the extract to perform decolorization, filtering, and obtaining a decolorized solution;

[0081] (T3) The decolorized liquid is mixed with 2% of a flocculant (cationic chitosan is first mixed with 5 times the mass of a 1% acetic acid aqueous solution and stirred until it becomes a paste, and then mixed with 50 times the mass of water and swelled for 12 hours) for flocculation treatment, centrifuged, filtered, and the filtrate is 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, and the preparation method is as follows:

[0084] (S1) crushing sorghum rice flour and mixing it with 5 times the mass of 90% ethanol aqueous solution, and then subjecting it to microwave-ultrasonic combined treatment at 55° C. for 20 min, with the microwave processing power of 250 W and the ultrasonic processing power of 250 W, to obtain a processing liquid and sorghum rice residue;

[0085] (S2) subjecting the treated liquid and sorghum rice residue to reflux extraction twice, each time for 1 h, solid-liquid separation, and combining the two extracts;

[0086] (S3) Concentrating the extract under reduced pressure and freeze-drying to obtain the kaoliang rice extract.

[0087] This preparation example also provides a wolfberry extract, and the preparation method is as follows:

[0088] (T1) Mix wolfberry with 5 times the mass of water, perform vacuum decompression reflux extraction twice at a vacuum degree of 0.05 MPa and a temperature of 40°C, each time for 1 h, and combine the two extracts;

[0089] (T2) adding 3% activated carbon to the extract to perform decolorization, filtering, and obtaining a decolorized solution;

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

[0091] Example 1

[0092] This embodiment provides a white tea fermentation product, and the preparation method is as follows:

[0093] (1) Dry white tea leaves were mixed with water at a solid-liquid ratio of 1:10 g / mL, soaked at 70°C for 3 h, and then heat extracted at 90°C for 2 h, filtered, and white tea extract was obtained;

[0094] (2) preparing seed solutions of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mixing them in a ratio of viable cell count of 1:2:1:1:1 to obtain a mixed seed solution;

[0095] Mix 100 parts of white tea extract, 2 parts of tryptone, 2 parts of glucose, 0.1 parts of NaCl and 0.1 parts of K2HPO4 to prepare a fermentation medium, and sterilize it under high temperature and high pressure;

[0096] The mixed seed liquid was inoculated into the sterilized fermentation medium at 3%, and a fermentation was carried out at 37°C for 48 h to obtain a primary fermentation product;

[0097] (3) preparing seed solutions of Bayer's yeast and Saccharomyces cerevisiae respectively, and mixing them in a ratio of 2:1 in terms of viable bacterial count to obtain a mixed seed solution;

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

[0099] (4) The secondary fermentation product is sterilized and then cooled, filtered through a 10 μm filter plate to obtain a white tea fermentation product, and the filtrate is freeze-dried to obtain a white tea fermentation product.

[0100] Example 2

[0101] This embodiment provides a white tea fermentation product, and the preparation method is as follows:

[0102] (1) Dry white tea leaves were mixed with water at a solid-liquid ratio of 1:12 g / mL, soaked at 60°C for 4 h, and then heat extracted at 95°C for 1.5 h, filtered, and white tea extract was obtained;

[0103] (2) preparing seed solutions of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mixing them in a ratio of viable cell count of 2:3:2:1:1 to obtain a mixed seed solution;

[0104] Mix 100 parts of white tea extract, 1 part of tryptone, 1 part of glucose, 0.1 part of NaCl, and 0.1 part of K2HPO4 to prepare a fermentation medium, and sterilize it under high temperature and high pressure;

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

[0106] (3) preparing seed solutions of Bayer's yeast and Saccharomyces cerevisiae respectively, and mixing them in a ratio of 1:1 with viable bacteria to obtain a mixed seed solution;

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

[0108] (4) The secondary fermentation product is sterilized and then cooled, filtered through a 10 μm filter plate to obtain a white tea fermentation product, and the filtrate is freeze-dried to obtain a white tea fermentation product.

[0109] Example 3

[0110] This embodiment provides a white tea fermentation product, and the preparation method is as follows:

[0111] (1) Dry white tea leaves were mixed with water at a solid-liquid ratio of 1:8 g / mL, soaked at 80°C for 2 h, and then heat extracted at 95°C for 1.5 h, filtered, and white tea extract was obtained;

[0112] (2) preparing seed solutions of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mixing them in a ratio of viable cell count of 1:2:1:2:2 to obtain a mixed seed solution;

[0113] Mix 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 to prepare a fermentation medium, and sterilize it under high temperature and high pressure;

[0114] The mixed seed liquid was inoculated into the sterilized fermentation medium at 4%, and a fermentation was carried out at 36°C for 42 h to obtain a primary fermentation product.

[0115] (3) preparing seed solutions of Bayer's yeast and Saccharomyces cerevisiae respectively, and mixing them in a ratio of 1:1 with viable bacteria to obtain a mixed seed solution;

[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 with 4% to carry out secondary fermentation at 28° C. for 48 hours to obtain a secondary fermentation product;

[0117] (4) The secondary fermentation product is sterilized and then cooled, filtered through a 10 μm filter plate to obtain a white tea fermentation product, and the filtrate is freeze-dried to obtain a white tea fermentation product.

[0118] Example 4

[0119] This embodiment provides a white tea fermentation product, and the preparation method thereof is different from that of the embodiment 1 only in step (2): preparing seed liquids of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius respectively, and mixing them in a ratio of viable cell count 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, and the preparation method thereof is different from that of the embodiment 1 only in that in step (3): seed liquids of bayer yeast and cerevisiae yeast are prepared separately and mixed at a live bacterial count 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, and the preparation method thereof differs from that of Example 1 only in that in step (2): seed liquids of Lactobacillus plantarum and Lactobacillus acidophilus are prepared separately and mixed at a live bacterial count 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, and the preparation method thereof differs from that of Example 1 only in that in step (2): seed solutions of Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius are prepared respectively, and mixed at a ratio of 1:1:1 of viable bacteria to obtain a mixed seed solution. Other conditions remain unchanged.

[0126] Comparative Example 3

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

[0128] Comparative Example 4

[0129] This comparative example provides a white tea fermentation product, and the preparation method thereof is different from that of Example 1 only in that a single brewer's yeast seed liquid is inoculated in step (3), and 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 is different from that of Example 1 only in that the sorghum rice extract of Preparation Example 1 is missing in step (3) and the wolfberry extract of Preparation 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 is different from that of Example 1 only in that the wolfberry extract of Preparation Example 1 is missing in step (3) and the sorghum rice extract of Preparation 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 is different from that of Example 1 only in that the wolfberry extract and the sorghum rice extract of Preparation Example 1 are missing in step (3), and the reduced amount is supplemented by the primary fermentation product. Other conditions remain unchanged.

[0136] Test Example 1

[0137] Safety performance evaluation:

[0138] The safety performance of the products of Examples 1-5 and Comparative Examples 1-7 was evaluated. Each group of products was diluted with PBS to a 10% sample solution to be tested. 30 social volunteers aged 20-45 years old who met the requirements were selected and randomly divided into 3 groups. Each group tried 4 different products. Placed in a spot tester chamber, the control well was not treated in any way. The spot tester with the test substance was applied to the inner side of the forearm of the subject with a low-allergenic tape, and gently pressed with the palm of the hand to evenly apply it to the skin for 24 hours. The skin reaction was observed according to the standards in Table 1 at 30 minutes, 24 hours and 48 hours after the removal of the test substance spot tester.

[0139] Table 1

[0140]

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

[0142] Test Example 2

[0143] Antioxidant effect evaluation test:

[0144] (1) Preparation of test samples: The products prepared in Examples 1-5 and Comparative Examples 1-7 were prepared into test samples with a concentration of 2% for later use.

[0145] (2) Test procedure: Add 1.0 mL of 40 mg / L DPPH test solution and 1.0 mL of 5% sample to a test tube, add anhydrous ethanol to the total volume to 3 mL, shake well, and react in the dark for 30 min. The absorbance at 517 nm is measured as Ai; add 1.0 mL of 40 mg / L DPPH test solution and 2 mL of anhydrous ethanol to a 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 5% sample to a test tube, shake well, and react in the dark for 30 min. The absorbance is measured as Aj.

[0146] The free radical scavenging rate was calculated according to the following formula.

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

[0148] Among them, Aj is the contribution of the reaction sample itself to the absorbance; Ai is the absorbance value of the sample after the action of DPPH; Ac is the absorption of DPPH itself at the measurement wavelength.

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

[0150] Test Example 3

[0151] Elastase inhibition test:

[0152] (1) Preparation of test samples: The products prepared in Examples 1-5 and Comparative Examples 1-7 were prepared into test samples with a concentration of 2% for later use.

[0153] (2) Test process: 10 μL of the sample solution to be tested was added to a 96-well plate, 10 μL of dilution solvent was added to the negative control group, and 20 μL of 0.1 U / mL elastase solution was added to the 96-well plate. The 96-well plate was placed in a 25°C constant temperature incubator for 15 min, and then 50 μL of 1 mg / mL substrate (98% N-succinyl-L-alanyl-L-alanine) was added. The absorbance of the sample at 410 nm was measured using an ELISA reader, and the inhibition rate of the sample to be tested on the elastase activity was calculated.

[0154] The inhibition rate of elastase activity was calculated by the following formula:

[0155] Elastase activity inhibition rate (%) = [(AB) / A] × 100%, 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] The higher the elastase inhibition rate, the better the anti-aging effect of the product. The results are shown in Table 2 (each group of samples was measured 5 times in parallel, and the data are presented as average values).

[0157] Table 2

[0158]

[0159] As shown in Table 2, compared with the comparative example product, the white tea fermented product of the present invention has excellent DPPH free radical scavenging activity and elastase inhibition activity by selecting specific strains for primary and secondary fermentation, and supplemented with sorghum rice extract and wolfberry extract as fermentation substrates, that is, has excellent skin antioxidant and anti-aging potential. And the ratio of fermented strains also affects the antioxidant and anti-aging effects of the product to a certain extent.

[0160] Test Example 4

[0161] Elastase inhibition test:

[0162] (1) Preparation of test samples: The products prepared in Examples 1-5 and Comparative Examples 1-7 were prepared into test samples with a concentration of 2% for later use.

[0163] (2) Test procedure: Add 0.1 mL of 2.5 mmol / L CaCl2 to 0.5 mL of hyaluronidase solution (600 U / mL) and keep warm at 37°C for 20 min; add 0.5 mL of the sample solution to be tested and keep warm at 37°C for 20 min; add 2 mL of 0.4 mg / mL sodium hyaluronate and keep warm at 37°C for 40 min; add 1 mL of acetylacetone solution, heat in a boiling water bath for 15 min, and immediately cool with ice water for 5 min; finally, add 1.0 mL of Ehrlich reagent and leave at room temperature for 60 min to develop color, and measure the absorbance at 530 nm.

[0164] The calculation formula of hyaluronidase activity inhibition rate is:

[0165]

[0166] Where A represents: the absorbance of the control solution (using acetate buffer instead of sample solution); B represents: the absorbance of the control blank solution (using acetate buffer instead of sample solution and enzyme solution); C represents: the absorbance of the sample solution to be tested; D represents: the absorbance of the sample blank solution (using acetate buffer instead of enzyme solution).

[0167] The higher the hyaluronidase activity inhibition rate, the better the soothing effect of the product. The results are shown in Table 3 (each group of samples was measured 3 times in parallel, and the data are presented as average values).

[0168] Table 3

[0169]

[0170] As shown in Table 3, compared with the comparative example product, the white tea fermented product of the present invention has excellent hyaluronidase activity inhibition activity, i.e., excellent skin soothing potential, by selecting specific strains for primary and secondary fermentation, and supplemented with sorghum rice extract and wolfberry extract as fermentation substrates. The ratio of fermented strains also affects the soothing effect of the product to a certain extent.

[0171] Test Example 5

[0172] Moisturizing evaluation test:

[0173] (1) Configuration 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, and the formulas thereof were composed of the following components: 5% of white tea fermentation products, 5% of squalane, 3% of dipotassium glycyrrhizinate, 2% of 1,2-pentanediol, 1% of cetyl esters, 1% of caprylyl glycol, and the balance of water. The components were mixed and dissolved uniformly at 45° C. to obtain the test essence, and a blank product of the same formula without the white tea fermentation product was used as a blank control white tea fermentation product.

[0175] (2) 120 female subjects were selected and divided into 12 groups, with 10 subjects in each group. A half-face control test was conducted at the same time. The test essence was used on the entire half of the face according to the usage instructions, and the blank control essence was used on the other half of the face. The test essence was applied twice a day for 28 consecutive days. Data on both sides of the face were collected on day 0 (before use) and after 28 consecutive days of use.

[0176] (3) Before the test, the subject cleans the face, air-dries it naturally, and marks the measurement area. Before the test, sit quietly in a standard room for at least 30 minutes, do not drink water, and stay relaxed. Use the probe Corneometer CM825 to measure the skin moisture content of the test area and the control area. Each area is measured three times in parallel and the average value is taken. First measure the blank value M0 of each test area, use it continuously for 28 days, and test the skin moisture content M1 on the 28th day. The test of the same subject is completed by the same measurement personnel. Calculate the growth rate (%) of the skin moisture content. The test results are shown in Table 4.

[0177] Table 4

[0178]

[0179] From the data results in Table 4, it can be seen that compared with the comparative example product, the white tea fermented product of the present invention has excellent skin moisturizing effect by selecting specific strains for primary fermentation and secondary fermentation in sequence, and supplemented with sorghum rice extract and wolfberry extract as fermentation substrates. The ratio of fermented strains also affects the moisturizing effect of the product to a certain extent.

[0180] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0181] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to 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 further describe various possible combinations.

Claims

1. A method for preparing a white tea fermentation product having moisturizing, anti-aging and soothing effects, characterized in that: The preparation method comprises the following steps: (1) Mixing dried white tea leaves with water, soaking them, performing heat extraction, and separating the solid from the liquid to obtain a white tea extract; (2) inoculating the composite lactobacillus into a sterilized culture medium containing white tea extract, performing a primary fermentation, and obtaining a primary fermentation product; The composite lactobacillus is a combination of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei and Lactobacillus salivarius; (3) mixing the primary fermentation product, sorghum rice extract, and wolfberry extract, sterilizing the mixture, and inoculating the mixture with a composite yeast to perform a secondary fermentation to obtain a secondary fermentation product; The composite yeast is a combination of Bayer's yeast and Saccharomyces cerevisiae; (4) sterilizing the secondary fermentation product, cooling it, and separating the solid from the liquid to obtain the white tea fermentation product.

2. The preparation method according to claim 1, characterized in that: The ratio of the 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); The ratio of the number of live bacteria of the Bayer binding yeast to that of brewer's yeast is (1-3):(1-3).

3. The preparation method according to claim 1, characterized in that: The immersion in step (1) is carried out at 40-80° C. for 1-4 h; The thermal extraction in step (1) is carried out at 90-100° C. for 1-3 h; The temperature of the first fermentation in step (2) is 35-38°C and the time is 30-48 hours; The culture medium in step (2) further contains an external nitrogen source, an external carbon source and an external inorganic salt; The culture medium in step (2) comprises, by weight, 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.

4. The preparation method according to claim 1, characterized in that: The mass ratio of the primary fermentation product, sorghum rice extract and wolfberry extract in step (3) is 100:(1-5):(1-5); The temperature of the secondary fermentation in step (3) is 28-35°C and the time is 30-48 hours.

5. The preparation method according to claim 1, characterized in that: The preparation method of the sorghum rice extract comprises the following steps: (S1) crushing sorghum rice powder, mixing it with an ethanol aqueous solution, and then subjecting it to a microwave-ultrasonic combined treatment to obtain a treatment liquid and sorghum rice residue; (S2) subjecting the treated liquid and sorghum rice residue to reflux extraction and solid-liquid separation to obtain an extract; (S3) concentrating and drying the extract to obtain the sorghum rice extract.

6. The preparation method according to claim 5, 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 carried out at 40-55° C. for 10-30 min; In step (S1), the processing power of the microwave is 100-300 W, and the processing power of the ultrasonic wave is 200-400 W; The number of reflux extractions in step (S2) is 1-3 times, and the time of each extraction is independently 0.5-3 h.

7. The preparation method according to claim 1, characterized in that: The preparation method of the wolfberry extract comprises the following steps: (T1) mixing wolfberry with water and subjecting the mixture to vacuum decompression reflux extraction, and performing solid-liquid separation to obtain an extract; (T2) decolorizing the extract and separating the solid from the liquid to obtain a decolorized solution; (T3) flocculating the decolorized liquid, separating the solid from the liquid, concentrating and drying the filtrate to obtain the wolfberry extract.

8. The preparation method according to claim 7, characterized in that: The number of vacuum decompression reflux extraction in step (T1) is 1-2 times, and the time of each extraction is independently 0.5-2 h; The decolorization treatment in step (T2) uses activated carbon added in an amount of 1-5%; The flocculation treatment in step (T3) uses a flocculant added in an amount of 1-5%, and the flocculant is selected from any one or a combination of at least two of water-soluble polysaccharides, cationic chitosan, guar gum, protein colloid, gelatin, animal glue, cationic starch or tannin.

9. A white tea fermentation product obtained by the preparation method according to any one of claims 1 to 8.

10. Use of the white tea fermentation product according to claim 9 in preparing a product having any one or at least two of the following effects; 1) Moisturizing; 2) Anti-aging; 3) Anti-oxidation; 4) Soothing; The products include skin care products, cosmetics, medicines or medical materials.

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