Preparation method and application of Cordyceps fermentation product

Abstract

The present invention relates to the field of daily chemical technology, and in particular to a method for preparing a cordyceps fermentation product and its application. The cordyceps fermentation product is obtained by fermenting cordyceps flowers as a solid culture medium with lactic acid bacteria and saccharomyces cerevisiae as strains. The cordyceps probiotic fermentation composition provided by the present invention significantly improves the content and stability of active substances such as cordycepin, cordyceps polysaccharides, adenosine, small molecule peptides, and amino acids in the fermentation product by constructing a composite solid-liquid dual-stage fermentation system using cordyceps flowers as raw materials and synergistically applying lactic acid bacteria and yeast for metabolic conversion, while giving the fermentation product good skin adaptability and skin care efficacy.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemicals, and in particular to a preparation method and application of a cordyceps fermentation product. Background Art

[0002] Cordyceps, a complex organism combining the properties of insects and fungi, is considered a rare and precious herbal ingredient in Traditional Chinese Medicine (TCM) for its diverse benefits, including nourishing the body, benefiting the lungs, relieving coughs, and boosting immunity. In recent years, the technology for cultivating cordyceps has matured. This cultivated form, particularly the Cordyceps flower (the fruiting body of Cordyceps militaris), has gained widespread application in healthcare, food, and cosmetics due to its high nutritional value and rich active ingredients.

[0003] Cordyceps flowers are rich in a variety of biologically active natural ingredients, including cordycepin, cordyceps polysaccharides, adenosine, mannitol, and amino acids. These ingredients possess significant antioxidant, anti-inflammatory, antibacterial, and immunomodulatory activities, which can improve skin condition and enhance skin barrier function. However, the natural content of active ingredients in cordyceps is limited, and traditional extraction processes often suffer from low efficiency, poor stability, and inadequate efficacy, making it difficult to meet the active ingredient content and stability requirements of high-performance functional skincare products.

[0004] As a natural and controllable biotransformation method, probiotic fermentation technology has been widely used in recent years to enhance the bioactivity of plant- and animal-derived extracts. Through targeted fermentation by specific strains, it not only releases the enclosed active ingredients in the raw materials but also produces functional metabolites such as active peptides, short-chain fatty acids, hyaluronic acid, and lactic acid, thereby synergistically enhancing the raw materials' anti-inflammatory, repair, and barrier-enhancing abilities.

[0005] Based on this, researchers are experimenting with incorporating functional probiotics such as lactic acid bacteria and yeast into the Cordyceps flower fermentation system. Through a two-stage, multi-strain complex fermentation, they aim to enrich its key active substances and impart new functional properties. Furthermore, combining this with plant enzymatic hydrolysates or traditional Chinese medicinal plants (such as wolfberry and astragalus) for complex fermentation can help introduce auxiliary ingredients such as natural polysaccharides, small molecule peptides, and amino acids, thereby achieving synergistic effects and improving the skin compatibility and activity of Cordyceps fermentation products.

[0006] Therefore, developing a Cordyceps-probiotic composite fermentation composition and its preparation method, by optimizing the fermentation process and component combination, to improve the content and stability of effective active substances from the source, has important application value, especially in the research and development of high-functional skin care products, showing broad application prospects. Summary of the Invention

[0007] To solve the above technical problems, the present invention provides a method for preparing a Cordyceps fermentation product, wherein the Cordyceps fermentation product is obtained by fermenting Cordyceps flowers as a solid culture medium with lactic acid bacteria and Saccharomyces cerevisiae as strains; the preparation method comprises the following steps:

[0008] (1) Crush the dried Cordyceps flowers, using a solid-liquid ratio of (1.6-2.5) ml: (0.5-1.5) g, mix the crushed Cordyceps residue with the solution, place it in a container and sterilize it. The total volume of the solid culture medium should occupy 10%-15% of the entire container.

[0009] (2) Inoculation of lactic acid bacteria into the cordyceps solid culture medium: inoculate lactic acid bacteria into the cordyceps solid culture medium at a rate of 3 wt% to 5 wt%, mix the bacterial solution and the solid culture medium evenly, and culture anaerobically at 37 °C for 48 to 72 h;

[0010] (3) Solid fermentation: Sterile liquid culture medium is added to the above Cordyceps solid culture medium, and 3 wt% to 5 wt% of Saccharomyces cerevisiae is added according to the total volume and cultured under microaerobic conditions at 28°C for 24 to 48 hours;

[0011] The formula of the sterile liquid culture medium is: 0.6 parts of peptone, 85-90 parts of water, sterilized at 115°C for 20 minutes, and added to the Cordyceps solid culture medium after cooling;

[0012] (4) Liquid fermentation: After the fermentation is completed, the solution is quickly heated to 72-75°C using a pasteurization method for 15-30 minutes, and then immediately cooled to 4-5°C. The fermentation solution is then filtered using an 8-10 μm filter plate to obtain the Cordyceps fermentation liquid.

[0013] The second aspect of the present invention provides a Cordyceps probiotic fermentation composition, which is prepared by the preparation method.

[0014] A third aspect of the present invention provides a soothing and moisturizing facial cream, wherein the raw materials for preparing the facial cream include the Cordyceps probiotic fermentation composition.

[0015] As an embodiment of the present invention, the raw materials for preparing the facial cream include, in parts by mass:

[0016] Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03;

[0017] Phase B: Ammonium Acryloyldimethyltaurate / VP Copolymer 0.50; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.80;

[0018] Phase C: OLIVEM1000 2.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00;

[0019] Phase D: 0.30 g of petrolatum; 0.20 g of tocopheryl acetate; 2.00 g of phytosteryl isostearate; 2.00 g of cyclodimethicone; 1.00 g of DC1403; 3.00 g of 1,3-butylene glycol; 0.50 g of p-hydroxyacetophenone; 0.50 g of 1,2-hexanediol; 2.00 g of β-glucan; and 2.00 g of the Cordyceps probiotic fermentation composition.

[0020] As an embodiment of the present invention, the steps of the preparation method of the soothing and moisturizing facial cream are as follows:

[0021] ① Disperse phase A, heat to about 85℃, and homogenize or stir to dissolve evenly;

[0022] ②Heat phase B to about 85℃ and homogenize or stir to dissolve evenly;

[0023] ③ Add phase A to phase B, homogenize for about 2 minutes, add phase C and phase D, continue homogenizing for about 5 minutes, and stir to cool down;

[0024] ④ Cool down to about 50℃, add phase E and phase F, stir evenly and discharge.

[0025] By adopting the above technical solution, the present invention has the following beneficial effects:

[0026] The cordyceps probiotic fermentation composition provided by the present invention constructs a composite solid-liquid dual-stage fermentation system using cordyceps flowers as raw materials, and synergistically applies lactic acid bacteria and yeast for metabolic transformation, thereby significantly improving the content and stability of active substances such as cordycepin, cordyceps polysaccharides, adenosine, small molecule peptides, and amino acids in the fermentation product, while giving it good skin adaptability and skin care efficacy.

[0027] The technical solution of the present invention has the following beneficial effects:

[0028] High efficiency of active ingredient enrichment: Through the metabolic action of probiotics and the synergistic enzymatic hydrolysis of plant coenzymes, the release rate and conversion efficiency of functional factors such as cordycepin, adenosine and polysaccharides are significantly improved. Compared with traditional water extraction or single-bacteria fermentation methods, the key ingredients can be increased by 1.3 to 3.5 times.

[0029] Synergistic enhancement of efficacy: Lactic acid bacteria and yeast produce active metabolites (such as lactic acid, hyaluronic acid, and glutathione) during the fermentation process, which work synergistically with the active ingredients of Cordyceps itself to exert comprehensive skin care effects such as soothing, repairing, anti-inflammatory, antioxidant, and barrier enhancement.

[0030] Better system stability: This fermentation system combines the advantages of solid-state pre-fermentation and liquid subsequent fermentation, which can effectively control the risk of bacterial contamination and form a microcolloid structure containing natural polysaccharides and mucilage polymers, making the final product easier to emulsify and more stable, suitable for application in skin care systems such as creams and essences.

[0031] Wide application adaptability: The fermentation product is mild, safe, and non-irritating, with a stable pH between 5.0 and 6.0. It has good skin tolerance and is especially suitable for sensitive skin, postoperative repair, barrier-damaged skin, etc., expanding its application in functional cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0033] Figure 1 This is the Cordyceps fermentation liquid provided in Example 1 of the present invention. DETAILED DESCRIPTION

[0034] The invention provides a cordyceps probiotic fermentation composition. The composition is obtained by fermenting cordyceps flowers as a solid culture medium with lactic acid bacteria and saccharomyces cerevisiae as strains.

[0035] As an embodiment of the present invention, the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0036] 1. Preparation of Cordyceps solid culture medium:

[0037] The dried Cordyceps flowers are crushed and the solid-liquid ratio is (1.6-2.5): (0.5-1.5) ml / g. The crushed Cordyceps residue is mixed with the solution and placed in a flask or fermentation tank for sterilization. The total volume of the solid culture medium occupies 10%-15% of the entire container.

[0038] The solution is a plant enzymatic hydrolysate, and the specific preparation method of the plant enzymatic hydrolysate is as follows:

[0039] (1) Crush Codonopsis pilosula and add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a 60-70°C water bath for 30 min to pre-soften the cell walls.

[0040] (2) Cool down to 45-50°C, adjust the pH to 5.5-6.0 with citric acid buffer, then add 0.2% (w / w) neutral protease, 0.2% (w / w) cellulase, and 0.1% (w / w) pectinase, and stir gently for 6 hours.

[0041] (3) Heat in an 85°C water bath for 10 minutes to terminate enzyme activity; filter the liquid and cool it to 35-37°C for later use.

[0042] 2. Inoculate lactic acid bacteria into Cordyceps solid culture medium:

[0043] Lactobacillus plantarum is inoculated into the Cordyceps solid culture medium at an addition rate of 3wt% to 5wt%, stirred evenly, and anaerobically cultured at 37°C for 48 to 72 hours.

[0044] 3. Add sterile liquid culture medium to the above Cordyceps solid culture medium, add 3wt%~5wt% of brewer's yeast according to the total volume, and culture under microaerobic conditions at 28℃ for 24~48h.

[0045] The formula of the sterile liquid culture medium is: 0.6 parts of peptone and 85-90 parts of plant enzyme hydrolysate, which is sterilized at 115° C. for 20 minutes and then added to the cordyceps solid culture medium after cooling.

[0046] 4. After fermentation, use pasteurization to quickly heat to 72-75°C for 15-30 minutes, then immediately cool to 4-5°C. Then, filter the fermentation solution through an 8-10μm filter plate and then filter it through a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

[0047] The present invention also provides a soothing and moisturizing facial cream. The raw materials for preparing the facial cream include, in parts by mass:

[0048] Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03;

[0049] Phase B: Ammonium Acryloyldimethyltaurate / VP Copolymer 0.50; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.80;

[0050] Phase C: OLIVEM10002.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00;

[0051] Phase D: 0.30 g of petrolatum; 0.20 g of tocopheryl acetate; 2.00 g of phytosteryl isostearate; 2.00 g of cyclodimethicone; 1.00 g of DC1403; 3.00 g of 1,3-butylene glycol; 0.50 g of p-hydroxyacetophenone; 0.50 g of 1,2-hexanediol; 2.00 g of β-glucan; and 2.00 g of the Cordyceps probiotic fermentation composition.

[0052] As a more preferred embodiment of the present invention, the raw materials for preparing the facial cream include, in parts by mass:

[0053] Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03;

[0054] Phase B: Ammonium Acryloyldimethyl Taurate / VP Copolymer 0.30; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.40; C14-22 Alcohol 0.50; C12-20 Alkyl Glucoside 0.50; PEG-20 Stearate 0.40; Polyglyceryl-6 Stearate 0.20;

[0055] Phase C: OLIVEM10001.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00;

[0056] Phase D: 0.30 g of petrolatum; 0.20 g of tocopheryl acetate; 2.00 g of phytosteryl isostearate; 2.00 g of cyclodimethicone; 1.00 g of DC1403; 3.00 g of 1,3-butylene glycol; 0.50 g of p-hydroxyacetophenone; 0.50 g of 1,2-hexanediol; 2.00 g of β-glucan; and 2.00 g of the Cordyceps probiotic fermentation composition.

[0057] As an embodiment of the present invention, the steps of preparing a soothing and moisturizing facial cream are as follows:

[0058] ① Disperse phase A, heat to about 85℃, and homogenize or stir to dissolve evenly;

[0059] ②Heat phase B to about 85℃ and homogenize or stir to dissolve evenly;

[0060] ③ Add phase A to phase B, homogenize for about 2 minutes, add phase C and phase D, continue homogenizing for about 5 minutes, and stir to cool down;

[0061] ④ Cool down to about 50℃, add phase E (pre-melted) and phase F, stir evenly, and discharge.

[0062] The present invention uses Cordyceps sinensis flowers as a fermentation substrate, leveraging the combined effects of hydration and fermentation enzymes to loosen the cell structure, facilitating the invasion of lactic acid bacteria and yeast, which secrete enzymes to act on the substrate. This results in more complete release. Furthermore, the use of brewer's yeast and the acidic environment enhance the release of cordycepin. Using commercially available solid culture media, while exogenous nutrients aid decomposition, creates a competitive substrate, eliminating the need for impurities such as rice flour or wheat bran. This results in a cleaner, more focused release of the active ingredient, which in turn inhibits cordycepin release. This also avoids the problem of strains focusing on growth while biased towards metabolizing non-Cordyceps sinensis-derived substrates.

[0063] During the preparation of the solid culture medium, the enzymatic hydrolysate contains small molecules such as oligosaccharides, monosaccharides, polyphenols, and amino acids. These components are preferentially utilized by lactic acid bacteria or yeast, activating metabolic pathways (glycolysis and the TCA cycle), thereby promoting the release and conversion of endogenous polysaccharides and cordycepin from the Cordyceps flower. The enzymatic hydrolysate often contains residual plant proteases and polysaccharidase enzymes, which aid in the degradation of the Cordyceps cell wall and improve the solubility of its contents. The plant enzymatic hydrolysate is rich in antioxidants such as polyphenols, flavonoids, and vitamin C, which can reduce the degradation of easily oxidized components such as cordycepin during the fermentation process, thereby improving retention. The plant itself contains a high proportion of water-soluble polysaccharides, which undergo structural modification (oligosaccharification and branch cleavage) after fermentation, enhancing overall yield and functionality. Furthermore, the Cordyceps polysaccharides are co-extracted, significantly increasing the overall polysaccharide content.

[0064] The present invention will be further explained below with reference to specific embodiments.

[0065] Example 1

[0066] This embodiment provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0067] 1. Preparation of Cordyceps solid culture medium:

[0068] The dried Cordyceps flowers are crushed and mixed with the solution at a solid-liquid ratio of 2:1 ml / g. The crushed Cordyceps residue is placed in a flask or fermentation tank for sterilization, and the total volume of the solid culture medium occupies 15% of the entire container.

[0069] The solution is a plant enzymatic hydrolysate, and the specific preparation method of the plant enzymatic hydrolysate is as follows:

[0070] (1) Crush Codonopsis pilosula and add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a 60°C water bath for 30 min to pre-soften the cell walls.

[0071] (2) Cool to 45°C, adjust the pH to 6.0 with citric acid buffer, then add 0.2% (w / w) neutral protease, 0.2% (w / w) cellulase, and 0.1% (w / w) pectinase, and stir gently for 6 hours.

[0072] (3) Heat in a water bath at 85°C for 10 minutes to terminate enzyme activity; filter the liquid and cool it to 35°C for later use.

[0073] 2. Inoculate lactic acid bacteria into Cordyceps solid culture medium:

[0074] Lactobacillus plantarum was inoculated into the Cordyceps solid culture medium at an addition rate of 3 wt %, stirred evenly, and cultured anaerobically at 37° C. for 72 h.

[0075] 3. Sterile liquid culture medium was added to the above Cordyceps solid culture medium (volume ratio 9:1), and 5 wt% of Saccharomyces cerevisiae was added to the total volume and cultured at 28°C under microaerobic conditions for 36 hours.

[0076] The formula of the sterile liquid culture medium is: 0.6 parts of peptone and 85 parts of plant enzymatic hydrolysate, which is sterilized at 115° C. for 20 minutes and then added to the cordyceps solid culture medium after cooling.

[0077] 4. After fermentation, use pasteurization to quickly heat to 72°C for 15 minutes, then immediately cool to 4°C. Then, filter the fermentation solution using an 8μm filter plate and then filter it using a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

[0078] This embodiment also provides a soothing and moisturizing facial cream. The raw materials for preparing the facial cream include, in parts by mass:

[0079] Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03;

[0080] Phase B: Ammonium Acryloyldimethyl Taurate / VP Copolymer 0.30; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.40; C14-22 Alcohol 0.50; C12-20 Alkyl Glucoside 0.50; PEG-20 Stearate 0.40; Polyglyceryl-6 Stearate 0.20;

[0081] Phase C: OLIVEM10001.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00;

[0082] Phase D: 0.30 g of petrolatum; 0.20 g of tocopheryl acetate; 2.00 g of phytosteryl isostearate; 2.00 g of cyclodimethicone; 1.00 g of DC1403; 3.00 g of 1,3-butylene glycol; 0.50 g of p-hydroxyacetophenone; 0.50 g of 1,2-hexanediol; 2.00 g of β-glucan; and 2.00 g of the Cordyceps probiotic fermentation composition.

[0083] The steps of preparing a soothing and moisturizing facial cream are as follows:

[0084] ① Disperse phase A, heat to about 85℃, and homogenize or stir to dissolve evenly;

[0085] ②Heat phase B to about 85℃ and homogenize or stir to dissolve evenly;

[0086] ③ Add phase A to phase B, homogenize for about 2 minutes, add phase C and phase D, continue homogenizing for about 5 minutes, and stir to cool down;

[0087] ④ Cool down to about 50℃, add phase E (pre-melted) and phase F, stir evenly, and discharge.

[0088] Figure 1 The invention provides a Cordyceps fermentation liquid.

[0089] Example 2

[0090] The difference between this embodiment and embodiment 1 is that this embodiment provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0091] 1. Preparation of Cordyceps solid culture medium:

[0092] The dried Cordyceps flowers are crushed and mixed with the solution at a solid-liquid ratio of 2:1 ml / g. The crushed Cordyceps residue is placed in a flask or fermentation tank for sterilization, and the total volume of the solid culture medium occupies 15% of the entire container.

[0093] The solution is distilled water.

[0094] 2. Inoculate lactic acid bacteria into Cordyceps solid culture medium:

[0095] Lactobacillus plantarum was inoculated into the Cordyceps solid culture medium at an addition rate of 3 wt %, stirred evenly, and cultured anaerobically at 37° C. for 72 h.

[0096] 3. Sterile liquid culture medium was added to the above Cordyceps solid culture medium (volume ratio 9:1), and 5 wt% of Saccharomyces cerevisiae was added to the total volume and cultured at 28°C under microaerobic conditions for 36 hours.

[0097] The formula of the sterile liquid culture medium is: 0.6 parts of peptone and 85 parts of distilled water, which is sterilized at 115° C. for 20 minutes and then added to the cordyceps solid culture medium after cooling.

[0098] 4. After fermentation, use pasteurization to quickly heat to 72°C for 15 minutes, then immediately cool to 4°C. Then, filter the fermentation solution using an 8μm filter plate and then filter it using a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

[0099] Example 3

[0100] The difference between this embodiment and embodiment 1 is that this embodiment provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0101] 1. Preparation of Cordyceps solid culture medium:

[0102] The dried cordyceps flowers are crushed and mixed with wheat bran in a ratio of 1:1, with a solid-liquid ratio of 2:1 ml / g. The crushed cordyceps residue is mixed with the solution and placed in a flask or fermentation tank for sterilization. The total volume of the solid culture medium accounts for 15% of the entire container.

[0103] The solution is a plant enzymatic hydrolysate, and the specific preparation method of the plant enzymatic hydrolysate is as follows:

[0104] (1) Crush Codonopsis pilosula and add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a 60°C water bath for 30 min to pre-soften the cell walls.

[0105] (2) Cool to 45°C, adjust the pH to 6.0 with citric acid buffer, then add 0.2% (w / w) neutral protease, 0.2% (w / w) cellulase, and 0.1% (w / w) pectinase, and stir gently for 6 hours.

[0106] (3) Heat in a water bath at 85°C for 10 minutes to terminate enzyme activity; filter the liquid and cool it to 35°C for later use.

[0107] 2. Inoculate lactic acid bacteria into Cordyceps solid culture medium:

[0108] Lactobacillus plantarum was inoculated into the Cordyceps solid culture medium at an addition rate of 3 wt %, stirred evenly, and cultured anaerobically at 37° C. for 72 h.

[0109] 3. Sterile liquid culture medium was added to the above Cordyceps solid culture medium (volume ratio 9:1), and 5 wt% of Saccharomyces cerevisiae was added to the total volume and cultured at 28°C under microaerobic conditions for 36 hours.

[0110] The formula of the sterile liquid culture medium is: 0.6 parts of peptone and 85 parts of plant enzymatic hydrolysate, which is sterilized at 115° C. for 20 minutes and then added to the cordyceps solid culture medium after cooling.

[0111] 4. After fermentation, use pasteurization to quickly heat to 72°C for 15 minutes, then immediately cool to 4°C. Then, filter the fermentation solution using an 8μm filter plate and then filter it using a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

[0112] Example 4

[0113] The difference between this embodiment and embodiment 1 is that this embodiment provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0114] 1. Crush the dried Cordyceps flowers and inoculate 3 wt% of Lactobacillus plantarum into a liquid culture medium (the mass ratio of liquid culture medium to Cordyceps flowers is 10:1). Stir evenly and culture anaerobically at 37°C for 72 hours. Add 5 wt% of Saccharomyces cerevisiae to the total volume and culture under microaerobic conditions at 28°C for 36 hours.

[0115] The liquid culture medium is formulated as follows: 0.6 parts of peptone and 85 parts of plant enzymatic hydrolysate, sterilized at 115° C. for 20 minutes, and used in the culture medium after cooling.

[0116] 2. After fermentation, use pasteurization method to quickly heat to 72℃ for 15 minutes, then immediately cool to 4℃, and then filter the fermentation solution through 8μm filter plate and then filter through 0.22μm PES membrane to obtain Cordyceps fermentation liquid.

[0117] Example 5

[0118] The difference between this embodiment and embodiment 1 is that this embodiment provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows:

[0119] 1. Preparation of Cordyceps solid culture medium:

[0120] Crush the dried Cordyceps sinensis flowers at a solid-liquid ratio of 2:1 ml / g. Add Codonopsis pilosula powder at a mass ratio of 1:24. Mix the crushed Cordyceps sinensis residue with the solution and place in a flask or fermentation tank for sterilization. The total volume of the solid culture medium should account for 15% of the entire container. The solution should be deionized water.

[0121] 2. Inoculate lactic acid bacteria into Cordyceps solid culture medium:

[0122] Lactobacillus plantarum was inoculated into the Cordyceps solid culture medium at an addition rate of 3 wt %, stirred evenly, and cultured anaerobically at 37° C. for 72 h.

[0123] 3. Sterile liquid culture medium was added to the above Cordyceps solid culture medium (volume ratio 9:1), and 5 wt% of Saccharomyces cerevisiae was added to the total volume. The culture was cultured at 28°C under microaerobic conditions for 36 hours.

[0124] The sterile liquid culture medium is formulated as follows: 0.6 parts of peptone and 85 parts of deionized water, which are sterilized at 115° C. for 20 minutes and then added to the cordyceps solid culture medium after cooling.

[0125] 4. After fermentation, use pasteurization to quickly heat to 72°C for 15 minutes, then immediately cool to 4°C. Then, filter the fermentation solution using an 8μm filter plate and then filter it using a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

[0126] Example 6

[0127] The difference between this embodiment and embodiment 1 is that this embodiment provides a soothing and moisturizing facial cream, and the raw materials for preparing the facial cream include:

[0128] Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03;

[0129] Phase B: Ammonium Acryloyldimethyltaurate / VP Copolymer 0.50; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.80;

[0130] Phase C: OLIVEM10002.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00;

[0131] Phase D: 0.30 g of petrolatum; 0.20 g of tocopheryl acetate; 2.00 g of phytosteryl isostearate; 2.00 g of cyclodimethicone; 1.00 g of DC1403; 3.00 g of 1,3-butylene glycol; 0.50 g of p-hydroxyacetophenone; 0.50 g of 1,2-hexanediol; 2.00 g of β-glucan; and 2.00 g of the Cordyceps probiotic fermentation composition.

[0132] Performance Testing

[0133] Test 1: The skin care products prepared in Examples 1 to 6 were placed at 0°C, 25°C, and 40°C for 90 days, and the product properties and color were checked. The test results are shown in Table 1.

[0134] Hot and cold cycle test: Place the skin care products prepared in Examples 1-6 into a sealed container, ensuring it is completely sealed to prevent interference from external moisture or gas. Place the sample in a thermostat set at 45°C for 12 hours. After removing the sample, quickly transfer it to a low-temperature box at -5°C and hold it for 12 hours. After completing one high and low temperature cycle, repeat six times; after each cycle, inspect the sample for physical properties such as appearance, color, odor, viscosity, stratification, and particle precipitation.

[0135] Table 1 Test results of performance test 1

[0136]

[0137] Test 2: Hyaluronidase inhibition rate

[0138] Hyaluronidase is a specific enzyme that cleaves hyaluronic acid and is a participant in allergic reactions, strongly correlated with the release of histamine from mast cells. The soothing effect of a test sample can be determined by its hyaluronidase inhibition rate. A higher hyaluronidase inhibition rate indicates a stronger soothing effect, and vice versa.

[0139] This test uses a reference laboratory method (HMC-WI-029 Hyaluronidase Inhibition Rate) to compare the hyaluronidase inhibition rate test results of the test sample with those of the negative control. If the inhibition rate of the test sample is significantly higher than that of the negative control (P < 0.05), the test sample is considered to have a certain soothing effect. This test method is an in vitro method and is suitable for evaluating cosmetics that claim to achieve soothing effects by inhibiting hyaluronidase.

[0140] Instruments and equipment: BSA224S analytical balance, L6s UV spectrophotometer.

[0141] Reagents: Hyaluronidase, BR, Sodium Hyaluronate, BR.

[0142] Test methods

[0143] (1) Treatment of control and test samples

[0144] Sample group: diluted with pure water to a sample concentration of 10%; positive control (dipotassium glycyrrhizate, purity ≥98%): diluted with pure water to a positive control concentration of 3%; negative control: pure water.

[0145] (2) Test operation steps

[0146] Set up sample group, sample background group, solvent group and solvent background group. Each group needs to set up 3 parallels. Add different reagent solutions to the four groups respectively, shake well, and leave at room temperature for 30 minutes for color development. Use ultraviolet spectrophotometer to measure the absorbance value at a wavelength of 528nm.

[0147] Calculation formula

[0148] Hyaluronidase inhibition rate (%) = (1-(CD) / (AB))*100%

[0149] Wherein: A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme; C is the absorbance of the reaction solution with sample and enzyme; D is the absorbance of the reaction solution with sample and no enzyme.

[0150] Table 2 Test 2 results

[0151]

[0152] The hyaluronidase inhibition rate was rounded to three decimal places, and P < 0.05 indicated that there were significant differences between the sample and the positive control compared with the negative control.

[0153] Test 3: Firming and anti-wrinkle test

[0154] Anti-wrinkle and firming efficacy is primarily characterized by evaluating the test sample's inhibition rate against elastase. Elastase, primarily synthesized and secreted by fibroblasts, degrades elastin in the skin, contributing to skin aging. The experimental principle for elastase inhibition is a catalytic reaction between porcine pancreatic elastase and its substrate. Adding an active substance causes a change in absorbance, which reflects the inhibition rate of the elastase inhibitor. This test follows a reference laboratory method (HMC-WI-028 Elastase Inhibition Rate). The elastase inhibition rate test results are compared between the test sample and the negative control. If the inhibition rate of the test sample is significantly higher than that of the negative control (P < 0.05), the test sample is considered to have anti-wrinkle and firming efficacy.

[0155] This test method is an in vitro method and is suitable for evaluating cosmetics that claim to achieve anti-wrinkle and firming effects by inhibiting elastase.

[0156] Instruments and equipment: BSA224S analytical balance, RT-6100 enzyme-labeled analyzer.

[0157] Reagents: Elastase (porcine pancreas), BR, N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%, epigallocatechin gallate (EGCG), 98%.

[0158] Test methods

[0159] (1) Treatment of control and test samples

[0160] Sample group: diluted with pure water to a sample concentration of 3%;

[0161] Positive control (EGCG): Dilute with pure water to a positive control concentration of 0.1%;

[0162] Negative control: pure water.

[0163] (2) Test operation steps

[0164] Set up sample group, sample background group, solvent group and solvent background group. Each group needs to set up 3 parallels. Add different reagent solutions into the 96-well plate, shake gently, incubate at 25℃ for 15 minutes, place it in the microplate reader, and measure the absorbance at 410nm.

[0165] Elastase inhibition rate (%) = (1-(CD) / (AB))*100%

[0166] Where: A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme; C is the absorbance of the reaction solution with sample and enzyme; D is the absorbance of the reaction solution with sample and no enzyme.

[0167] Table 3 Test 3 results

[0168]

[0169] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A soothing and moisturizing cream, characterized in that In parts by mass, the raw materials for preparing the facial cream include: Phase A: Water 62.77; Allantoin 0.20; Lubrica Gum 1.00; Water-soluble Silicone Wax 2.00; Glycerin 4.00; Sodium Hyaluronate 0.03; Phase B: Ammonium Acryloyldimethyl Taurate / VP Copolymer 0.30; Sodium Stearoyl Glutamate 0.20; M68 0.50; ARLACEL 165 0.40; C14-22 Alcohol 0.50; C12-20 Alkyl Glucoside 0.50; PEG-20 Stearate 0.40; Polyglyceryl-6 Stearate 0.20; Phase C: OLIVEM10001.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00; Phase D: Vaseline 0.30; Tocopheryl Acetate 0.20; Phytosteryl Isostearate 2.00; Cyclodimethicone 2.00; DC1403 1.00; 1,3-Butanediol 3.00; Parahydroxyacetophenone 0.50; 1,2-Hexanediol 0.50; β-Glucan 2.00; Cordyceps Probiotic Fermentation Composition 2.00; The preparation method of the Cordyceps probiotic fermentation composition is as follows: Preparation of Cordyceps solid culture medium: Crush the dried Cordyceps flowers, mix the crushed Cordyceps residue with the solution at a solid-liquid ratio of 2:1 ml / g, and place it into a flask or fermentation tank for sterilization. The total volume of the solid culture medium should account for 15% of the entire container. The solution is a plant enzymatic hydrolysate, and the specific preparation method of the plant enzymatic hydrolysate is as follows: (1) Crush Codonopsis pilosula and add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a 60°C water bath for 30 min to pre-soften the cell walls; (2) Cooling to 45°C, adjusting the pH to 6.0 with citric acid buffer, then adding 0.2% w / w neutral protease, 0.2% w / w cellulase, and 0.1% w / w pectinase, and stirring for 6 hours; (3) Heat in a water bath at 85°C for 10 minutes to terminate enzyme activity; filter the liquid and cool it to 35°C for later use; Cordyceps solid culture medium inoculated with lactic acid bacteria: Lactobacillus plantarum was inoculated into the Cordyceps solid culture medium at a rate of 3 wt %, stirred evenly, and cultured anaerobically at 37°C for 72 h. Sterile liquid culture medium was added to the above Cordyceps solid culture medium at a volume ratio of 9:1, and 5 wt% of Saccharomyces cerevisiae was added to the total volume and cultured at 28°C under microaerobic conditions for 36 hours; The formula of the sterile liquid culture medium is: 0.6 parts of peptone, 85 parts of plant enzyme hydrolysate, sterilized at 115°C for 20 minutes, and added to the Cordyceps solid culture medium after cooling; After the fermentation is completed, the pasteurization method is used to quickly heat the solution to 72°C for 15 minutes, and then immediately cooled to 4°C. The fermentation solution is then filtered using an 8μm filter plate and then filtered using a 0.22μm PES membrane to obtain the Cordyceps fermentation liquid.

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