A preparation method of kapok fermentation filtrate and its product and application

Abstract

The present invention relates to a method for preparing kapok flower fermentation filtrate, a product and an application thereof, wherein the method for preparing kapok flower fermentation filtrate comprises: (1) preparing kapok flower powder and water into a yeast culture medium; (2) inoculating yeast fermentation seed liquid into the yeast culture medium for fermentation to obtain a fermentation product, and centrifuging and filtering the fermentation product to obtain the kapok flower fermentation filtrate. The present invention extracts active ingredients from kapok flowers by fermentation, and the method has the advantages of energy saving, environmental protection, safety and the like compared with traditional extraction technology, and the obtained product has better anti-aging and moisturizing effects.

Description

Technical Field

[0001] The invention belongs to the technical field of daily cosmetics, and particularly relates to a preparation method of kapok flower fermentation filtrate, a product thereof and an application thereof. Background Art

[0002] Kapok flowers can effectively whiten the skin. Its petals contain a large amount of natural organic acids and vitamin C. These substances can penetrate deep into the skin, promote the metabolism of skin cells, make the skin smoother and more delicate, and prevent the skin from being damaged while maintaining skin elasticity. In addition, kapok flowers also contain a variety of antioxidants, such as anthocyanins and volatile oils, which can fight against free radical damage and further delay skin aging. Therefore, many beauty and skin care products choose to add kapok flowers and their extracts. In order to maximize the role of kapok flowers, the extraction method of kapok flower active ingredients has received more and more attention from manufacturers.

[0003] CN103202784A discloses a method for preparing a kapok flower extract having whitening and anti-aging activity. The method is characterized in that dried kapok flowers are crushed and sieved, extracted twice by ethanol immersion, the two extracts are combined, a suspension is obtained by vacuum concentration, the suspension is extracted with petroleum ether and ethyl acetate in turn, the ethyl acetate is recovered by vacuum concentration, and the kapok flower extract is obtained by vacuum drying into a paste. The kapok flower extract obtained by the method has the functions of resisting free radical oxidation, inhibiting tyrosinase activity and resisting ultraviolet radiation, and can be used for skin whitening and anti-aging, especially anti-photoaging. The kapok flower extract can be used as a cosmetic additive or a pharmaceutical active ingredient, and is applied to whitening and anti-aging cosmetics.

[0004] CN114931535A discloses a preparation method and application of kapok flower extract with moisturizing and soothing effects, the method comprising the following steps: crushing fresh kapok flowers to obtain crushed kapok flowers; stirring and mixing the crushed kapok flowers with ethanol aqueous solution to obtain a stirred mixed solution, wherein the mass concentration of the ethanol aqueous solution is 70% to 80%; filtering the stirred mixed solution, collecting the filtrate, and obtaining the kapok flower extract. An application of kapok flower extract with moisturizing and soothing effects includes application in gel, spray and facial mask liquid.

[0005] However, the above-mentioned prior art is not very effective in extracting and converting the active ingredients of kapok flowers, and thus cannot fully exert the efficacy of kapok flowers. Therefore, there is an urgent need to provide a method with simple steps and better extraction effect of kapok flower active ingredients to meet application needs. Summary of the invention

[0006] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for preparing kapok flower fermentation filtrate, its products and applications. The present invention extracts the active ingredients of kapok flowers by fermentation. Compared with traditional extraction technology, the method has the advantages of energy saving, environmental protection and safety, and the obtained product has better anti-aging and moisturizing effects.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing kapok flower fermentation filtrate, the method for preparing kapok flower fermentation filtrate comprising:

[0009] (1) Prepare a yeast culture medium by mixing kapok flower powder and water;

[0010] (2) The fermented seed liquid of yeast is inoculated into the yeast culture medium for fermentation to obtain a fermentation product, and the fermentation product is centrifuged and filtered to obtain the kapok flower fermentation filtrate.

[0011] The method of the present invention uses yeast as a starting strain to ferment kapok flowers, which can promote the growth of yeast and effectively increase the content of polyphenols in the fermentation product. At the same time, combined with the antioxidant capacity of the kapok flower matrix, the fermentation product can achieve anti-aging and moisturizing effects by removing excess free radicals and inhibiting the activity of related enzymes.

[0012] Preferably, the particle size of the kapok flower powder in step (1) is 20-50 mesh, for example, 25 mesh, 30 mesh, 35 mesh, 40 mesh or 45 mesh.

[0013] The yeast in step (2) includes any one of wine yeast, beer yeast or brewer's yeast, or a combination of at least two of them.

[0014] Preferably, the yeast is a combination of wine yeast, beer yeast and cerevisiae yeast.

[0015] The present invention adopts a mixed bacteria method, using wine yeast, beer yeast and brewer's yeast to coordinately regulate the fermentation process, which can effectively improve the fermentation efficiency and fully extract the active ingredients of kapok flowers. In addition, the yeast can produce a variety of prebiotic factors during the fermentation metabolism process, thereby better enhancing the anti-aging, moisturizing and other effects of the product.

[0016] Preferably, the wine yeast is numbered GDM2.95.

[0017] Preferably, the brewer's yeast is numbered as GDM2.9.

[0018] Preferably, the Saccharomyces cerevisiae is numbered as GDM2.200.

[0019] Preferably, the volume ratio of the wine yeast, beer yeast and brewer's yeast is (1-2): (0.1-1): (0.5-2).

[0020] Wherein "1-2" can be 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 or 1.9, etc.;

[0021] “0.1-1” can be 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 or 0.9, etc.;

[0022] “0.5-2” can be 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 or 1.9, etc.

[0023] Preferably, the method for preparing the fermentation seed liquid of the yeast in step (2) is: culturing the yeast in a solid culture medium to obtain a yeast strain, and then transferring the yeast strain to a liquid culture medium to obtain a fermentation seed liquid.

[0024] Preferably, the formula of the solid culture medium is: yeast extract powder 10 g / L, glucose 20 g / L, peptone 10 g / L, agar 20 g / L.

[0025] Preferably, the formula of the liquid culture medium is: yeast extract powder 10 g / L, glucose 20 g / L, and peptone 10 g / L.

[0026] Preferably, the yeast is cultured in a solid culture medium at a temperature of 25-35°C, for example, 26°C, 28°C, 30°C, 32°C or 34°C.

[0027] Preferably, the liquid culture medium is cultured at a temperature of 25-35°C for 10-15 h, for example, 26°C, 28°C, 30°C, 32°C or 34°C, etc., for 11 h, 12 h, 13 h or 14 h, etc.

[0028] Preferably, the yeast culture medium in step (2) comprises 1-10% kapok powder and 90-99% water by mass percentage.

[0029] In the present invention, kapok flowers are configured as yeast culture medium, and there is no need to supplement the nitrogen source and carbon source required for fermentation, which can reduce the fermentation cost. The kapok flowers are slowly fermented by yeast to release anthocyanins, volatile oils, natural organic acids, vitamin C and other active ingredients in the kapok flowers to the greatest extent, thereby improving the fermentation efficiency. The structure is modified, transformed or decomposed by enzymes in microorganisms to obtain better anti-aging and moisturizing effects.

[0030] The mass percentage of the kapok powder can be 2%, 3%, 4%, 5%, 6%, 7%, 8% or 9%;

[0031] The mass percentage of the water can be 91%, 92%, 93%, 94%, 95%, 96%, 97% or 98%, etc.

[0032] Preferably, in step (2), the volume ratio of the fermentation seed liquid to the yeast culture medium is 1:(7-12), for example, it can be 1:8, 1:9, 1:10, 1:11, 1:12, etc.

[0033] Preferably, the fermentation temperature in step (2) is 25-35°C and the fermentation time is 108-144h. For example, the temperature may be 26°C, 28°C, 30°C, 32°C or 34°C, and the fermentation time may be 110h, 115h, 120h, 125h, 130h, 135h or 140h.

[0034] Preferably, stirring is performed during the fermentation in step (2).

[0035] Preferably, the stirring speed is 100-150 rpm, for example, it can be 105 rpm, 110 rpm, 115 rpm, 120 rpm, 125 rpm, 130 rpm, 135 rpm, 140 rpm or 145 rpm, etc.

[0036] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0037] In a second aspect, the present invention provides a kapok fermentation filtrate, which is prepared by the method described in the first aspect.

[0038] In a third aspect, the present invention provides a use of the kapok flower fermentation filtrate as described in the second aspect in the preparation of an anti-aging and moisturizing product.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] (1) The positive effect of the present invention is that fermentation of kapok substrate with yeast as the starting strain can promote the growth of yeast and effectively increase the content of polyphenols in the fermentation product;

[0041] (2) The fermentation product prepared by the present invention contains polyphenols as an active ingredient and combines the antioxidant capacity of the kapok matrix to achieve the effects of anti-aging and moisturizing by removing excess free radicals and inhibiting the activity of related enzymes.

[0042] (3) The preparation method provided by the present invention has simple operation steps, low cost, and is easy to industrialize and mass produce. DETAILED DESCRIPTION

[0043] 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.

[0044] The wine yeast used in the examples of the present invention is numbered GDM2.95, the beer yeast is numbered GDM2.9, and the brewer's yeast is numbered GDM2.200, all of which were purchased from Guangdong Microbiological Culture Collection Center.

[0045] The rest of the raw materials can be used as long as they are purchased from regular dealers.

[0046] Example 1

[0047] This embodiment provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0048] (1) The kapok flowers are dried and crushed to a particle size of 40 mesh to obtain kapok flower powder, which is sterilized at 110°C for 10 min and then used for later use;

[0049] (2) First, transfer wine yeast, beer yeast and brewer's yeast to solid culture medium respectively and place them at 28°C for activation until single colonies grow; then select single colonies from solid culture medium and transfer them to liquid culture medium, and expand the culture at 28°C and 120 rpm for 12 h; adjust the OD of the three fermentation seed liquids to 1, mix them in equal volumes, and obtain mixed yeast fermentation seed liquid;

[0050] (3) Prepare yeast culture medium according to the following mass percentages: 6% kapok powder and 94% water;

[0051] (4) The mixed yeast fermentation seed liquid is inoculated into the above-mentioned culture medium (the volume ratio of the fermentation seed liquid to the yeast culture medium is 1:9), and fermentation is carried out at 28°C and 120 rpm. After 120 hours, a fermentation product is obtained. The fermentation product is centrifuged and filtered to remove the bacterial cells and kapok flower powder to obtain the kapok flower fermentation filtrate.

[0052] Example 2

[0053] This embodiment provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0054] (1) The kapok flowers are dried and crushed to a particle size of 30 mesh to obtain kapok flower powder, which is sterilized at 120°C for 5 min and then used for later use;

[0055] (2) First, wine yeast, beer yeast and brewing yeast were transferred to solid culture medium respectively and left to stand at 25°C for activation until single colonies grew; then, single colonies on solid culture medium were transferred to liquid culture medium and expanded for 15 h at 35°C and 100 rpm; the OD of the three fermentation seed liquids was adjusted to 1, and the volume ratio of wine yeast, beer yeast and brewing yeast was 2:0.5:0.5 to obtain mixed yeast fermentation seed liquid;

[0056] (3) Prepare a yeast culture medium according to the following mass percentages: 3% kapok powder and 97% water;

[0057] (4) The mixed yeast fermentation seed liquid is inoculated into a yeast culture medium (the volume ratio of the fermentation seed liquid to the yeast culture medium is 1:7), and fermentation is carried out at 35° C. and 100 rpm. After 108 hours, a fermentation product is obtained. The fermentation product is centrifuged and filtered to remove the bacterial cells and kapok flower powder, thereby obtaining the kapok flower fermentation filtrate.

[0058] Example 3

[0059] This embodiment provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0060] (1) The kapok flowers are dried and crushed to a particle size of 50 mesh to obtain kapok flower powder, which is sterilized at 100°C for 8 min and then used for later use;

[0061] (2) First, wine yeast, beer yeast and brewing yeast were transferred to solid culture medium respectively and left to stand at 35°C for activation until single colonies grew; then, single colonies on solid culture medium were transferred to liquid culture medium and expanded for 10 h at 25°C and 110 rpm; the OD of the three fermentation seed liquids was adjusted to 1, and the volume ratio of wine yeast, beer yeast and brewing yeast was 1:0.8:2 to obtain mixed yeast fermentation seed liquid;

[0062] (3) Prepare yeast culture medium according to the following mass percentages: 1% kapok powder and 99% water;

[0063] (4) The mixed yeast fermentation seed liquid is inoculated into the yeast culture medium (the volume ratio of the fermentation seed liquid to the yeast culture medium is 1:10), and the fermentation is carried out at 25° C. and 150 rpm. After 144 hours, a fermentation product is obtained. The fermentation product is centrifuged and filtered to remove the bacterial cells and kapok flower powder to obtain the kapok flower fermentation filtrate.

[0064] Example 4

[0065] This embodiment provides a method for preparing kapok flower fermentation filtrate. The method for preparing kapok flower fermentation filtrate is different from that of embodiment 1 only in that the volume ratio of fermentation seed liquid to yeast culture medium in step (4) is 1:5, and the remaining steps and process parameters are consistent with those of embodiment 1.

[0066] Example 5

[0067] This embodiment provides a method for preparing kapok flower fermentation filtrate. The method for preparing kapok flower fermentation filtrate is different from that of embodiment 1 only in that the volume ratio of fermentation seed liquid to yeast culture medium in step (4) is 1:15, and the remaining steps and process parameters are consistent with those of embodiment 1.

[0068] Example 6

[0069] This embodiment provides a method for preparing kapok flower fermentation filtrate. The method for preparing kapok flower fermentation filtrate is different from that of embodiment 1 only in that the yeast in step (2) is wine yeast and beer yeast in a volume ratio of 1:1, and the yeast inoculation amount is kept unchanged. The remaining steps and process parameters are consistent with those of embodiment 1.

[0070] Example 7

[0071] This embodiment provides a method for preparing kapok flower fermentation filtrate. The method for preparing kapok flower fermentation filtrate is different from that of Embodiment 1 only in that the yeast in step (2) is wine yeast and brewing yeast in a volume ratio of 1:1, and the yeast inoculation amount is kept unchanged. The remaining steps and process parameters are consistent with those of Embodiment 1.

[0072] Example 8

[0073] This embodiment provides a method for preparing kapok flower fermentation filtrate. The method for preparing kapok flower fermentation filtrate is different from that of embodiment 1 only in that the yeast in step (2) is brewer's yeast and cerevisiae yeast in a volume ratio of 1:1, and the yeast inoculation amount is kept unchanged. The remaining steps and process parameters are consistent with those of embodiment 1.

[0074] Comparative Example 1

[0075] This comparative example provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0076] (1) The kapok flowers are dried and crushed to a particle size of 40 mesh to obtain kapok flower powder, which is sterilized at 110°C for 10 min and then used for later use;

[0077] (2) Prepare yeast culture medium according to the following mass percentages: 6% kapok powder and 94% water;

[0078] (3) The yeast is inoculated into a yeast culture medium (the mass ratio of the yeast to the yeast culture medium is 1:9), and fermentation is carried out at 28° C. and 120 rpm. After 120 hours, a fermentation product is obtained. The fermentation product is centrifuged and filtered to remove the bacterial body and kapok flower powder, and then the kapok flower fermentation filtrate is obtained.

[0079] Comparative Example 2

[0080] This comparative example provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0081] (1) The kapok flowers are dried and crushed to a particle size of 40 mesh to obtain kapok flower powder, which is sterilized at 110°C for 10 min and then used for later use;

[0082] (2) Transfer the yeast to a solid culture medium and place it at 28°C for activation until yeast colonies grow into strains; then transfer the strains on the solid culture medium to a liquid culture medium and expand the culture at 28°C and 120 rpm for 12 hours to obtain a fermentation seed liquid;

[0083] (3) Prepare a yeast culture medium according to the following mass percentages: 1% yeast extract, 2% peptone, 2% glucose and the balance water;

[0084] (4) The fermentation seed liquid was inoculated into a yeast culture medium (the volume ratio of the fermentation seed liquid to the yeast culture medium was 1:9), and the fermentation was carried out at 28°C and 120 rpm for 24 h. Kapok cotton powder was added to the fermentation system at a ratio of 6 g of kapok cotton powder / 100 mL of yeast culture medium, and the fermentation was continued for 120 h to obtain a fermentation product. The fermentation product was centrifuged and filtered to remove the bacterial cells and the kapok cotton powder to obtain the kapok cotton fermentation filtrate.

[0085] Comparative Example 3

[0086] This comparative example provides a method for preparing kapok flower fermentation filtrate, and the method for preparing kapok flower fermentation filtrate is as follows:

[0087] (1) The kapok flowers were dried and crushed to a particle size of 40 mesh to obtain kapok powder, and the kapok powder was mixed with water 20 times the weight of the kapok powder, and ultrasonic-assisted extraction was performed at 380W and 60°C for 1 h. After centrifugation, the supernatant was taken and fixed to volume with a certain amount of water (kapok powder: water = 1:25) to obtain kapok water extract (40 g / L), which was sterilized at 110°C for 10 min and then used for later use;

[0088] (2) Transfer the yeast to a solid culture medium and place it at 28°C for activation until yeast colonies grow into strains; then transfer the strains on the solid culture medium to a liquid culture medium and expand the culture at 28°C and 120 rpm for 12 hours to obtain a fermentation seed liquid;

[0089] (3) Prepare a yeast culture medium according to the following mass percentages: 1% yeast extract, 2% peptone, 2% glucose and the balance water;

[0090] (4) The fermentation seed liquid is inoculated into a yeast culture medium (the volume ratio of the fermentation seed liquid to the yeast culture medium is 1:9) to obtain a fermentation system, the kapok flower water extract from step (1) is added to the fermentation system (the amount of kapok flower water extract added is 15% of the total volume of the fermentation system), and fermentation is carried out at 28° C. and 120 rpm. After 120 hours, a fermentation product is obtained. The fermentation product is centrifuged and filtered to remove the bacterial cells and kapok flower powder to obtain the kapok flower fermentation filtrate.

[0091] Test Example 1 Determination of polyphenol content

[0092] Polyphenols react specifically with Folin-Ciocalteu reagent, and the reaction product has the best maximum absorption at a specific wavelength. The absorbance value is linearly related to the amount of polyphenols within a certain concentration range. Based on this, the content of polyphenols in the sample to be tested can be calculated.

[0093] The specific experimental steps are as follows:

[0094] (1) Standard stock solution: Accurately weigh 0.011 g of gallic acid standard (relative molecular weight 188.14, containing monohydrate), dissolve it in water, and make up to 10 mL to obtain a 1.0 mg / mL standard stock solution;

[0095] (2) 7.5% Na2CO3 solution: weigh 37.50 g Na2CO3, add appropriate amount of water to dissolve, transfer to a 500 mL volumetric flask, dilute to the mark, and shake well;

[0096] (3) 10% Folin phenol reagent: Take 20 ml of 1 mol / L Folin phenol reagent in a 200 ml volumetric flask, dilute to volume with water and shake well;

[0097] (4) Standard working solution: Take 1.0mL, 2.0mL, 3.0mL, 4.0mL, and 5.0mL of the standard stock solution in a 100mL volumetric flask, dilute to the mark with water, and shake well. The concentrations are 10μg / mL, 20μg / mL, 30μg / mL, 40μg / mL, and 50μg / mL, respectively.

[0098] (5) Sample determination: 1.0 ml of the standard working solution, water (for blank control), and the kapok fermentation filtrate obtained in Examples 1-8 and Comparative Examples 1-3 were respectively transferred into stoppered test tubes. 5.0 ml of 10% Folin phenol reagent was added to each test tube, and the mixture was shaken and reacted for 5 min. 4.0 ml of 7.5% sodium carbonate solution was added and shaken. The mixture was allowed to stand at room temperature for 60 min. The absorbance was measured using a 10 mm cuvette and a spectrophotometer at a wavelength of 765 nm. The concentration of the gallic acid reference substance was taken as the horizontal coordinate and the corresponding absorbance value was taken as the vertical coordinate. The two were subjected to linear regression to obtain a first linear equation, and a method for determining the content of polyphenols was established.

[0099] Each group of samples was measured 4 times in parallel and the average value was taken.

[0100] The test results are shown in Table 1.

[0101] Table 1

[0102]

[0103] From the data in Table 1, it can be seen that the kapok fermentation filtrate prepared in Examples 1-3 of the present invention has a relatively high polyphenol content of more than 1.35 mg / mL;

[0104] It can be seen from Examples 4 and 5 that when the volume ratio of the fermentation seed liquid to the yeast culture medium is too high or too low, the final polyphenol content will be affected;

[0105] It can be seen from Examples 6-8 that when the fermentation is a double-bacteria mixed fermentation, the fermentation effect of kapok is reduced, which in turn affects the final polyphenol content;

[0106] It can be seen from Comparative Example 1 that when there is no step of activating the strain in the preparation method, the polyphenol content in the final fermentation liquid is low;

[0107] From Comparative Examples 2 and 3, it can be seen that the method of the present invention can significantly increase the polyphenol content of the product by making kapok flower powder into yeast culture medium.

[0108] Test Example 2 Antioxidant Efficacy Test

[0109] (1) DPPH free radical scavenging experiment

[0110] Steps: Weigh 0.0079g DPPH accurately and dilute to 100.0mL with anhydrous ethanol to obtain DPPH test solution (0.2mmol / L). Add 200μL of test solution to each 96-well plate according to the sequence shown in Table 2, shake well, react at room temperature in the dark for 30min, and detect absorbance at 517nm with a microplate reader.

[0111] Table 2

[0112]

[0113] DPPH clearance rate (%) = [1-(A1-A2) / A0] × 100%

[0114] The results are shown in Table 4 (each group of samples was measured 5 times in parallel, and the data are presented as average values).

[0115] (2) ABTS free radical scavenging experiment

[0116] Steps: Weigh 0.066g potassium persulfate, dilute to 100mL with water, then accurately weigh 0.0386g ABTS, dilute to 10mL with the above potassium persulfate solution, and stand in the dark for 12 to 16 hours. Dilute with 10mmol / L PBS (pH=7.4) to make the absorbance between 0.7-0.8 to obtain the working solution. Add 200μL of test solution to each 96-well plate as shown in Table 3, shake well, incubate at room temperature for 6 minutes, and detect the absorbance at 734nm with an enzyme reader.

[0117] Table 3

[0118]

[0119] The scavenging rate of the sample solution for ABTS free radicals was calculated according to the following formula.

[0120] ABTS free radical scavenging rate (%) = [A0-(A1-A2)] / A0×100%.

[0121] The results are shown in Table 4 (each group of samples was measured 5 times in parallel, and the data are presented as average values).

[0122] Table 4

[0123]

[0124] As shown in Table 4, the kapok fermentation filtrate prepared in Examples 1-3 of the present invention has a scavenging rate of DPPH free radicals of more than 85%, and a scavenging rate of ABTS free radicals of more than 99%, indicating that the kapok fermentation filtrate described in Examples 1-3 has a good antioxidant effect.

[0125] Test Example 3 Moisturizing and anti-aging efficacy test

[0126] PCR experiments were used to detect the expression levels of the anti-aging gene COL1 and the moisturizing gene SPTLC1.

[0127] Test method: Human skin fibroblasts were inoculated into a 6-well plate, with about 300,000 cells per well. After culturing in a cell culture incubator for 24 hours, the culture medium was discarded, and a culture medium containing the kapok fermentation filtrate obtained in Examples 1-8 and Comparative Examples 1-3 was added. An equal volume of DMEM complete culture medium was added to the control well. After culturing in a cell culture incubator for 24 hours, the cells were collected into sterile centrifuge tubes. For each group of collected cells, they were washed twice with sterile PBS, centrifuged using a centrifuge (1000rpm, 10min), the supernatant was discarded, and the cell pellet was stored at -80°C. From the collected cells, RNA was extracted and purified using a column extraction method, and the RNA was reverse transcribed according to the instructions of the kit to obtain cDNA. Subsequently, primers for the moisturizing gene SPTLC1 and the anti-aging gene COL1 were added for RT-PCR amplification, and 2 -ΔΔCT The relative expression levels of the two genes were analyzed by the method. The results are shown in Table 5 (expression level of moisturizing gene SPTLC1) and Table 6 (expression level of anti-aging gene COL1).

[0128] Table 5

[0129]

[0130] Table 6

[0131]

[0132] It can be seen from Tables 5 and 6 that the kapok fermentation filtrate prepared by the present invention can promote the expression of the anti-aging gene COL1 and the moisturizing gene SPTLC1, and the expression values ​​of Examples 1-3 are relatively high, having relatively good anti-aging and moisturizing effects.

[0133] The applicant declares that the present invention illustrates the preparation method of kapok fermentation filtrate and its product and application 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.

[0134] 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.

[0135] 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 kapok fermentation filtrate, characterized in that: The method for preparing kapok fermentation filtrate comprises: (1) Prepare a yeast culture medium by mixing kapok flower powder and water; (2) inoculating the fermented seed liquid of yeast into a yeast culture medium for fermentation to obtain a fermentation product, and centrifuging and filtering the fermentation product to obtain the kapok flower fermentation filtrate; The method for preparing the fermentation seed liquid of the yeast comprises: culturing the yeast in a solid culture medium to obtain a yeast strain, and then transferring the yeast strain to a liquid culture medium to obtain a fermentation seed liquid; The yeast is a combination of wine yeast, beer yeast and brewer's yeast; The volume ratio of the fermentation seed liquid to the yeast culture medium is 1:(7-12).

2. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The particle size of the kapok flower powder in step (1) is 20-50 meshes.

3. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The volume ratio of the wine yeast, beer yeast and brewing yeast is (1-2): (0.1-1): (0.5-2).

4. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The yeast is cultured in a solid culture medium at a temperature of 25-35°C.

5. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The liquid culture medium is cultured at a temperature of 25-35° C. for 10-15 hours.

6. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The yeast culture medium in step (2) comprises, by mass percentage, 1-10% kapok powder and 90-99% water.

7. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: The fermentation temperature in step (2) is 25-35°C and the fermentation time is 108-144h.

8. The method for preparing kapok fermentation filtrate according to claim 1, characterized in that: During the fermentation in step (2), stirring is performed.

9. The method for preparing kapok fermentation filtrate according to claim 8, characterized in that: The stirring speed is 100-150 rpm.

10. A kapok fermentation filtrate, characterized in that: The kapok fermentation filtrate is prepared by the method according to any one of claims 1 to 8.

11. Use of the kapok fermentation filtrate according to claim 10 in the preparation of anti-aging and moisturizing products.

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