Preparation method and application of gallnut fermentation extract

Abstract

The present invention discloses a preparation method and application of a gallnut fermentation extract. The preparation method comprises the following steps: S1, dissolving glucose, fermentation nutrient salts, and phosphate buffer in water, and adding a gallnut extract solution to prepare a gallnut culture medium; S2, inoculating a lactobacillus seed solution in the gallnut culture medium, and performing anaerobic fermentation at a constant pH of 5.4-6.4 to obtain a gallnut fermentation extract. The present invention finds that the gallnut extract prepared by the above method has the characteristics of enhancing mitochondrial function and anti-aging, and can be applied to the preparation of cosmetics with enhanced mitochondrial function, antioxidant function, or anti-aging function.

Description

Technical Field

[0001] The present invention relates to a gallnut fermentation extract, in particular to a preparation method and application of the gallnut fermentation extract. Background Art

[0002] There is a close connection between skin aging and mitochondria. Mitochondria, as the "energy factory" of cells, are responsible for producing the energy needed by cells and are essential for maintaining normal cell function and metabolism. However, with age, the function of mitochondria gradually declines, resulting in a decrease in the energy supply to skin cells, which in turn triggers a series of aging phenomena. Specifically, a decline in mitochondrial function can lead to slower skin cell turnover, reduced collagen synthesis, and the skin becoming dull, dull, and losing elasticity. In addition, mitochondrial damage will increase the production of free radicals, further exacerbating oxidative stress in the skin, thereby accelerating the aging process. Therefore, specific skin care interventions targeting mitochondria to improve mitochondrial activity and enhance mitochondrial function can effectively improve skin condition and delay skin aging.

[0003] Elastase, also known as elastin hydrolase, primarily breaks down elastin, a key component in maintaining skin elasticity. Due to endocrine disorders, aging, and environmental factors (such as UV exposure), elastase may over-break down elastin in the skin, leading to a decrease in skin elasticity. Therefore, inhibiting elastase can have anti-aging effects.

[0004] Galla chinensis (Galla rhois), mainly derived from galls on the leaves of Rhus chinensis, Populus chinensis, or Populus rubra (all belonging to the Anacardiaceae family), has the effects of astringing the lungs and reducing internal heat, stopping bleeding, and absorbing dampness and healing sores. As a traditional Chinese medicine, Galla chinensis has been reported to have the potential to treat skin diseases such as psoriasis. It is also often used in combination with other traditional Chinese medicines or plant extracts in cosmetics to achieve skin firming, repairing, and whitening effects. Invention patent CN115671021A discloses a Galla chinensis extract repairing and firming essence and its preparation method and application. The Galla chinensis extract, when combined with Angelica sinensis extract, Polygonum multiflorum extract, Borneolum, Artemisia argyi extract, etc., has the effects of whitening, repairing, firming the skin, and nourishing and moisturizing.

[0005] However, in the prior art, there is no public report on the effects of Chinese gallnut and its fermentation products (extracts) on mitochondrial activity and function, proving that they have anti-aging effects. Summary of the Invention

[0006] The present invention aims to provide a preparation method and application of a fermented extract of Chinese gallnut. The present invention finds that the Chinese gallnut extract has the characteristics of enhancing mitochondrial function and anti-aging.

[0007] The technical solution of the present invention is a method for preparing a gallnut fermentation extract, comprising the following steps:

[0008] S1. Dissolving glucose, fermentation nutrient salts, and phosphate buffer in water, and adding the gallnut extract solution to prepare a gallnut culture medium;

[0009] S2. Inoculating Lactobacillus seed liquid into a gallnut culture medium, and performing anaerobic fermentation at a constant pH of 5.4-6.4 to obtain a gallnut fermentation extract.

[0010] In the aforementioned method for preparing a gallnut fermentation extract, in step S1, the gallnut extract solution is prepared by dissolving the gallnut extract in a 0.08-0.12 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 8-10 mg / mL.

[0011] In the aforementioned method for preparing a gallnut fermentation extract, in step S1, the gallnut extract is a gallnut extract having an ellagic acid content of ≥75% obtained by extracting gallnuts with hot water.

[0012] In the aforementioned method for preparing a gallnut fermentation extract, in step S1, the mass percentage of glucose in the gallnut culture medium is 0.5-1%, the mass percentage of fermentation nutrient salt is 0.5-1%, the mass percentage of phosphate buffer is 0.4-0.5%, and the mass percentage of the gallnut extract solution is 0.5-1%.

[0013] In the aforementioned method for preparing the gallnut fermentation extract, in step S2, the mass percentage of the lactobacillus seed liquid is 2-4%.

[0014] In the aforementioned method for preparing the gallnut fermentation extract, in step S2, the conditions for anaerobic fermentation are a rotation speed of 100-200 rpm, a fermentation temperature of 35-40° C., and a fermentation time of 36-48 h.

[0015] In the aforementioned method for preparing a gallnut fermentation extract, the gallnut fermentation extract is the gallnut fermentation liquid obtained after anaerobic fermentation in step S2.

[0016] In the aforementioned method for preparing the gallnut fermentation extract, the gallnut fermentation extract is the gallnut fermentation supernatant obtained by high-pressure homogenization and centrifugation of the gallnut fermentation liquid.

[0017] In the aforementioned method for preparing a gallnut fermentation extract, the gallnut fermentation extract is a gallnut fermentation concentrated supernatant obtained by rotary evaporation and concentration of the gallnut fermentation supernatant.

[0018] The gallnut fermentation extract is used in the preparation of cosmetics with enhanced mitochondrial function, antioxidant function or anti-aging function.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention found that the fermentation supernatant obtained by constant pH fermentation of gallnut has an increased soluble solid content and improved antioxidant capacity compared with the fermentation supernatant obtained by natural fermentation, and has a better inhibition rate on elastase. In addition, the concentrated supernatant of constant pH fermentation of gallnut has the best inhibitory effect on elastase and improvement on HDF mitochondrial function, and has good anti-aging potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a diagram showing the effects of different treatment groups on the mitochondrial oxygen consumption rate (OCR) of human fibroblasts (HDF). DETAILED DESCRIPTION

[0022] The present invention will be further described below with reference to the examples, but they are not intended to limit the present invention.

[0023] A method for preparing a gallnut fermentation extract comprises the following steps:

[0024] S1. Preparation of Gallnut Extract Solution:

[0025] The gallnut extract with ellagic acid content of ≥75% obtained by extracting the gallnut with hot water is dissolved in a 0.08-0.12 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 8-10 mg / mL.

[0026] S2. Preparation of Gallnut Culture Medium:

[0027] Dissolve 0.5-1% by mass of glucose, 0.5-1% by mass of fermentation nutrient salt, and 0.4-0.5% by mass of phosphate buffer in water, add 0.5-1% by mass of an 8-10 mg / mL gallnut extract solution, sterilize at a temperature of 115-125° C. for 15-30 minutes to prepare a gallnut culture medium, and cool it for later use;

[0028] The fermentation nutrient salts are existing substances purchased from Angel Yeast, including ammonium dihydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride, glucose, and yeast extract powder.

[0029] S3. Preparation of Lactobacillus seeds:

[0030] The plant lactobacillus is inoculated into a seed culture medium, which is an MRS culture medium. The culture conditions are a temperature of 35-40° C. and a fermentation time of 16-20 hours to obtain a lactobacillus seed liquid.

[0031] S4. Fermentation:

[0032] Inoculating a Lactobacillus seed solution accounting for 2-4% of the total mass of the gallnut culture medium into a gallnut culture medium, and performing anaerobically fermentation at 37° C., 100-200 rpm, and a constant pH of 5.4-6.4 for 36-48 hours to obtain a gallnut fermentation liquid;

[0033] During the constant pH fermentation process, 8-12 mol / L NaOH solution was added to adjust the pH to remain constant at 5.4-6.4.

[0034] S5. Clarification:

[0035] The gallnut fermentation liquid is subjected to high-pressure homogenization at 800-1500 bar and 4-6° C. for 3-5 cycles; and then centrifuged at 3000-8000 rpm and 4-6° C. for 15-30 minutes to obtain the gallnut fermentation supernatant.

[0036] S6, Concentration:

[0037] The gallnut fermentation supernatant was concentrated by rotary evaporation to 3-4 times to obtain the gallnut fermentation concentrated supernatant.

[0038] The present invention also provides a use of the gallnut fermentation extract in preparing cosmetics with mitochondrial function enhancement, antioxidant function or anti-aging function.

[0039] The gallnut fermentation extract is gallnut fermentation liquid, gallnut fermentation supernatant or gallnut fermentation concentrated supernatant, which are prepared by the above-mentioned preparation methods respectively.

[0040] Example 1:

[0041] A gallnut fermentation extract, and a preparation method thereof, comprising the following steps:

[0042] S1. Preparation of Gallnut Extract Solution:

[0043] 100 mg of gallnut extract (ellagic acid content ≥ 75%) obtained by hot water extraction of gallnut was dissolved in 10 mL of 0.1 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 10 mg / mL.

[0044] S2. Preparation of Gallnut Culture Medium:

[0045] 10 g of glucose, 10 g of fermentation nutrient salt, and 4.19 g of phosphate buffer were dissolved in 990 mL of water, and 10 mL of a 10 mg / mL gallnut extract solution was added, and the mixture was sterilized at 121° C. for 20 min to prepare a gallnut culture medium.

[0046] S3. Preparation of Lactobacillus seeds:

[0047] Lactobacillus plantarum was inoculated into a seed culture medium, which was an MRS culture medium. The culture conditions were a temperature of 37° C. and fermentation for 18 hours to obtain a lactobacillus seed liquid.

[0048] S4. Fermentation:

[0049] 2% by mass of Lactobacillus seed liquid was inoculated into a gallnut culture medium, and anaerobically fermented at 37° C. and 150 rpm for 48 h. During the fermentation process, 10 mol / L NaOH solution was added to adjust the pH to be constant at 5.5±0.1 to obtain a gallnut fermentation liquid.

[0050] S5. Clarification:

[0051] The gallnut fermentation broth was subjected to high-pressure homogenization at 1000 bar and 4° C.; then, the broth was centrifuged at 5000 rpm and 4° C. for 20 min. The obtained supernatant was passed through a 0.22 μm membrane to obtain the gallnut fermentation supernatant 1.

[0052] Example 2:

[0053] A gallnut fermentation extract, the preparation method of which is as follows:

[0054] S1. Preparation of Gallnut Extract Solution:

[0055] 100 mg of gallnut extract (ellagic acid content ≥ 75%) obtained by hot water extraction of gallnut was dissolved in 10 mL of 0.1 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 10 mg / mL.

[0056] S2. Preparation of Gallnut Culture Medium:

[0057] 10 g of glucose, 10 g of fermentation nutrient salt, and 4.19 g of phosphate buffer were dissolved in 990 mL of water, and 10 mL of a 10 mg / mL gallnut extract solution was added, and the mixture was sterilized at 121° C. for 20 min to prepare a gallnut culture medium.

[0058] S3. Preparation of Lactobacillus seeds:

[0059] Lactobacillus plantarum was inoculated into a seed culture medium, which was an MRS culture medium. The culture conditions were a temperature of 37° C. and fermentation for 18 hours to obtain a lactobacillus seed liquid.

[0060] S4. Fermentation:

[0061] 2% by mass of Lactobacillus seed liquid was inoculated into a gallnut culture medium, and anaerobically fermented at 37° C. and 150 rpm for 48 h. During the fermentation process, 10 mol / L NaOH solution was added to adjust the pH to be constant at 6.0±0.1 to obtain a gallnut fermentation liquid.

[0062] S5. Clarification:

[0063] The gallnut fermentation broth was subjected to high-pressure homogenization at 1000 bar and 4° C., and then centrifuged at 5000 rpm and 4° C. for 20 min. The obtained supernatant was passed through a 0.22 μm membrane to obtain the gallnut fermentation supernatant 2.

[0064] Example 3:

[0065] A gallnut fermentation extract, and a preparation method thereof, comprising the following steps:

[0066] S1. Preparation of Gallnut Extract Solution:

[0067] 100 mg of gallnut extract (ellagic acid content ≥ 75%) obtained by hot water extraction of gallnut was dissolved in 10 mL of 0.1 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 10 mg / mL.

[0068] S2. Preparation of Gallnut Culture Medium:

[0069] 10 g of glucose, 10 g of fermentation nutrient salt, and 4.19 g of phosphate buffer were dissolved in 990 mL of water, and 10 mL of a 10 mg / mL gallnut extract solution was added. The mixture was sterilized at 121° C. for 20 min and cooled to prepare a gallnut culture medium.

[0070] S3. Preparation of Lactobacillus seeds:

[0071] Lactobacillus plantarum was inoculated into a seed culture medium, which was an MRS culture medium. The culture conditions were a temperature of 37° C. and fermentation for 18 hours to obtain a lactobacillus seed liquid.

[0072] S4. Fermentation:

[0073] 2% by mass of Lactobacillus seed liquid was inoculated into a gallnut culture medium, and anaerobically fermented at 37° C. and 150 rpm for 48 h. During the fermentation process, 10 mol / L NaOH solution was added to adjust the pH to be constant at 5.5±0.1 to obtain a gallnut fermentation liquid.

[0074] S5. Clarification:

[0075] The gallnut fermentation liquid was subjected to high-pressure homogenization for three cycles at 1000 bar and 4° C.; the liquid was then centrifuged at 5000 rpm and 4° C. for 20 minutes, and the obtained supernatant was passed through a 0.22 μm membrane to obtain the gallnut fermentation supernatant.

[0076] S6, Concentration:

[0077] The gallnut fermentation supernatant was concentrated by rotary evaporation at 55° C. to a Brix of 7.5% to obtain a concentrated gallnut fermentation supernatant.

[0078] Comparative Example 1:

[0079] A gallnut fermentation extract, the preparation method of which is as follows:

[0080] S1. Preparation of Gallnut Extract Solution:

[0081] 100 mg of gallnut extract (ellagic acid content ≥ 75%) obtained by hot water extraction of gallnut was dissolved in 10 mL of 0.1 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 10 mg / mL.

[0082] S2. Preparation of Gallnut Culture Medium:

[0083] 10 g of glucose, 10 g of fermentation nutrient salt, and 4.19 g of phosphate buffer were dissolved in 990 mL of water, and 10 mL of a 10 mg / mL gallnut extract solution was added. The mixture was sterilized at 121° C. for 20 min to prepare a gallnut culture medium.

[0084] S3. Preparation of Lactobacillus seeds:

[0085] Lactobacillus plantarum was inoculated into a seed culture medium, which was an MRS culture medium. The culture conditions were a temperature of 37° C. and fermentation for 18 hours to obtain a lactobacillus seed liquid.

[0086] S4. Fermentation:

[0087] 2% by mass of Lactobacillus seed liquid was inoculated into a gallnut culture medium, and the culture medium was anaerobically fermented at 37° C. and 150 rpm for 48 hours to obtain a gallnut fermentation liquid with a pH of 3.49.

[0088] S5. Clarification:

[0089] The gallnut fermentation broth was subjected to high-pressure homogenization at 1000 bar and 4° C.; then, the broth was centrifuged at 5000 rpm and 4° C. for 20 min. The obtained supernatant was passed through a 0.22 μm membrane to obtain the gallnut fermentation supernatant 3.

[0090] Comparative Example 2:

[0091] A gallnut unfermented concentrated supernatant, the preparation method of which is as follows:

[0092] S1. Weigh 100 mg of gallnut extract (ellagic acid content ≥ 75%) obtained by hot water extraction of gallnuts, and dissolve it in 10 mL of 0.1 mol / L NaOH solution to prepare a 10 mg / mL gallnut extract solution.

[0093] S2. Weigh 10 g of glucose, 10 g of fermentation nutrient salt, and 4.19 g of phosphate buffer solution and dissolve them in 990 mL of water. Add 10 mL of a 10 mg / mL gallnut extract solution and sterilize at 121° C. for 20 min to prepare a gallnut culture medium.

[0094] S3. Concentrate the Chinese gall culture medium to Brix = 7.5%, centrifuge at 5000 rpm, 4° C. for 20 min, and pass the obtained supernatant through a 0.22 μm membrane to obtain the Chinese gall unfermented concentrated supernatant.

[0095] Test method:

[0096] 1) The DPPH free radical scavenging ability of the supernatant samples of the Chinese gallnut fermentation under different fermentation conditions was determined by a kit method. The specific determination method is as follows:

[0097] 1. Preparation of DPPH solution: Weigh 20 mg of DPPH, dissolve it in anhydrous ethanol and dilute it to a 250 mL volumetric flask to obtain 2 × 10 -4 1 mol / L DPPH ethanol solution.

[0098] 2. Preparation of the sample to be tested: Dilute the sample 5 times with distilled water to obtain the sample to be tested.

[0099] 3. Assay: Mix an equal volume (750 μL) of the sample to be tested with the DPPH solution (tube A1); mix an equal volume (750 μL) of distilled water with the DPPH solution (tube A2); and mix an equal volume (750 μL) of the sample to be tested with anhydrous ethanol (tube A3). Perform three replicates for each sample. After incubation in the dark for 30 minutes, measure the absorbance of tubes A1, A2, and A3 at 517 nm using a microplate reader. Repeat the assay for three replicates.

[0100] 4. Calculation of results: Clearance rate (%) = [(A2+A3)-A1] / A2.

[0101] The test results are shown in Table 1:

[0102] Table 1 pH, Brix and DPPH clearance of the supernatant of gallnut fermentation under different fermentation conditions

[0103] Sample solution pH Brix DPPH clearance rate (%) Gallnut culture medium 6.36 2.5 77.33 Example 1 5.54 2.5 94.06 Example 2 6.03 2.6 89.99 Comparative Example 1 3.49 2.1 86.17

[0104] Comparison of Comparative Example 1 and Galla chinensis culture medium shows that natural fermentation significantly reduces Birx compared to Galla chinensis culture medium, indicating that the natural fermentation process metabolizes a portion of the soluble solids in the culture medium. Comparison of Examples 1-2 and Comparative Example 1 shows that constant pH fermentation increases Brix and DPPH clearance in the Galla chinensis fermentation supernatant compared to natural fermentation, indicating that constant pH fermentation can increase the soluble solids content in the fermentation broth and enhance antioxidant capacity.

[0105] As can be seen from Table 1, Example 1 obtained by fermentation at a constant pH = 5.5 has the strongest ability to scavenge DPPH free radicals (p < 0.05). Compared with Example 2 fermented at a constant pH = 6.0, its antioxidant capacity is increased by 4.07%, and compared with the unfermented gallnut culture medium, its antioxidant capacity is increased by 16.73%; compared with the naturally fermented comparative example 1, its antioxidant capacity is increased by 7.89%; therefore, the gallnut fermentation supernatant obtained by maintaining a constant pH at 5.5 ± 0.1 during the fermentation process has the best antioxidant activity and capacity.

[0106] 2) Determination of the content of urolithin A and its derivatives in the supernatant of the gallnut fermentation liquid, specifically as follows:

[0107] Qualitative and quantitative analysis was performed using an Agilent 6470 triple quadrupole liquid spectrometer (Agilent Technologies (China) Co., Ltd.). The chromatographic column was a Waters Acquity Premier CSH Phenyl-Hexyl column (1.7 μm × 2.1 mm × 100 mm); the column temperature was 40°C; the mobile phase was: A-0.1% formic acid in water, B-acetonitrile; the flow rate was 0.3 min / L; the injection volume was 10 μL; the gradient elution was: 5%-60% B from 0 to 7.0 min, 60%-95% B from 7 to 9 min, 95% B from 9 to 12 min, 95% B from 12 to 12.5 min, 95%-5% B, and 5% B from 12.5 to 16 min. The ion source was an ESI source with a drying gas temperature of 325°C, a sheath gas temperature of 350°C, a drying gas flow rate of 5 L / min, a sheath gas flow rate of 11 L / min, and a capillary voltage of 3500 V. Negative ion scanning mode was used, and the monitoring mode was multiple reaction monitoring (MRM). Standards urolithin M5, urolithin M6, urolithin C, urolithin A, and isourolitin A were used for qualitative and quantitative analysis.

[0108] The experimental results are shown in Table 2.

[0109] Table 2 Determination of urolithin A and its derivatives in the supernatant of gallnut fermentation

[0110]

[0111] The results showed that the unfermented Galla chinensis culture medium did not contain urolithin A and its derivatives, while the Galla chinensis fermentation supernatant of Example 1 contained 1.33 ng / mL of urolithin M5 and 0.41 ng / mL of urolithin A / isourolithin A, indicating that fermentation produced urolithin A and its derivatives. Studies have shown that urolithin A and its derivatives have certain effects on improving mitochondrial function and delaying aging.

[0112] 3) The inhibitory effect of the Chinese gall fermentation supernatant on elastase was determined as follows:

[0113] 1.600 mU / mL elastase solution and 150 μL of 1.015 mmol / L AAAPAN (N-succinyl-alanine-alanine-alanine-p-nitroaniline) solution were both prepared by dissolving in 50 mmol / L Tris-HCl.

[0114] 2. Take 420 μL of 50 mmol / L Tris-HCl buffer (PH=8.0) and mix it with 180 μL of water (solvent), add 150 μL of elastase solution, and incubate at 25°C for 15 minutes as A1.

[0115] 3. Take 420 μL of 50 mmol / L Tris-HCl buffer (PH=8.0) and mix it with 180 μL of sample solution, add 150 μL of elastase solution, and incubate at 25°C for 15 minutes as A2.

[0116] 4. Take 600 μL of 50 mmol / L Tris-HCl buffer (PH=8.0) and mix it with 180 μL of sample solution, add 150 μL of elastase solution, and incubate at 25°C for 15 minutes as A3.

[0117] 5. Take 600 μL of 50 mmol / L Tris-HCl buffer (PH=8.0) and mix it with 180 μL of water (solvent), add 150 μL of elastase solution, and incubate at 25°C for 15 minutes as A0.

[0118] 6. Add 150 μL of AAAPAN solution to tubes A1, A2, and A3, and measure the absorbance at 410 nm after 20 minutes.

[0119] 7. Calculation of results: Inhibition rate (%) = [1-(A2-A3) / (A1-A0)] x 100%.

[0120] The calculation results are shown in Table 3.

[0121] Table 3 Inhibitory effect of gallnut fermentation supernatant on elastase

[0122] Sample solution Elastase inhibition rate (%) Gallnut culture medium 15.78％ Example 1 30.79％ Example 3 58.12％ Comparative Example 2 40.30％

[0123] As shown in Table 3, the inhibitory effect of the gallnut fermentation supernatant on elastase is increased by about 15% compared with the unfermented gallnut culture medium, indicating that the gallnut fermentation supernatant has anti-aging ability. In addition, the inhibitory effect of the gallnut fermentation concentrated supernatant of Example 3 on elastase is increased by about 18% compared with the inhibitory effect of the gallnut unfermented concentrated supernatant sample solution of Comparative Example 2, proving that fermentation can enhance the anti-aging ability of the gallnut extract.

[0124] 4) Detect the effects of gallnut extract on mitochondrial oxygen consumption rate (OCR) and mitochondrial function of human fibroblasts (HDF).

[0125] The CCK-8 method was used to detect the cytotoxicity of the fermentation concentrated supernatant of Example 3 and the unfermented concentrated supernatant of gallnut of Comparative Example 2 on human fibroblasts (HDF). The detection found that the samples of Example 3 and Comparative Example 2 at a mass concentration of 1% had no cytotoxicity to HDF cells.

[0126] Four experimental groups were set up: a blank group, a model group, a non-fermented group, and a fermented group. The experimental groups and treatments are shown in Table 4. The seahorse XF-96 method was used to measure the mitochondrial OCR of HDF cells. The seahorse XF-96 method was used to measure the basal respiration, ATP production, maximal respiration, and spare respiratory capacity of HDF cells to explore the effects of the concentrated supernatants of unfermented and fermented Chinese gallnut on mitochondrial function.

[0127] Table 4 Experimental groups and treatments

[0128]

[0129] Calculation formula: Change rate (%) = measured value of each group of indicators / measured value of the experimental group * 100%.

[0130] The results of HDF cell mitochondrial OCR detection are shown in Figure 1 .

[0131] Depend on Figure 1 It can be seen that UVB post-irradiation caused the OCR of HDF cells to decrease. Both the non-fermentation group and the fermentation group could alleviate the effect of UVB irradiation on the OCR of HDF cells, and the OCR of the fermentation group was the highest at 40-50 min, indicating that both the non-fermented concentrated supernatant of gallnut and the fermented concentrated supernatant of gallnut could improve the acute response of cell mitochondria caused by UVB.

[0132] The results of the mitochondrial function test of HDF cells are shown in Table 5.

[0133] Table 5 Effects of different gallnut extracts on mitochondrial function of human fibroblasts (HDF)

[0134]

[0135] As shown in Table 5, compared with the experimental group, the unfermented group and the fermented group improved the basal respiration, ATP production, maximum respiration and spare respiratory capacity of HDF cells, indicating that both the unfermented concentrated supernatant and fermented concentrated supernatant of Galla chinensis can improve the mitochondrial function of HDF.

[0136] Comparing the non-fermented group with the fermented group, the fermented group showed an improvement of 20.99%, 25.80%, 30.13% and 21.41% in the basal respiration, ATP production, maximal respiration and spare respiratory capacity of HDF cells, respectively, compared with the non-fermented group, indicating that the concentrated supernatant of gallnut fermentation has a better effect on improving the mitochondrial function of HDF.

[0137] In conclusion, the concentrated supernatant of Galla chinensis fermentation can improve mitochondrial function and has good anti-aging potential.

[0138] Application Example 1:

[0139] Application of concentrated supernatant of gallnut fermentation in preparing lotion with enhanced mitochondrial function.

[0140] The formula of the lotion is shown in Table 6 below.

[0141] Table 6: Formula of lotion

[0142]

[0143]

[0144] The preparation process of lotion is:

[0145] 1. Add A01 into the reactor, add A02 and disperse evenly, then add A03-A11 into the reactor, stir and heat to 85°C, keep stirring for 20 minutes until completely dissolved, and vacuum cool down;

[0146] 2. Cool down to 55°C, add the pre-mixed and dissolved Phase B ingredients, and stir evenly;

[0147] 3. Cool down to below 37°C, add Phase C raw materials into the pot, homogenize at 1800 rpm for 2 minutes, stir evenly, and vacuum degas;

[0148] 4. After passing the inspection, filter the material through a 300-mesh filter to obtain a lotion with mitochondrial enhancement, anti-oxidation and anti-aging functions.

[0149] Application Example 2:

[0150] Application of concentrated supernatant of Galla chinensis fermentation in preparing emulsion with enhanced mitochondrial function.

[0151] The formula of the emulsion is shown in Table 7 below.

[0152] Table 7 Emulsion formula

[0153]

[0154]

[0155] The preparation process of the emulsion is:

[0156] 1. Add A01 into the reactor, sprinkle A02 in and disperse evenly, then add A03-A07 into the reactor, stir and heat to 85℃, keep stirring for 20 minutes until completely dissolved;

[0157] 2. Accurately weigh the raw materials of phase B, heat to 85℃ and stir evenly;

[0158] 3. Pump phase A into the emulsifying pot, then pump into phase B, homogenize at 85°C, 2800 rpm for 5 minutes, and cool down;

[0159] 4. Cool down to 55°C, add the pre-mixed and dissolved Phase C raw materials, and stir evenly;

[0160] 5. Cool to below 37°C, add phase D raw materials, homogenize at 2500rpm for 2 minutes, stir evenly, and after passing the inspection, filter the material with an 80-mesh filter to obtain an emulsion with enhanced mitochondrial function, antioxidant and anti-aging functions.

Claims

1. A method for preparing a gallnut fermentation extract, characterized in that: The following steps are involved: S1. Dissolving glucose, fermentation nutrient salts, and phosphate buffer in water, and adding the gallnut extract solution to prepare a gallnut culture medium; S2, inoculating a Lactobacillus seed solution into a gallnut culture medium, and performing anaerobic fermentation at a constant pH of 5.4-6.4 to obtain a gallnut fermentation broth, wherein the gallnut fermentation extract is a gallnut fermentation supernatant obtained by high-pressure homogenization and centrifugation of the gallnut fermentation broth; In step S1, the gallnut extract solution is prepared by dissolving the gallnut extract in a 0.08-0.12 mol / L NaOH solution to obtain a gallnut extract solution with a concentration of 8-10 mg / mL.

2. The method for preparing a gallnut fermentation extract according to claim 1, wherein: In step S1, the gallnut extract is a gallnut extract with an ellagic acid content of ≥75% obtained by extracting gallnut with hot water.

3. The method for preparing a gallnut fermentation extract according to claim 1, wherein: In step S1, the mass percentage of glucose in the gallnut culture medium is 0.5-1%, the mass percentage of fermentation nutrient salt is 0.5-1%, the mass percentage of phosphate buffer is 0.4-0.5%, and the mass percentage of the gallnut extract solution is 0.5-1%.

4. The method for preparing a gallnut fermentation extract according to claim 1, wherein: In step S2, the mass percentage of the lactobacillus seed liquid is 2-4%.

5. The method for preparing the Chinese gallnut fermentation extract according to claim 1, wherein: In step S2, the conditions for anaerobic fermentation are a rotation speed of 100-200 rpm, a fermentation temperature of 35-40° C., and a fermentation time of 36-48 h.

6. The method for preparing a gallnut fermentation extract according to claim 1, characterized in that: The gallnut fermentation extract is a gallnut fermentation concentrated supernatant obtained by rotary evaporation and concentration of the gallnut fermentation supernatant.

7. Use of the gallnut fermentation extract according to any one of claims 1 to 6 in the preparation of cosmetics having antioxidant or anti-aging functions.

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