Preparation method and application of garden burnet leavening

Abstract

The invention discloses a preparation method and application of a garden burnet leavening, and the preparation method specifically comprises the following steps: mixing garden burnet root powder with water to obtain a garden burnet aqueous solution with the concentration of 1%-3%, and then carrying out ultrasonic treatment and filtration on the garden burnet aqueous solution to obtain a garden burnet ultrasonic extracting solution; 0.5%-1% of glucose, 0.5%-1% of fermented nutritive salt and 0.4%-0.5% of phosphate are dissolved in the sanguisorba officinalis ultrasonic extracting solution, then the mixed solution is sterilized, and a sanguisorba officinalis culture medium is obtained; inoculating lactobacillus plantarum into a seed culture medium for culturing to obtain a lactobacillus seed solution; inoculating a lactobacillus seed solution into the sanguisorba officinalis culture medium to obtain a sanguisorba officinalis fermentation solution; and finally, carrying out centrifugal filtration and pH value adjustment on the garden burnet fermentation liquid to obtain the finished garden burnet fermentation product. The invention has the effects of improving the mitochondrial activity and enhancing the mitochondrial function.

Description

Technical Field

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

[0002] As the core of cell energy metabolism, the ATP generated by mitochondrial respiration is crucial for maintaining various physiological functions of cells, such as material transport, signal transduction, gene expression, etc. In addition, the energy generated also plays an important role in maintaining cell homeostasis. When cells are stimulated by the outside world or have metabolic imbalance, mitochondria enhance energy metabolism to maintain normal cell operation. However, when mitochondria are damaged and unable to respond to external stimuli, cell homeostasis is disrupted and metabolic imbalance is exacerbated; taking skin cells as an example, when cells are stimulated by ultraviolet radiation or other factors causing mitochondrial damage, cell energy metabolism is imbalanced, and reactive oxygen species accumulate, disrupting cell homeostasis and leading to a series of problems such as rough, loose, lack of firmness, and dull skin.

[0003] On this basis, sanguisorba officinalis is the root of the plant sanguisorba officinalis or sanguisorba officinalis var. longifolia. Due to the presence of a large amount of tannins, flavonoids, steroids and other chemical components in the root, it can have anti-inflammatory, analgesic, and leukocyte-elevating effects. Currently, it is often used to treat burns, scalds, inflammation, bleeding and other diseases, and is also often combined with other traditional Chinese medicines or plant extracts and applied in cosmetics to achieve skin moisturizing, skin rejuvenation, anti-wrinkle, sunscreen, whitening and other effects.

[0004] However, the current defect in the application method of sanguisorba officinalis in cosmetics is that current technicians have not studied and explored the application of sanguisorba officinalis fermentation products in improving mitochondrial activity and function. The application method of sanguisorba officinalis in cosmetics is usually to use sanguisorba officinalis extract as a simple whitening and anti-wrinkle ingredient in cosmetics, or to mix it with other ingredients to form a fermentation product for use.

[0005] For example, Patent 201410828632.7 mentions the separation and purification of the active ingredients in the root of sanguisorba officinalis to obtain sanguisorba officinalis extract and use it in cosmetics to delay skin aging. Patent 202210588923.8 mentions using sanguisorba officinalis extract as an antioxidant; however, none of these application methods explore the efficacy of sanguisorba officinalis fermentation products in improving mitochondrial activity and function.

[0006] Patent 201911271571.8 mentions that Sanguisorba officinalis is fermented and mixed with other cosmetics to form an ointment for repairing skin wounds and hyperplastic scars; Patent 201680059039.8 mentions that Sanguisorba officinalis and other ingredients are fermented together so that the prepared cosmetics have the effect of improving wrinkles; Patent 202410202996.8 mentions that Sanguisorba officinalis and other ingredients are fermented together to obtain fermented puree, and the fermented puree is added to cosmetics to achieve whitening effect. The above patents only prove that the composite fermentation extract has the effects of whitening and improving wrinkles, but because the ingredients in the composite fermentation extract are very complex, technicians cannot clearly understand which substances produce the improvement effect, or whether the new ingredients produced in the mixed fermentation process have the above effects, which is not conducive to the large-scale production of cosmetics and the research on the efficacy mechanism of Sanguisorba officinalis extract by technicians.

[0007] Therefore, there is currently no single fermentation of Sanguisorba officinalis and no technical ideas to explore the fermentation products in improving mitochondrial activity and enhancing mitochondrial function. Summary of the invention

[0008] The purpose of the present invention is to provide a preparation method and application of Sanguisorba officinalis fermented product, which has the effects of improving mitochondrial activity and enhancing mitochondrial function.

[0009] The technical solution of the present invention is a method for preparing a fermented product of Sanguisorba officinalis, comprising the following steps:

[0010] ① Mixing the Sanguisorba officinalis root powder with water to obtain a 1% to 3% Sanguisorba officinalis aqueous solution, and then subjecting the Sanguisorba officinalis aqueous solution to ultrasonic treatment and filtration to obtain a Sanguisorba officinalis ultrasonic extract A;

[0011] ② Dissolving 0.5% to 1% glucose, 0.5% to 1% fermentation nutrient salt and 0.4 to 0.5% phosphate in the Sanguisorba officinalis ultrasonic extract A by mass percentage, and then sterilizing the mixed solution to obtain Sanguisorba officinalis culture medium B;

[0012] ③ Inoculating Lactobacillus plantarum into a seed culture medium for culturing to obtain Lactobacillus C seed liquid;

[0013] ④ Inoculate the B Sanguisorba officinalis culture medium with Lactobacillus C seed solution, the inoculation amount of Lactobacillus C seed solution is 5% to 10% of the total mass of the B Sanguisorba officinalis culture medium, to obtain D Sanguisorba officinalis fermented liquid;

[0014] ⑤ Centrifuge and filter the fermentation liquid of Sanguisorba officinalis D to obtain the fermentation supernatant of Sanguisorba officinalis E;

[0015] ⑥ The pH value of the fermentation supernatant of E Sanguisorba officinalis was adjusted to 4.5-6.0 to obtain the finished Sanguisorba officinalis fermentation product.

[0016] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the fermentation nutrient salts of the B sanguisorba officinalis culture medium in step ② include ammonium dihydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride, glucose and yeast extract powder. The temperature during sterilization of the mixed solution is 110-125°C, and the sterilization time is 15-30 minutes.

[0017] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the seed culture medium in step ③ is MRS culture medium. The culture temperature of lactobacillus plantarum during cultivation is 35-40°C, and the fermentation time is 15-20 hours.

[0018] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the stirring speed of the C lactobacillus seed liquid during fermentation in step ④ is 50-100 rpm, the fermentation temperature is 35-40°C, and the fermentation time is 8-24 hours.

[0019] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the sanguisorba officinalis aqueous solution in step ① is ultrasonically treated at an ultrasonic frequency of 50-100% for 30-50 minutes, and then filtered through a 100-300 mesh filter bag to obtain the A sanguisorba officinalis ultrasonic extract

[0020] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the D sanguisorba officinalis fermentation broth is centrifuged at a speed of 5000-10000 rpm and a centrifugation temperature of 10-30°C for 10-30 minutes in step ⑤, and then passed through a 100-300 mesh filter bag to obtain the E sanguisorba officinalis fermentation supernatant.

[0021] In the preparation method of the above-mentioned sanguisorba officinalis ferment, the pH value of the E sanguisorba officinalis fermentation supernatant is adjusted by a 0.2%-0.5% NaOH solution in step ⑥ to make the pH value of the E sanguisorba officinalis fermentation supernatant reach 4.5-6.0.

[0022] Application of the above-mentioned sanguisorba officinalis ferment in cosmetics.

[0023] Application of the above-mentioned sanguisorba officinalis ferment as a component for improving mitochondrial activity in cosmetics.

[0024] Compared with the prior art, by limiting the preparation method of sanguisorba officinalis, the present invention enables sanguisorba officinalis to have the effects of improving mitochondrial activity and enhancing mitochondrial function after single fermentation, thereby improving the functionality of the sanguisorba officinalis ferment and facilitating the large-scale production of manufacturers; at the same time, by comparing the sanguisorba officinalis ferment with the unfermented sanguisorba officinalis extract, it can also be seen that the sanguisorba officinalis ferment prepared by the present invention can effectively improve its antioxidant performance, anti-aging performance and mitochondrial function compared with the unfermented sanguisorba officinalis extract, thereby improving the addition effect of the sanguisorba officinalis component in cosmetics;

[0025] On the other hand, through the improvement of the extraction and fermentation process of sanguisorba root powder, the present invention makes the process have lower energy consumption and no environmental pollution compared with the existing process, thereby improving the preparation effect of the present invention. Therefore, the present invention has the effects of improving mitochondrial activity and enhancing mitochondrial function. Description of the Drawings

[0026] Figure 1 are the top 20 tannin substances with significant changes in the two groups of samples in Experimental Example 2;

[0027] Figure 2 are the top 20 phenolic acid substances with significant changes in the two groups of samples in Experimental Example 2;

[0028] Figure 3 is the result graph of the effects of the three experimental groups on the mitochondrial function of human fibroblasts in Experimental Example 4. Detailed Embodiments

[0029] The present invention will be further described below in conjunction with the embodiments, but it shall not be used as the basis for limiting the present invention.

[0030] Embodiment 1. A preparation method of sanguisorba ferment includes the following steps:

[0031] ① Dissolve 10 g of sanguisorba root powder in 1 L of ultrapure water to obtain a sanguisorba aqueous solution. The sanguisorba is purchased from Bozhou Jingwan Chinese Herbal Pieces Factory. Then, ultrasonically treat the sanguisorba aqueous solution at an ultrasonic frequency of 80% for 45 minutes, and then filter it through a 100-mesh filter bag to obtain A sanguisorba ultrasonic extract;

[0032] ② Dissolve 10 g of glucose, 5 g of fermentation nutrient salts, and 4 g of phosphates in 1 L of A sanguisorba ultrasonic extract. Then, sterilize the mixed solution at 115°C for 20 minutes, and cool it to obtain B sanguisorba culture medium. The fermentation nutrient salts are conventional commercially purchased fermentation nutrient salts, and the phosphates include disodium hydrogen phosphate anhydrous and potassium dihydrogen phosphate;

[0033] ③ Inoculate Lactobacillus plantarum in MRS culture medium and ferment at a temperature of 35°C for 15 hours to obtain C lactic acid bacteria seed liquid. The OD600 in the C lactic acid bacteria seed liquid is 7.0 - 9.0, and the viable bacteria count is 1 - 2×10^9 CFU / mL;

[0034] ④ Inoculate 5% of the C lactic acid bacteria seed liquid into the B sanguisorba culture medium and carry out anaerobic fermentation. During the fermentation process, stir at a speed of 50 rpm, the fermentation temperature is 37°C, and the fermentation time is 12 hours to obtain D sanguisorba fermentation liquid;

[0035] ⑤ Centrifuge the D sanguisorba fermentation liquid at a speed of 8000 rpm and a centrifugation temperature of 15°C for 15 minutes, and then filter it through a 200-mesh filter bag to obtain E sanguisorba fermentation supernatant;

[0036] ⑥Adjust the pH value of the fermentation supernatant of Sanguisorba officinalis from area E to 5.0 with a 0.4% NaOH solution to obtain the finished Sanguisorba officinalis fermentation product.

[0037] Example 2. A method for preparing a Sanguisorba officinalis fermentation product, comprising the following steps:

[0038] ①Dissolve 20 g of Sanguisorba officinalis root powder in 1 L of ultrapure water to obtain a Sanguisorba officinalis aqueous solution. The Sanguisorba officinalis was purchased from Bozhou Jingwan Chinese Herbal Pieces Factory. Then, ultrasonically treat the Sanguisorba officinalis aqueous solution at an ultrasonic frequency of 100% for 35 minutes, and then filter it through a 200-mesh filter bag to obtain the ultrasonic extract of Sanguisorba officinalis A.

[0039] ②Dissolve 15 g of glucose, 8 g of fermentation nutrient salts, and 5 g of phosphate in 1 L of the ultrasonic extract of Sanguisorba officinalis A, and then sterilize the mixture at 120 °C for 30 minutes. After cooling, obtain the Sanguisorba officinalis medium B.

[0040] ③Inoculate Lactobacillus plantarum in MRS medium and ferment at 40 °C for 20 hours to obtain the C Lactobacillus seed liquid.

[0041] ④Add 10% of the C Lactobacillus seed liquid to the Sanguisorba officinalis medium B and ferment anaerobically. During the fermentation process, stir at a speed of 80 rpm, the fermentation temperature is 37 °C, and the fermentation time is 24 hours to obtain the Sanguisorba officinalis fermentation liquid D.

[0042] ⑤Centrifuge the Sanguisorba officinalis fermentation liquid D at a speed of 10000 rpm and a centrifugation temperature of 25 °C for 20 minutes, and then filter it through a 300-mesh filter bag to obtain the Sanguisorba officinalis fermentation supernatant E.

[0043] ⑥Adjust the pH value of the Sanguisorba officinalis fermentation supernatant E to 5.5 with a 0.5% NaOH solution to obtain the finished Sanguisorba officinalis fermentation product.

[0044] Example 3. A method for preparing a Sanguisorba officinalis fermentation product, comprising the following steps:

[0045] ①Dissolve 30 g of Sanguisorba officinalis root powder in 1 L of ultrapure water to obtain a Sanguisorba officinalis aqueous solution. The Sanguisorba officinalis was purchased from Bozhou Jingwan Chinese Herbal Pieces Factory. Then, ultrasonically treat the Sanguisorba officinalis aqueous solution at an ultrasonic frequency of 100% for 30 minutes, and then filter it through a 300-mesh filter bag to obtain the ultrasonic extract of Sanguisorba officinalis A.

[0046] ②Dissolve 8 g of glucose, 10 g of fermentation nutrient salts, and 4.19 g of phosphate in 1 L of the ultrasonic extract of Sanguisorba officinalis A, and then sterilize the mixture at 121 °C for 15 minutes. After cooling, obtain the Sanguisorba officinalis medium B.

[0047] ③Inoculate Lactobacillus plantarum in MRS medium and ferment at 38 °C for 18 hours to obtain the C Lactobacillus seed liquid.

[0048] ④ Inoculate 8% of Lactobacillus ciferrii seed liquid into the Sanguisorba officinalis L. medium and carry out anaerobic fermentation. During the fermentation process, stir at a speed of 100 rpm, the fermentation temperature is 40 °C, and the fermentation time is 8 hours to obtain the Sanguisorba officinalis L. fermentation broth D;

[0049] ⑤ Centrifuge the Sanguisorba officinalis L. fermentation broth D at a speed of 8500 rpm and a centrifugation temperature of 30 °C for 10 minutes, and then filter it through a 100-mesh filter bag to obtain the Sanguisorba officinalis L. fermentation supernatant E;

[0050] ⑥ Adjust the pH value of the Sanguisorba officinalis L. fermentation supernatant E to 6.0 with 0.34% NaOH solution to obtain the finished Sanguisorba officinalis L. fermented product.

[0051] Example 4. Apply the Sanguisorba officinalis L. fermented product prepared in Example 1, Example 2 or Example 3 in lotion or emulsion, and use it to improve mitochondrial activity and enhance mitochondrial function.

[0052] Experimental Example 1: Prepare the Sanguisorba officinalis L. extract as Comparative Example 1 according to the following steps:

[0053] ① Dissolve 60 g of Sanguisorba officinalis L. root powder in 1 L of ultrapure water to obtain the Sanguisorba officinalis L. aqueous solution. This Sanguisorba officinalis L. is purchased from Bozhou Jingwan Chinese Herbal Pieces Factory. Then, ultrasonically treat the Sanguisorba officinalis L. aqueous solution at an ultrasonic frequency of 100% for 30 minutes, and then filter it through a 300-mesh filter bag to obtain the Sanguisorba officinalis L. ultrasonic extract A;

[0054] ② Dissolve 10 g of glucose, 10 g of fermentation nutrient salts and 4.19 g of phosphate in 990 mL of water, and add 10 mL of the Sanguisorba officinalis L. ultrasonic extract A. The concentration of the Sanguisorba officinalis L. ultrasonic extract A is 10 mg / mL. After sterilization and cooling, prepare the Sanguisorba officinalis L. medium B. The fermentation nutrient salts and phosphates are the same as those in Example 1;

[0055] ③ Centrifuge the Sanguisorba officinalis L. medium B at a speed of 10000 rpm and a centrifugation temperature of 25 °C for 20 minutes to obtain the Sanguisorba officinalis L. extract.

[0056] Then, use the Sanguisorba officinalis L. fermented product prepared in Example 1 and the Sanguisorba officinalis L. extract of Comparative Example 1 as samples. After diluting both groups of samples 200 times, use the kit method to measure the ability of both to scavenge DPPH free radicals. The specific measurement method is as follows:

[0057] a. Weigh 8 mg of DPPH, dissolve it with absolute ethanol and make up the volume to 100 mL in a volumetric flask to obtain the DPPH solution;

[0058] b. Dissolve and dilute the samples with absolute ethanol, and use the concentration that can dissolve in the system reagent as the high concentration. Set 3 - 5 concentration gradients for each sample to obtain the sample solution;

[0059] c. Mix 750 mL of the sample solution with the DPPH solution and name it Tube A1; mix 750 mL of absolute ethanol with the DPPH solution and name it Tube A2; mix 750 mL of the sample solution evenly with absolute ethanol and name it Tube A3; perform three replicates for each sample. After reacting in the dark for 30 minutes, use a microplate reader to measure the absorbance values of Tubes A1, A2, and A3 at 517 nm respectively, and then calculate the DPPH scavenging rate of the sample according to the scavenging rate (%) = [(A2 + A3) - A1] / A2.

[0060] Then, use the tannin content determination method to measure the content of non-adsorbed phenolic acids and tannin content in the two groups of samples. The specific measurement method is as follows:

[0061] a. Take 25 mL of the sample and water, add 0.6 g of casein to each, keep it warm in a water bath at 30 °C for 1 h, shake it from time to time, and obtain the adsorption solution, with water as the blank control;

[0062] b. Weigh 50 mg of gallic acid, dissolve it with 5 mL of methanol, and then make up the volume to 100 mL with deionized water to obtain the gallic acid preparation solution;

[0063] c. Dilute the gallic acid preparation solution respectively to obtain gallic acid working solutions with concentrations of 0, 10, 20, 30, 40, and 50 μg / mL. Then, take 10 mL EP tubes, add 0.5 mL of the gallic acid working solution, 0.25 mL of Folin-Ciocalteu phenol, 1 mL of sodium carbonate, and 3.25 mL of deionized water to each tube. After reacting in a water bath at 70 °C for 30 minutes, measure the absorbance at 750 nm;

[0064] d. Place 0.5 mL of the adsorption solution, 0.25 mL of Folin-Ciocalteu phenol, 1 mL of 7.5% sodium carbonate, and 3.25 mL of deionized water in a water bath at 70 °C for 30 minutes respectively, and then measure the absorbance at 750 nm.

[0065] Among them, the content of non-adsorbed polyphenols = adsorption solution - blank, and the tannin content = total phenol content - content of non-adsorbed polyphenols. The measurement results are shown in Table 1:

[0066] Table 1: pH, Brix, and DPPH scavenging rate of Example 1 and Comparative Example 1

[0067]

[0068] As can be seen from Table 1, the content of non-adsorbable phenolic acids in Example 1 increased by 0.11 mg / mL compared with that in Comparative Example 1, and the tannin content decreased by 0.76 mg / mL, indicating that a part of tannins were metabolized into non-adsorbable phenolic acids. Moreover, the DPPH scavenging effect of Example 1 was better than that of Comparative Example 1, suggesting that the sanguisorba officinalis ferment prepared by the present invention has better antioxidant efficacy than the unfermented sanguisorba officinalis extract.

[0069] Experimental Example 2: Weigh the sanguisorba officinalis ferment of Example 2 and the sanguisorba officinalis extract of Comparative Example 1 as samples, add the extraction solution for metabolite extraction treatment in a low-temperature environment, and then centrifuge to take the supernatant metabolite solvent for liquid chromatography-mass spectrometry detection. The downloaded data is processed by the metabolomics software Progenesis QI (Waters Corporation, Milford, USA) for peak extraction, alignment, identification, etc., and finally a data matrix containing retention time, peak area, mass-to-charge ratio, and identification information is obtained for substance composition analysis.

[0070] The top 20 tannins and phenolic acids with significant changes in the two groups of samples are as Figure 1 and Figure 2 shown, where Figure 1 represents tannin substances, and Figure 2 represents phenolic acid substances. From the test results, it can be seen that in Example 2 compared with Comparative Example 1, the contents of 13 tannins such as Pedunculagin, Vitilagin, Beta-Glucogallin, Castanin, Granatin A, Strictinin, Chebulinic Acid, Casuarinin, Brevilagin I, Pterocaryanin B, and 2-O-(4-Hydroxycinnamoyl)-1-O-Galloyl-β-D-Glucopyranoside decreased significantly; the contents of 10 phenolic acids such as Veratricacid, Normetanephrine, Catechol, and Epi gallocatechin Gallate increased significantly. This corroborates the experimental results of Experimental Example 1 and confirms that the sanguisorba officinalis ferment prepared by the present invention has better antioxidant efficacy than the unfermented sanguisorba officinalis extract.

[0071] Experimental Example 3: Take the sanguisorba officinalis ferment of Example 2 and the sanguisorba officinalis extract of Comparative Example 1, dilute them 2-fold with water as the solvent as samples, and measure the inhibitory effects of the two groups of samples on elastase through the elastase inhibition rate experiment. The specific measurement method is as follows:

[0072] a. Prepare an elastase solution at 600 mU / mL, an AAAPAN (N-succinyl-alanine-alanine-alanine-p-nitroaniline) solution at 1.015 mmol / L, and a 50 mmol / L Tris-HCl buffer solution;

[0073] b. Mix 420 μL of Tris-HCl buffer (pH 8.0) with 180 μL of water, then add 150 μL of the elastase solution, and incubate at 25 °C for 15 minutes to obtain Sample A1;

[0074] c. Mix 420 μL of Tris-HCl buffer (pH 8.0) with 180 μL of the sample, then add 150 μL of the elastase solution, and incubate at 25 °C for 15 minutes to obtain Sample A2;

[0075] d. Mix 570 μL of Tris-HCl buffer (pH 8.0) with 180 μL of water or the sample, and incubate at 25 °C for 15 minutes as the non-enzyme control group to obtain Sample A3;

[0076] e. Add 150 μL of the AAAPAN solution to Samples A1, A2, and A3 respectively. After 20 minutes, measure the absorbance value at 410 nm, and then calculate the inhibition rate of the sample according to the formula: Inhibition rate (%) = [1 - (A2 - A3) / (A1 - A3)] × 100%.

[0077] The measurement results are shown in Table 2:

[0078] Table 2: Inhibitory effects of examples and comparative examples on elastase

[0079] Sample Elastase inhibition rate (%) Comparative Example 1 80.45 Example 2 88.56

[0080] It can be seen from the measurement results that the inhibition rate of Example 2 on elastase is 88.56%, which is 8.11% higher than that of Comparative Example 1, indicating that the fermentation process of the present invention can improve the inhibitory effect of Sanguisorba officinalis on elastase, and the Sanguisorba officinalis fermentation product prepared by the present invention has certain anti-aging potential.

[0081] Experimental Example 4: First, use the CCK-8 method to detect the cytotoxicity of three groups of examples and Comparative Example 1 on HDF, and determine that 1% of Comparative Example 1 and the three groups of examples have no cytotoxicity to HDF cells. Then, set up four experimental groups, namely the blank group, the model group, the comparative example group, and the example group; the grouping and treatment methods of the four experimental groups are shown in Table 3.

[0082] Table 3 Grouping and treatment methods of four experimental groups

[0083]

[0084] Then, the change rate (%) of the four experimental groups was calculated according to the formula: change rate (%) = measured value of each group of indicators / measured value of the experimental group * 100%, and the calculation results are as follows Figure 3 shown. Then, the Seahorse XF-96 method was used to measure the oxygen consumption rate (OCR) of basal respiration, ATP production, maximal respiration, and spare respiratory capacity of cell mitochondria to evaluate its effect on mitochondrial function. The test results are shown in Table 4:

[0085] Table 4 Effects of different treatments on the mitochondrial function of human dermal fibroblasts (HDF)

[0086]

[0087]

[0088] Through Figure 2 and the test results in Table 4, it can be seen that at the same concentration, Comparative Example 1 and Example 3 improved the basal respiration, ATP production, maximal respiration, and spare respiratory capacity of HDF cells; and the indicators of Comparative Example 1 increased by 110%, 107%, 223%, and 354%, while the indicators of Example 3 increased by 163%, 160%, 281%, and 424%, indicating that both Example 3 and Comparative Example 1 can improve the mitochondrial function of HDF. By comparing the data of the two, it shows that Example 3 improved the indicators of basal respiration, ATP production, maximal respiration, and spare respiratory capacity of HDF cells by 53%, 53%, 58%, and 70% respectively compared with Comparative Example 1, indicating that the sanguisorba ferment obtained by the present invention can improve the improvement effect on the mitochondrial function of HDF compared with the unfermented sanguisorba extract, thereby enhancing cell energy metabolism, improving cell respiratory metabolism ability, and maintaining cell homeostasis.

Claims

1. A preparation method of sanguisorba officinalis fermentation product, characterized in that, It includes the following steps: ① Mix the sanguisorba root powder with water to obtain a sanguisorba aqueous solution with a concentration of 1% - 3%, and then perform ultrasonic treatment and filtration on the sanguisorba aqueous solution to obtain ultrasonic extract A of sanguisorba; ② Dissolve 0.5% - 1% glucose, 0.5% - 1% fermentation nutrient salts, and 0.4 - 0.5% phosphate in ultrasonic extract A of sanguisorba, and then sterilize the mixture to obtain culture medium B of sanguisorba; ③ Inoculate Lactobacillus plantarum in a seed culture medium for cultivation to obtain seed solution C of lactic acid bacteria; ④ Inoculate seed solution C of lactic acid bacteria into culture medium B of sanguisorba, and the inoculation amount of seed solution C of lactic acid bacteria is 5% - 10% of the total mass of culture medium B of sanguisorba to obtain fermentation broth D of sanguisorba; ⑤ Centrifuge and filter fermentation broth D of sanguisorba to obtain supernatant E of fermentation broth of sanguisorba; ⑥ Adjust the pH value of supernatant E of fermentation broth of sanguisorba to 4.5 - 6.0 to obtain the finished product of fermented sanguisorba; 2. The preparation method of a sanguisorba officinalis fermentation product according to claim 1, characterized in that: The fermentation nutrient salts of culture medium B of sanguisorba in step ② include ammonium dihydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, calcium chloride, glucose, and yeast extract powder. The temperature during sterilization of the mixture is 110 - 125°C, and the sterilization time is 15 - 30 minutes.

3. The preparation method of a sanguisorba officinalis ferment according to claim 1, characterized in that: The seed culture medium in step ③ is MRS culture medium, and the cultivation temperature of Lactobacillus plantarum during cultivation is 35 - 40°C, and the fermentation time is 15 - 20 hours.

4. The preparation method of a sanguisorba officinalis fermentation product according to claim 1, characterized in that: The stirring speed of seed solution C of lactic acid bacteria during fermentation in step ④ is 50 - 100 rpm, the fermentation temperature is 35 - 40°C, and the fermentation time is 8 - 24 hours.

5. The preparation method of a sanguisorba officinalis fermentation product according to claim 1, characterized in that: In step ①, the sanguisorba aqueous solution is ultrasonically treated at an ultrasonic frequency of 50 - 100% for 30 - 50 minutes, and then filtered through a filter bag with 100 - 300 meshes to obtain ultrasonic extract A of sanguisorba.

6. The preparation method of a sanguisorba officinalis fermentation product according to claim 1, characterized in that: In step ⑤, fermentation broth D of sanguisorba is centrifuged at a speed of 5000 - 10000 rpm and a centrifugation temperature of 10 - 30°C for 10 - 30 minutes, and then filtered through a filter bag with 100 - 300 meshes to obtain supernatant E of fermentation broth of sanguisorba.

7. The preparation method of a sanguisorba officinalis fermentation product according to claim 1, characterized in that: In step ⑥, the pH value of supernatant E of fermentation broth of sanguisorba is adjusted by a 0.2% - 0.5% NaOH solution to make the pH value of supernatant E of fermentation broth of sanguisorba reach 4.5 - 6.

0.

8. Application of the fermented sanguisorba described in any one of claims 1 - 7 in cosmetics.

9. Application of the fermented sanguisorba described in any one of claims 1 - 7 as a component for improving mitochondrial activity in cosmetics.

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