Preparation method and application of rose extract

Abstract

The invention belongs to the technical field of biology, and particularly relates to a preparation method and application of a rose extract, and the content of water-soluble vitamins in roses is increased by utilizing a microbial fermentation technology. According to the method, strains including lactobacillus plantarum, saccharomyces cerevisiae and bacillus subtilis are selected, the proportion of the strains is adjusted, the viable count proportion of the lactobacillus plantarum to the saccharomyces cerevisiae to the bacillus subtilis is (1-3): (1-2): (1-2), a complex microbial inoculant is formed, and pretreated roses are added for segmented fermentation. And finally, the content of water-soluble vitamins in the rose extract is increased, so that the total antioxidant activity is increased by about 35% while rose polysaccharide substances are retained. The rose extract is suitable for providing raw materials for functional food, health care products and cosmetics.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to a preparation method and application of a rose extract. Background Art

[0002] Roses are easy to be planted on a large scale, such as double - petal roses and Damascus roses, and they have a long flowering period and a large biomass, with a stable raw material supply. Roses are rich in active ingredients such as phenols (such as gallic acid and ellagic acid), polysaccharides, and flavonoids, and have antioxidant and anti - inflammatory properties. At the same time, these active substances can be used as substrates for microbial fermentation, and can be converted into water - soluble vitamins (such as vitamin B group and vitamin C) or their derivatives after microbial metabolism. Water - soluble vitamins have the effects of improving skin barrier function, reducing inflammation, and inhibiting melanin production. These vitamins can synergistically act with the active ingredients of roses themselves, enhancing the biological activity of rose extracts, so that they have better whitening, anti - aging and other effects. Therefore, "rose extract" has a high recognition in the food and cosmetic fields.

[0003] However, existing fermentation processes are mostly optimized for the functional components of roses (such as polyphenols, flavonoids, etc.) rather than vitamin synthesis. Moreover, water - soluble vitamins are easily damaged by factors such as heat, light, and oxygen. Existing fermentation processes such as high - temperature sterilization and long - time fermentation may reduce the content of water - soluble vitamins in rose extracts, thereby affecting the use effect of rose extracts in cosmetics. In addition, the traditional preparation methods of rose extracts have problems such as low preparation efficiency, unstable yield, chemical reagent residues and safety risks, high energy consumption and environmental pollution. Using strong acids / alkalis or long - time heating during extraction may damage the active ingredients in rose extracts. These problems have limited the application of rose extracts in the food and health product fields to a certain extent.

[0004] In order to overcome the above problems, there is an urgent need for a new method for preparing rose extracts, which can increase the content of water - soluble vitamins in rose extracts while retaining polysaccharides, so as to expand the application of rose extracts from a single beauty function to the field of nutritional supplements and expand the application of roses in the food, health product and cosmetic fields. Summary of the Invention

[0005] In order to overcome the problems in the background art, the present application provides a preparation method and application of a rose extract, which improve the content of water - soluble vitamins in the rose extract through microbial fermentation without damaging the active substances in the roses themselves.

[0006] To achieve the above object, on the one hand, the present invention provides a preparation method of a rose extract, and the specific steps include: (1) Prepare a composite bacterium agent: Lactobacillus plantarum (L. plantarum ), Lactobacillus plantarum ( S. cerevisiae ), Saccharomyces cerevisiae ( B. subtilis ) and Bacillus subtilis ( ) are proportioned according to the viable count ratio of (1 - 3):(1 - 2):(1 - 2) to obtain a compound microbial agent; (2) Prepare rose matrix: The rose flowers are crushed and mixed with deionized water to obtain a mixture. Cellulase is added to the mixture for enzymatic hydrolysis, and after heating and sterilization and cooling, a rose matrix is formed; (3) Inoculation: The compound microbial agent is inoculated into the rose matrix, and the mass - volume ratio of the compound microbial agent to the rose matrix is (0.2 - 2):100 (g / mL) to form a rose flower fermentation substrate; (4) Fermentation: The rose flower fermentation substrate obtained in (3) is fermented for 48 - 72 hours. The dissolved oxygen content in the first 12 hours is 50% - 70%, and the dissolved oxygen content is controlled at 0 - 10% during the subsequent fermentation time. After fermentation is completed, rose flower residues are formed, and the rose flower residues are filtered to obtain a rose flower fermentation product;

[0007] Further preferably, in the step (1), the viable count of Lactobacillus plantarum is 1×10 7 cfu / g; the viable count of Saccharomyces cerevisiae is 2.8×10 7 cfu / g; the viable count of Bacillus subtilis is 1×10 7 cfu / g.

[0008] Further preferably, in the step (2), the material - liquid ratio of the crushed rose flowers to deionized water is 1:(5 - 10) (g / mL).

[0009] Further preferably, in the step (2), the material - liquid ratio of the cellulase to the mixture is (1 - 5):1000 (g / mL).

[0010] Further preferably, in the step (2), the temperature of the enzymatic hydrolysis is 40 - 50 °C, and the time of the enzymatic hydrolysis is 1 - 2 hours.

[0011] Further preferably, in the step (2), the temperature of the sterilization is 80 °C, and the time is 15 minutes; the temperature of the cooling is 25 - 40 °C.

[0012] Further preferably, in the step (4), the temperature of the fermentation is 30 - 37 °C, and the pH is 5.5 - 6.5.

[0013] Further preferably, in the step (4), the filtration method is to first filter the rose flower residues with a filter medium of 100 - 200 meshes to obtain a filtrate, and then filter and sterilize the filtrate with a 0.2 μm sterilizing filter membrane.

[0014] Further preferably, in step (5), the pore size of the ultrafiltration membrane of the ultrafiltration is 0.01 μm; the rotation speed of the centrifugation is 8000-12000 rpm, and the time is 10-20 minutes.

[0015] On the other hand, the present invention also provides the use of the rose extract obtained according to the above preparation method in food, health products or cosmetics.

[0016] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present application.

[0017] Beneficial effects of the present invention: 1. This application provides a method for preparing a rose extract, which significantly increases the water-soluble vitamin and polysaccharide contents in the resulting extract. This is because the method gently releases vitamin synthesis precursors, such as sugars and amino acids, from the rose through enzymatic hydrolysis. The method then uses a composite bacterial strain for synergistic fermentation to increase the vitamin content. Staged fermentation is used during the fermentation process to improve fermentation efficiency. Furthermore, a combined centrifugation and membrane separation technique is used during extraction to enrich the water-soluble vitamins. This method improves the yield of water-soluble vitamins in the rose extract while effectively retaining the polysaccharides.

[0018] 2. This application provides a method for preparing a rose extract. The resulting extract significantly increases the content of vitamins B1, B2, B6, and C compared to the control. This is achieved by using a composite bacterial strain for synergistic fermentation, employing both acid-producing bacteria (Lactobacillus plantarum) and vitamin-synthesizing bacteria (Saccharomyces cerevisiae and Bacillus subtilis). The acidic environment created by the former protects the vitamins' stability, while the latter selectively synthesizes water-soluble vitamins, thereby achieving the aforementioned effects.

[0019] 3. This application improves the efficiency of microbial fermentation through staged fermentation, thereby promoting an increase in the content of water-soluble vitamins in rose extract. This is because the dissolved oxygen level is controlled in stages. In the early stages of fermentation, when the oxygen content is high, i.e., 50%-70%, it is conducive to bacterial growth and rapid formation of bacterial colonies. In the later stages, when the dissolved oxygen level is 0-10%, anaerobic bacteria enhance metabolism, increase product accumulation, and avoid oxidative loss of water-soluble vitamins.

[0020] 4. This application utilizes an ultrafiltration process to retain rose polysaccharides and vitamins, thereby enhancing the overall antioxidant activity of the rose extract. This is because conventional rose extraction involves solvent heating, which can lead to degradation of polysaccharides and vitamins and the risk of solvent residue. This application, however, utilizes high-speed centrifugation to separate the supernatant, ultrafiltration to trap macromolecular impurities, and collection of the filtrate for extraction. This overall process is environmentally friendly and could improve the application of rose extract in food, health supplements, and cosmetics or skincare products. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a comparison chart of vitamin content of rose extracts in this application; Figure 2 This is a comparison chart of the polysaccharide content of rose extracts in this application; Figure 3 This is a diagram showing changes in human wrinkles using the formulation in Table 2 of Example 3 of this application; Figure 4 This is a diagram showing changes in human wrinkles using the formulation in Table 3 of Example 3 of this application; Figure 5 This is a comparison chart of the antioxidant activity of Example 4 of the present application. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solutions and beneficial effects of the present invention clearer, the preferred embodiments of the present invention will be described in detail below to facilitate understanding by technicians.

[0023] The embodiments of the present invention provide a preparation method and application of a rose extract, which can effectively increase the content of water-soluble vitamins in the rose extract without destroying the active substances in the rose itself, thereby expanding its application rate in the fields of food, health products and cosmetics.

[0024] Unless otherwise specified, all reagents and materials used in the following examples were purchased from the market.

[0025] Example 1: (A method for preparing rose extract) (1) Preparation of composite bacterial agent: The strain selected is Lactobacillus plantarum ( L. plantarum )(1×10 7 cfu / g, purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.), Saccharomyces cerevisiae ( S. cerevisiae )(2.8×10 7 cfu / g, purchased from Angel Yeast Co., Ltd.) and Bacillus subtilis ( B. subtilis )(1×10 7cfu / g, purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.), accurately weighed and prepared with a balance of one ten-thousandth according to the viable count ratio of Lactobacillus plantarum, Saccharomyces cerevisiae and Bacillus subtilis of 2:1:1; (2) Preparation of rose matrix: The rose petals were crushed to 40 mesh, mixed with deionized water at a solid-liquid ratio of 1:7 (g / mL) to form a mixture, and cellulase was added at a ratio of 2:1000 (g / mL) of cellulase to the mixture, enzymolyzed at 45 °C for 2 hours to release the substrate, heated and sterilized at 80 °C for 15 minutes, and cooled to 25 °C to form a rose matrix; (3) Inoculation: Inoculated into the rose matrix described in step (2) at a mass-volume ratio of the compound bacterium agent to the rose matrix of 1:100 (g / mL) to form a rose flower fermentation substrate; (4) Fermentation: The rose flower fermentation substrate obtained in (3) was fermented at 30-37 °C and pH 5.5-6.5 for 72 hours. Sterile air was introduced into the rose flower fermentation substrate in the first 12 hours to make the dissolved oxygen content 50%-70%, and it was in a closed system in the next 60 hours, controlling the dissolved oxygen content below 10%. After fermentation, rose flower residues were formed. The rose flower residues were first filtered through a 200-mesh filter material to obtain a filtrate, and then the filtrate was filtered and sterilized with a 0.2 μm sterilizing filter membrane to obtain a rose flower fermentation product; (5) Collection: The rose flower fermentation product obtained in step (4) was filtered through an ultrafiltration membrane (SUEZ GK8040F-30D) to retain macromolecular impurities, and the filtrate was collected by high-speed centrifugation at a speed of 8000 rpm for 15 minutes. The filtrate was the rose flower extract.

[0026] Example 2: (A method for preparing rose flower extract) (1) Preparation of compound bacterium agent: The selected strains were Lactobacillus plantarum ( L. plantarum )(1×10 7 cfu / g, purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.), Saccharomyces cerevisiae ( S. cerevisiae )(2.8×10 7 cfu / g, purchased from Angel Yeast Co., Ltd.) and Bacillus subtilis ( B. subtilis )(1×10 7 cfu / g, purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.), accurately weighed and prepared with a balance of one ten-thousandth according to the viable count ratio of Lactobacillus plantarum, Saccharomyces cerevisiae and Bacillus subtilis of 1:1:1; (2) Preparation of rose matrix: Crush rose petals to 60 mesh, mix them with deionized water at a solid-liquid ratio of 1:7 (g / mL) to form a mixture, add cellulase to the mixture at a ratio of 2:1000 (g / mL), enzymolyze at 45 °C for 2 hours to release the substrate, heat and sterilize at 80 °C for 15 minutes, and cool to 40 °C to form a rose matrix; (3) Inoculation: Inoculate the rose matrix described in step (2) at a mass-volume ratio of the compound bacterial agent to the rose matrix of 1:100 (g / mL) to form a rose flower fermentation substrate; (4) Fermentation: Ferment the rose flower fermentation substrate obtained in (3) at 30 - 37 °C and pH 5.5 - 6.5 for 48 hours. In the first 12 hours, introduce sterile air into the rose flower fermentation substrate to make the dissolved oxygen content 50% - 70%. In the next 36 hours, keep it in a closed system and control the dissolved oxygen content below 10%. After fermentation, rose flower residue is formed. First, filter the rose flower residue with a 200-mesh filter material to obtain a filtrate, and then filter and sterilize the filtrate with a 0.2-μm sterilizing filter membrane to obtain a rose flower fermentation product; (5) Collection: Filter and retain macromolecular impurities from the rose flower fermentation product obtained in step (4) using an ultrafiltration membrane (SUEZ GK8040F-30D), collect the filtrate by high-speed centrifugation at a speed of 8000 rpm for 15 minutes, and the filtrate is the rose flower extract.

[0027] Comparative Example 1: (1) Crush rose petals to 40 mesh, mix them with deionized water at a solid-liquid ratio of 1:7 (g / mL) to form a mixture, add cellulase to the mixture at a ratio of 2:1000 (g / mL), and enzymolyze at 45 °C for 2 hours to release the substrate; (2) Perform ultrasonic treatment at 50 °C for 30 minutes, with an ultrasonic power of 300 W and a frequency of 40 kHz; (3) Centrifuge to obtain the supernatant at a speed of 8000 rpm for 15 minutes, and repeat the extraction once; (4) Combine the supernatants to obtain the rose flower extract.

[0028] Comparative Example 2: A rose flower extract, in the preparation method, except for not adding the compound bacterial agent, directly ferment the rose matrix in step (5), and the other steps are the same as in Example 1.

[0029] Comparative Example 3: A rose flower extract, in the preparation method, except that the viable cell number ratio of Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis in the compound bacterial agent in step (2) is 4:1:1, the other steps are the same as in Example 1.

[0030] Comparative Example 4: A rose extract, except that the compound bacterium agent in steps (1) and (2) is set as Lactobacillus plantarum and Saccharomyces cerevisiae, and the viable bacteria number ratio is 2:1, the remaining steps are the same as in Example 1.

[0031] Control Example 5: A rose extract, except that the fermentation conditions in step (5) are changed to: fermenting for 60 hours under the conditions of 30 - 37 °C, pH 5.5 - 6.5, and the dissolved oxygen content being constantly 0% - 10%, the remaining steps are the same as in Example 1.

[0032] Example 3 (Determination and Data Analysis of Vitamin and Polysaccharide Contents) 1. HPLC Detection of Vitamin Content A. Chromatographic Conditions: Chromatographic column: Alltima C18 chromatographic column (250 mm × 4.6 mm, 5 μm); Mobile phase: Acetonitrile - 50 mmol / L (A), ammonium dihydrogen phosphate solution (adjusted to pH 3.0 with phosphoric acid) (B), gradient elution (0 - 8 min, 5% A; 8 - 23 min, 5% - 35% A; 23 - 28 min, 35% - 40% A); Flow rate: 0.5 mL / min; Detection wavelength: 275 nm; Column temperature: 30 °C.

[0033] B. Sample Preparation: Weigh appropriate amounts of reference substances of vitamin B1, vitamin B2, vitamin B6, and vitamin C accurately, and prepare a mixed reference substance stock solution with each concentration of about 2000 μg / mL using acetonitrile - water (5∶95). Accurately measure 1, 2, 2.5, 3, 5 mL of the mixed reference substance stock solution into 50 mL volumetric flasks, dilute to the scale with acetonitrile - water (5∶95), and shake well to obtain mixed reference substance solutions with the concentrations of each component being about 40, 80, 100, 120, 200 μg / mL.

[0034] C. Preparation of Test Solution: Accurately measure 5 mL of the rose extract, place it in a 100 mL volumetric flask, add acetonitrile - water (5∶95) to volume, shake well, filter through a 0.22 μm microporous membrane, and take the subsequent filtrate as the test solution.

[0035] D. Content Calculation: Determine according to the chromatographic conditions described in A. Taking the peak area as the ordinate (Y) and the concentration as the abscissa (X, mg / mL), draw a standard curve to obtain a regression equation. The vitamin content in the rose extract is calculated based on dry matter (%), and the calculation method is as follows: : The detected concentration (μg / mL) calculated according to HPLC determination; 20: The dilution factor during the detection process; : The moisture content (%) in the rose extract, and the moisture content can be quickly detected by a rapid moisture analyzer (sartorius, MA35M - 000230V1).

[0036] 2. Determine the polysaccharide content in the rose extract Implement according to the polysaccharide content detection method described in the specification of Patent - ZL202410312477.7 - Example 6. The polysaccharide content in the rose extract is calculated based on dry matter (%), and the calculation method is as follows: : The polysaccharide concentration in the test sample, mg / mL; 100: The dilution factor during the detection process; 0.9: The correction coefficient of glucose for polysaccharide; : The moisture content (%) in the rose extract, and the moisture content can be quickly detected by a rapid moisture analyzer (sartorius, MA35M - 000230V1).

[0037] Data analysis: From Table 1 and Figure 1 、 Figure 2 it can be seen that: Overall, the contents of vitamin B, vitamin C, and polysaccharide in Comparative Example 1 and Comparative Example 2 are relatively low. No compound bacterial agent is added in both groups, indicating that the use of compound bacterial agent can increase the contents of vitamin B, vitamin C, and polysaccharide in the rose extract. At the same time, Comparative Example 1 adopts the traditional preparation process of rose extract. Except for not adding compound bacterial agent, the extraction method used is also the traditional ultrasonic extraction process. Therefore, there are significant differences in the polysaccharide content between Comparative Example 1 and the two examples. This shows that the ultrafiltration extraction process adopted in this application can better retain the polysaccharide components in the rose extract and plays an important role in improving the activity of the rose extract.

[0038] The contents of vitamin B and vitamin C in Example 1 are the highest, followed by Example 2 and Comparative Example 3. The difference between Example 2, Comparative Example 3 and Example 1 lies in the viable count ratio of the compound bacterial agent, indicating that the difference in viable count ratio will affect the contents of vitamin B and vitamin C. Moreover, the vitamin contents in the extracts of Comparative Example 3 are lower than those of the two examples, indicating that the ratio of the compound bacterial agent selected in this application is relatively excellent. In addition, the contents of vitamin B and vitamin C in Comparative Example 4 are significantly lower than those in Example 1, indicating that the types of bacteria in the compound bacterial agent will affect the contents of vitamin B and vitamin C in the rose extract. Therefore, the above data show that the types of bacteria and their ratios in the compound bacterial agent selected in this application play an important role in increasing the content of water-soluble vitamins in the rose extract.

[0039] The contents of vitamin B and vitamin C in Comparative Example 5 are significantly lower than those of the two examples, indicating that the process control conditions during the fermentation of the compound bacterial agent will affect the contents of vitamin B, vitamin C and polysaccharides in the rose extract, further proving that the segmented fermentation process adopted in this application can greatly improve the fermentation efficiency of the compound bacterial agent and increase the synthesis efficiency of vitamins B and C in the product.

[0040] In summary, the comparison of the contents of vitamins B1, B2, B6, C and polysaccharides in the rose extracts of Comparative Examples 1-5 and Examples 1-2 shows that the compound bacterial agent, segmented fermentation and ultrafiltration extraction processes adopted in this application all play important roles in increasing the contents of vitamins and polysaccharides in the rose extract.

[0041] Table 1 Comparison table of vitamin and polysaccharide contents in rose extracts Note: For the same component content group, the same letter indicates no significant difference (P>0.05), and different letters indicate significant difference (P<0.05).

[0042] Example 4: (An example of applying a rose extract to skin care products) Apply the rose extract obtained in Example 1 to skin care products according to Table 2; Table 2 Rose skin care essence formula table (1) Apply the rose extract obtained in Comparative Example 2 to skin care products according to Table 3; Table 3 Rose skin care essence formula table (2) Randomly select 30 people and let them try the rose skin care essence (formula in Table 2) and the rose skin care essence (formula in Table 3) respectively. 15 people try each formula. Let the subjects self-evaluate through a questionnaire survey, and detect the change of wrinkles of the testers through the MAX version of Mojit AI intelligent imager. Conduct a product use evaluation survey on the subjects at the time of testing on the 0th day and the 28th day; (3) Subjective evaluation results: As shown in Table 4, 100% of the people think that the overall face is improved after using the rose skin care essence with the formula in Table 1, and 47% of the people think that the overall face is improved after using the rose skin care essence with the formula in Table 2.

[0043] Table 4 Subjective evaluation results of trying the essence for 28 days It can be seen from Table 4 that the satisfaction of using the rose skin care essence with the formula in Table 2 is higher than that with the formula in Table 3. From Figure 3 and Figure 4 the human wrinkle change diagrams, it can be seen that the degree of wrinkle reduction in the forehead part of the human body using the formula in Table 2 is higher than that using the formula in Table 3. The formula in Table 2 adds the rose extract prepared in Example 1, and the formula in Table 3 adds the rose extract of Comparative Example 2. The rose extract of Example 1 has higher contents of vitamin B, vitamin C and polysaccharides than that of Comparative Example 2. These substances have the functions of firming and anti-wrinkle, moisturizing, repairing, etc. Therefore, with the same addition amount of rose extract, the improvement degree of the skin using the formula in Table 2 of Example 1 will be more obvious.

[0044] Example 5 (Antioxidant activity comparison experiment) Experimental principle: Reactive oxygen species (ROS) are oxygen-containing chemically reactive chemical substances. It includes peroxides, superoxides, hydroxyl free radicals, etc. ROS is a natural by-product of the normal metabolism of oxygen and plays an important role in cell signaling and homeostasis. However, during ultraviolet exposure, the level of ROS will increase sharply. This may cause serious damage to the cell structure, which is called oxidative stress.

[0045] CellROX® reagent is a DNA dye with weak fluorescence itself. It can be oxidized by ROS in cells, and the oxidation product binds to DNA to produce bright green fluorescence. CellROX® is soluble in organic solvents (such as dimethyl sulfoxide). Since the main component of zebrafish yolk sac is fat, CellROX® has strong permeability in the yolk sac and obvious staining in this part.

[0046] Menadione can produce reactive oxygen free radicals. When the amount of free radicals produced is greater than the body's scavenging ability, an oxidative stress reaction occurs. Evaluate the antioxidant efficacy of the sample through the fluorescence intensity of the zebrafish yolk sac.

[0047] The rose extracts provided in Example 1, Comparative Example 1, and Comparative Example 2 were selected for antioxidant activity detection, and the method was as follows: (1) Drug preparation: Prepare a menadione solution (model group) at 0.2 mg / mL, 0.5% N-acetyl-L-cysteine (NAC, positive group), and the rose extract samples provided in Example 1, Comparative Example 1, and Comparative Example 2; (2) Aspirate zebrafish into a 6-well plate, make up the volume to 3 mL with the standard diluent, and add drugs to each well after deducting the drug addition volume except for the blank group. Add 3 μL of menadione solution to the model group, add 60 μL of NAC solution and 3 μL of menadione solution to the positive group, and add 60 μL of the solution of Example 1 or Comparative Example 1 or Comparative Example 2 and 3 μL of menadione solution to the sample group; (3) After mixing, wrap it with tin foil and place it in the dark at 28 °C for 24 h; (4) Stain, cover the cover plate, wrap it with tin foil and place it in the dark in an incubator at 28 °C for 1-3 h; (5) Perform statistical processing using Dunnett ’s T-test. Taking the blank group as the standard, compare the yolk sac staining intensity of each experimental group. P<0.05 indicates a significant difference; (6) Calculate: Antioxidant efficacy (%) = ×100%; (7) Results: As shown in Table 5, the antioxidant activity of the rose extract provided in Example 1 was comparable to that of the positive group (containing 0.5% NAC, antioxidant) (P>0.05), 37.78% higher than that of Comparative Example 1 (P<0.01), and 35.21% higher than that of Comparative Example 2 (P<0.01).

[0048] From Figure 5 It can be seen that in the zebrafish antioxidant experiment, compared with the blank group, there was a significant difference in the fluorescence intensity of the yolk sac of zebrafish in the model group (P<0.05), indicating that the model was successfully established. Compared with the model group, there were significant differences in the fluorescence intensity of the yolk sac of zebrafish in the positive group and the sample group of Example 1 (P<0.05). The fluorescence intensity of the yolk sac of zebrafish in the sample groups of Comparative Example 1 and Comparative Example 2 was significantly higher than that of Example 1 (P<0.01), indicating that in the zebrafish antioxidant experiment, the rose extract provided in Example 1 had better antioxidant efficacy than Comparative Example 1 and Comparative Example 2. This was because the contents of vitamin B, vitamin C, and polysaccharides in the rose extract provided in Example 1 were higher, and these components had antioxidant effects. Therefore, under the condition of the same concentration of rose extract, the antioxidant activity of the rose extract provided in Example 1 was much greater than that of Comparative Example 1 and Comparative Example 2.

[0049] Therefore, the above data indicate that using the method provided in this application can significantly enhance the antioxidant activity of rose extract, which is more conducive to its application in the cosmetic field.

[0050] Table 5 Comparison Table of Antioxidant Activity of Rose Extract The embodiments of the present application have been described above. The above description is exemplary and not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The choice of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing a rose extract, characterized in that, the preparation method comprises the following steps: (1) Preparation of compound microbial agent: Lactobacillus plantarum ( L. plantarum ), Saccharomyces cerevisiae ( S. cerevisiae ), and Bacillus subtilis ( B. subtilis ) are proportioned according to the viable bacteria count ratio of (1-3):(1-2):(1-2) to obtain the compound microbial agent; (2) Prepare a rose matrix: Crush roses and mix them with deionized water to obtain a mixture. Add cellulase to the mixture for enzymatic hydrolysis, heat and sterilize, and then cool to form a rose matrix; (3) Inoculation: Inoculate the composite bacterium agent into the rose matrix, and the mass-volume ratio of the composite bacterium agent to the rose matrix is (0.2-2):100 (g / mL) to form a rose fermentation substrate; (4) Fermentation: Ferment the rose fermentation substrate obtained in (3) for 48-72 hours. The dissolved oxygen content in the first 12 hours is 50%-70%, and the dissolved oxygen content is controlled at 0-10% during the subsequent fermentation time. After fermentation is completed, rose dregs are formed, and the rose dregs are filtered to obtain a rose fermentation product; (5) Collection: Ultrafilter the rose fermentation product, and then centrifuge to collect the filtrate, which is the rose extract.

2. The preparation method according to claim 1, wherein In the step (1), the viable count of Lactobacillus plantarum is 1×10 7 cfu / g; the viable count of Saccharomyces cerevisiae is 2.8×10 7 cfu / g; the viable count of Bacillus subtilis is 1×10 7 cfu / g.

3. The preparation method according to claim 1, characterized in that, In the step (2), the material-liquid ratio of the crushed roses to deionized water is 1:(5-10) (g / mL).

4. The preparation method according to claim 1, characterized in that, In step (2), the material-liquid ratio of the cellulase to the mixture is (1-5):1000 (g / mL).

5. The preparation method according to claim 1, characterized in that, In step (2), the temperature of the enzymatic hydrolysis is 40-50°C, and the time of the enzymatic hydrolysis is 1-2 hours.

6. The preparation method according to claim 1, wherein In step (2), the temperature of the sterilization is 80°C, and the time is 15 minutes; the temperature of the cooling is 25-40°C.

7. The preparation method according to claim 1, characterized in that, In step (4), the temperature of the fermentation is 30-37°C, and the pH is 5.5-6.

5.

8. The preparation method according to claim 1, characterized in that, In step (4), the filtering method is to first filter the rose dregs with a 100-200 mesh filter material to obtain a filtrate, and then filter and sterilize the filtrate with a 0.2 μm sterilizing filter membrane.

9. The preparation method according to claim 1, wherein In step (5), the pore size of the ultrafiltration membrane for ultrafiltration is 0.01 μm; the rotation speed of the centrifugation is 8000-12000 rpm, and the time is 10-20 minutes.

10. Use of the rose extract obtained by the preparation method according to any one of claims 1-9 in food, health products or cosmetics.

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