Preparation method and application of rose extract

Abstract

This application belongs to the field of biotechnology, specifically to a preparation method and application of rose extract, which uses microbial fermentation technology to increase the content of water-soluble vitamins in rose. The strain selected in this method is Lactobacillus plantarum ( L. plantarum ), Saccharomyces cerevisiae ( S. cerevisiae ) and Bacillus subtilis ( B. subtle ), adjusting the strain ratio to achieve a viable bacterial count ratio of Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis of (1-3):(1-2):(1-2), forming a composite inoculum, which was then added to pretreated roses for staged fermentation. This ultimately increased the water-soluble vitamin content in the rose extract, while retaining rose polysaccharides and increasing the total antioxidant activity by approximately 35%. This rose extract is suitable for use as a raw material for functional foods, health products, and cosmetics.

Description

Technical Field

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

[0002] Roses, such as double roses and Damascus roses, are easy to cultivate on a large scale, have a long flowering period, produce a large biomass, and offer a stable supply of raw materials. Roses are rich in active ingredients such as phenolics (such as gallic acid and ellagic acid), polysaccharides, and flavonoids, which possess antioxidant and anti-inflammatory properties. Furthermore, these active substances can serve as substrates for microbial fermentation and, after microbial metabolism, be converted into water-soluble vitamins (such as B vitamins and vitamin C) or their derivatives. Water-soluble vitamins have the potential to improve skin barrier function, reduce inflammation, and inhibit melanin production. These vitamins can synergize with the active ingredients in roses to enhance the biological activity of rose extracts, resulting in enhanced whitening and anti-aging benefits. Consequently, "rose extract" is highly recognized in the food and cosmetics sectors.

[0003] However, existing fermentation processes are primarily optimized for the functional components of roses (such as polyphenols and flavonoids) rather than vitamin synthesis. Furthermore, water-soluble vitamins are easily destroyed by factors such as heat, light, and oxygen. Existing fermentation processes, such as high-temperature sterilization and prolonged fermentation, can reduce the content of water-soluble vitamins in rose extracts, thereby affecting the effectiveness of rose extracts in cosmetics. Furthermore, traditional methods for preparing rose extracts suffer from low efficiency and unstable yields, as well as chemical reagent residues and safety risks, high energy consumption, and environmental pollution. The use of strong acids / bases or prolonged heating during extraction can destroy the active ingredients in rose extracts. These issues have, to a certain extent, limited the application of rose extracts in food and health supplements.

[0004] In order to overcome the above problems, a new method for preparing rose extract is urgently needed, which can increase the content of water-soluble vitamins in rose extract while retaining polysaccharides, thereby expanding the function of rose extract from a single beauty function to the field of nutritional supplementation, and expanding the application of rose in food, health products and cosmetics. Summary of the Invention

[0005] In order to overcome the problems existing in the background technology, the present application provides a preparation method and application of rose extract, which increases the content of water-soluble vitamins in the rose extract through microbial fermentation without destroying the active substances in the rose itself.

[0006] To achieve the above object, the present invention provides a method for preparing a rose extract, the specific steps comprising:

[0007] (1) Preparation of composite bacterial agent: Lactobacillus plantarum ( L. plantarum ), Saccharomyces cerevisiae ( S. cerevisiae ) and Bacillus subtilis ( B. subtilis ) The ratio of viable bacteria count is (1-3): (1-2): (1-2) to obtain a composite bacterial agent;

[0008] (2) Preparation of rose matrix: crush rose flowers and mix them with deionized water to obtain a mixture, add cellulase to the mixture for enzymatic hydrolysis, heat sterilize, and then cool to form a rose matrix;

[0009] (3) Inoculation: inoculating the composite bacterial agent into the rose matrix, with the mass volume ratio of the composite bacterial agent to the rose matrix being (0.2-2):100 (g / mL), to form a rose fermentation substrate;

[0010] (4) Fermentation: The rose fermentation substrate obtained in (3) is fermented for 48-72 hours, with the dissolved oxygen content being 50%-70% in the first 12 hours and the dissolved oxygen content being controlled at 0-10% during the subsequent fermentation period. After the fermentation is completed, rose residue is formed, and the rose residue is filtered to obtain a rose fermentation product;

[0011] (5) Collection: The rose fermentation product is subjected to ultrafiltration, and the filtrate is collected by centrifugation to obtain the rose extract.

[0012] More preferably, in step (1), the number of viable Lactobacillus plantarum is 1×10 7 cfu / g; the number of viable bacteria of Saccharomyces cerevisiae is 2.8×10 7 cfu / g; the number of viable bacteria of Bacillus subtilis is 1×10 7 cfu / g.

[0013] Further preferably, in step (2), the material-liquid ratio of the crushed rose flowers to deionized water is 1:(5-10) (g / mL).

[0014] Further preferably, in step (2), the material-liquid ratio of the cellulase to the mixture is (1-5):1000 (g / mL).

[0015] More preferably, in step (2), the enzymatic hydrolysis temperature is 40-50° C., and the enzymatic hydrolysis time is 1-2 hours.

[0016] Further preferably, in step (2), the sterilization temperature is 80°C and the time is 15 minutes; and the cooling temperature is 25-40°C.

[0017] More preferably, in step (4), the fermentation temperature is 30-37°C and the pH is 5.5-6.5.

[0018] Further preferably, in step (4), the filtering method is to first filter the rose residue 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.

[0019] 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.

[0020] 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.

[0021] 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.

[0022] Beneficial effects of the present invention:

[0023] 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.

[0024] 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.

[0025] 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.

[0026] 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

[0027] Figure 1 This is a comparison chart of vitamin content of rose extracts in this application;

[0028] Figure 2 This is a comparison chart of the polysaccharide content of rose extracts in this application;

[0029] Figure 3 This is a diagram showing changes in human wrinkles using the formulation in Table 2 of Example 3 of this application;

[0030] Figure 4 This is a diagram showing changes in human wrinkles using the formulation in Table 3 of Example 3 of this application;

[0031] Figure 5 This is a comparison chart of the antioxidant activity of Example 4 of the present application. DETAILED DESCRIPTION

[0032] 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.

[0033] 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.

[0034] Unless otherwise specified, all reagents and materials used in the following examples were purchased from the market.

[0035] Example 1: (A method for preparing rose extract)

[0036] (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.), according to the viable count ratio of Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis of 2:1:1, accurately weighed on a 1 / 10,000 balance;

[0037] (2) Preparation of rose matrix: crush rose petals to 40 mesh, mix with deionized water at a material-liquid ratio of 1:7 (g / mL) to form a mixture, add cellulase at a ratio of cellulase to the mixture of 2:1000 (g / mL), enzymolysis at 45°C for 2 hours to release the substrate, heat sterilize at 80°C for 15 minutes, and cool to 25°C to form the rose matrix;

[0038] (3) Inoculation: inoculating the rose matrix described in step (2) at a mass volume ratio of the composite bacterial agent to the rose matrix of 1:100 (g / mL) to form a rose fermentation substrate;

[0039] (4) Fermentation: The rose fermentation substrate obtained in (3) is fermented at 30-37°C and pH 5.5-6.5 for 72 hours. Sterile air is introduced into the rose fermentation substrate for the first 12 hours to make the dissolved oxygen content 50%-70%. The substrate is kept in a closed system for the next 60 hours to control the dissolved oxygen content below 10%. After the fermentation is completed, rose residue is formed. The rose residue is first filtered through a 200-mesh filter material to obtain a filtrate, and the filtrate is then filtered and sterilized through a 0.2 μm sterilizing filter membrane to obtain a rose fermentation product.

[0040] (5) Collection: The rose fermentation product obtained in step (4) was filtered using 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 extract.

[0041] Example 2: (A method for preparing rose extract)

[0042] (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 7 cfu / g, purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.), according to the viable count ratio of Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis of 1:1:1, accurately weighed on a 1 / 10,000 balance;

[0043] (2) Preparation of rose matrix: crush rose petals to 60 mesh, mix with deionized water at a material-liquid ratio of 1:7 (g / mL) to form a mixture, add cellulase at a ratio of cellulase to the mixture of 2:1000 (g / mL), enzymolysis at 45°C for 2 hours to release the substrate, heat sterilize at 80°C for 15 minutes, and cool to 40°C to form the rose matrix;

[0044] (3) Inoculation: inoculating the rose matrix described in step (2) at a mass volume ratio of the composite bacterial agent to the rose matrix of 1:100 (g / mL) to form a rose fermentation substrate;

[0045] (4) Fermentation: The rose fermentation substrate obtained in (3) is fermented at 30-37°C and pH 5.5-6.5 for 48 hours. Sterile air is introduced into the rose fermentation substrate for the first 12 hours to make the dissolved oxygen content 50%-70%. The substrate is kept in a closed system for the next 36 hours to control the dissolved oxygen content below 10%. After the fermentation is completed, rose residue is formed. The rose residue is first filtered through a 200-mesh filter material to obtain a filtrate, and the filtrate is then filtered and sterilized through a 0.2 μm sterilizing filter membrane to obtain a rose fermentation product.

[0046] (5) Collection: The rose fermentation product obtained in step (4) was filtered using 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 extract.

[0047] Comparative Example 1:

[0048] (1) Rose petals were crushed into 40 mesh and mixed with deionized water at a material-liquid ratio of 1:7 (g / mL) to form a mixture. Cellulase was added at a ratio of 2:1000 (g / mL) to the mixture, and enzymatic hydrolysis was performed at 45°C for 2 hours to release the substrate.

[0049] (2) Ultrasonic treatment at 50°C for 30 min, ultrasonic power 300 W, frequency 40 kHz;

[0050] (3) Centrifuge the supernatant at 8000 rpm for 15 minutes and repeat the extraction once;

[0051] (4) Combine the supernatants to obtain rose extract.

[0052] Comparative Example 2:

[0053] A rose extract is prepared by directly fermenting the rose matrix in step (5) without adding a composite bacterial agent, and the remaining steps are the same as those in Example 1.

[0054] Comparative Example 3:

[0055] A rose extract is prepared in the same manner as in Example 1, except that the ratio of the viable bacteria of the composite bacterial agent Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis in step (2) is 4:1:1.

[0056] Comparative Example 4:

[0057] A rose extract is prepared in the same manner as in Example 1, except that the composite bacterial agents in steps (1) and (2) are set to be Lactobacillus plantarum and Saccharomyces cerevisiae, with the live bacteria count ratio being 2:1.

[0058] Comparative Example 5:

[0059] A rose extract, wherein the preparation method is the same as that of Example 1 except that the fermentation conditions in step (5) are changed to: fermentation at 30-37°C, pH 5.5-6.5, and a constant dissolved oxygen content of 0%-10% for 60 hours.

[0060] Example 3 (Determination of vitamin and polysaccharide content and data analysis)

[0061] 1. HPLC detection of vitamin content

[0062] A. Chromatographic conditions:

[0063] Chromatographic column: Alltima C18 column (250 mm × 4.6 mm, 5 μm);

[0064] Mobile phase: 50 mmol / L acetonitrile (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);

[0065] Flow rate: 0.5 mL / min;

[0066] Detection wavelength: 275 nm;

[0067] Column temperature: 30℃.

[0068] B. Sample preparation:

[0069] Accurately weigh appropriate amounts of vitamin B1, vitamin B2, vitamin B6, and vitamin C reference substances and prepare mixed reference substance stock solutions at approximately 2000 μg / mL each using acetonitrile-water (5:95). Accurately measure 1, 2, 2.5, 3, and 5 mL of the mixed reference substance stock solution into a 50 mL volumetric flask, dilute to the mark with acetonitrile-water (5:95), and shake well to obtain mixed reference substance solutions with concentrations of approximately 40, 80, 100, 120, and 200 μg / mL for each component.

[0070] C. Preparation of test solution:

[0071] Accurately measure 5 mL of rose extract and place it in a 100 mL volumetric flask. Add acetonitrile and water (5:95) to the volume, shake well, filter through a 0.22 μm microporous membrane, and take the filtrate as the test solution.

[0072] D. Content calculation:

[0073] The standard curve was plotted using the chromatographic conditions described in A, with peak area as the ordinate (Y) and concentration as the abscissa (X, mg / mL). The regression equation was obtained. The vitamin content in the rose extract was calculated as dry matter (%) using the following method:

[0074]

[0075] : Detection concentration calculated based on HPLC determination (μg / mL);

[0076] 20: dilution factor of the detection process;

[0077] : The moisture content (%) in rose extract can be quickly tested using a rapid moisture analyzer (sartorius, MA35M-000230V1).

[0078] 2. Determination of polysaccharide content in rose extract

[0079] The polysaccharide content detection method described in Example 6 of the patent specification ZL202410312477.7 was used. The polysaccharide content in the rose extract was calculated as dry matter (%) as follows:

[0080]

[0081] : polysaccharide concentration in the test sample, mg / mL;

[0082] 100: dilution factor of the detection process;

[0083] 0.9: Glucose to polysaccharide correction factor;

[0084] : The moisture content (%) in rose extract can be quickly tested using a rapid moisture analyzer (sartorius, MA35M-000230V1).

[0085] Data Analysis:

[0086] From Table 1 and Figure 1 、 Figure 2 It can be seen that:

[0087] The vitamin B, vitamin C, and polysaccharide contents in Comparative Examples 1 and 2 were relatively low overall. Neither group added a composite bacterial agent, indicating that the use of a composite bacterial agent can increase the vitamin B, vitamin C, and polysaccharide contents in rose extracts. Furthermore, Comparative Example 1 employed a traditional rose extract preparation process. In addition to not adding a composite bacterial agent, the extraction method used was also a traditional ultrasonic extraction process. Therefore, the polysaccharide content in Comparative Example 1 was significantly different from that in both examples. This demonstrates that the ultrafiltration extraction process employed in this application effectively retains the polysaccharide components in the rose extract and plays a significant role in increasing the activity of the rose extract.

[0088] Example 1 had the highest levels of vitamin B and vitamin C, followed by Example 2 and Comparative Example 3. The difference between Example 2 and Comparative Example 3 and Example 1 was the different ratios of viable bacterial counts in the composite bacterial inoculum, indicating that differences in viable bacterial count ratios can affect the levels of vitamin B and vitamin C. Furthermore, the vitamin content in the extract of Comparative Example 3 was lower than in both examples, demonstrating that the composite bacterial inoculum ratio selected in this application is superior. Furthermore, the vitamin B and vitamin C contents in Comparative Example 4 were significantly lower than those in Example 1, indicating that the species of the composite bacterial inoculum can affect the levels of vitamin B and vitamin C in the rose extract. Therefore, these data demonstrate that the species and ratio of the composite bacterial inoculum selected in this application play an important role in increasing the content of water-soluble vitamins in the rose extract.

[0089] The contents of vitamin B and vitamin C in Comparative Example 5 were significantly lower than those in the two examples, indicating that the process control conditions of the composite bacterial agent fermentation process affect the contents of vitamin B, vitamin C and polysaccharides in the rose extract, further proving that the staged fermentation process adopted in this application can greatly improve the fermentation efficiency of the composite bacterial agent and improve the synthesis efficiency of vitamins B and C in the product.

[0090] 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 composite bacterial agent, staged fermentation and ultrafiltration extraction processes adopted in this application all play an important role in increasing the content of vitamins and polysaccharides in the rose extract.

[0091] Table 1 Comparison of vitamin and polysaccharide contents in rose extracts

[0092]

[0093] Note: The same letters in the same component content groups indicate no significant difference (P>0.05), and different letters indicate significant difference (P<0.05).

[0094] Example 4: (An example of using rose extract in skin care products)

[0095] The rose extract obtained in Example 1 was applied to skin care products according to Table 2;

[0096] Table 2 Rose Skin Care Essence Formula

[0097]

[0098] (1) The rose extract obtained in Comparative Example 2 was applied to skin care products according to Table 3;

[0099] Table 3 Rose Skin Care Essence Formula

[0100]

[0101] (2) 30 people were randomly selected to try Rose Skin Care Essence (Formula in Table 2) and Rose Skin Care Essence (Formula in Table 3), with 15 people trying each formula. The subjects were asked to self-evaluate through a questionnaire survey, and the changes in wrinkles of the trial subjects were detected using the MAX version of Moji AI intelligent imager. The product usage evaluation survey was conducted on the subjects on the 0th day and the 28th day;

[0102] (3) Subjective evaluation results: As shown in Table 4, 100% of the respondents believed that their overall facial appearance improved after using the rose skin care essence formulated in Table 1, and 47% of the respondents believed that their overall facial appearance improved after using the rose skin care essence formulated in Table 2.

[0103] Table 4 Subjective evaluation results of the serum after 28-day trial

[0104]

[0105] As shown in Table 4, the satisfaction of the rose skin care essence using the formula in Table 2 is higher than that using the formula in Table 3. Figure 3 and Figure 4 As can be seen from the wrinkle change chart, the formula in Table 2 shows a greater reduction in forehead wrinkles than the formula in Table 3. The formula in Table 2 incorporates the rose extract prepared in Example 1, while the formula in Table 3 incorporates the rose extract from Comparative Example 2. The rose extract in Example 1 contains higher levels of vitamin B, vitamin C, and polysaccharides than the rose extract in Comparative Example 2. These substances have firming, anti-wrinkle, moisturizing, and repairing properties. Therefore, at the same rose extract dosage, the formula in Table 2 in Example 1 produces more pronounced skin improvements.

[0106] Example 5 (Comparative Experiment on Antioxidant Activity)

[0107] Principle: Reactive oxygen species (ROS) are chemically reactive species containing oxygen. They include peroxides, superoxides, and hydroxyl radicals. ROS are natural byproducts of normal oxygen metabolism and play an important role in cell signaling and homeostasis. However, during UV exposure, ROS levels increase dramatically. This can cause severe damage to cellular structures, a condition known as oxidative stress.

[0108] CellROX® reagent is a DNA dye with weak intrinsic fluorescence. It can be oxidized by intracellular ROS. The oxidation products bind to DNA, producing bright green fluorescence. CellROX® is soluble in organic solvents (such as dimethyl sulfoxide). Since the zebrafish yolk sac is primarily composed of fat, CellROX® has strong permeability in the yolk sac, resulting in distinct staining in this area.

[0109] Menadione can produce reactive oxygen free radicals. When the amount of free radicals produced is greater than the body's clearance capacity, an oxidative stress reaction occurs. The antioxidant efficacy of the sample was evaluated by the fluorescence intensity of the zebrafish yolk sac.

[0110] The rose extracts provided in Example 1, Comparative Example 1, and Comparative Example 2 were used to test the antioxidant activity, and the method was as follows:

[0111] (1) Drug preparation: Prepare 0.2 mg / mL menadione solution (model group), 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;

[0112] (2) The zebrafish were aspirated into a 6-well plate, and the standard dilution solution was adjusted to 3 mL. The drugs were added to each well after deducting the drug volume except for the blank group. 3 μl of menadione solution was added to the model group, 60 μl of NAC solution and 3 μl of menadione solution were added to the positive group, and 60 μl of Example 1 or Comparative Example 1 or Comparative Example 2 solution and 3 μl of menadione solution were added to the sample group;

[0113] (3) After mixing, wrap with tin foil and incubate at 28°C in the dark for 24 h;

[0114] (4) Staining: Cover the plate, wrap it with tin foil and place it in a 28°C incubator to avoid light and stain for 1-3 hours;

[0115] (5) Dunnett's T-test was used for statistical analysis. The blank group was used as the standard to compare the yolk sac staining intensity of each experimental group. P < 0.05 was considered to be a significant difference.

[0116] (6) Calculation: Antioxidant efficacy (%) = ×100%;

[0117] (7) Results:

[0118] 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).

[0119] from Figure 5 As can be seen in the zebrafish antioxidant experiment, the zebrafish yolk sac fluorescence intensity in the model group was significantly different from that in the blank group (P < 0.05), indicating that the model was successfully established. The zebrafish yolk sac fluorescence intensity in the positive group and the sample group in Example 1 was significantly different from that in the model group (P < 0.05). The zebrafish yolk sac fluorescence intensity in the sample groups in Comparative Examples 1 and 2 was significantly higher than that in Example 1 (P < 0.01), indicating that the rose extract provided in Example 1 exhibited superior antioxidant efficacy compared to Comparative Examples 1 and 2 in the zebrafish antioxidant experiment. This is due to the higher content of vitamin B, vitamin C, and polysaccharides in the rose extract provided in Example 1, which have antioxidant effects. Therefore, at the same rose extract concentration, the rose extract provided in Example 1 exhibited significantly greater antioxidant activity than those in Comparative Examples 1 and 2.

[0120] Therefore, the above data indicate that the method provided in this application can significantly enhance the antioxidant activity of rose extract, which is more conducive to its application in the cosmetics field.

[0121] Table 5 Comparison of antioxidant activity of rose extracts

[0122]

[0123] The embodiments of the present application have been described above. The above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is selected to best explain the principles of the embodiments, their practical applications, or improvements to the technology in the market, or to enable other persons skilled 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 composite bacterial agent: Lactobacillus plantarum ( L. plantarum ), Saccharomyces cerevisiae ( S. cerevisiae ) and Bacillus subtilis ( B. subtilis ) The ratio of viable bacteria count is (1-3): (1-2): (1-2) to obtain a composite bacterial agent; (2) Preparation of rose matrix: crush rose flowers and mix them with deionized water to obtain a mixture, add cellulase to the mixture for enzymatic hydrolysis, heat sterilize, and then cool to form a rose matrix; (3) Inoculation: inoculating the composite bacterial agent into the rose matrix, with the mass volume ratio of the composite bacterial agent to the rose matrix being (0.2-2):100 (g / mL), to form a rose fermentation substrate; (4) Fermentation: fermenting the rose fermentation substrate obtained in (3) at 30-37° C. and pH 5.5-6.5 for 48-72 hours, with the dissolved oxygen content being 50%-70% for the first 12 hours and then being controlled at 0-10% for the subsequent fermentation period. After the fermentation is completed, rose residue is formed, and the rose residue is filtered to obtain a rose fermentation product; (5) Collection: The rose fermentation product is subjected to ultrafiltration, and the filtrate is collected by centrifugation to obtain the rose extract.

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

3. The preparation method according to claim 1, characterized in that In step (2), the material-liquid ratio of the crushed rose flowers 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 enzymatic hydrolysis temperature is 40-50° C., and the enzymatic hydrolysis time is 1-2 hours.

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

7. The preparation method according to claim 1, characterized in that In step (4), the filtration method is to first filter the rose residue 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.

8. The preparation method according to claim 1, characterized in that 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.

9. Use of the rose extract obtained by the preparation method according to any one of claims 1 to 8 in food, health products or cosmetics.

Images