A preparation method of Rosa roxburghii Tratt fermentation broth for relieving hyperuricemia

Abstract

The present invention relates to the technical field of Rosa roxburghii Tratt processing. In order to solve the problem that it is impossible to simultaneously improve the SOD activity and flavonoid content in Rosa roxburghii Tratt and reduce the fructose content under the existing technology, a preparation method of a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia is provided, which includes the following steps: mixing Rosa roxburghii Tratt with water and then juicing, removing solids and adjusting the pH with edible alkali to obtain a Rosa roxburghii Tratt blended juice; after sterilizing the Rosa roxburghii Tratt blended juice, adding a cell suspension of a compound Lactobacillus for fermentation, and removing solids after fermentation to obtain a Rosa roxburghii Tratt fermentation broth; the compound Lactobacillus is a mixture of Lactobacillus rhamnosus and Lactobacillus plantarum. This preparation method can improve the SOD activity and flavonoid content in Rosa roxburghii Tratt and reduce the fructose content, and the obtained Rosa roxburghii Tratt fermentation broth has the effect of alleviating hyperuricemia.

Description

Technical Field

[0001] The present invention relates to the technical field of Rosa roxburghii Tratt processing, and particularly relates to a method for preparing a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia. Background Art

[0002] With the rapid economic development and the change of dietary structure, the increasing intake of purine-rich foods has led to an increasing prevalence of hyperuricemia, bringing a great burden to society. At present, the drugs for treating hyperuricemia clinically include allopurinol, probenecid, benzbromarone, and febuxostat, but long-term use can cause serious side effects such as liver and kidney damage and allergic reactions. Therefore, it is necessary to develop more effective and non-toxic natural anti-hyperuricemia products.

[0003] Rosa roxburghii Tratt is a plant with both medicinal and edible properties, rich in various active substances such as vitamin C, superoxide dismutase (SOD), polysaccharides, flavonoids, and triterpenoids. SOD is an antioxidant enzyme that can catalyze the dismutation of superoxide radicals into hydrogen peroxide and oxygen, thereby scavenging free radicals and protecting cells or tissues from oxidative damage. Flavonoids are the second largest bioactive substances in Rosa roxburghii Tratt after vitamin C. They are important highly active natural substances with antioxidant, antibacterial, anti-inflammatory, and antiviral effects. Some studies have also shown that flavonoids can be used to treat hyperuricemia. However, Rosa roxburghii Tratt also contains a large amount of fructose. Studies have confirmed that excessive intake of fructose will have adverse effects on the cardiovascular system and metabolism, leading to obesity and metabolic syndrome, and will also increase the serum uric acid level and induce hyperuricemia. Therefore, untreated Rosa roxburghii Tratt cannot fully exert its anti-hyperuricemia effect.

[0004] In the currently disclosed domestic research on the preparation and application of Rosa roxburghii Tratt fermentation broth, CN109105889A discloses a preparation process and application of fermented Rosa roxburghii Tratt juice. Using a composite lactic acid bacteria including Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus casei, and Acetobacter to ferment Rosa roxburghii Tratt juice, the DPPH free radical scavenging ability and the immunity of mice of the Rosa roxburghii Tratt fermentation broth are significantly enhanced. CN108523109B discloses a method for fermenting Rosa roxburghii Tratt. Using a strain mixture including Lactobacillus casei strains, Bifidobacterium longum strains, and Saccharomyces cerevisiae strains to ferment Rosa roxburghii Tratt samples, a Rosa roxburghii Tratt fermentation broth with improved antioxidant ability can be obtained. However, in the existing Rosa roxburghii Tratt fermentation methods, only the fermentation ability of the strains is considered, and the safety and functions of the strains are not evaluated. Moreover, the Rosa roxburghii Tratt fermentation broth prepared by the existing strains and fermentation methods cannot simultaneously improve the SOD activity and flavonoid content, reduce the fructose content, and show the effect of alleviating hyperuricemia. Summary of the Invention

[0005] In order to overcome the problem in the prior art that it is impossible to simultaneously increase the SOD activity and flavonoid content in Rosa roxburghii tratt while reducing the fructose content, the present invention provides a method for preparing a Rosa roxburghii tratt fermentation broth for alleviating hyperuricemia. This preparation method can increase the SOD activity and flavonoid content in Rosa roxburghii tratt and reduce the fructose content, and the obtained Rosa roxburghii tratt fermentation broth has the effect of alleviating hyperuricemia.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for preparing a Rosa roxburghii tratt fermentation broth for alleviating hyperuricemia, comprising the following steps:

[0008] (1) Mix Rosa roxburghii tratt with water, juice, remove solids, and adjust the pH with edible alkali to obtain a Rosa roxburghii tratt blended juice;

[0009] (2) After sterilizing the Rosa roxburghii tratt blended juice, add a cell suspension of compound Lactobacillus and ferment, and remove solids after fermentation to obtain a Rosa roxburghii tratt fermentation broth.

[0010] The present invention uses compound probiotics to ferment Rosa roxburghii tratt, reducing the fructose content in Rosa roxburghii tratt, while increasing the SOD activity and flavonoid content in Rosa roxburghii tratt. The obtained Rosa roxburghii tratt fermentation broth can alleviate hyperuricemia, significantly reduce the levels of blood uric acid, urea nitrogen, malondialdehyde and xanthine oxidase, and has the effects of antioxidation and improvement of kidney injury.

[0011] Preferably, the compound Lactobacillus in (2) is a mixture of Lactobacillus rhamnosus and Lactobacillus plantarum.

[0012] Both Lactobacillus rhamnosus and Lactobacillus plantarum are derived from acidic foods and are probiotics with health effects evaluated in vitro and in vivo. On the basis of screening out Lactobacillus rhamnosus and Lactobacillus plantarum, the present invention uses Lactobacillus rhamnosus and Lactobacillus plantarum in combination to further improve the fermentation effect.

[0013] Preferably, the Lactobacillus rhamnosus is Lactobacillus rhamnosus 1155 CGMCC NO.11955, and the Lactobacillus plantarum is Lactobacillus plantarum 581 CGMCC No.13121.

[0014] Preferably, (1) is to mix Rosa roxburghii tratt with water, juice, filter, centrifuge and concentrate to obtain a Rosa roxburghii tratt raw juice, add water to the Rosa roxburghii tratt raw juice and adjust the pH with edible alkali to obtain a Rosa roxburghii tratt blended juice.

[0015] The Rosa roxburghii tratt raw juice can be stored frozen for later use after sterilization.

[0016] Preferably, (1) is to mix Rosa roxburghii Tratt with water that is 2 - 16 times the mass of Rosa roxburghii Tratt, then juice, filter through a 60 - 100 mesh filter, centrifuge, and concentrate to obtain Rosa roxburghii Tratt raw juice. Mix the Rosa roxburghii Tratt raw juice with water at a volume ratio of 1:(0.5 - 4), and adjust the pH with edible alkali.

[0017] Preferably, in (1), the pH is adjusted to 4.0 - 8.0 with edible alkali, and the edible alkali is food - grade Na2CO3 and / or food - grade NaHCO3.

[0018] Preferably, in (2), the sterilization process is to sterilize the Rosa roxburghii Tratt blended juice at 80 - 95°C for 5 - 20 minutes and then cool it.

[0019] Preferably, in (2), the cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum are obtained by the following steps: Inoculate Lactobacillus rhamnosus and Lactobacillus plantarum into sterilized MRS liquid medium respectively, incubate at 30 - 37°C for 12 - 24 h, activate for 2 generations, then centrifuge at 5000 r / min for 10 - 20 min, discard the supernatant, collect the cells respectively, and finally adjust the cell concentration of the cell suspension to 10 6 CFU / mL or more to obtain the cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum. Then mix the cell suspension of Lactobacillus rhamnosus and the cell suspension of Lactobacillus plantarum at a volume ratio of 1:(0.5 - 4) to obtain the cell suspension of the compounded lactobacilli.

[0020] Preferably, in (2), after sterilizing the Rosa roxburghii Tratt blended juice, add the cell suspension of the compounded lactobacilli to make the initial inoculation amount greater than 10 6 CFU and then carry out fermentation.

[0021] Preferably, the fermentation condition in (2) is constant - temperature fermentation at 37°C for 24 - 120 hours.

[0022] More preferably, the fermentation condition in (2) is constant - temperature fermentation at 37°C for 24 - 120 hours, and then transfer to 4°C for after - ripening for 1 - 4 days.

[0023] After fermentation, transferring to 4°C for after - ripening can further improve the flavor of the Rosa roxburghii Tratt fermentation broth.

[0024] Preferably, in (2), after fermentation, centrifuge at a speed of 4000 - 8000 rpm / min and collect the supernatant to obtain the Rosa roxburghii Tratt fermentation broth.

[0025] Therefore, the present invention has the following beneficial effects: (1) Applying the fermentation technology of specific compound probiotics to the fermentation of Rosa roxburghii tratt, optimizing the fermentation conditions to increase the SOD activity, improve the flavonoid content and reduce the fructose content in the fermented Rosa roxburghii tratt juice; (2) Enhancing the antioxidant capacity and uric acid-lowering capacity of the obtained Rosa roxburghii tratt juice, showing the effect of alleviating hyperuricemia in in vivo animal experiments. Specific embodiments

[0026] The present invention will be further described below in conjunction with specific implementation methods.

[0027] Example 1

[0028] A Rosa roxburghii tratt fermentation broth is prepared by the following steps:

[0029] (1) Preparation of Rosa roxburghii tratt raw juice

[0030] Select Rosa roxburghii tratt without rotten fruits, then wash and remove impurities, cut the washed Rosa roxburghii tratt into small pieces, add water at twice the mass of Rosa roxburghii tratt, put it into a juicer for juicing, then filter through a 100-mesh filter screen, centrifuge and concentrate, and finally sterilize to obtain Rosa roxburghii tratt raw juice, which is stored frozen for later use;

[0031] (2) Preparation of Rosa roxburghii tratt blended juice

[0032] Mix the Rosa roxburghii tratt raw juice and water at a volume ratio of 1:1, adjust the pH to 4 with food-grade Na2CO3, sterilize at 80 °C for 15 minutes, and cool to room temperature as Rosa roxburghii tratt blended juice for later use;

[0033] (3) Activation of strains and preparation of bacterial suspensions

[0034] Inoculate Lactobacillus rhamnosus (Lactobacillus rhamnosus 1155) and Lactobacillus plantarum (Lactobacillus plantarum 581) into sterilized MRS liquid medium respectively, incubate at 37 °C for 16 h respectively, activate for 2 generations, then centrifuge at 5000 r / min for 15 min, discard the supernatant, collect the bacterial cells respectively, and finally adjust the bacterial liquid concentration to 10 6 CFU / mL or more to obtain the bacterial cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum respectively;

[0035] (4) Fermentation

[0036] Mix the bacterial cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum at a volume ratio of 1:0.5 to prepare a compound bacterial cell suspension, then inoculate it into the Rosa roxburghii tratt blended juice, ensure that the initial inoculation amount is 10 6 CFU or more, then ferment at a constant temperature of 37 °C for 24 hours, transfer to 4 °C for after-ripening for 1 day to obtain Rosa roxburghii tratt fermentation broth;

[0037] (5) Clarification treatment

[0038] The Rosa roxburghii Tratt fermentation broth was centrifuged at 5000 rpm / min, and the supernatant was collected to obtain a clarified Rosa roxburghii Tratt fermentation broth.

[0039] Example 2

[0040] A Rosa roxburghii Tratt fermentation broth is prepared by the following steps:

[0041] (1) Preparation of Rosa roxburghii Tratt raw juice

[0042] Select Rosa roxburghii Tratt fruits without rot, then wash to remove impurities, cut the washed Rosa roxburghii Tratt fruits into small pieces, add water at 4 times the mass of Rosa roxburghii Tratt, put them into a juicer for juicing, then filter through a 100-mesh sieve, centrifuge and concentrate, and finally sterilize to obtain Rosa roxburghii Tratt raw juice, which is stored frozen for later use;

[0043] (2) Preparation of Rosa roxburghii Tratt blended juice

[0044] Mix the Rosa roxburghii Tratt raw juice and water at a volume ratio of 1:1, and adjust the pH to 5 with food-grade Na2CO3; sterilize at 80 °C for 10 minutes and cool to room temperature for use as Rosa roxburghii Tratt blended juice;

[0045] (3) Activation of strains and preparation of bacterial suspensions

[0046] Lactobacillus rhamnosus (Lactobacillus rhamnosus 1155) and Lactobacillus plantarum (Lactobacillus plantarum 581) were respectively inoculated into sterilized MRS liquid medium, cultured at 37 °C for 16 h and activated for 2 generations, then centrifuged at 5000 r / min for 15 min, the supernatant was discarded, the bacterial cells were collected respectively, and finally the concentration of the bacterial suspension was adjusted to 10 6 CFU / mL or more to obtain the bacterial suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum respectively;

[0047] (4) Fermentation

[0048] The bacterial suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum were mixed at a volume ratio of 1:1 to prepare a composite bacterial suspension, which was then inoculated into Rosa roxburghii Tratt blended juice to ensure that the initial inoculation amount was 10 6 CFU or more, and then fermented at a constant temperature of 37 °C for 48 hours, transferred to 4 °C for after-ripening for 2 days to obtain Rosa roxburghii Tratt fermentation broth;

[0049] (5) Clarification treatment

[0050] The Rosa roxburghii Tratt fermentation broth was centrifuged at 6000 rpm / min, and the supernatant was collected to obtain a clarified Rosa roxburghii Tratt fermentation broth.

[0051] Example 3

[0052] A Rosa roxburghii Tratt fermentation broth is prepared by the following steps:

[0053] (1) Preparation of Rosa roxburghii Tratt raw juice

[0054] Select Rosa roxburghii Tratt fruits without rot, then wash and remove impurities. Cut the washed Rosa roxburghii Tratt fruits into small pieces, add water at 8 times the mass of Rosa roxburghii Tratt, put them into a juicer for juicing, then filter through a 100-mesh sieve, centrifuge and concentrate. Finally, perform sterilization treatment to obtain Rosa roxburghii Tratt raw juice, which is stored frozen for later use;

[0055] (2) Preparation of Rosa roxburghii Tratt blended juice

[0056] Mix the Rosa roxburghii Tratt raw juice and water at a volume ratio of 1:1, adjust the pH to 4 with food-grade Na2CO3, sterilize at 95 °C for 5 minutes, and cool to room temperature for later use as Rosa roxburghii Tratt blended juice;

[0057] (3) Activation of strains and preparation of bacterial suspensions

[0058] Inoculate Lactobacillus rhamnosus (Lactobacillus rhamnosus 1155) and Lactobacillus plantarum (Lactobacillus plantarum 581) into sterilized MRS liquid medium respectively, incubate at 37 °C for 24 h respectively, activate for 2 generations, then centrifuge at 5000 r / min for 15 min, discard the supernatant, collect the bacterial cells respectively, and finally adjust the bacterial suspension concentration to 10 6 CFU / mL or more to obtain the bacterial cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum respectively;

[0059] (4) Fermentation

[0060] Mix the bacterial cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum at a volume ratio of 1:2 to obtain a composite bacterial cell suspension, then inoculate it into the Rosa roxburghii Tratt blended juice, ensure that the initial inoculation amount is 10 6 CFU or more, then ferment at a constant temperature of 37 °C for 72 hours, transfer to 4 °C for after-ripening for 3 days to obtain Rosa roxburghii Tratt fermentation broth;

[0061] (5) Clarification treatment

[0062] Centrifuge the Rosa roxburghii Tratt fermentation broth at a speed of 7000 rpm / min and collect the supernatant to obtain clarified Rosa roxburghii Tratt fermentation broth.

[0063] Example 4

[0064] A Rosa roxburghii Tratt fermentation broth is prepared by the following steps:

[0065] (1) Preparation of Rosa roxburghii Tratt raw juice

[0066] Select Rosa roxburghii fruits without rotten ones, then wash and remove impurities. Cut the washed Rosa roxburghii fruits into small pieces, add water at 16 times the mass of Rosa roxburghii fruits, put them into a juicer for juicing, then filter through a 100-mesh sieve, centrifuge and concentrate. Finally, perform sterilization treatment to obtain Rosa roxburghii raw juice, which is stored frozen for later use;

[0067] (2) Preparation of Rosa roxburghii blended juice

[0068] Mix the Rosa roxburghii raw juice and water at a volume ratio of 1:1, adjust the pH to 7 with food-grade Na2CO3, sterilize at 95°C for 10 minutes, and cool to room temperature for use as Rosa roxburghii blended juice;

[0069] (3) Activation of strains and preparation of bacterial suspensions

[0070] Inoculate Lactobacillus rhamnosus (Lactobacillus rhamnosus 1155) and Lactobacillus plantarum (Lactobacillus plantarum 581) into sterilized MRS liquid medium respectively, incubate at 37°C for 24 hours each, activate for 2 generations, then centrifuge at 5000 r / min for 15 minutes, discard the supernatant, collect the bacterial cells respectively, and finally adjust the bacterial suspension concentration to above 10 6 CFU / mL with sterile normal saline to obtain bacterial suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum respectively;

[0071] (4) Fermentation

[0072] Mix the bacterial suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum at a volume ratio of 1:3 to obtain a composite bacterial suspension, then inoculate it into Rosa roxburghii blended juice, ensuring the initial inoculation amount is above 10 6 CFU, then ferment at a constant temperature of 37°C for 96 hours, transfer to 4°C for after-ripening for 1 day to obtain Rosa roxburghii fermented liquid;

[0073] (5) Clarification treatment

[0074] Centrifuge the Rosa roxburghii fermented liquid at 8000 rpm / min and collect the supernatant to obtain clarified Rosa roxburghii fermented liquid.

[0075] Comparative example 1

[0076] A kind of Rosa roxburghii blended juice is prepared by the following steps:

[0077] (1) Preparation of Rosa roxburghii raw juice

[0078] Select Rosa roxburghii fruits without rotten ones, then wash and remove impurities. Cut the washed Rosa roxburghii fruits into small pieces, add water at 16 times the mass of Rosa roxburghii fruits, put them into a juicer for juicing, then filter through a 100-mesh sieve, centrifuge and concentrate. Finally, perform sterilization treatment to obtain Rosa roxburghii raw juice, which is stored frozen for later use;

[0079] (2) Preparation of Rosa roxburghii blended juice

[0080] Mix the raw juice of Rosa roxburghii tratt with water at a volume ratio of 1:1, and store it refrigerated as the Rosa roxburghii tratt blended juice for standby.

[0081] Comparative Example 2

[0082] A Rosa roxburghii tratt fermentation broth, the difference in its preparation steps from those of Example 4 lies in:

[0083] (3) Activation of the strain and preparation of the bacterial suspension

[0084] Inoculate Lactobacillus fermentum 2644 CGMCC NO.16754 into the sterilized MRS liquid medium, incubate at 37 °C for 24 h respectively, activate for 2 generations, then centrifuge at 5000 r / min for 15 min, discard the supernatant, collect the bacterial cells, and finally adjust the concentration of the bacterial suspension to 10 6 CFU / mL or more to obtain the bacterial cell suspension of Lactobacillus plantarum;

[0085] (4) Fermentation

[0086] Inoculate the bacterial cell suspension of Lactobacillus fermentum into the Rosa roxburghii tratt blended juice, ensure that the initial inoculation amount is 10 6 CFU or more, then ferment at a constant temperature of 37 °C for 96 hours, transfer to 4 °C for after-ripening for 1 day to obtain the Rosa roxburghii tratt fermentation broth.

[0087] Comparative Example 3

[0088] A Rosa roxburghii tratt fermentation broth, the difference in its preparation steps from those of Example 4 lies in:

[0089] (3) Activation of the strain and preparation of the bacterial suspension

[0090] Inoculate Lactobacillus rhamnosus (Lactobacillus rhamnosus 1155) into the sterilized MRS liquid medium, incubate at 37 °C for 24 h respectively, activate for 2 generations, then centrifuge at 5000 r / min for 15 min, discard the supernatant, collect the bacterial cells, and finally adjust the concentration of the bacterial suspension to 10 6 CFU / mL or more to obtain the bacterial cell suspension of Lactobacillus rhamnosus;

[0091] (4) Fermentation

[0092] Inoculate the bacterial cell suspension of Lactobacillus rhamnosus into the Rosa roxburghii tratt blended juice, ensure that the initial inoculation amount is 10 6 CFU or more, then ferment at a constant temperature of 37 °C for 96 hours, transfer to 4 °C for after-ripening for 1 day to obtain the Rosa roxburghii tratt fermentation broth.

[0093] Comparative Example 4

[0094] A Rosa roxburghii Tratt fermentation broth, the difference in its preparation steps from Example 4 lies in:

[0095] (3) Strain activation and preparation of bacterial suspension

[0096] Inoculate Lactobacillus plantarum (Lactobacillus plantarum 581) into the sterilized MRS liquid medium, cultivate at 37°C for 24 hours respectively, activate for 2 generations, then centrifuge at 5000 r / min for 15 minutes, discard the supernatant, collect the bacterial cells, and finally adjust the concentration of the bacterial suspension to 10 6 CFU / mL or more to obtain a bacterial cell suspension of Lactobacillus plantarum;

[0097] (4) Fermentation

[0098] Inoculate the bacterial cell suspension of Lactobacillus plantarum into the Rosa roxburghii Tratt blended juice, inoculate into the Rosa roxburghii Tratt blended juice, ensure that the initial inoculation amount is 10 6 CFU or more, then ferment at a constant temperature of 37°C for 96 hours, transfer to 4°C for after-ripening for 1 day to obtain the Rosa roxburghii Tratt fermentation broth.

[0099] I. Nutritional component detection

[0100] Test the content of nutritional components in the Rosa roxburghii Tratt fermentation broth obtained in the above examples and comparative examples, and the results are shown in Table 1.

[0101] Table 1: Test results of nutritional component content

[0102]

[0103] As can be seen from Table 1, in Examples 1 to 4, the method of the present invention is adopted to ferment Rosa roxburghii Tratt with compound probiotics, which can effectively improve the SOD activity, vitamin C, polyphenol and total flavonoid contents in the Rosa roxburghii Tratt fermentation broth and reduce the fructose content. In Comparative Example 1, Rosa roxburghii Tratt is not fermented. Compared with other examples, the SOD activity, vitamin C, polyphenol and flavonoid contents in the Rosa roxburghii Tratt blended juice are significantly reduced, and the fructose content is significantly increased. Comparative Examples 2 to 4 use a single lactic acid bacterium. Among them, the improvement effect of Comparative Example 2 on the SOD and flavonoid contents is weaker than that of Comparative Example 3 and Comparative Example 4, which indicates that the fermentation effects of Lactobacillus rhamnosus and Lactobacillus plantarum are better than that of Lactobacillus fermentum belonging to the same lactic acid bacteria. At the same time, comparing Comparative Example 3, Comparative Example 4 and Example 4, it can be seen that the SOD activity and total flavonoid contents in the Rosa roxburghii Tratt fermentation broth obtained in Comparative Example 3 and Comparative Example 4 are both lower than those in Example 4 fermented with compound lactic acid bacteria, while the fructose content is significantly higher than that in Example 4, which indicates that Lactobacillus rhamnosus and Lactobacillus plantarum have a synergistic effect.

[0104] II. Bioactivity detection

[0105] 1. Xanthine oxidase (XOD) inhibitory activity

[0106] XOD is a key enzyme in uric acid formation. For patients with hyperuricemia, inhibiting the activity of XOD can inhibit the elevated serum uric acid concentration. Therefore, the XOD inhibitory activity of the Rosa roxburghii Tratt fermentation broth obtained in Example 2 and the comparative examples was tested to evaluate its uric acid-lowering activity. The test method is as follows:

[0107] 1.1. Prepare 50 mM PBS solution at pH 7.8: Weigh 358.14 g of disodium hydrogen phosphate dodecahydrate and make up to 1 L to obtain 1 M disodium hydrogen phosphate solution. Weigh 156.01 g of sodium dihydrogen phosphate and make up to 1 L to obtain 1 M sodium dihydrogen phosphate solution. Mix 896 mL of disodium hydrogen phosphate solution with 104 mL of sodium dihydrogen phosphate solution, and dilute the mixture 2-fold to 50 mM PBS solution, then adjust the pH to 7.8 with 2 M NaOH;

[0108] 1.2. 110 μM xanthine solution:

[0109] Weigh 0.016 g of xanthine, first dissolve it in 0.02 M sodium hydroxide in a water bath at 50 °C, then make up to 100 mL with PBS, and dilute it 10-fold to obtain 110 μM xanthine solution;

[0110] 1.3. 0.027 U / mL XOD solution

[0111] Centrifuge at 12000 rpm at 4 °C for 10 min, discard the supernatant, resuspend the precipitate in 5 mL of PBS and mix well to prepare 1 U / mL, and then dilute it to obtain 0.027 U / Ml XOD solution;

[0112] 1.4. 3% perchloric acid solution

[0113] Dilute 3 mL of perchloric acid to 100 mL with ultrapure water;

[0114] 1.5. Standard uric acid 20 mg / mL

[0115] Weigh 2 g of uric acid standard, first dissolve it in 1 M sodium hydroxide, and then make up to 100 mL with PBS;

[0116] 1.6. Positive control allopurinol 10 mg / mL

[0117] Weigh 1 g of allopurinol, first dissolve it in 1 M sodium hydroxide, and then make up to 100 mL with PBS;

[0118] 1.7. Operating steps: First, add 270 μL of xanthine solution, then add 30 μL of the sample to be tested. For the blank, add 10 μL of PBS solution, and incubate at 37 °C for 10 min; add 60 μL of xanthine oxidase at 0.027 U / mL, and incubate at 37 °C for 10 min; then add 75 μL of 3% perchloric acid to terminate the reaction. Filter the above reaction solution through a 0.45 μm membrane and detect by HPLC injection. The HPLC detection conditions are as follows: the mobile phase is methanol: 1% phosphoric acid solution = 5:9, the flow rate is 0.5 mL / min, the injection volume is 20 μL, the temperature is room temperature, a C18 column is used, and the detection wavelength is 290 nm;

[0119] Calculate the XOD inhibition ability (%) according to the following formula = [(uric acid peak area of the blank group - uric acid peak area of the sample group) / uric acid peak area of the blank group] × 100%

[0120] The test results are shown in Table 2.

[0121] 2. Antioxidant activity test

[0122] Test the DPPH radical scavenging activity of the Rosa roxburghii Tratt fermentation broth obtained in Example 2 and Comparative Example 1 to evaluate its antioxidant activity. The test method is as follows:

[0123] 2.1. 0.1 mM DPPH ethanol solution: Weigh 0.0039432 g of DPPH and dissolve it in ethanol and make up the volume to 100 mL.

[0124] 2.2. Operating steps: Add 25 μL of the sample together with 475 μL of the DPPH solution, shake well, and measure the absorbance A1 at 540 nm after reacting in the dark for 30 min. At the same time, measure the absorbance A2 after mixing 25 μL of the sample with 475 μL of the ethanol solution and reacting in the dark for 30 min. And measure the absorbance A0 after reacting 25 μL of the sample solvent with 475 μL of the DPPH solution in the dark for 30 min. Calculate the DPPH radical scavenging ability (%) according to the following formula = [1 - (A1 - A2) / A0] × 100%.

[0125] Table 2: Test results of xanthine oxidase (XOD) inhibition ability and DPPH radical scavenging ability

[0126] Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 XOD Inhibitory Ability (%) 80.57 63.46 66.55 70.06 73.72 DPPH Free Radical Scavenging Ability (%) 88.61 69.46 72.33 79.93 81.14

[0127] As can be seen from Table 2, compared with Comparative Example 1, the fermented Rosa roxburghii Tratt juice exhibited stronger antioxidant activity and uric acid-lowering activity. The XOD inhibitory activity and DPPH free radical scavenging ability of the Rosa roxburghii Tratt juice fermented by different lactic acid bacteria were improved. Among them, the DPPH free radical scavenging ability and XOD inhibitory activity of Example 2 were the highest. And it can be seen from Table 2 that the antioxidant activity and uric acid-lowering activity of the Rosa roxburghii Tratt fermentation broth were positively correlated with its flavonoid content and SOD activity, and negatively correlated with the fructose content.

[0128] III. Animal experiment for hyperuricemia Establish a hyperuricemia rat model to analyze the effect of Rosa roxburghii Tratt fermentation broth on alleviating hyperuricemia: Healthy SPF-grade male SD rats (6 - 8 weeks old, 200 - 220 g) were randomly divided into 5 groups of 10 rats each after a 7-day adaptation period; the animals were kept in an environment with a temperature of 21 ± 2 °C, a humidity of 30 - 70%, 12 h light-dark alternation, free access to water and feed;

[0129] The grouping and treatment methods are as follows:

[0130] Control group 1: Normal control group, fed with basal diet;

[0131] Control group 2: Model group, fed with high-purine diet + intragastric administration of 300 mg / kg / d potassium oxonate for modeling;

[0132] Control group 3: Positive drug group, on the basis of feeding with high-purine diet + intragastric administration of 300 mg / kg / d potassium oxonate for modeling, intragastric administration of 20 mg / kg / d allopurinol;

[0133] Experimental group 1: Intragastric administration of the unfermented Rosa roxburghii Tratt juice obtained from Comparative Example 1, on the basis of feeding with high-purine diet + intragastric administration of 300 mg / kg / d potassium oxonate for modeling, the intragastric administration dose was 3 mL / kg / d;

[0134] Experimental group 2: Intragastric administration of the Rosa roxburghii Tratt fermentation juice of Example 2 in the present invention, on the basis of feeding with high-purine diet + intragastric administration of 300 mg / kg / d potassium oxonate for modeling, the intragastric administration dose was 3 mL / kg / d;

[0135] During the experiment, the animal body weight and food intake were recorded. At the end of 4 weeks of the experiment, the whole blood was collected by cardiac puncture under ether anesthesia. The taken blood was allowed to stand at room temperature for 30 min, then centrifuged at 4000 rpm for 15 minutes, and the serum was collected. The levels of uric acid, malondialdehyde (MDA) and xanthine oxidase (XOD) in the serum were measured using a kit. After the rats were sacrificed by decapitation and dissected, the liver tissue was taken, and the XOD level in the liver was detected using a kit after tissue homogenization.

[0136] Table 3: Effect of Rosa roxburghii Tratt fermentation broth (Experimental group 2) on the body weight and food intake of hyperuricemic rats

[0137] Control Group 1 Control Group 2 Control Group 3 Experimental Group 1 Experimental Group 2 Body Weight (g) 429.6±51.5 408.0±18.4 397.6±16.7 405.5±11.7 405.0±19.7 Food Intake (g / d / rat) 25.28±0.17 <![CDATA[28.19±0.53 ## > <![CDATA[27.05±0.19 *** > <![CDATA[26.28±0.33 *** > <![CDATA[26.30±0.33 *** >

[0138] Data were expressed as mean ± standard deviation. Compared with control group 1, # indicating p < 0.05, ## indicating p < 0.01, ### indicating p < 0.001; compared with control group 2, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

[0139] During the experiment, the body weight and food intake of rats were recorded. The results are shown in Table 3. There was no significant difference in body weight among different groups. Compared with control group 1, the food intake of the model group (control group 2) was significantly increased, and fermented Rosa roxburghii tratt (experimental group 2) could significantly reduce its food intake.

[0140] Table 4: Effects of fermented Rosa roxburghii tratt (experimental group 2) on serum biochemical indexes of hyperuricemic rats

[0141] Control Group 1 Control Group 2 Control Group 3 Experimental Group 1 Experimental Group 2 Serum Uric Acid (μmol / L) 151.3±39.6 <![CDATA[221.7±27.6 ## > <![CDATA[146.8±23.6 *** > 219.0±16.8 <![CDATA[185.7±20.8 * > Blood Urea Nitrogen (mmol / L) 5.24±0.61 <![CDATA[6.14±0.42 # > 5.70±0.49 5.84±0.27 <![CDATA[4.96±0.94 ** > MDA (nmol / mL) 7.54±1.88 <![CDATA[10.56±1.86 ### > <![CDATA[8.79±1.75 * > <![CDATA[8.98±1.72 * > <![CDATA[7.97±0.79 ** >

[0142] Data were expressed as mean ± standard deviation. Compared with control group 1, # indicating p < 0.05, ## indicating p < 0.01, ### indicating p < 0.001; compared with control group 2, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

[0143] Uric acid is the end product of purine metabolism. When serum uric acid exceeds the normal concentration, hyperuricemia will occur. The level of serum urea nitrogen is an important indicator of kidney injury. The higher the content, the greater the damage. Hyperuricemia is closely related to high levels of oxidative stress injury in the body. When the balance between ROS and the antioxidant system is lost, the accumulation of ROS will cause oxidative stress reactions and attack macromolecules (such as lipids, proteins, and DNA) and cell organelles, thus forming lipid peroxidation products (such as MDA), damaging the human body. The content of MDA can reflect the degree of lipid peroxidation.

[0144] The specific results are shown in Table 4. The serum uric acid level in the model group (Control Group 2) was significantly higher than that in the normal control group (Control Group 1), indicating successful modeling. The intragastric administration of Rosa roxburghii Tratt fermentation broth (Experimental Group 2) significantly prevented the increase in serum uric acid, suggesting a strong effect against hyperuricemia. The serum urea nitrogen level in the model group (Control Group 2) was relatively high, showing a significant difference compared with the normal control group (Control Group 1), indicating that hyperuricemia modeling caused kidney damage. In addition, the content of urea nitrogen decreased significantly after intragastric administration of Rosa roxburghii Tratt fermentation broth, showing a certain degree of kidney protection. Compared with Control Group 1, the serum MDA level also increased significantly. The intragastric administration of unfermented Rosa roxburghii Tratt (Experimental Group 1) and Rosa roxburghii Tratt fermentation broth (Experimental Group 2) could significantly reduce the MDA level and relieve oxidative stress related to hyperuricemia, with the Rosa roxburghii Tratt fermentation broth showing a stronger effect.

[0145] Table 5: Effects of Rosa roxburghii Tratt Fermentation Broth (Experimental Group 2) on the Activity of XOD in the Serum and Liver of Hyperuricemic Rats

[0146] Control Group 1 Control Group 2 Control Group 3 Experimental Group 1 Experimental Group 2 Serum XOD (U / L) 13.06±2.39 <![CDATA[16.48±2.99 ### > <![CDATA[13.44±1.98 ** > <![CDATA[14.26±0.84 * > <![CDATA[13.06±1.98 ** > Liver XOD (U / g prot) 5.40±0.75 <![CDATA[6.57±1.08 # > <![CDATA[4.78±0.61 *** > <![CDATA[5.52±0.96 * > <![CDATA[4.67±0.57 *** >

[0147] The data are presented as mean ± standard deviation. Compared with Control Group 1, # indicates p < 0.05, ## indicates p < 0.01, ### indicates p < 0.001; compared with Control Group 2, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

[0148] The results are shown in Table 5. Compared with the normal control group (Control Group 1), the activity of XOD in the serum and liver of rats in the model group (Control Group 2) increased significantly. After intragastric administration of unfermented Rosa roxburghii Tratt (Experimental Group 1) and Rosa roxburghii Tratt fermentation broth (Experimental Group 2), the activity of XOD decreased significantly, with the Rosa roxburghii Tratt fermentation broth showing a more significant effect. Thus, it can be seen that Rosa roxburghii Tratt fermentation broth can relieve hyperuricemia by inhibiting the activity of XOD, thereby effectively reducing the production of uric acid.

[0149] Thus, it can be seen that the Rosa roxburghii Tratt fermentation broth obtained by fermenting Rosa roxburghii Tratt with compound Lactobacillus in the present invention has increased flavonoid content and SOD activity and decreased fructose content, and has the effects of reducing uric acid, improving kidney damage and antioxidation, and its effect is stronger than that of unfermented Rosa roxburghii Tratt.

Claims

1. A preparation method of Rosa roxburghii Tratt fermentation broth for relieving hyperuricemia, characterized in that, It includes the following steps: (1) Mix the Rosa roxburghii tratt with water and then juice it. After removing the solids, adjust the pH with edible alkali to obtain the Rosa roxburghii tratt blended juice; (2) After sterilizing the Rosa roxburghii tratt blended juice, add the cell suspension of the compound Lactobacillus for fermentation. After fermentation, remove the solids to obtain the Rosa roxburghii tratt fermentation broth. The compound Lactobacillus is a mixture of Lactobacillus rhamnosus with the preservation number of CGMCC NO.11955 and Lactobacillus plantarum with the preservation number of CGMCC No.13121.

2. The preparation method of a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 1, characterized in that, In (1), the Rosa roxburghii tratt is mixed with water and then juiced. After filtration, centrifugation and concentration, the Rosa roxburghii tratt raw juice is obtained. Add water to the Rosa roxburghii tratt raw juice and adjust the pH with edible alkali to obtain the Rosa roxburghii tratt blended juice.

3. The preparation method of the Rosa roxburghii Tratt fermentation broth for relieving hyperuricemia according to claim 2, wherein, In (1), the Rosa roxburghii tratt is mixed with water that is 2 - 16 times the mass of the Rosa roxburghii tratt and then juiced. Then it is filtered through a 60 - 100 mesh sieve, centrifuged and concentrated to obtain the Rosa roxburghii tratt raw juice. Mix the Rosa roxburghii tratt raw juice with water at a volume ratio of 1:(0.5 - 4) and adjust the pH with edible alkali.

4. A method for preparing a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 1 or 2 or 3, characterized in that, In (1), adjust the pH to 4.0 - 8.0 with edible alkali. The edible alkali is food - grade Na2CO3 and / or food - grade NaHCO3.

5. The preparation method of the rosa roxburghii tratt fermentation broth for relieving hyperuricemia according to claim 1, characterized in that, In (2), the sterilization process is to sterilize the Rosa roxburghii tratt blended juice at 80 - 95 °C for 5 - 20 minutes and then cool it.

6. The preparation method of the Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 1, characterized in that, The cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum in (2) are obtained by the following steps: inoculate Lactobacillus rhamnosus and Lactobacillus plantarum into the sterilized MRS liquid medium respectively, incubate at 30 - 37 °C for 12 - 24 h respectively, activate for 2 generations, then centrifuge at 5000 r / min for 10 - 20 min, discard the supernatant, collect the cells respectively, and finally adjust the cell suspension concentration to 10 6 CFU / mL or more to obtain the cell suspensions of Lactobacillus rhamnosus and Lactobacillus plantarum, and then mix the cell suspension of Lactobacillus rhamnosus and the cell suspension of Lactobacillus plantarum according to a volume ratio of 1:(0.5 - 4) to obtain the cell suspension of the compound Lactobacillus.

7. A method for preparing a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 1 or 6, characterized in that After the sterilization treatment of the Rosa roxburghii Tratt blended juice, add the cell suspension of the compound Lactobacillus to make the initial inoculation amount greater than 10 6 CFU and then carry out fermentation.

8. A method for preparing a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 7, characterized in that, In (2), the fermentation conditions are constant - temperature fermentation at 37 °C for 24 - 120 hours, and then transfer to 4 °C for after - ripening for 1 - 4 days.

9. A method for preparing a Rosa roxburghii Tratt fermentation broth for alleviating hyperuricemia according to claim 1, characterized in that, In (2), after fermentation, centrifuge at a speed of 4000 - 8000 r / min and collect the supernatant to obtain the Rosa roxburghii tratt fermentation broth.

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