Liver-protecting antioxidant health-care functional beverage and preparation method thereof

Abstract

The invention belongs to the technical field of health-care food, mainly relates to a health-care functional beverage, and in particular relates to a liver-protecting antioxidant health-care functional beverage and a preparation method thereof. The invention aims to provide a beverage with liver-protecting and anti-oxidation effects, the beverage is prepared by selecting Chinese yam, medlar, coix seed, dandelion root, red date and honey purple perilla as raw materials and carrying out synergistic fermentation modification through bifidobacterium breve and lactobacillus acidophilus, and the beverage has the effects of resisting oxidation, assisting in protecting chemical liver injury and improving haemorrhoids.

Description

Technical Field

[0001] The invention belongs to the technical field of health food, and mainly relates to a health functional beverage, in particular to a liver-protecting and anti-oxidation health functional beverage and a preparation method thereof. Background Art

[0002] As the pace of modern life accelerates, people are facing increasing pressure. Unhealthy lifestyles such as irregular eating habits and excessive drinking have a serious impact on liver health. Chemical liver damage may lead to serious health problems such as cirrhosis and hepatitis. Therefore, developing a health drink that can protect the liver and has an antioxidant effect is of great significance for improving and enhancing the quality of life of modern people.

[0003] There are not many liver-protecting health drinks on the market, and these products often have limited effects, cannot quickly decompose chemicals such as alcohol, and may cause additional metabolic burdens on the liver and kidneys. In addition, some products contain chemical ingredients, which can temporarily relieve liver damage caused by chemical substances, but long-term use may cause side effects.

[0004] The present invention aims to develop a new type of liver-protecting antioxidant health-care functional beverage through innovative formulas and processes. The beverage uses yam, wolfberry, coix seed, dandelion root, red dates, and perilla as raw materials, all of which are natural plants and rich in a variety of bioactive compounds, such as flavonoids, phenols, saponins, alkaloids, etc. These compounds have powerful antioxidant, anti-inflammatory, antibacterial and antiviral effects. At the same time, the present invention optimizes the preparation process, and by accurately controlling the time, temperature, pressure and other conditions during the processing process, the nutrients in the raw materials are retained to the maximum extent, thereby improving the health-care effect of the product. Summary of the invention

[0005] In view of the above-mentioned problems, the present invention aims to provide a beverage with liver protection and antioxidant effects. Chinese yam, wolfberry, coix seed, dandelion root, red dates and honey basil are selected and modified through the synergistic fermentation of Bifidobacterium breve and Lactobacillus acidophilus to obtain a beverage. The beverage has the effects of antioxidant, auxiliary protection against chemical liver damage and improvement of hemorrhoids.

[0006] S1. Remove the core of red dates and peel yam, wash and cut into pieces; wash dandelion root, coix seed and wolfberry with clean water and dry them. Then, weigh 40-50g of yam, 10-15g of wolfberry, 20-30g of coix seed, 10-30g of dandelion root, 15-20g of red dates and 8-12g of perilla, crush them separately and set aside;

[0007] S2, put the perilla prepared in step S1 into a beaker, add 1-1.2g refined honey and 15-20mL water into the beaker, stir evenly, put it in a pot and heat it over low heat until it is dark brown and not sticky, take it out and dry it to obtain honey perilla;

[0008] S3. Inoculate Bifidobacterium breve CICC-6183 onto glucose blood liver agar medium and culture at 37°C for 12-18 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500-1000 mL glucose blood liver liquid medium shake flask and fermented for another 8-14 hours to obtain 2.2-2.5×10 7 cfu / mL of bacterial solution;

[0009] S4. Inoculate Lactobacillus acidophilus CICC-6095 onto MRS agar medium and culture at 37°C for 15-20 hours to activate the strain. Transfer the activated Lactobacillus acidophilus to a 500-1000 mL MRS liquid medium shake flask and ferment for another 8-14 hours to obtain 1.4-1.6×10 8 cfu / mL of bacterial solution;

[0010] S5. After uniformly mixing the yam, wolfberry, coix seed, dandelion root, and red dates prepared in step S1 and the honey basil prepared in step S2, 18-20 mL of the Bifidobacterium breve CICC-6183 bacterial solution prepared in step S4 and 18-20 mL of the Lactobacillus acidophilus CICC-6095 bacterial solution prepared in step S3 were added, and anaerobically fermented at 37° C. for 18-24 h;

[0011] S6. After the fermentation is completed, the Bifidobacterium breve and Lactobacillus acidophilus in the fermentation product are inactivated with a 3% H2O2 solution to obtain a fermentation product, and then CaCl2 is added to stand for 15-30 minutes and then centrifuged to obtain a supernatant, and then 50-75% ethanol is added and centrifuged again to collect the supernatant, and then filtered, and then reflux extraction is performed through a condensation reflux device for 30-60 minutes. After the extraction is completed, the desired fermentation liquid is obtained;

[0012] S7, pouring the fermentation liquid prepared in step S5 into a homogenizer and homogenizing it twice at 30 MPa;

[0013] S8. High temperature instantaneous sterilization is performed, the sterilization temperature is 135° C., and the time is 10-20 seconds. After the sterilization is completed, the beverage of the present invention is obtained.

[0014] Preferably: in step S1 of the present invention, the raw material is crushed and filtered through a 120-mesh sieve to obtain a 120-mesh powder.

[0015] Preferably, the method for preparing refined honey in step S2 of the present invention is: pour the honey into a clean pot, heat it over a low fire for 10-15 minutes, stir it continuously and remove the foam.

[0016] Preferably: the components of the glucose blood-liver agar medium used in step S3 of the present invention are: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g plant peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (10 g K2HPO4, 10 g KH2PO4, 100 mL distilled water), 5 mL solution B (4 g MgSO4·7H2O, 0.2 g NaCI, 0.2 g FeSO4·7H2O, 0.2 g MnSO4·2H2O, 100 mL distilled water), 10 mL 5% L-cysteine ​​hydrochloride solution, 1 g Tween-80, 15 g agar, 50 mL defibrinated sheep blood, and 825 mL distilled water.

[0017] Preferably: the components of the MRS agar medium used in step S4 of the present invention are: 10g peptone, 10g beef extract powder, 5g yeast extract powder, 20g glucose, 1g Tween, 5g sodium acetate, 2g triammonium citrate, 2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate heptahydrate, 0.05g sulfuric acid tetrahydrate, 15g agar, 1000mL distilled water, and the pH is adjusted to 6.2-6.5; the components of the glucose blood liver agar medium are: 2.5g peptone, 1.5g beef extract, 10g glucose blood liver, 18g agar, and 1000mL distilled water.

[0018] Preferably, in step S5 of the present invention, the volumes of the Bifidobacterium breve bacterial solution and the Lactobacillus acidophilus bacterial solution are both 18 mL.

[0019] Preferably, the time of anaerobic fermentation in step S5 of the present invention is 20 hours.

[0020] In the second aspect, the beverage of the present invention has the functions of auxiliary protection against chemical liver damage, anti-oxidation and improvement of hemorrhoids.

[0021] The present invention has the following beneficial effects:

[0022] 1. In the present invention, under the fermentation conditions of Bifidobacterium breve CICC-6183 and Lactobacillus acidophilus CICC-6095, wolfberry, dandelion root and yam synergistically have an auxiliary protective effect on chemical liver damage.

[0023] 2. The yam fermentation liquid, coix seed fermentation liquid and red date fermentation liquid in the present invention have a synergistic effect on anti-oxidation.

[0024] 3. The wolfberry fermentation liquid, coix seed fermentation liquid and honey basil fermentation liquid in the present invention have a synergistic effect on improving hemorrhoids.

[0025] 4. The beverages of the present invention are all derived from natural plants and have multiple biological activities. Compared with Western medicine, the beverages have more functions, higher safety and fewer side effects.

[0026] 5. The beverage of the present invention is rich in nutrition and contains various vitamins, trace elements, minerals and alkaloids, etc. These nutrients are essential for maintaining good health.

[0027] 6. The beverage of the present invention uses all natural plants as raw materials, does not contain artificial additives, preservatives and sweeteners, and is a better choice for people who pursue clean diet. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0029] Figure 1 It is a comparison chart of the antioxidant effects of Example 2 and Comparative Examples 8-10.

[0030] Figure 2 The acne areas on the back skin of mice in Example 3 and Comparative Examples 11-14 groups after 7 days of drug intervention.

[0031] Figure 3 The following are the changes in the back skin of mice in Example 3 and Comparative Example 11-14 groups after 7 days of drug intervention.

[0032] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0033] Example 1

[0034] This example demonstrates that under the fermentation conditions of Bifidobacterium breve CICC-6183 and Lactobacillus acidophilus CICC-6095, wolfberry, dandelion root and yam synergistically have an auxiliary protective effect on chemical liver injury.

[0035] This embodiment provides a beverage, and the specific steps are as follows:

[0036] S1. Remove the core of red dates and peel yam, clean them and cut them into pieces; then wash dandelion root, coix seed and wolfberry with clean water respectively, dry them, weigh 40g yam, 10g wolfberry, 20g coix seed, 10g dandelion root, 15g red dates and 8g perilla, grind them respectively, filter them through a 120-mesh sieve to obtain 120-mesh powder, and set aside;

[0037] S2, put the perilla prepared in step S1 into a beaker, then mix 1.0g of refined honey and 15mL of water and pour them into the beaker together, stir evenly, put them in a pot and heat them over low heat until they are dark brown and not sticky, take them out and dry them, and obtain honey perilla;

[0038] S3. Bifidobacterium breve CICC-6183 (purchased from China Industrial Microbiological Culture Collection Center) was inoculated on glucose blood liver agar medium and cultured at 37°C for 12 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to obtain 2.2×10 7 cfu / mL of bacterial solution;

[0039] S4. Inoculate Lactobacillus acidophilus CICC-6095 (purchased from China Industrial Microbiological Culture Collection Center) onto MRS agar medium and culture at 37°C for 15 h to activate the strain. The activated Lactobacillus acidophilus was transferred to a 500 mL MRS liquid medium shake flask and fermented for another 8 h to obtain 1.4×10 8 cfu / mL of bacterial solution;

[0040] S5. After uniformly mixing the yam, wolfberry, coix seed, dandelion root, and red dates prepared in step S1 and the honey basil prepared in step S2, 18 mL of the Bifidobacterium breve CICC-6183 bacterial solution prepared in step S4 and 18 mL of the Lactobacillus acidophilus CICC-6095 bacterial solution prepared in step S3 were added, and anaerobically fermented at 37° C. for 20 h;

[0041] S6. After the fermentation is completed, the Bifidobacterium breve and Lactobacillus acidophilus in the fermentation product are inactivated with a 3% H2O2 solution to obtain a fermentation product, and then CaCl2 is added to stand for 15 minutes and centrifuged to obtain the supernatant, and then 50% ethanol is added and centrifuged again to collect the supernatant, and then filtered, and then reflux extraction is performed through a condensation reflux device for 30 minutes. After the extraction is completed, the desired fermentation liquid is obtained;

[0042] S7, pouring the fermentation liquid prepared in step S5 into a homogenizer and homogenizing it twice at 30 MPa;

[0043] S8. High temperature instantaneous sterilization treatment is performed, with the sterilization temperature being 135° C. and the time being 10 seconds. After the sterilization is completed, the beverage of the present invention is obtained.

[0044] Comparative Example 1: In step S5, wolfberry is not added, and the remaining steps are consistent with Example 1.

[0045] Comparative Example 2: In step S5, dandelion root was not added, and the remaining steps were kept consistent with Example 1.

[0046] Comparative Example 3: In step S5, yam is not added, and the remaining steps are consistent with Example 1.

[0047] Comparative Example 4: In step S3, Bifidobacterium breve CICC-6183 was not added, and the remaining steps were the same as those in Example 1.

[0048] Comparative Example 5: In step S4, Lactobacillus acidophilus CICC-6095 was not added, and the remaining steps were kept consistent with Example 1.

[0049] Comparative Example 6: In step S3, Bifidobacterium breve CICC-6183 was replaced with CICC-6204 (purchased from China Industrial Microbiological Culture Collection Center), and the remaining steps were kept consistent with Example 1.

[0050] Comparative Example 7: In step S4, Lactobacillus acidophilus CICC-6095 was replaced by CICC-6091 (purchased from China Industrial Microbiological Culture Collection Center), and the remaining steps were kept consistent with Example 1.

[0051] Study on the protective effect of beverages on acute liver injury caused by CCl4 in mice

[0052] Beijing Weitonglihua Experimental Animal Technology Co., Ltd. purchased 88 BALB / c male mice, weighing 18-22 g. Before the experiment officially began, they were kept in a constant temperature and oxygen supply environment at 25±2℃ for one week. The entire experimental process strictly followed the national experimental animal management regulations.

[0053] 88 mice were randomly divided into 11 groups, namely, a blank group, a model group, a positive control group, Example 1 group, and Comparative Example 1-7 groups.

[0054] The Example 1 group and the Comparative Examples 1-7 groups were gavaged with the beverage corresponding to the present invention, with a dose of 20 mL / (kg·d); the positive control group was gavaged with bifendate, with a dose of 100 mg / (kg·d); the blank group and the model group were gavaged with purified water, with a dose of 20 mL / (kg·d); the gavage was continued for 3 weeks, and 1 hour after the last gavage, the blank group was intraperitoneally injected with 10 mL / kg of peanut oil, and the other groups were intraperitoneally injected with 10 mL / kg of peanut oil containing 2 g / L CCl4, and then fasted but not watered for 16 hours, after which the mice were killed and the mouse livers were removed. The livers were placed in a culture dish filled with 4°C physiological saline, the surface blood was washed off, the surface water was blotted dry, weighed, and the liver coefficient was calculated.

[0055] Liver coefficient = liver mass / mouse mass x 100%

[0056] After killing the mice, the eyeballs were removed and blood was collected. The blood samples were centrifuged at 4°C and 3500 r / min for 10 min, and serum was collected to determine the concentrations of ALT, AST, and TBIL according to the operating specifications of the kit.

[0057] Table 1

[0058] Group Number of mice Liver coefficient (%) Blank Group 8 4.55±0.09 Model Group 8 5.00±0.14 Positive control group 8 4.53±0.13 Example 1 Group 8 4.16±0.10 Comparative Example 1 8 4.76±0.18 Comparative Example 2 8 4.81±0.17 Comparative Example 3 8 4.74±0.16 Comparative Example 4 8 4.66±0.15 Comparative Example 5 8 4.65±0.12 Comparative Example 6 8 4.58±0.10 Comparative Example 7 8 4.63±0.11

[0059] Table 2

[0060]

[0061]

[0062] Table 1 is a comparison table of liver coefficients of Example 1 and Comparative Examples 1-7. As can be seen from the table, the liver coefficients of mice in the model group are significantly different from those in the blank group, which indicates that CCl4 can induce hepatocyte edema, thereby promoting an increase in the liver coefficient; compared with the model group, the liver coefficients of mice in Comparative Examples 1-7 are relatively reduced, while the liver coefficient of mice in Example 1 is the smallest, indicating that Comparative Examples 1-7 and Example 1 have a certain alleviating effect on liver enlargement caused by CCl4 liver poisoning, and the effect of Example 1 is the best.

[0063] Table 2 is a comparison table of ALT, AST activity and TBI concentration in serum of Example 1 group and Comparative Examples 1-7 groups. As can be seen from the table, after CCl4 was intraperitoneally injected into the model group mice, the ALT, AST activity and TBI concentration in the serum were significantly increased, indicating that the liver cells had suffered serious damage; compared with the model group, the serum ALT, AST activity and TBI concentration of the mice in the Comparative Examples 1-7 groups were relatively reduced, while the serum ALT, AST activity and TBI concentration of the mice in the Example 1 group were the smallest, indicating that the Comparative Examples 1-7 groups and the Example 1 group can inhibit the increase of ALT, AST activity and TBIL concentration to a certain extent, reduce the degree of damage to the liver of mice by CCl4, and the effect of the Example 1 group is the best.

[0064] In summary, the protective effect of Example 1 on acute liver injury caused by CCl4 is the best, which confirms that under fermentation conditions, the synergistic fermentation of wolfberry, dandelion root and yam has a significant auxiliary protective effect on chemical liver injury.

[0065] Example 2

[0066] This example proves that the yam fermentation liquid, the coix seed fermentation liquid and the red date fermentation liquid have synergistic antioxidant effects.

[0067] This embodiment provides a beverage, and the specific steps are as follows:

[0068] S1. Remove the core of red dates and peel yam, clean them and cut them into pieces; then wash the dandelion root, coix seed and wolfberry with clean water and dry them. Then, weigh 45g of yam, 12g of wolfberry, 25g of coix seed, 20g of dandelion root, 17g of red dates and 10g of perilla, grind them respectively and filter them through a 120-mesh sieve to obtain a 120-mesh powder for standby use;

[0069] S2, put the perilla prepared in step S1 into a beaker, and then mix 1.1g of refined honey and 17mL of water and pour them into the beaker together, stir evenly, put them in a pot and heat them over low heat until they are dark brown and not sticky, take them out and dry them, and obtain honey perilla;

[0070] S3. Bifidobacterium breve CICC-6183 (purchased from China Industrial Microbiological Culture Collection Center) was inoculated on glucose blood liver agar medium and cultured at 37°C for 15 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 750 mL glucose blood liver liquid medium shake flask and fermented for another 11 hours to finally obtain 2.3×10 7 cfu / mL of bacterial solution;

[0071] S4. Inoculate Lactobacillus acidophilus CICC-6095 (purchased from China Industrial Microbiological Culture Collection Center) onto MRS agar medium and culture at 37°C for 17 hours to activate the strain. The activated Lactobacillus acidophilus was transferred to a 750 mL MRS liquid medium shake flask and fermented for another 11 hours to obtain 1.5×10 8 cfu / mL of bacterial solution;

[0072] S5. After uniformly mixing the yam, wolfberry, coix seed, dandelion root, and red dates prepared in step S1 and the honey basil prepared in step S2, 18 mL of the Bifidobacterium breve CICC-6183 bacterial solution prepared in step S4 and 18 mL of the Lactobacillus acidophilus CICC-6095 bacterial solution prepared in step S3 were added, and anaerobically fermented at 37° C. for 20 h;

[0073] S6. After the fermentation is completed, the Bifidobacterium breve and Lactobacillus acidophilus in the fermentation product are inactivated with a 3% H2O2 solution to obtain a fermentation product, and then CaCl2 is added to stand for 22 minutes and centrifuged to obtain the supernatant, and then 60% ethanol is added and centrifuged again to collect the supernatant, and then filtered, and then reflux extraction is performed through a condensation reflux device for 40 minutes, and the desired fermentation liquid is obtained after the extraction is completed;

[0074] S7, pouring the fermentation liquid prepared in step S5 into a homogenizer and homogenizing it twice at 30 MPa;

[0075] S8. High temperature instantaneous sterilization treatment is performed, with the sterilization temperature being 135° C. and the time being 15 seconds. After the sterilization is completed, the beverage of the present invention is obtained.

[0076] Comparative Example 8: In step S5, it is chosen not to add yam, and the remaining steps are kept consistent with Example 2.

[0077] Comparative Example 9: In step S5, it is chosen not to add coix seeds, and the remaining steps are kept consistent with Example 2.

[0078] Comparative Example 10: In step S5, red dates are not added, and the remaining steps are consistent with Example 2.

[0079] Studying the antioxidant effects of beverages

[0080] (1) Determination of DPPH free radical scavenging rate: The beverage of the present invention was diluted with water to 1 mg / mL, 200uL of the beverage dilution was taken, and 200uL of 0.2mmol / L·DPPH anhydrous ethanol solution was added. After mixing evenly, the mixture was reacted at room temperature in the dark for 30 minutes, and then the absorbance of the sample at 517nm was measured by an ELISA instrument. The control group was a mixture of 200uL sample solution and 200uL anhydrous ethanol, and the blank group was a mixture of 200uL distilled water and 200uL DPPH-anhydrous ethanol solution. VC was the positive control. The DPPH free radical scavenging rate (R1, %) was calculated according to the formula. The results are shown in the figure. Figure 1 shown.

[0081] R1 / %=[1-(A 样 -A 对 ) / A 空 ]x100

[0082] Where: A 样 is the absorbance of the sample group; A 对 is the absorbance of the control group; A 空 is the absorbance of the blank group.

[0083] (2) Determination of OH radical scavenging rate: dilute the beverage of the present invention with water to 1 mg / mL, take 200uL of the beverage dilution and mix it with 200uL of ferrous sulfate solution (9mmol / L), 200uL of salicylic acid ethanol solution (9mmol / L), and 200uL of hydrogen peroxide solution (9mmoL), and mix them evenly, and place in a water bath at 37°C for 30 minutes. After standing for 20 minutes, use an enzyme marker to measure the absorbance of the sample at 510nm. Replace the sample with distilled water as the blank group, and a mixed solution of 200uL sample solution, 400uL distilled water and 200uL anhydrous ethanol as the control group. VC is the positive control. Calculate the OH radical scavenging rate (R2, %) according to the formula. The results are as follows: Figure 1 shown.

[0084] R2 / % = [1-(A 样 -A 对 ) / A 空 ]x100

[0085] Where: A 样 is the absorbance of the sample group; A 对 is the absorbance of the control group; A 空 is the absorbance of the blank group. Figure 1 As shown:

[0086] Figure 1 This is a comparison chart of the antioxidant effects of Example 2 and Comparative Examples 8-10. As can be seen from the figure, the elimination rates of DPPH radicals and ·OH radicals in Example 2 are 88.9% and 93.5% respectively; the elimination rates of DPPH radicals and ·OH radicals in Comparative Example 1 are 64.2% and 69.5% respectively; and the elimination rates of DPPH radicals and ·OH radicals in Comparative Example 2 are 76.2% and 79.5% respectively. It can be seen that compared with Comparative Examples 1 and 2, Example 2 has a higher elimination rate of DPPH radicals and ·OH radicals and a better antioxidant effect, which confirms that the ginseng fermented liquid and the ganoderma lucidum fermented liquid have good antioxidant capacity under the synergistic effect.

[0087] Example 3

[0088] This example proves that the synergistic effect of wolfberry fermentation liquid, coix seed fermentation liquid and honey perilla fermentation liquid has the effect of improving hemorrhoids.

[0089] This embodiment provides a beverage, and the specific steps are as follows:

[0090] S1. Remove the core of red dates and peel yam, clean them and cut them into pieces; then wash the dandelion root, coix seed and wolfberry with clean water and dry them. Then, weigh 50g of yam, 15g of wolfberry, 30g of coix seed, 30g of dandelion root, 20g of red dates and 12g of perilla, grind them separately and filter them through a 120-mesh sieve to obtain a 120-mesh powder for standby use;

[0091] S2, put the perilla prepared in step S1 into a beaker, and then mix 1.2g of refined honey and 20mL of water and pour them into the beaker together, stir evenly, put them in a pot and heat them over low heat until they are dark brown and not sticky, take them out and dry them, and obtain honey perilla;

[0092] S3. Bifidobacterium breve CICC-6183 (purchased from China Industrial Microbiological Culture Collection Center) was inoculated on glucose blood liver agar medium and cultured at 37°C for 18 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 1000 mL glucose blood liver liquid medium shake flask and fermented for another 14 hours to finally obtain 2.5×10 7 cfu / mL of bacterial solution;

[0093] S4. Inoculate Lactobacillus acidophilus CICC-6095 (purchased from China Industrial Microbiological Culture Collection Center) onto MRS agar medium and culture at 37°C for 20 h to activate the strain. The activated Lactobacillus acidophilus was transferred to a 1000 mL MRS liquid medium shake flask and fermented for another 14 h to obtain 1.6×10 8 cfu / mL of bacterial solution;

[0094] S5. After uniformly mixing the yam, wolfberry, coix seed, dandelion root, and red dates prepared in step S1 and the honey basil prepared in step S2, 18 mL of the Bifidobacterium breve CICC-6183 bacterial solution prepared in step S4 and 18 mL of the Lactobacillus acidophilus CICC-6095 bacterial solution prepared in step S3 were added, and anaerobically fermented at 37° C. for 20 h;

[0095] S6. After the fermentation is completed, the Bifidobacterium breve and Lactobacillus acidophilus in the fermentation product are inactivated with a 3% H2O2 solution to obtain a fermentation product, and then CaCl2 is added to stand for 30 minutes and centrifuged to obtain the supernatant, and then 75% ethanol is added and centrifuged again to collect the supernatant, and then filtered, and then reflux extraction is performed through a condensation reflux device for 60 minutes. After the extraction is completed, the desired fermentation liquid is obtained;

[0096] S7, pouring the fermentation liquid prepared in step S5 into a homogenizer and homogenizing it twice at 30 MPa;

[0097] S8, adopting high temperature instantaneous sterilization treatment, the sterilization temperature is 135° C., the time is 20 seconds, and the beverage of the present invention is obtained after the sterilization is completed.

[0098] Comparative Example 11: In step S4, wolfberry is not added, and the remaining steps are consistent with Example 3.

[0099] Comparative Example 12: In step S4, it is chosen not to add coix seeds, and the remaining steps are kept consistent with Example 3.

[0100] Comparative Example 13: In step S4, it is chosen not to add honey basil, and the remaining steps are kept consistent with Example 3.

[0101] Comparative Example 14: In step S4, it is chosen not to modify the perilla by refining honey, and the perilla is directly used for fermentation, and the remaining steps are kept consistent with Example 3.

[0102] Study on the effect of beverage on improving hemorrhoids in mice

[0103] Beijing Weitonglihua Experimental Animal Technology Co., Ltd. purchased 56 BALB / c male mice. Before the experiment officially started, they were kept in a constant temperature and oxygen supply environment at 25±2℃ for one week. The entire experimental process strictly followed the national experimental animal management regulations.

[0104] 56 mice were randomly divided into 7 groups, 8 mice in each group, namely normal group, model group, Example 3 group, and Comparative Examples 11-14 group. All mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (0.01 mL / g), and then the back was depilated. After depilation, except for the mice in the normal group, the mice in other groups were intradermally injected with 50 μL of P. acnes suspension on the back. When small lumps appeared on the back skin of the mice, the top of the lumps was white, the base was red, and accompanied by symptoms of inflammation and swelling, it indicated that the modeling was successful. After the modeling was successful, Example 3 group and Comparative Examples 11-14 group were gavaged with the beverage corresponding to the present invention, with a dose of 20 mL / (kg·d); the blank group and the model group were gavaged with the same amount of purified water as that of Example 1 group. The gavage was continued for 7 days. After 7 days, the redness, swelling, small lumps, ulcers and other phenomena on the back skin of each group of mice were observed, and the acne area on the back skin was measured with a vernier caliper. The experimental results are shown in Table 1. Figure 2 and Figure 3 .

[0105] Figure 2 The acne areas on the back skin of mice in Example 3 and Comparative Examples 11-14 groups after 7 days of drug intervention. Figure 3The results show that the back skin of mice in Example 3 and Comparative Examples 11-14 changed after 7 days of drug intervention. The results show that the back of mice in the blank group was smooth and there was no acne area. Compared with the blank group mice, the back skin of mice in the model group was obviously raised with small bags, the base was red and there were symptoms of inflammation and swelling, and the acne area increased significantly. Compared with the model group, the skin changes and acne area of ​​mice in Comparative Examples 11-14 did not change much, the small bags on the back skin of mice in Example 3 group were significantly reduced, the base redness and inflammation and swelling symptoms were significantly alleviated, and the acne area was also significantly reduced, confirming that the synergistic effect of Polygonum multiflorum fermentation, ginseng fermentation, and saponin fermentation has the effect of treating hemorrhoids.

[0106] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A liver-protecting and antioxidant health beverage. Features: The beverage is obtained by extracting 6 kinds of natural plants through composite bacterial strain fermentation, and the prepared beverage has the effects of anti-oxidation, auxiliary protection against chemical liver damage and improvement of hemorrhoids.

2. A liver-protecting and antioxidant health-care functional beverage according to claim 1, Features: The natural plants include 40-50g of yam, 10-15g of wolfberry, 20-30g of coix seed, 10-30g of dandelion root, 15-20g of red dates and 8-12g of perilla by weight.

3. A liver-protecting and antioxidant health-care functional beverage according to claim 2, Features: The composite strain includes Bifidobacterium breve CICC-6183 and Lactobacillus acidophilus CICC-6095.

4. A liver-protecting and antioxidant health-care functional beverage according to claim 3, Features: The perilla needs to be processed with refined honey to make honey perilla, and the specific preparation method is: dry the cleaned perilla, weigh 8-12g and crush it and put it into a beaker, then add 1-1.2g of refined honey and 15-20mL of water to the beaker, stir evenly, put it in a pot and heat it over low heat to stir-fry until it turns dark brown and is not sticky, take it out and dry it, and get the honey perilla.

5. A liver-protecting, antioxidant health-care functional beverage and a preparation method thereof according to claims 1-4, wherein the specific preparation method is as follows: S1. Weigh 40-50g of Chinese yam, 10-15g of wolfberry, 20-30g of coix seed, 10-30g of dandelion root, 15-20g of red dates, and 8-12g of perilla, and crush them separately for later use; S2, put the perilla prepared in step S1 into a beaker, add 1-1.2g refined honey and 15-20mL water into the beaker, stir evenly, put it in a pot and heat it over low heat until it is dark brown and not sticky, take it out and dry it to obtain honey perilla; S3. Bifidobacterium breve CICC-6183 was inoculated on glucose blood liver agar medium and cultured at 37°C for 12-18 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500-1000 mL glucose blood liver liquid medium shake flask and fermented for another 8-14 hours to obtain 2.2-2.5×10 7 cfu / mL of bacterial solution; S4. Lactobacillus acidophilus CICC-6095 was inoculated on MRS agar medium and cultured at 37°C for 15-20 hours to activate the strain. The activated Lactobacillus acidophilus was transferred to a 500-1000 mL MRS liquid medium shake flask and fermented for another 8-14 hours to obtain 1.4-1.6×10 8 cfu / mL of bacterial solution; S5. After uniformly mixing the yam, wolfberry, coix seed, dandelion root, and red dates prepared in step S1 and the honey basil prepared in step S2, 18-20 mL of the Bifidobacterium breve CICC-6183 bacterial solution prepared in step S4 and 18-20 mL of the Lactobacillus acidophilus CICC-6095 bacterial solution prepared in step S3 were added, and anaerobically fermented at 37° C. for 18-24 h; S6. After fermentation, 3% H 2 O 2 The solution is used to inactivate Bifidobacterium breve and Lactobacillus acidophilus in the fermentation product, and then the fermentation product is obtained, and CaCl is added. 2 After standing for 15-30 minutes, centrifuge to obtain the supernatant, then add 50-75% ethanol, centrifuge again, collect the supernatant, filter, and perform reflux extraction for 30-60 minutes through a condensation reflux device. After the extraction is completed, the desired fermentation liquid is obtained; S7, pouring the fermentation liquid prepared in step S5 into a homogenizer and homogenizing it twice at 30 MPa; S8. High temperature instantaneous sterilization is performed, the sterilization temperature is 135° C., and the time is 10-20 seconds. After the sterilization is completed, the beverage of the present invention is obtained.

6. A liver-protecting and antioxidant health-care functional beverage and a preparation method thereof according to claim 3, Features: In the step S1, the raw material is crushed and filtered through a 120-mesh sieve to obtain a 120-mesh powder.

7. A liver-protecting and antioxidant health-care functional beverage and a preparation method thereof according to claim 3, Features: The method for preparing refined honey in step S2 of the present invention is: pour the honey into a clean pot, heat it over a slow fire for 10-15 minutes, stir it continuously and remove the foam during the heating.

8. A liver-protecting and antioxidant health-care functional beverage and a preparation method thereof according to claim 3, Features: The glucose blood liver agar medium used in step S3 of the present invention comprises: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g plant peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (K 2 HPO 4 10g, KH 2 PO 4 10 g, distilled water 100 mL), solution B 5 mL (MgSO 4 7H 2 O 4g, NaCl 0.2g, FeSO 4 7H 2 O 0.2g, MnSO 4 ·2H 2 O 0.2g, distilled water 100mL), 5% L-cysteine ​​hydrochloride solution 10mL, Tween-80 1g, agar 15g, defibrinated sheep blood 50mL, distilled water 825mL.

9. A liver-protecting and antioxidant health-care functional beverage and a preparation method thereof according to claim 3, Features: The components of the MRS agar medium used in step S4 of the present invention are: 10g peptone, 10g beef extract powder, 5g yeast extract powder, 20g glucose, 1g Tween, 5g sodium acetate, 2g triammonium citrate, 2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate heptahydrate, 0.05g sulfuric acid tetrahydrate, 15g agar, 1000mL distilled water, and the pH is adjusted to 6.2-6.5; the components of the glucose blood liver agar medium are: 2.5g peptone, 1.5g beef extract, 10g glucose blood liver, 18g agar, and 1000mL distilled water.

10. A liver-protecting and antioxidant health-care functional beverage and a preparation method thereof according to claim 3, Features: In step S5 of the present invention, the volumes of the Bifidobacterium breve bacterial solution and the Lactobacillus acidophilus bacterial solution are both 18 mL.

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