Healthy food composition for protecting liver and relieving fatigue and preparation method thereof

Abstract

The invention relates to the technical field of healthy food, and discloses a healthy food composition for protecting liver and relieving fatigue and a preparation method of the healthy food composition for protecting liver and relieving fatigue. 5 to 10 parts of propolis; 5-7 parts of a chrysanthemum extract; 2-4 parts of a wolfberry extract; 7-10 parts of a cassia seed extract; 2-5 parts of a ginseng extract; 1.3 to 1.7 parts of a saffron crocus extract; the royal jelly and the propolis are combined according to the formula proportion, the prepared composition finished product has the effect of effectively relieving physical fatigue under the specific dosage, metabolite after exercise can be removed, the composition has the effect of increasing glycogen reserve, and the composition can be used for treating alcoholic liver injury model mice, and has the advantages that the composition finished product can be used for treating alcoholic liver injury; the composition can reduce the content of malondialdehyde and triglyceride in liver tissue and increase the content of glutathione, so that the composition provided by the invention not only has a liver protection effect, but also can relieve physical fatigue, reduce generation of serum urea and lactic acid after exercise and increase in-vivo glycogen reserve.

Description

Technical Field

[0001] The present invention relates to the technical field of healthy food, in particular to a healthy food composition for protecting liver and relieving fatigue and a preparation method thereof. Background Art

[0002] Alcohol has a significant impact on the health of the liver and stomach. Long-term drinking or excessive drinking can cause liver damage and trigger a series of liver diseases, such as fatty liver, hepatitis, and cirrhosis. When the liver is damaged, its detoxification and metabolic functions will decline, which in turn affects the overall health of the body. In addition, alcohol can irritate the gastric mucosa, leading to gastric diseases such as gastritis and gastric ulcers, and in severe cases may even cause gastric bleeding.

[0003] There is also a close relationship between the liver and physical fatigue. The liver is an important metabolic organ in the human body, responsible for breaking down and converting nutrients to provide energy for the body. When liver function is impaired, its metabolic capacity is weakened and it cannot effectively provide enough energy to the body, leading to symptoms such as physical fatigue and weakness. Therefore, protecting liver health is of great significance to relieving physical fatigue.

[0004] Currently, the liver protection products on the market are mainly thistle liver protection products (such as milk thistle, milk thistle) and medicinal and edible liver protection products. However, the functions of these two types of products are relatively simple, almost all of them only have the effect of protecting the liver, and lack the effect of relieving fatigue and nourishing the stomach. In particular, thistle products, long-term use will irritate the gastrointestinal tract, causing abdominal distension, abdominal pain and diarrhea; if they cannot be completely metabolized, they may also increase the burden on the liver. In addition, milk thistle has the effect of cooling blood and stopping bleeding. Women should not take it during their menstrual period. At the same time, some people will experience abdominal discomfort after taking it.

[0005] Therefore, developing a healthy food that can sober up, protect the liver, nourish the stomach and relieve physical fatigue is of great significance to human health. This healthy food can not only meet the health needs of modern people in a fast-paced life, but also help improve liver and stomach problems caused by bad living habits and improve the quality of life. Summary of the invention

[0006] 1. Technical issues to be solved

[0007] In view of the deficiencies in the prior art, the present invention provides a healthy food composition for protecting the liver and relieving fatigue and a preparation method thereof, which has the advantages of sobering up, protecting the liver, nourishing the stomach and relieving physical fatigue, and solves the problem that traditional healthy foods only have the effect of protecting the liver but lack the effects of relieving fatigue and nourishing the stomach.

[0008] (II) Technical solution

[0009] To achieve the above object, the present invention provides the following technical solution: a health food composition for protecting the liver and relieving fatigue, wherein the raw materials of the composition and their mass fractions are: 90-100 parts of royal jelly; 5-10 parts of propolis; 5-7 parts of chrysanthemum extract; 2-4 parts of wolfberry extract; 7-10 parts of cassia seed extract; 2-5 parts of ginseng extract; 1.3-1.7 parts of saffron extract; 8-11 parts of licorice extract.

[0010] Preferably, a preparation method of a health food composition for protecting the liver and relieving fatigue is prepared according to the raw materials and mass fractions of the above-mentioned health food composition for protecting the liver and relieving fatigue, and comprises the following preparation steps:

[0011] Step 1, preparation of royal jelly freeze-dried powder: Pretreat, pre-freeze, sublimation dry, analytical dry and freeze-dry the royal jelly, and then pulverize the freeze-dried royal jelly to obtain royal jelly freeze-dried powder;

[0012] Step 2, preparation of propolis powder: Freeze, pulverize, ethanol extract, filter the extract, concentrate the extract and recover ethanol from the raw propolis, and finally obtain pure propolis. Then, put the pure propolis into a freezer at -18°C for freezing, and then pulverize to obtain propolis powder;

[0013] Step 3, mixing of medicinal materials and spray-drying process: Pour the chrysanthemum extract, wolfberry extract, cassia seed extract, ginseng extract, saffron extract and licorice extract in the formula ratio into a glass beaker containing 100 ml of pure water, heat at a temperature of 60-80°C for 30-40 minutes until the solution is concentrated to 10 ml to obtain a mixed medicinal material solution, and then spray-dry the medicinal material solution into medicinal material particles by spray drying after concentration;

[0014] Step 4, sterilization: Spread the royal jelly freeze-dried powder, propolis powder and medicinal material particles on the baking trays respectively, irradiate and sterilize under ultraviolet light for 30 minutes, turn over and flatten the powder, and then irradiate under ultraviolet light for 30 minutes, and repeat the irradiation 2 times for sterilization;

[0015] Step 5, mixing, filling capsules and polishing: After the operating environment reaches the 100,000-class cleanliness standard, mix, fill capsules and polish the sterilized royal jelly freeze-dried powder, propolis powder and medicinal material particles according to the prescription amount to obtain the finished product. The room temperature is controlled at 18-24°C during the whole production process, and the environmental relative humidity is maintained at 45% - 55%.

[0016] Preferably, the pretreatment of the royal jelly in Step 1: First, add fresh royal jelly to be freeze-dried to sterile distilled water in a ratio of 1:1, filter through a 100-mesh filter screen to remove impurities, then rinse the filter residue with distilled water 1-2 times, separate the 10-hydroxy-2-decenoic acid crystals, grind them to the required mesh number for filtration, and then return them to the royal jelly filtrate and mix well.

[0017] Preferably, in the step 1, the royal jelly is pre-frozen: the pre-treated royal jelly is transferred into a square plate and rapidly frozen into a solid at a low temperature of -18°C to -20°C for later use.

[0018] Preferably, in the step 1, the royal jelly is dried by sublimation: the royal jelly frozen into a solid is moved into a freeze drying chamber, and the royal jelly is dried by sublimation in an environment of -25°C and 13.33 Pa for 12-14 hours.

[0019] Preferably, in the step 1, the royal jelly is dried by sublimation: after sublimation drying, the water content of the royal jelly is 10%, and drying is continued to the water content of 1.8% to 2%, the temperature of the drying is 30°C-40°C, the vapor pressure in the drying oven is maintained at 13.33-14.00 Pa, and the drying time is 4-6 hours.

[0020] Preferably, in the pulverizing process in step 1: the freeze-dried royal jelly is in the form of loose blocks, which are pulverized by a ball mill and then sieved through a 100-mesh sieve to obtain freeze-dried royal jelly powder, and the relative humidity of the operating room is set below 20%.

[0021] Preferably, in the freezing and crushing process of raw propolis in step 2, the propolis is placed in a freezer at -18°C to -20°C, frozen for more than 2 hours, crushed by a ball mill, and the temperature of the working environment is maintained below 15°C.

[0022] Preferably, the step 2 comprises:

[0023] S1.1, ethanol extraction of raw propolis: raw propolis powder and 95% ethanol in a ratio of 1:4 are placed in a stainless steel stirring barrel, and a countercurrent multi-stage extraction operation is performed at room temperature. After the operation is completed, the extraction is placed in a storage tank;

[0024] S1.2, Filtration of propolis ethanol extract: Filter the propolis filtrate to be concentrated in the storage tank through a 100-mesh screen, immerse the coiled pipe passing refrigerated brine into the storage tank, lower the storage tank temperature to below 5°C, let it stand for 3 to 4 hours, take the supernatant, filter it through a 100-mesh screen, and pump it into the concentration tank;

[0025] S1.3. Concentration of propolis ethanol extract and recovery of ethanol: The propolis ethanol liquid in the concentration tank is passed into a distiller for condensation and concentration to make propolis paste, which is then cooled to obtain pure propolis.

[0026] Preferably, in the pure propolis crushing process in step 2, the pure propolis is placed in a freezer at -18°C to -20°C and frozen, the pure propolis blocks are crushed using a ball mill, and filtered through a 100-mesh sieve to obtain pure propolis powder, and the temperature of the operating environment is maintained below 15°C.

[0027] Compared with the prior art, the present invention provides a healthy food composition and preparation method for protecting liver and relieving fatigue, which has the following beneficial effects:

[0028] The present invention combines royal jelly and propolis according to a formula ratio to prepare a finished composition, which can effectively prolong the weight-bearing swimming time of mice at a specific dosage, indicating that the composition has the effect of relieving physical fatigue, and by reducing the urea and lactic acid levels in the serum of mice after exercise, it shows a positive effect on the clearance of post-exercise metabolites. The composition can also increase the content of liver glycogen in mice or reduce its consumption, further proving its effect in increasing glycogen reserves. For mice with alcoholic liver damage model, the composition can reduce the content of malondialdehyde and triglyceride in liver tissue while increasing the content of glutathione, indicating that it has a certain liver protective effect. In summary, the composition provided by the present invention not only has a liver protective effect, but also can relieve physical fatigue, reduce the production of serum urea and lactic acid after exercise, and increase glycogen reserves in the body. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 A flow chart was prepared for the present invention. DETAILED DESCRIPTION

[0030] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] See also Figure 1 A healthy food composition for protecting the liver and relieving fatigue. The raw materials of the composition and their mass proportions are: 90-100 parts of royal jelly; 5-10 parts of propolis; 5-7 parts of chrysanthemum extract; 2-4 parts of wolfberry extract; 7-10 parts of cassia seed extract; 2-5 parts of ginseng extract; 1.3-1.7 parts of saffron extract; and 8-11 parts of licorice extract.

[0032] Specifically, a method for preparing a healthy food composition for protecting liver and relieving fatigue is prepared according to the raw materials and mass fractions of the healthy food composition for protecting liver and relieving fatigue, comprising the following preparation steps:

[0033] Step 1, preparation of royal jelly freeze-dried powder: pre-treating, pre-freezing, sublimation drying, analytical drying and freeze drying the royal jelly, and then crushing the freeze-dried royal jelly to obtain royal jelly freeze-dried powder;

[0034] Step 2, preparation of propolis powder: freezing, crushing, ethanol extraction, filtering the extract, concentrating the extract and recovering the ethanol to obtain pure propolis, and then freezing the pure propolis in a refrigerator at a temperature of -18°C, and then crushing to obtain propolis powder;

[0035] Step 3, mixing medicinal materials and spray drying process: pouring the chrysanthemum extract, wolfberry extract, cassia seed extract, ginseng extract, saffron extract and licorice extract in a formula ratio into a glass beaker filled with 100 ml of pure water, heating at a temperature of 60-80° C. for 30-40 minutes until the solution is concentrated to 10 ml to obtain a mixed medicinal material solution, and after concentration, spray drying the medicinal material solution into medicinal material particles;

[0036] Step 4, sterilization: Spread the royal jelly freeze-dried powder, propolis powder and medicinal material granules on a baking tray respectively, sterilize under ultraviolet light for 30 minutes, turn the powder over and spread it flat, and then irradiate under ultraviolet light for 30 minutes, repeat irradiation twice for sterilization;

[0037] Step 5. Mixing, capsule filling and polishing: After the operating environment reaches the 100,000-level cleanliness standard, the sterilized royal jelly freeze-dried powder, propolis powder and medicinal granules are mixed, capsule filled and polished according to the prescribed amount to obtain the finished product. The room temperature of the entire production process is controlled at 18-24°C, and the relative humidity of the environment is maintained at 45% to 55%.

[0038] Specifically, the royal jelly is pretreated in step 1: first, fresh royal jelly to be freeze-dried is added with sterile distilled water at a ratio of 1:1, filtered through a 100-mesh filter to remove impurities, and then the filter residue is rinsed with distilled water for 1-2 times to separate 10-hydroxy-2-decenoic acid crystals, and ground according to the mesh number required for filtration, and then returned to the royal jelly filtrate for mixing. Through dilution and filtration, large particles of impurities and some microorganisms in the royal jelly can be effectively removed, thereby improving the purity of the product; at the same time, separating and recovering 10-hydroxy-2-decenoic acid crystals helps to retain the active ingredients in the royal jelly.

[0039] Specifically, in step 1, the royal jelly is pre-frozen: the pre-treated royal jelly is transferred into a square plate and quickly frozen into a solid at a low temperature of -18°C to -20°C for standby use. Rapid freezing can reduce the formation of ice crystals, protect the biologically active substances in the royal jelly from being destroyed, and provide a good foundation for subsequent sublimation drying.

[0040] Specifically, in step 1, the royal jelly is sublimated and dried: the frozen royal jelly is moved into a freeze drying box, and the royal jelly is sublimated and dried in an environment of -25°C and 13.33 Pa for 12-14 hours. Sublimation drying (i.e. freeze drying) is a gentle drying method that can be carried out at low temperature and low pressure, avoiding the influence of high temperature on heat-sensitive components, thereby maximizing the retention of nutrients and biological activity in the royal jelly. In addition, the freeze-dried product has a long shelf life and is easy to store and transport.

[0041] Specifically, in step 1, the royal jelly is dried by sublimation: after drying, the water content of the royal jelly is 10%. (Royal jelly with such a water content cannot be preserved for a long time. It is necessary to increase the heating temperature and maintain a high vacuum degree so that the adsorbed water molecules can be decomposed from the loose royal jelly under a larger decomposition driving force) and continue to dry until the water content is 1.8% to 2%. The temperature of the decomposition drying is 30°C-40°C, the vapor pressure in the drying oven is maintained at 13.33-14.00 Pa, and the drying time is 4-6 hours.

[0042] Specifically, in the crushing process of step 1: the royal jelly after freeze-drying is a loose block, which is crushed by a ball mill and then sieved through a 100-mesh sieve to obtain royal jelly freeze-dried powder (the freeze-dried powder has strong hygroscopicity. In order to prevent pollution and water vapor intrusion, the air entering the operating room must be purified), and the relative humidity of the operating room is set below 20%.

[0043] Specifically, in step 2, the freezing and crushing process of the original propolis is as follows: the propolis is placed in a freezer at -18°C to -20°C, frozen for more than 2 hours, and crushed using a ball mill, and the temperature of the working environment is maintained below 15°C.

[0044] Specifically, step 2 includes:

[0045] S1.1, ethanol extraction of raw propolis: raw propolis powder and 95% ethanol are mixed in a ratio of 1:4, placed in a stainless steel stirring barrel, and subjected to countercurrent multi-stage extraction at room temperature. After the extraction, the raw propolis powder is placed in a storage tank (to separate the effective components in propolis to the maximum extent and to reduce the energy consumption of concentrating the propolis ethanol extract);

[0046] S1.2, Filtration of propolis ethanol extract: Filter the propolis filtrate to be concentrated in the storage tank through a 100-mesh screen (after the filtration, some cold ethanol insoluble matter still exists), immerse the storage tank with a coil of refrigerated brine, lower the storage tank temperature to below 5°C (to accelerate the precipitation of this part of cold ethanol insoluble matter), let it stand for 3 to 4 hours, then take the supernatant, filter it through a 100-mesh screen, and pump it into the concentration tank;

[0047] S1.3. Concentration of propolis ethanol extract and recovery of ethanol: The propolis ethanol liquid in the concentration tank is passed into a distiller for condensation and concentration to make propolis paste, which is then cooled to obtain pure propolis.

[0048] Specifically, the pure propolis crushing process in step 2 is as follows: the pure propolis is placed in a freezer at -18°C to -20°C, the pure propolis blocks are crushed using a ball mill, and filtered through a 100-mesh sieve to obtain pure propolis powder, and the temperature of the operating environment is maintained below 15°C.

[0049] Example 1

[0050] The finished product of the composition was subjected to a test for its function of relieving physical fatigue:

[0051] Animals and groups: A total of 160 male Kunming mice weighing 18-22g were randomly divided into 16 groups according to their weight, and each of the 4 groups was used for the test of one indicator, i.e. the normal control group was gavaged with distilled water and the three dose groups were gavaged with the composition at 125mg / kg.BW / d, 250mg / kg.BW / d, and 500mg / kg.BW / d, respectively, which were equivalent to 5 times, 10 times, and 20 times the recommended intake of the human body, respectively. All experimental animals were fed with a complete nutritional formula. The animals were free to eat and drink.

[0052] Preparation of instruments and reagents:

[0053] Instruments: swimming box (50cmX50cmX40cm), 722-type spectrophotometer, water bath, stopwatch, SL-1001-type electronic scale.

[0054] Reagents: serum urea assay kit, liver glycogen assay kit, whole blood lactate assay kit.

[0055] Methods: According to the "Health Food Functional Evaluation Procedure and Test Method Specification", the weighted swimming test, serum urea determination (diacetyl monoxime method), liver glycogen determination (anthrone method) and blood lactate determination in the fatigue relief function test method were carried out. The gavage volume was 0.4ml / 20g body weight, once a day, for 30 consecutive days.

[0056] Weighted swimming test: 30 minutes after the last administration of the test substance, place the mice in a swimming box with a water depth of 30 cm and a water temperature of 25℃±0.5℃, and a lead sheet of 5% of the body weight is loaded on the base of the mouse's tail. The time from the start of swimming to the death of the mouse is recorded as the mouse's swimming time.

[0057] Serum urea determination (diacetyl monoxime method): 30 minutes after the last administration of the test substance, swim without weight in water at 30℃±0.5℃ for 90 minutes, rest for 60 minutes, remove the eyeball and collect 0.8ml of blood (without anticoagulant). After the blood coagulates, centrifuge and collect serum, which is determined using the urea determination kit.

[0058] Determination of liver glycogen (anthrone method): The animals were killed 30 minutes after the last sampling, and the liver was taken out and the liver glycogen was determined according to the instructions of the kit.

[0059] Blood lactate determination: 30 minutes after the last animal sample was given, 20 μl of blood was collected from the medial canthal vein of the eyeball using a capillary glass tube, and the lactate content was determined using a whole blood lactate determination kit. After blood collection, the animal was allowed to swim in water at a temperature of 30°C ± 0.5°C for 10 minutes without any weight, and then stopped. 20 μl of blood was immediately collected for blood lactate determination, and 20 μl of blood was collected again after a 20-minute rest for blood lactate determination.

[0060] Data statistical processing method: The experimental data were compared between the experimental groups using one-way analysis of variance in the SAS statistical software package, and Dunnett's T test was used to compare the experimental groups with the control group.

[0061] Mouse weighted swimming test:

[0062] The results are shown in Table 1

[0063] Table 1 Effect of finished composition on weight-bearing swimming time of mice (mean ± Standard Deviation

[0064] Group Number of animals <![CDATA[Negative weight swimming time (min) a > Normal control group 10 4.72±1.78 Low dose group 10 4.58±1.96 Medium dose group 10 7.01±1.82 High dose group 10 <![CDATA[8.14±3.36 * >

[0065] a One-way ANOVA among the experimental groups, F=5.68, P<0.01

[0066] *Compared with the normal control group, P<0.05.

[0067] As shown in Table 1, the difference in the mean value of the weighted swimming time of each group was significant by one-way analysis of variance (F=5.68, P<0.01). By Dunnett's T test, the mean value of the weighted swimming time of the high-dose group of the finished composition was different from that of the normal control group (P<0.05). This indicates that the finished composition has the effect of prolonging the weighted swimming time of mice at a dose of 500 mg / kg.BW / d (equivalent to 20 times the recommended intake of the human body).

[0068] Table 2 Effect of finished composition on serum urea in mice after exercise (mean ± Standard Deviation

[0069] Group Number of animals <![CDATA[Serum urea (mmol / L) a > Normal control group 10 9.36±1.01 Low dose group 10 8.88±1.02 Medium dose group 10 <![CDATA[7.39±0.97 * ]]> High dose group 10 <![CDATA[7.14±0.89 * ]]>

[0070] a One-way ANOVA among the experimental groups, F=12.62, P<0.01

[0071] *Compared with the normal control group, P<0.05.

[0072] Serum urea determination The results of serum urea determination of each group of animals are shown in Table 2. As shown in Table 2, the difference in the mean values ​​of serum urea in each group was polar (F=12.62, P<0.01) by one-way analysis of variance. By Dunnett's T test, the mean values ​​of urea in the medium and high dose groups of the finished composition were different from those in the normal control group (P<0.05). This indicates that the finished composition can reduce the production of serum urea in mice after exercise at a dose of 250 mg / kg.BW / d and 500 mg / kg.BW / d (equivalent to 10 times and 20 times the recommended intake of the human body).

[0073] The results of liver glycogen determination are shown in Table 3.

[0074] Table 3 Effect of finished composition on mouse liver glycogen (mean ± Standard Deviation

[0075] Group Number of animals <![CDATA[Hepatic glycogen (mg / g liver tissue) a > Normal control group 10 5.10±1.56 Low dose group 10 6.45±1.68 Medium dose group 10 6.16±2.41 High dose group 10 <![CDATA[10.40±2.34 * ]]>

[0076] a One-way ANOVA among the experimental groups, F=13.02, P<0.01*Difference compared with the normal control group, P<0.05.

[0077] As shown in Table 3, the differences in the mean values ​​of liver glycogen in each group of animals were analyzed by one-way ANOVA (F=13.02, P<0.01). The differences in the mean values ​​of liver glycogen in the high-dose group of the finished composition and the normal control group were compared by Dunnett's T test (P<0.05). This indicates that the finished composition has the effect of increasing the glycogen reserve or reducing the glycogen consumption of mice at a dose of 500 mg / kg.BW / d (equivalent to 20 times the recommended intake of humans).

[0078] Blood lactate determination The blood lactate levels of each group of mice at rest, immediately after swimming, after rest, and the area under the blood lactate curve are shown in Table 4.

[0079] Table 4 Effects of the finished composition on blood lactate levels and area under the curve of mice before and after swimming (mean ± Standard Deviation

[0080]

[0081]

[0082] A Blood lactate before swimming: B Blood lactate immediately after swimming: C Blood lactate 20 minutes after swimming a One-way ANOVA among the experimental groups, F=3.30, P<0.05, *Difference compared with the normal control group, P<0.05.

[0083] After one-way analysis of variance, there was no significant difference in the mean area under the blood lactate curve of each group of mice (P>0.05), indicating that the finished composition had no obvious effect on the production of blood lactate in mice after exercise at doses of 125 mg / kg.BW / d, 250 mg / kg.BW / d and 500 mg / kg.BW / d (equivalent to 5 times, 10 times and 20 times the recommended human intake).

[0084] Example 2

[0085] A trial report on the auxiliary protection function against chemical liver damage (alcoholic liver damage model)

[0086] Animals and groups: A total of 50 male Kunming mice, weighing 18-22g, were provided by the Shanghai Experimental Animal Center of the Chinese Academy of Sciences. Production license number: SCXK (Shanghai) 2002-0010; use license number: SYXK (Suzhou) 2002-0057. The animals were randomly divided into 5 groups according to their body weight, namely, a blank control group, a model control group and three dose groups (125mg / kg.BW / d, 250mg / kg.BW / d and 500mg / kg.BW / d of the finished composition were administered by gavage, which are equivalent to 5 times, 10 times and 20 times the recommended intake of the human body, respectively), and the gavage volume was 20ml / kg.BW. All experimental animals were fed with complete nutritional formula feed, and the animals were free to eat and drink.

[0087] Experimental method: The test method for auxiliary protection of chemical liver damage (alcoholic liver damage model) in the "Health Food Functional Evaluation Procedure and Test Method Specification" was followed. The test samples were given by oral gavage every day, and the blank control group and model control group were given distilled water. The animals were weighed twice a week, and the dose of the test samples was adjusted according to body weight. The test samples were given for 30 consecutive days. At the end, the model control group and each sample group were gavaged with 50% ethanol 12ml / kg.BW once, and the blank control group was given distilled water. The animals were fasted for 16 hours and killed, and various indicators were tested and pathological histological examinations were performed.

[0088] Instruments and reagents

[0089] Instruments: SL-1001 electronic scale, 722 spectrophotometer, water bath, ordinary centrifuge, vortexer, tissue homogenizer, A0820 pathology slicer, OLYMPUS biological microscope.

[0090] Reagents: physiological saline; anhydrous ethanol (analytical grade); MDA assay kit, GSH assay kit.

[0091] Detection indicators

[0092] Determination of MDA content in liver tissue: Take mouse liver and prepare 10% liver tissue homogenate. Take 0.1 ml of tissue homogenate, add detection reagents according to the steps described in the MDA determination kit instructions, and then use a 722 spectrophotometer to measure the absorbance of the blank tube, standard tube and measurement tube. Calculate the MDA content in liver tissue with the following formula: MDA content in liver tissue = (absorbance of measurement tube - absorbance of blank tube) / (absorbance of standard tube - absorbance of blank tube) x concentration of standard product ÷ concentration of tissue homogenate.

[0093] Determination of GSH content in liver tissue: Take 0.03 ml of 10% liver tissue homogenate, add the detection reagents according to the steps described in the GSH determination kit manual, and then use a 722 spectrophotometer to measure the absorbance of the blank tube, standard tube, measurement tube and measurement blank tube. Calculate the GSH content in liver tissue using the following formula: GSH content in liver tissue = (measurement tube absorbance - measurement blank tube absorbance) / (standard tube absorbance - blank tube absorbance) x standard concentration ÷ tissue homogenate concentration.

[0094] Determination of TG content in liver tissue: Take 0.03 ml of 10% liver tissue homogenate, add the detection reagents according to the steps described in the TG determination kit instructions, and then use a 722 spectrophotometer to measure the absorbance of the standard tube and the determination tube. Calculate the TG content in liver tissue using the following formula: TG content in liver tissue = absorbance of the determination tube / absorbance of the standard tube X concentration of the standard product ÷ concentration of tissue homogenate.

[0095] Pathological histological examination of the liver: a cross-section was made from the middle of the left lobe of the liver, frozen sections were made, and pathological histological examination was performed by Sudan III staining. The entire tissue section was continuously observed with a 40x objective lens, mainly to observe the distribution, range, and area of ​​fat droplets in the liver.

[0096] Data statistical processing method: The experimental data were compared between the experimental groups using one-way analysis of variance in the SAS statistical software package, and the Q test was used to compare the two experimental groups.

[0097] Determination of MDA content in liver tissue: The results are shown in Table 5. The difference of the mean value of MDA in the liver tissue of each group of animals was analyzed by one-way ANOVA (F=9.92, P<0.01). By Q test, the mean value of MDA in the liver tissue of the model control group was different from that of the normal control group (P<0.05), and the mean value of MDA in the liver tissue of the medium and high dose groups of the finished composition was different from that of the model control group (P<0.05). It shows that the finished composition has the effect of reducing the MDA content in the liver tissue of the alcoholic liver injury model mice at a dose of 250mg / kg.BW / d and 500mg / kg.BW / d (equivalent to 10 times and 20 times the recommended intake of the human body).

[0098] Table 5 Effect of finished composition on MDA content in mouse liver tissue

[0099] Group Number of animals <![CDATA[MDA(nmol / mg) a ]]> Blank control group 10 <![CDATA[421.1±82.9 * ]]> Model control group 10 631.2±84.5 Low dose group 10 577.7±65.8 Medium dose group 10 <![CDATA[542.3±63.9 * ]]> High dose group 10 <![CDATA[533.3±87.9 * >

[0100] Note: a One-way ANOVA among the experimental groups, F=9.92, P<0.01, * Compared with the model control group, P<0.05.

[0101] Determination of GSH content in liver tissue: The results are shown in Table 6.

[0102] Table 6 Effect of finished composition on GSH content in mouse liver tissue

[0103] Group Number of animals <![CDATA[GSH(μmol / g) a ]]> Blank control group 10 <![CDATA[12.38±1.02 * ]]> Model control group 10 8.50±0.81 Low dose group 10 8.55±0.82 Medium dose group 10 9.18±1.08 High dose group 10 <![CDATA[10.16±1.12 * ]]>

[0104] Note: a One-way ANOVA among the experimental groups, F=27.15, P<0.01, * Compared with the model control group, P<0.05.

[0105] As shown in Table 6, the difference of the mean value of GSH in the liver tissue of each group of animals was analyzed by one-way ANOVA (F=27.15, P<0.01). By Q test, the mean value of GSH in the liver tissue of the model control group was different from that of the normal control group (P<0.05), and the mean value of GSH in the liver tissue of the high-dose group of the finished composition was different from that of the model control group (P<0.05). This shows that the finished composition has the effect of increasing the content of GSH in the liver tissue of the alcoholic liver injury model mice at a dose of 500 mg / kg.BW / d (equivalent to 20 times the recommended intake of the human body).

[0106] Determination of TG content in liver tissue: The results are shown in Table 7. The difference of the mean TG value in the liver tissue of each group of animals was analyzed by one-way ANOVA (F=11.26, P<0.01). By Q test, the mean TG value in the liver tissue of the model control group was different from that of the normal control group (P<0.05), and the mean TG value in the liver tissue of the high-dose group of the finished composition was different from that of the model control group (P<0.05). This shows that the finished composition has the effect of reducing the TG content in the liver tissue of the alcoholic liver injury model mice at a dose of 500 mg / kg.BW / d (equivalent to 20 times the recommended intake of the human body).

[0107] Table 7 Effect of finished composition on TG content in mouse liver tissue

[0108] Group Number of animals <![CDATA[TG(μmol / g) a ]]> Blank control group 10 <![CDATA[7.63±1.07 * <!-- 8 -->]]> Model control group 10 11.83±1.22 Low dose group 10 10.73±1.66 Medium dose group 10 10.25±1.64 High dose group 10 <![CDATA[9.31±1.75 * ]]>

[0109] Note: a One-way ANOVA among the experimental groups, F=11.26, P<0.01, * Compared with the model control group, P<0.05.

[0110] Pathological examination of liver tissue: The results are shown in Table 8. The difference in the level of fatty degeneration of liver tissue of each group of animals was analyzed by one-way ANOVA (F=29.59, P<0.01). The Q test showed that the level of fatty degeneration of liver tissue in the model control group was different from that in the normal control group (P<0.05), but the level of fatty degeneration of liver tissue in each dose group of the finished composition was not different from that in the model control group (P>0.05). This indicates that the finished composition had no significant effect on the fatty degeneration of liver in the alcoholic liver injury model mice at doses of 125 mg / kg.BW / d, 250 mg / kg.BW / d and 500 mg / kg.BW / d (equivalent to 5 times, 10 times, and 20 times the recommended intake of the human body).

[0111] Table 8 Effect of finished composition on fatty degeneration of mouse liver tissue (quantitative value a , )

[0112] Group Number of animals <![CDATA[Steatosis a > Blank control group 10 <![CDATA[0.10±0.32 * ]]> Model control group 10 2.60±0.70 Low dose group 10 2.70±0.48 Medium dose group 10 2.40±0.70 High dose group 10 2.30±0.82

[0113] Note: a The degree of fatty degeneration of liver tissue was scored, with 0 points for scattered and scarce fat droplets in hepatocytes, 1 point for no more than 1 / 4 of hepatocytes containing fat, 2 points for no more than 1 / 2 of hepatocytes containing fat droplets, 3 points for no more than 3 / 4 of hepatocytes containing fat droplets, and 4 points for quantification: b One-way ANOVA among the experimental groups, F=29.59, P<0.01, *Difference compared with the model control group, P<0.05.

[0114] The experimental results of Example 1 and Example 2 show:

[0115] The finished composition, at a dosage of 500 mg / kg.BW / d, can prolong the weight-bearing swimming time of mice, indicating that the composition has the effect of alleviating physical fatigue.

[0116] The finished composition can also reduce the production of urea in the serum of mice after exercise at a dosage of 500 mg / kg.BW / d, indicating that it has a significant effect on reducing the serum urea of ​​mice after exercise.

[0117] The finished composition, at a dosage of 250 mg / kg.BW / d and 500 mg / kg.BW / d, can reduce the lactic acid content in the serum of mice after exercise, indicating that it has a significant effect on reducing blood lactic acid after exercise.

[0118] The finished composition can increase the content of liver glycogen or reduce the consumption of glycogen in mice at a dosage of 500 mg / kg.BW / d, indicating that it is effective in increasing glycogen reserves or reducing glycogen consumption.

[0119] The finished composition had no significant effect on hepatic fatty degeneration in mice with alcoholic liver injury model at dosages of 125 mg / kg.BW / d, 250 mg / kg.BW / d and 500 mg / kg.BW / d, but was able to reduce the MDA and TG contents in liver tissue and increase the GSH content, indicating that it has a certain liver protective effect.

[0120] In summary, the finished product of the composition shows positive effects on relieving physical fatigue, reducing the production of serum urea and lactic acid after exercise, increasing glycogen reserves and protecting the liver at a specific dosage.

[0121] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A healthy food composition for protecting liver and relieving fatigue, characterized in that: The raw materials of the composition and their mass proportions are: 90-100 parts of royal jelly; 5-10 parts of propolis; 5-7 parts of chrysanthemum extract; 2-4 parts of wolfberry extract; 7-10 parts of cassia seed extract; 2-5 parts of ginseng extract; 1.3-1.7 parts of saffron extract; and 8-11 parts of liquorice extract.

2. A method for preparing a healthy food composition for protecting liver and relieving fatigue, characterized in that: The raw materials and mass fractions of a healthy food composition for protecting liver and relieving fatigue according to claim 1 are prepared, comprising the following preparation steps: Step 1, preparation of royal jelly freeze-dried powder: pre-treating, pre-freezing, sublimation drying, analytical drying and freeze drying the royal jelly, and then crushing the freeze-dried royal jelly to obtain royal jelly freeze-dried powder; Step 2, preparation of propolis powder: freezing, crushing, ethanol extraction, filtering the extract, concentrating the extract and recovering the ethanol, finally obtaining pure propolis, freezing the pure propolis in a refrigerator at a temperature of -18°C, and then crushing to obtain propolis powder; Step 3, mixing medicinal materials and spray drying process: pouring the chrysanthemum extract, wolfberry extract, cassia seed extract, ginseng extract, saffron extract and licorice extract in a formula ratio into a glass beaker filled with 100 ml of pure water, heating at a temperature of 60-80° C. for 30-40 minutes until the solution is concentrated to 10 ml to obtain a mixed medicinal material solution, and after concentration, spray drying the medicinal material solution into medicinal material particles; Step 4, sterilization: Spread the royal jelly freeze-dried powder, propolis powder and medicinal material granules on a baking tray respectively, sterilize under ultraviolet light for 30 minutes, turn the powder over and spread it flat, and then irradiate under ultraviolet light for 30 minutes, repeat irradiation twice for sterilization; Step 5. Mixing, capsule filling and polishing: After the operating environment reaches the 100,000-level cleanliness standard, the sterilized royal jelly freeze-dried powder, propolis powder and medicinal granules are mixed, capsule filled and polished according to the prescribed amount to obtain the finished product. The room temperature of the entire production process is controlled at 18-24°C, and the relative humidity of the environment is maintained at 45% to 55%.

3. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: The royal jelly is pretreated in step 1 as follows: first, fresh royal jelly to be freeze-dried is added with sterile distilled water at a ratio of 1:1, filtered through a 100-mesh filter to remove impurities, and then the filter residue is rinsed 1-2 times with distilled water to separate 10-hydroxy-2-decenoic acid crystals, which are ground into fine powder according to the mesh number required for filtration and then returned to the royal jelly filtrate for mixing.

4. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: Pre-freezing of royal jelly in step 1: the pre-treated royal jelly is transferred into a square plate and rapidly frozen into a solid at a low temperature of -18°C to -20°C for later use.

5. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: In the step 1, the royal jelly is sublimated and dried: the royal jelly frozen into a solid is moved into a freeze drying box, and the royal jelly is sublimated and dried in an environment of -25°C and 13.33 Pa for 12-14 hours.

6. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: The royal jelly is dried by sublimation in step 1: after sublimation drying, the water content of the royal jelly is 10%, and the royal jelly is dried further until the water content is 1.8% to 2%. The temperature of the drying is 30° C. to 40° C., the vapor pressure in the drying oven is maintained at 13.33 to 14.00 Pa, and the drying time is 4 to 6 hours.

7. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: In the crushing process of step 1, the royal jelly after freeze-drying is in the form of loose blocks, which are crushed by a ball mill and then sieved through a 100-mesh sieve to obtain freeze-dried royal jelly powder, and the relative humidity of the operating room is set to be less than 20%.

8. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: The freezing and crushing process of the raw propolis in the step 2 is as follows: the propolis is placed in a freezer at -18°C to -20°C, frozen for more than 2 hours, and crushed using a ball mill, and the temperature of the working environment is kept below 15°C.

9. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: The second step comprises: S1.1, ethanol extraction of raw propolis: raw propolis powder and 95% ethanol are mixed in a ratio of 1:4, placed in a stainless steel stirring barrel, and subjected to countercurrent multi-stage extraction at room temperature. After the extraction, the mixture is placed in a storage tank; S1.2, Filtration of propolis ethanol extract: Filter the propolis filtrate to be concentrated in the storage tank through a 100-mesh screen, immerse the coiled pipe passing refrigerated brine into the storage tank, lower the storage tank temperature to below 5°C, let it stand for 3 to 4 hours, take the supernatant, filter it through a 100-mesh screen, and pump it into the concentration tank; S1.

3. Concentration of propolis ethanol extract and recovery of ethanol: The propolis ethanol liquid in the concentration tank is passed into a distiller for condensation and concentration to make propolis paste, which is then cooled to obtain pure propolis.

10. The method for preparing a healthy food composition for protecting liver and relieving fatigue according to claim 2, characterized in that: The pure propolis crushing process in step 2 is as follows: the pure propolis is placed in a freezer at -18°C to -20°C and frozen, the pure propolis blocks are crushed using a ball mill, and filtered through a 100-mesh sieve to obtain pure propolis powder, and the temperature of the operating environment is maintained below 15°C.

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