Compound herbal functional health-care beverage for protecting liver and removing fat and preparation method of compound herbal functional health-care beverage

Abstract

The invention discloses a liver-protecting and fat-reducing compound herbal functional health-care beverage and a preparation method thereof, and relates to preparation of a compound extract healthy herbal tea beverage which has Mongolian medicine characteristics, liver-protecting and fat-reducing effects and pure natural herbal taste. The liver-protecting and fat-removing functional beverage which is fresh in taste and sweet in aftertaste is obtained by mixing medicinal materials such as fructus gardeniae, fructus lycii, herba taraxaci, flos lonicerae and stigma croci according to a certain proportion and performing extraction, filtration, sterilization, filling and secondary sterilization. Animal experiments and cell experiments prove that the functional beverage capable of protecting liver and removing fat has the effects of protecting liver and reducing fat.

Description

Technical Field

[0001] The invention relates to the field of food and beverages, and in particular to a liver-protecting and lipid-reducing health beverage. Background Art

[0002] The Mongolian medicine formula for protecting liver and clearing fat, called eligehamagalagqi in Mongolian, is a new formula based on the combination of traditional Chinese medicine data mining technology and Mongolian medicine efficacy compatibility method. The Mongolian medicine formula for protecting liver and clearing fat is composed of gardenia, wolfberry, dandelion, honeysuckle, saffron and other compatibility, and has the effects of protecting liver and lowering fat.

[0003] With the continuous development of medical science and technology, the research and development of new liver protection drugs is also making continuous progress. Some new drugs have better efficacy and lower side effects, providing new options for the treatment and prevention of liver diseases. At present, the market for liver protection drugs and health products has reached a certain scale and still has a lot of room for development in the future. As people's needs increase, the types of drugs and health products on the market for preventing non-alcoholic fatty liver disease are gradually increasing. There are many liver protection drugs and health products on the market, such as bifendate dripping pills, diammonium glycyrrhizinate enteric-coated capsules, reduced glutathione tablets, polyene phosphatidylcholine capsules, liver protection tablets, silybin, etc.

[0004] Therefore, the development of a compound beverage based on gardenia can solve the shortcomings in taste and function, and at the same time, it can inherit the essence of the health-preserving theory in traditional medicine, expand the user group of the product, and enrich the types of non-alcoholic fatty liver health products on the market. However, there are no literature or patent reports on compound herbal functional health beverages that use traditional compound formulas to protect liver and reduce lipids. Summary of the invention

[0005] The purpose of the present invention is to provide a compound herbal functional health drink with liver protection and lipid reduction effects and a preparation method thereof. The health drink of the present invention is composed of five Mongolian medicinal materials with medicinal and edible properties, has the effects of liver protection and lipid reduction, and can effectively alleviate symptoms such as liver lipid deposition.

[0006] The liver-protecting and lipid-clearing compound herbal functional health beverage of the present invention mainly contains gardenia, wolfberry, dandelion, honeysuckle and saffron as raw materials, which are mixed in proportions of 0.5-1% of gardenia, 0.25-0.75% of wolfberry, 0.2-0.5% of honeysuckle, 0.2-0.5% of dandelion, 0.01-0.1% of saffron, 2-4% of xylitol, 0.05-0.15% of citric acid and 0.05-0.1% of l-carrageenan, and are obtained by diluting with 90-95% of purified water.

[0007] On the basis of the above technical solution, the following technical features can be added for further limitation.

[0008] The more preferred weight percentages are 0.75% gardenia, 0.5% wolfberry, 0.25% dandelion, 0.25% honeysuckle, 0.05% saffron, 3.0% xylitol, 0.1% citric acid, 0.08% l-carrageenan, and the pure water dilution ratio is 95.02%.

[0009] The preparation method of the liver-protecting and lipid-clearing compound herbal functional health beverage of the present invention comprises the steps of raw material pretreatment, extract preparation, blending, canning and sterilization, taking out and cooling, and the like.

[0010] On the basis of the above technical solution, the following steps can be added for further limitation.

[0011] In the raw material pretreatment step, mud, weeds, pests and other parts of the raw materials are sorted and removed.

[0012] In the extract preparation step, water 10 times the weight of the raw materials is added into a cooking pot, and gardenia, wolfberry, dandelion, honeysuckle and saffron are added in sequence, stirred and heated to boiling, and kept warm for 120-180 minutes. During the insulation process, stirring is performed intermittently and slowly; after the insulation is completed, coarse filtration is performed to obtain filtrate A; the filtrate A is cooled to 50-60° C., water and citric acid are added respectively, and passed through an ultrafiltration device to obtain a clear and transparent filtrate, which is recorded as feed liquid B.

[0013] In the preparation step, 2-4% xylitol and 0.08% l-carrageenan are added to liquid B in a weight ratio, and the mixture is stirred evenly to obtain liquid C.

[0014] In the filling and sterilization step, liquid C is filled into bottles and placed in a high-temperature and high-pressure autoclave for sterilization.

[0015] The method also includes a sugaring step, which includes the following steps before mixing and preparing: adding the remaining water of the formula into the sugaring tank, heating to 40-50° C., adding 2-4% xylitol by weight, filtering through an 80-100 mesh pipeline, and pumping to a mixing tank.

[0016] It also includes a sterilization step before canning, and after mixing is completed, ultra-high temperature instantaneous sterilization is used.

[0017] The present invention proves through animal experiments and cell experiments that the liver-protecting and lipid-clearing compound herbal functional health beverage has the effects of resisting non-alcoholic fatty liver and alleviating liver lipid deposition, has a herbal, pure and natural taste, and its preparation method is simple in steps and low in cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Effects of liver-protecting and lipid-clearing compound herbal beverage on body weight in HFD-induced NAFLD rats

[0019] Figure 2 The effect of liver-protecting and lipid-clearing compound herbal beverage on the pathological morphology of liver in rats with NAFLD induced by HFD was shown

[0020] Figure 3 The effect of liver-protecting and lipid-clearing compound herbal beverage on serum ALT and AST levels in HFD-induced NAFLD rats

[0021] Figure 4 Effects of liver-protecting and lipid-clearing compound herbal beverage on GLU content in HFD-induced NAFLD rats

[0022] Figure 5 Effects of liver-protecting and lipid-clearing compound herbal beverage on TG content in HFD-induced NAFLD rats

[0023] Figure 6 Effects of liver-protecting and lipid-clearing compound herbal beverage on MDA content in HFD-induced NAFLD rats

[0024] Figure 7 The effect of liver-protecting and lipid-clearing compound on the content of TG and TC in HepG cells induced by OA is shown

[0025] Figure 8 The effect of liver protection and lipid-clearing compound on the content of ALT and AST in HepG cells induced by OA is shown

[0026] Fig. 9 The effect of liver-protecting and lipid-clearing compound on the content of TNF-α and IL-6 in the supernatant of HepG cells induced by OA is shown. DETAILED DESCRIPTION

[0027] The specific examples, animal experiment verification, cell experiment verification, safety, health care function effectiveness experiment verification and other contents of the present invention are described in detail below.

[0028] 1. Preparation method of liver-protecting and lipid-clearing compound herbal health functional beverage

[0029] Taking the preparation of 2kg of finished product as an example, the laboratory preparation is as follows:

[0030] (1) Raw material pretreatment: According to the formula composition, select and remove the mud, weeds, pests and other parts in the plant raw materials.

[0031] (2) Preparation of extract: According to the formula composition of 2 kg of finished product, add 0.36 kg of water (about 10 times the total amount of water in the raw materials) into a cooking pot, and then add 1 g of saffron, 15 g of gardenia (gardenia can be crushed appropriately), 10 g of wolfberry, 5 g of dandelion, and 5 g of honeysuckle in sequence, stir and heat to 90°C, and keep warm for 150 minutes (during the insulation process, stir slowly and intermittently); after the insulation is completed, coarse filter (300 mesh) to obtain filtrate A; cool the filtrate A to 50-60°C, add 1776 g of water and 2 g of citric acid respectively, make up the water to 2 kg, and filter through an ultrafiltration device (0.05 micron) to obtain a clear and transparent filtrate, which is recorded as feed liquid B.

[0032] (3) Preparation: According to the formula, add 3% xylitol and 0.08% l-carrageenan (relative to the amount of liquid B) to liquid B and stir evenly.

[0033] (4) Filling and sterilization: After preparation, fill into glass bottles, place in a high temperature and high pressure sterilizer, sterilize at 105°C for 30 minutes, take out, and cool.

[0034] Taking the production of 2000kg finished products as an example, the factory production process is as follows:

[0035] (1) Raw material pretreatment: According to the formula composition, select and remove the mud, weeds, pests and other parts in the plant raw materials.

[0036] (2) Preparation of extract: According to the formula of 2000 kg of finished product, add 360 kg of water (about 10 times the total amount of water in the raw materials) into the extraction tank; continue to add 1 kg of saffron, 15 kg of gardenia (gardenia can be appropriately crushed), 10 kg of wolfberry, 5 kg of dandelion, and 5 kg of honeysuckle in sequence, start stirring and heat to 90°C, stop heating, and keep warm for 150 minutes (during the insulation process, stir slowly and intermittently); after the insulation is completed, coarse filter (horizontal screw centrifugation / disc centrifugation) to obtain filtrate A; cool filtrate A to 40-50°C, add 2 kg of citric acid, stir evenly until dissolved, and pass through an ultrafiltration device (0.05 micron) to obtain a clear and transparent filtrate, which is recorded as feed liquid B.

[0037] (3) Sugar dissolving: Add the remaining water (1540.4 kg) in the formula into the sugar dissolving tank, heat to 40-50°C, add all the xylitol (60 kg) and 1.6 kg l-carrageenan, filter through an 80-100 mesh pipe, and pump to the batching tank.

[0038] (4) Preparation: Continue to add all the liquid B to the batching tank and stir evenly; add water to make the weight or volume to 2000 kg (the amount of water added is mainly the actual loss during the extraction of liquid B), stir evenly, and measure the indicators.

[0039] (5) Sterilization: After preparation, ultra-high temperature instantaneous sterilization is adopted, and the sterilization conditions are 121℃ and 20s.

[0040] (6) Filling: After sterilization, cool to 85-90℃ for hot filling.

[0041] (7) Secondary sterilization: After filling, the secondary sterilization conditions are 90°C and 20 minutes.

[0042] (8) Blow dry and code: Blow away the water on the can and spray the code on the bottom of the can.

[0043] (9) Packing, coding, palletizing and warehousing.

[0044] Product physical and chemical properties:

[0045] Appearance: Light yellow to dark yellow liquid.

[0046] PH value: 3.5-4.5,

[0047] Brix:6.5-7.5

[0048] 2. Animal experiment on the liver protection effect of liver-protecting and lipid-clearing compound herbal functional health drink

[0049] 1. Experimental materials and instruments:

[0050] 1.1 Experimental animals: 40 SPF male SD rats weighing 220±20 g were selected and provided by Shanghai Jiesijie Experimental Animal Co., Ltd. Certificate of qualification: SCXK(Shanghai)2018-0004.

[0051] 1.2 Experimental materials: liver-protecting and lipid-reducing beverage, high-fat feed, anhydrous ethanol, xylene, hematoxylin-eosin stain, hydrochloric acid, ammonia water, normal saline, malondialdehyde (MDA) content determination kit (Shanghai Youxuan Biotechnology Co., Ltd., product number: YX-W-A401), neutral gum, silybin (Shanghai McLean Biochemical Technology Co., Ltd., product number: S817884).

[0052] 1.3 Experimental instruments: round-bottom flask, condenser, surgical scissors, blood drawing needle, blood drawing core, electronic balance, Sorval lST 40 desktop high-speed refrigerated centrifuge (Thermo, USA), TECAN Infinite M200Pro multi-function fluorescence microplate reader, constant temperature water bath, high-speed tissue grinder, vortex mixer, fully automatic biochemical analyzer, horizontal freezer, paraffin embedding machine, tissue dehydrator, freezing table, pathology slicer, tissue spreader, oven, upright optical microscope, imaging system.

[0053] 2. Experimental methods:

[0054] 2.1 Experimental animal grouping, modeling and drug dosage

[0055] SPF male SD rats were randomly divided into 5 groups: blank control group (B), model group (M), positive control group (Y), low-dose liver-protecting and fat-clearing beverage group (L), and high-dose liver-protecting and fat-clearing beverage group (H).

[0056] High-fat diet (HFD) is a widely used inducer. In view of the literature reports of the use of HFD to induce non-alcoholic fatty liver disease (NAFLD) in animals, this experiment chose the HFD-induced NAFLD model. During the experiment, the method of building the model and administering the drug was adopted. There were 8 rats in the normal control group, which were fed with ordinary feed and given normal saline by gavage. The remaining rats were randomly divided into 4 groups, namely the model control group, the low-dose beverage group (75 mg / kg·d), the high-dose beverage group (150 mg / kg·d) and the silybin group (50 mg / kg·d), with 8 rats in each group, fed with a high-fat diet, and given normal saline or beverages of corresponding concentrations by gavage every day.

[0057] (1) Calculation of dosage of liver-protecting and fat-clearing beverage

[0058] According to the "Chinese Pharmacological Experimental Methodology", the conversion rule of human and animal body surface area is adopted. First, the dosage of an adult weighing 70kg is used as the benchmark to calculate the equivalent dose for rats. Set this dose as the dosage for the high-dose beverage group, and at the same time, halve it as the dosage for the low-dose group. The dosage for the high-dose beverage group and the low-dose beverage group was determined to be 150mg / kg·d and 75mg / kg·d, respectively. The low-dose beverage group and the high-dose beverage group were gavaged with 1mL and 2mL of beverage per day, respectively, for 8 weeks.

[0059] (2) Calculation of the dosage of the positive control drug silybin

[0060] For the western medicine group, the equivalent dose for rats was calculated according to the clinical daily dosage of 210 mg, which was used as the therapeutic dosage, i.e. 50 mg / kg·d. Specifications of Silybin Capsules: Prepare a 10 mg / mL solution with cooled boiling water and store at 4°C for later use.

[0061] 2.2 Effects of liver-protecting and lipid-clearing compound herbal beverage on body weight in HFD-induced NAFLD rats

[0062] The rats were raised for 8 weeks, and the body weight changes of rats in each group were observed and recorded every week.

[0063] As shown in Table 1, after oral administration, the body weight of rats in the model control group increased significantly compared with the normal control group (P < 0.01); compared with the model control group, the body weight of each experimental group decreased significantly, especially the high-dose beverage group (P < 0.01); there was no difference in body weight change among the experimental groups. This result shows that the beverage can significantly reduce the body weight of obese rats and has a promoting effect on inhibiting the formation of NAFLD. (See Table 1, Figure 1 ).

[0064] Table 1 Comparison of body weight of rats in the eighth week

[0065]

[0066] 2.3 Effects of liver-protecting and lipid-clearing compound herbal beverage on the pathological morphology of liver in rats with HFD-induced NAFLD

[0067] like Figure 2 As shown in the figure, the normal control group B: the liver lobule structure is clear and complete, the cells are similar in size and arranged neatly, the hepatic cords are arranged radially around the central vein, the cell nucleus is located in the center, the cells have no fatty degeneration, and no inflammatory cell infiltration is observed. Model control group M: the liver lobule structure is destroyed, the cells are swollen and deformed, and diffuse fatty degeneration is observed, vacuoles of varying sizes are observed, the hepatic sinusoids disappear, the cell nuclei are located at the edge, similar to rings, and punctate necrosis and inflammatory cell infiltration can be seen. Positive control group Y: the liver lobule structure is blurred, the hepatocytes are slightly edematous, vacuoles of varying sizes are observed, the hepatic cords are arranged slightly regularly, the cytoplasm is still rich, the cell nucleus is not centered, but the morphology is normal, and inflammatory cell infiltration is occasionally observed. High-dose beverage group H: the liver lobule structure is blurred, the hepatocytes are larger in size, and they have mild fatty degeneration, vacuoles of varying sizes are observed, the hepatic cords are slightly disordered, the hepatic sinusoids are narrowed, and inflammatory cell infiltration is occasionally observed. Low-dose beverage group L: The liver lobules had a disordered structure with unclear boundaries, the cells showed mild to moderate fatty degeneration, vacuoles of varying sizes were seen, the liver sinusoids disappeared, and scattered punctate necrosis and inflammatory cell infiltration were seen.

[0068] 2.4 Effect of liver-protecting and lipid-clearing compound herbal beverage on serum ALT and AST levels in HFD-induced NAFLD rats ALT and AST levels were determined using an automatic biochemical analyzer I FCC method:

[0069] After intragastric administration, compared with the normal control group, the serum ALT and AST levels of rats in the model control group increased significantly (P < 0.01); compared with the model control group, the above indicators of each experimental group decreased significantly (P < 0.01), especially the high-dose beverage group (P < 0.01); compared with the experimental groups, the above indicators of the high-dose beverage group were significantly lower than those of the low-dose beverage group (P < 0.01). This result shows that taking the beverage can significantly reduce the levels of alanine aminotransferase and aspartate aminotransferase in NAFLD rats, and follow a certain concentration-dependent relationship. (See Table 2, Figure 3 ).

[0070] Table 2 Comparison of ALT and AST levels in rat serum

[0071]

[0072]

[0073] 2.5 Effects of liver-protecting and lipid-clearing compound herbal beverage on GLU, TG, and MDA levels in HFD-induced NAFLD rats A. GLU content was determined by enzymatic method using an automatic biochemical analyzer:

[0074] Compared with the normal control group, the blood glucose level of rats in the model control group increased significantly (P < 0.01); compared with the model control group, the blood glucose level of each experimental group decreased significantly (P < 0.01). This result shows that taking the beverage can significantly reduce the blood glucose level in NAFLD rats and restore it to the normal range. (See Table 3, Figure 4 ).

[0075] Table 3 Comparison of serum GLU content in rats of each group

[0076]

[0077] B. TG content was determined by enzymatic method using fully automatic biochemical analyzer:

[0078] Compared with the normal control group, the serum TG level of rats in the model control group increased significantly (P < 0.01); compared with the model control group, the TG level of each experimental group decreased significantly (P < 0.01), especially the high-dose beverage group (P < 0.01); there was no difference between the positive control group and the experimental group; there was no difference between the experimental groups; the TG level of the high-dose beverage group was significantly lower than that of the normal control group (P < 0.01). This result shows that taking high-concentration beverages can effectively reduce the level of triglycerides in NAFLD rats. (See Table 4, Figure 5 ).

[0079] Table 4 Comparison of serum TG content in rats of each group

[0080]

[0081] CE li sa method to measure liver MDA level:

[0082] Compared with the normal control group, the MDA level of blood lipids in the model control group was significantly increased (P < 0.01); compared with the model control group, the MDA level of each experimental group was significantly reduced (P < 0.01); there was a significant difference between the experimental group and the positive control group (P < 0.01); there was no significant difference between the experimental groups. This result shows that taking high-concentration beverages can restore the MDA content in NAFLD rats to normal levels. (See Table 5, Figure 6 ).

[0083] Table 5 Comparison of MDA content in blood lipids of rats in each group

[0084]

[0085] 2. Cell experiment on the liver protection effect of liver-protecting and lipid-clearing compound herbal functional health drink

[0086] 1. Experimental materials and instruments:

[0087] 1.1 Cell line: human hepatocellular carcinoma HepG2 cells.

[0088] 1.2 Experimental materials: liver protection and lipid-clearing compound, DMEM high glucose culture medium, fetal bovine serum (FBS), penicillin-streptomycin double antibody solution, 0.25% Trypsin-EDTA (1X) trypsin, PBS cell buffer, oleic acid, triglyceride (TG) determination kit, total cholesterol (TC) determination kit, BCA protein quantitative test kit (enhanced), human tumor necrosis factor (TNF-a) Elisa kit, human interleukin 6 (IL-6) Elisa kit, alanine aminotransferase test kit, aspartate transferase test kit.

[0089] 1.3 Experimental instruments: biological safety cabinet, multifunctional microplate reader, CO2 constant temperature cell culture incubator, inverted microscope, high pressure sterilizer, constant temperature water bath, automatic fluorescence cell counter.

[0090] 2. Experimental methods:

[0091] Take the LPLC storage solution, dissolve it in a constant temperature water bath, and then use complete medium to gradient dilute to the corresponding concentration (low dose 1mg / ml, medium dose 2.5mg / ml, high dose 5mg / ml), pre-treat for 48h, positive control group 25μm silybin, pre-treat for 24h. Prepare 120μg / ml oleic acid modeling solution in advance. After administration, take the cell supernatant and use Elisa detection kit to detect TNF-α and IL-6 components in the supernatant. Add 2% Tritonx-100 lysis solution to the cell pellet, lyse on ice for 30-40min, and operate according to TG, TC, ALT, AST detection kits, and detect protein with BCA protein kit.

[0092] Based on the reference to books and literature reports on Mongolian medicine prescriptions, the present invention has developed a liver-protecting and lipid-clearing compound herbal functional health beverage in combination with experimental research, and has proved the liver-protecting and health-care function of the beverage through animal experiments and cells. At the same time, the product pilot test was also carried out to determine the production process.

[0093] In summary, the development of the composite beverage mainly based on gardenia involved in the technology of the present invention can solve the insufficiency of the taste and function of the simple gardenia beverage. At the same time, the present invention is based on the traditional Mongolian medicine formula theory, combined with the best formula obtained by scientific experiments, and has the effect of resisting non-alcoholic fatty liver and removing liver lipid deposition. The gardenia compound herbal beverage has a refreshing taste, can assist the health needs of the respiratory system, has certain effects in preventing and treating non-alcoholic fatty liver, is suitable for a wide range of people, and is a nutritious tea beverage with definite health care functions. In addition, the preparation cost of the liver-protecting and lipid-clearing compound herbal functional health beverage in the present invention is low.

Claims

1. A liver-protecting and lipid-clearing compound herbal functional health beverage, characterized in that: The invention contains gardenia, wolfberry, honeysuckle, dandelion, saffron, xylitol, citric acid and l-carrageenan as raw materials, which are mixed in proportions of 0.5-1% of gardenia, 0.25-0.75% of wolfberry, 0.2-0.5% of honeysuckle, 0.2-0.5% of dandelion, 0.01-0.1% of saffron, 2-4% of xylitol, 0.05-0.15% of citric acid and 0.05-0.1% of l-carrageenan by weight, and then diluted with 90-95% purified water to obtain the product.

2. The liver-protecting and lipid-clearing compound herbal functional health beverage according to claim 1, characterized in that: The weight percentage is 0.75% of gardenia, 0.5% of wolfberry, 0.25% of dandelion, 0.25% of honeysuckle, 0.05% of saffron, 3.0% of xylitol, 0.1% of citric acid, 0.08% of l-carrageenan, and the pure water dilution ratio is 95.02%.

3. A method for preparing the liver-protecting and lipid-clearing compound herbal functional health beverage according to any one of claims 1-2, characterized in that: The method comprises the following steps: raw material pretreatment, extract preparation, blending, canning and sterilization, and taking out and cooling.

4. The preparation method according to claim 3, characterized in that: In the raw material pretreatment step, mud, weeds and pests are sorted and removed from the raw materials.

5. The preparation method according to claim 4, characterized in that: In the extract preparation step, 8-12 times the weight of the raw material water is added into a cooking pot or an extraction tank, and gardenia, wolfberry, honeysuckle, dandelion and saffron are added in sequence, stirred and heated to boiling, and kept warm for 120-180 minutes. During the insulation process, the mixture is slowly stirred intermittently; after the insulation is completed, the filtrate A is obtained by coarse filtration; the filtrate A is cooled to 40-60° C., water and citric acid are added respectively, and the filtrate is passed through an ultrafiltration device to obtain a clear and transparent filtrate, thereby obtaining a feed liquid B.

6. The preparation method according to claim 5, characterized in that: In the preparation step, 2-4% xylitol and 0.05-0.1% l-carrageenan are added to the liquid B in a weight ratio, and the mixture is stirred evenly to obtain the liquid C.

7. The preparation method according to claim 6, characterized in that: In the filling and sterilization step, liquid C is filled into bottles and placed in a high-temperature and high-pressure autoclave for sterilization.

8. The preparation method according to claim 3, characterized in that: The preparation method also includes a sugaring step, which includes the following steps before preparation: adding the remaining water of the formula into the sugaring tank, heating to 40-50° C., adding 2-4% xylitol by weight, filtering through an 80-100 mesh pipeline, and pumping to a mixing tank.

9. The preparation method according to claim 3, characterized in that The preparation method also includes a sterilization step before canning, and after the preparation is completed, ultra-high temperature instantaneous sterilization is adopted.

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