Plant composite extract for inhibiting helicobacter pylori as well as preparation method and application of plant composite extract

Abstract

The invention discloses a plant composite extract for inhibiting helicobacter pylori as well as a preparation method and application of the plant composite extract, and belongs to the field of plant extracts. The plant compound extract comprises 7%-45% of sulfide volatile oil, 1%-9% of mushroom terpenoids and 10%-33% of compound polysaccharide; the plant compound extract is extracted from garlic, Chinese chive seeds, hericium erinaceus and lucid ganoderma; wherein the sulfide volatile oil is extracted from garlic and Chinese chive seeds, and the sulfide volatile oil comprises allicin; the mushroom terpenoids are extracted from hericium erinaceus and lucid ganoderma; the compound polysaccharide is extracted from garlic, Chinese chive seeds, hericium erinaceus and lucid ganoderma. Efficient bacteriostasis and mucous membrane repair are achieved through the synergistic effect of natural components, the drug resistance and hepatorenal toxicity of antibiotics are avoided, the possibility that helicobacter pylori relapses in the stomach can be effectively reduced, the traditional Chinese medicine composition is suitable for drug-resistant patients or chronic gastritis patients needing long-term conditioning, and the curative effect is good. The health-care food is a safe and effective health-care food raw material and a dietary supplement.

Description

Technical Field

[0001] The present invention belongs to the field of plant extracts, and in particular relates to a plant composite extract for inhibiting Helicobacter pylori, and a preparation method and application thereof. Background Art

[0002] Helicobacter pylori is a Gram-negative spiral bacterium that is adaptable to various environments and possesses a strong survival ability. Unlike most bacteria that struggle to survive in the highly acidic environment of the stomach, H. pylori secretes urease, which breaks down urea into ammonia. This creates an "ammonia cloud" around itself, neutralizing stomach acid and enabling it to survive in the stomach's acidic environment for extended periods. H. pylori's spiral, S-shaped, or curved shape helps it penetrate the mucus layer of the gastric mucosa, allowing it to approach and adhere to the surface of gastric epithelial cells, thereby avoiding being washed away by gastric peristalsis and digestive juices. H. pylori is a major cause of various gastric diseases, such as chronic gastritis, peptic ulcers, and gastric cancer. Promptly suppressing H. pylori can reduce medical expenses incurred by patients, as well as indirect economic losses such as work and life restrictions caused by the disease, thereby alleviating the burden of disease on both individuals and society.

[0003] However, Helicobacter pylori is a Gram-negative bacterium with strong survival ability in the stomach, and it is difficult to completely eliminate it once infected. Even if Helicobacter pylori is temporarily eliminated, if the gastric environment is not fundamentally improved, such as abnormal gastric acid secretion or damaged gastric mucosal barrier function, it still provides favorable conditions for the re-colonization and growth of Helicobacter pylori. The classic clinical treatment of Helicobacter pylori infection is triple therapy (proton pump inhibitors plus two antibiotics) and quadruple therapy (bismuth + PPI + two antibacterial drugs). However, with the widespread use of antibiotics, Helicobacter pylori's resistance to antibiotics has gradually increased, resulting in unsatisfactory results and increased recurrence rates. At the same time, these therapies may also cause side effects such as intestinal flora imbalance.

[0004] With the increasing problem of antibiotic resistance and the side effects of traditional antibiotic treatments, plant extracts have gradually become a research hotspot for the treatment of Helicobacter pylori infection due to their natural, safe, and minimally toxic side effects. However, there are many types of plant extracts containing multiple active ingredients. How to select active ingredients with synergistic effects and ensure that they can effectively prevent and treat Helicobacter pylori infection and reduce recurrence rates has become a major challenge. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the technical problem to be solved by the present invention is to overcome the technical problems in the existing technology that Helicobacter pylori has strong drug resistance, resulting in unsatisfactory effects and increased recurrence rate, and proposes to provide a plant composite extract that can effectively inhibit Helicobacter pylori, reduce the risk of drug resistance, and reduce the recurrence rate, as well as its preparation method and application.

[0006] In order to solve the technical problem, the technical solution adopted by the present invention is:

[0007] The first aspect of the present invention provides a plant composite extract for inhibiting Helicobacter pylori, wherein the plant composite extract comprises 7%-45% of sulfide volatile oil, 1%-9% of mushroom terpenoid compounds, and 10%-33% of complex polysaccharides;

[0008] The plant compound extract is obtained from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum;

[0009] Among them, sulfide volatile oil is extracted from garlic and leek seeds, and sulfide volatile oil includes allicin;

[0010] Mushroom terpenoids are extracted from Hericium erinaceus and Ganoderma lucidum;

[0011] The complex polysaccharide is extracted from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum.

[0012] Preferably, the plant composite extract is obtained by extracting 1-3 parts of garlic, 0.1-1 parts of leek seeds, 1-3 parts of Hericium erinaceus and 6-12 parts of Ganoderma lucidum.

[0013] Preferably, the plant composite extract is obtained by extracting 2 parts of garlic, 0.3 parts of leek seeds, 1 part of Hericium erinaceus and 6 parts of Ganoderma lucidum.

[0014] The second aspect of the present invention provides a method for preparing the above-mentioned plant composite extract for inhibiting Helicobacter pylori, comprising the following steps:

[0015] Mixing garlic and leek seeds and crushing them, adding water and keeping warm for 1-2 hours, adding n-hexane for extraction, and filtering after extraction to obtain a first filtrate and a first filter residue;

[0016] The solvent is recovered from the first filtrate to obtain garlic / leek seed volatile oil, and an aqueous solution of sodium starch octenylsuccinate is added and stirred for 0.5-2 hours for emulsification to obtain an emulsion, which is freeze-dried and pulverized to obtain sulfide volatile oil.

[0017] Furthermore, the method further includes the following steps:

[0018] The crushed Ganoderma lucidum and Hericium erinaceus are mixed, ethanol is added, and the mixture is heated under reflux for extraction. After extraction, the mixture is filtered to obtain a second filtrate and a second filter residue. The solvent is recovered from the second filtrate, and the mixture is concentrated and dried to obtain mushroom terpenoid compounds.

[0019] Furthermore, the method further includes the following steps:

[0020] The first filter residue and the second filter residue are mixed to obtain a mixed filter residue, water is added to perform microwave extraction, and after extraction, an extract is obtained by filtering, and the extract is concentrated and dried to obtain a complex polysaccharide.

[0021] Preferably, the mass ratio of garlic, leek seeds and n-hexane is (1-3): (0.1-1): (11-80);

[0022] The ratio of the added amount of garlic / leek seed volatile oil to the sodium starch octenylsuccinate aqueous solution is 1g:6-10mL, and the concentration of the sodium starch octenylsuccinate aqueous solution is 0.5-1.5g / mL.

[0023] Preferably, the mass ratio of the ganoderma lucidum shreds, the hericium erinaceus shreds and the ethanol is (6-12): (1-3): (42-150);

[0024] The volume fraction of ethanol is 70%-80%, and the heating reflux extraction time is 1-2 hours.

[0025] Preferably, the mass ratio of the mixed filter residue to water is 1:(6-10), the power of the microwave extraction is 500-1000W, and the time is 0.5-1h.

[0026] The third aspect of the present invention provides use of the above-mentioned plant composite extract for inhibiting Helicobacter pylori in the preparation of a product for inhibiting Helicobacter pylori.

[0027] Preferably, the dosage form of the product is selected from at least one of powder, tablet, granule and capsule.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] 1. The present invention provides a plant compound extract for inhibiting Helicobacter pylori. The plant compound extract is obtained by composite extraction of raw medicinal materials consisting of garlic, leek seeds, ganoderma lucidum and hericium erinaceus. The plant compound extract includes 7%-45% of sulfide volatile oil, 1%-9% of mushroom terpenoid compounds and 10%-33% of complex polysaccharides. The sulfides in garlic and leek seeds synergistically destroy Hp biofilm and reduce drug resistance. The terpenoid compounds in hericium erinaceus and ganoderma lucidum can reduce the release of inflammatory factors, enhance macrophage activity, and improve the body's ability to clear Hp. The mushroom terpenoid compounds can reduce the release of inflammatory factors, enhance macrophage activity, and improve The body's ability to clear H. pylori. Ganoderma lucidum terpenoids can regulate the function of immune cells, promote the proliferation and differentiation of lymphocytes, and enhance the body's cellular and humoral immunity. Complex polysaccharides accelerate mucosal regeneration, preventing Helicobacter pylori from drilling into the mucus layer on the surface of the gastric mucosa and adhering to the surface of gastric epithelial cells. Through the synergistic effect of natural ingredients, efficient antibacterial and mucosal repair are achieved, avoiding antibiotic resistance and liver and kidney toxicity, and can effectively reduce the possibility of Helicobacter pylori recurrence in the stomach. It is suitable for drug-resistant patients or patients with chronic gastritis who require long-term conditioning. It is a safe and effective health food raw material and dietary supplement.

[0030] 2. In the embodiment of the present invention, garlic and leek seeds are subjected to composite extraction, and the yield of allicin is greater than 1.9mg / g, while comparative example 1 only extracts garlic, and the yield of allicin is only 1.08mg / g, and comparative example 2 only extracts leek seeds, and the yield of allicin in sulfide volatile oil is 0.91mg / g. It can be seen from this that garlic and leek seeds are subjected to composite extraction, and the two can promote dissolution mutually, and have a synergistic effect for improving the yield of allicin. When comparative example 10 extracts garlic and leek seeds respectively, the yield of allicin in volatile oil is 1.10mg / g, which further verifies the mutual promotion dissolution effect between garlic and leek seeds. Comparative example 11 is in traditional water extraction process, and the yield of allicin is 1.29mg / g, which illustrates that the extraction process of the present application further promotes the dissolution of allicin. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The extraction process flow chart provided for Example 1 and Example 2 of the present invention. DETAILED DESCRIPTION

[0032] The following is a detailed and complete description of the technical solutions in specific embodiments of the present invention, with reference to the accompanying drawings. It should be understood that the described embodiments are merely partial implementations of the overall technical solution of the present invention, and are not exhaustive. All other embodiments derived by those skilled in the art based on the overall concept of the present invention are intended to fall within the scope of protection of the present invention.

[0033] On the one hand, the present invention provides a plant composite extract for inhibiting Helicobacter pylori, which comprises 7%-45% of sulfide volatile oil, 1%-9% of mushroom terpenoid compounds and 10%-33% of complex polysaccharides; the plant composite extract is extracted from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum; wherein the sulfide volatile oil is extracted from garlic and leek seeds, and the sulfide volatile oil comprises allicin; the mushroom terpenoid compounds are extracted from Hericium erinaceus and Ganoderma lucidum; and the complex polysaccharides are extracted from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum.

[0034] It should be noted that the classic regimens for clinical treatment of Helicobacter pylori infection in the prior art are triple therapy (proton pump inhibitor plus two antibiotics) and quadruple therapy (bismuth + PPI + two antibacterial drugs). However, with the widespread use of antibiotics, Helicobacter pylori's resistance to antibiotics has gradually increased, resulting in increased treatment failure rate and relapse rate. At the same time, these therapies may also cause side effects such as intestinal flora imbalance. With the aggravation of the problem of antibiotic resistance and the side effects of traditional antibiotic treatments, plant extracts have gradually become a research hotspot for the treatment of Helicobacter pylori infection because of their natural, safe, and less toxic and side effects. However, there are many types of plant extracts, which contain a variety of active ingredients. How to select active ingredients with synergistic effects and ensure that they can effectively treat Helicobacter pylori infection and reduce relapse rate has become an important problem currently faced.

[0035] In response to the above problems, the present invention fully extracts and refines garlic, leek seeds, ganoderma lucidum, and hericium erinaceus to obtain a variety of effective ingredients such as sulfide volatile oil, mushroom terpenoids, and complex polysaccharides. The sulfides in garlic and leek seeds synergistically destroy Hp biofilms and reduce drug resistance. The sulfide allicin in garlic has broad-spectrum antibacterial properties and has a direct inhibitory and killing effect on Helicobacter pylori. It can destroy the cell wall and cell membrane of bacteria, allowing the bacterial contents to leak out, thereby achieving a bactericidal effect, reducing the number of Helicobacter pylori, and reducing the possibility of its re-multiplication in the stomach and causing recurrence. Ganoderma lucidum terpenoids and Hericium erinaceus terpenoids can reduce the release of inflammatory factors, enhance macrophage activity, and improve the body's ability to clear Hp. Ganoderma lucidum terpenoids can promote the proliferation and differentiation of lymphocytes by regulating the function of immune cells, thereby enhancing the body's cellular immunity and humoral immunity. After H. pylori infection, the body can establish more effective immune defenses, preventing H. pylori reinvasion and colonization, and reducing the risk of recurrence. Hericium erinaceus terpenoids can promote the regeneration and repair of gastric mucosal cells, increase gastric mucosal thickness and mucus secretion, and enhance the gastric mucosal barrier function. This effectively prevents direct contact between H. pylori and the gastric mucosa, reducing damage to the mucosa and the chance of H. pylori reinfection and colonization. The polysaccharides in the hericium accelerate mucosal regeneration, preventing H. pylori from burrowing through the mucus layer on the gastric mucosal surface and adhering to the surface of gastric epithelial cells. Thus, through direct and effective bactericidal action, immune suppression, and enhanced gastric mucosal barrier function, H. pylori is inhibited and the likelihood of recurrence is reduced.

[0036] In a preferred embodiment, the plant composite extract is obtained by extracting 1-3 parts of garlic, 0.1-1 parts of leek seeds, 1-3 parts of Hericium erinaceus and 6-12 parts of Ganoderma lucidum.

[0037] In a preferred embodiment, the plant composite extract is obtained by extracting 2 parts of garlic, 0.3 parts of leek seeds, 1 part of Hericium erinaceus and 6 parts of Ganoderma lucidum.

[0038] Another aspect of the present invention provides a method for preparing the above-mentioned plant composite extract, comprising the following steps:

[0039] Garlic and leek seeds are mixed and crushed, then water is added and kept warm for 1-2 hours. Then, n-hexane is added for extraction. After extraction, the mixture is filtered to obtain a first filtrate and a first filter residue. The solvent is recovered from the first filtrate to obtain garlic / leek seed volatile oil, which is then emulsified by adding an aqueous solution of sodium starch octenylsuccinate for 0.5-2 hours to obtain an emulsion. The emulsion is freeze-dried and crushed to obtain an allicin complex. Preferably, the garlic is fresh garlic or dried garlic slices. Preferably, the amount of water added is one times the total mass of the garlic and leek seeds, and the water temperature is 50°C. Preferably, the mass ratio of garlic, leek seeds, and n-hexane is (1-3):(0.1-1):(11-80); the ratio of the garlic / leek seed volatile oil to the aqueous solution of sodium starch octenylsuccinate is 1 g:6-10 mL, and the concentration of the aqueous solution of sodium starch octenylsuccinate is 0.5-1.5 g / mL.

[0040] It should be noted that the yield of allicin extracted directly from garlic in the prior art is low. To address this problem, the present invention combines garlic and leek seeds for extraction, as leek seeds can promote the dissolution of allicin. Specifically, allicin is obtained by the conversion of alliin and alliinase, and leek seeds contain a large amount of alliinase, which promotes the production of allicin. The above steps also require that the obtained garlic / leek seed volatile oil be emulsified. The reason is that the emulsification of the volatile oil with the auxiliary ingredient, sodium starch octenylsuccinate aqueous solution, is a conventional volatile oil inclusion process, the purpose of which is to allow the volatile oil to be completely included. The included volatile oil will have improved water solubility, bioavailability, and ingredient stability.

[0041] In a preferred embodiment, the following steps are also included:

[0042] Ganoderma lucidum and Hericium erinaceus minced are mixed, ethanol is added, and the mixture is heated under reflux for extraction. After extraction, the mixture is filtered to obtain a second filtrate and a second filter residue. The solvent is recovered from the second filtrate, and the mixture is then concentrated and dried to obtain mushroom terpenoid compounds. The mass ratio of Ganoderma lucidum minced, Hericium erinaceus minced, and ethanol is (6-12):(1-3):(42-150); the volume fraction of ethanol is 70%-80%, and the heating and reflux extraction time is 1-2 hours.

[0043] This technical solution limits the composite extraction of Ganoderma lucidum and Hericium erinaceus because it can save time and extraction costs and is more environmentally friendly.

[0044] In a preferred embodiment, the following steps are also included:

[0045] The first filter residue and the second filter residue are mixed to obtain a mixed filter residue, water is added to perform microwave extraction, and after extraction, the extract is filtered to obtain an extract, which is concentrated and dried to obtain a complex polysaccharide. Preferably, the mass ratio of the mixed filter residue to water is 1:(6-10), the microwave extraction power is 500-1000W, and the time is 0.5-1h.

[0046] The technical solution specifies that the first filter residue obtained by extracting garlic and leek seeds, and the second filter residue obtained by extracting Hericium erinaceus and Ganoderma lucidum are mixed, and water is added for mixed extraction to obtain a composite polysaccharide, wherein garlic polysaccharide can selectively promote the growth and reproduction of beneficial intestinal bacteria, such as bifidobacteria and lactic acid bacteria, inhibit the growth of harmful bacteria, maintain the balance of intestinal microecology, and have a positive effect on improving intestinal function and preventing intestinal diseases; leek seed polysaccharide can activate immune cells such as macrophages, T lymphocytes and B lymphocytes, enhance their phagocytic ability and immune response, thereby improving the body's immune defense function. At the same time, it can also promote the secretion of cytokines, such as interleukins and interferons, which play an important role in regulating immune responses; Ganoderma lucidum polysaccharides and Hericium erinaceus polysaccharides accelerate mucosal regeneration, preventing Helicobacter pylori from drilling in the mucus layer on the surface of the gastric mucosa and adhering to the surface of gastric epithelial cells. The reason why the present invention extracts the polysaccharide by compounding the four medicinal materials is that it can save time and extraction costs and is more environmentally friendly.

[0047] Another aspect of the present invention provides the use of the above plant compound extract in the preparation of a product for inhibiting Helicobacter pylori. Preferably, the dosage form of the product is selected from at least one of powder, tablet, granule, and capsule.

[0048] In order to more clearly and in detail introduce the plant composite extract for inhibiting Helicobacter pylori provided by the embodiments of the present invention, its preparation method and application, the following description will be made in conjunction with specific examples.

[0049] The test materials used in the examples of the present invention are all conventional test materials in the art and can be purchased through commercial channels.

[0050] The content of the active ingredient in the plant extract of the present invention is tested by the following method:

[0051] 1.1 Allicin content test:

[0052] 1.1.1 Materials and Equipment

[0053] Methanol, volumetric flask, weighing bottle, electronic balance, 0.45 μm microporous filter membrane, Dipropylene trisulfide (Diallyl Trisulfide) reference substance, Dipropylene disulfide (Diallyl disulfide) reference substance;

[0054] Ultrasonic processor, HPLC system, chromatographic injection;

[0055] 1.1.2 Experimental methods

[0056] HPLC conditions:

[0057] Mobile phase: methanol / 0.17% formic acid solution = 80 / 20, flow rate: 1.0 ml / min, column temperature: 28°C, detection wavelength: 240 nm, injection volume: 25 ul, running time: 40 min.

[0058] Preparation of standard solution:

[0059] Accurately weigh appropriate amounts of Diallyl Trisulfide reference substance and Diallyl Disulfide reference substance, add methanol to dissolve and prepare a reference solution containing approximately 0.1 mg of Diallyl Trisulfide and Diallyl Disulfide per 1 mL.

[0060] Preparation of sample solution:

[0061] Accurately weigh an appropriate amount of the sample to be tested, add methanol to a 50 mL volumetric flask, ultrasonically oscillate for 30 min, cool to room temperature, dilute to volume with methanol, and filter with a 0.45 μm microporous membrane.

[0062] Determination and calculation

[0063] Under the above chromatographic conditions, the sample was injected and measured, and the contents of allyl trisulfide and allyl disulfide were calculated respectively by the external standard method. The allicin content was the sum of the two.

[0064] It should be noted that, in the following embodiments and comparative examples of the present invention:

[0065] The extraction rate of allicin in garlic = allicin mass / garlic raw medicinal material mass;

[0066] The extraction rate of allicin in leek seeds = allicin mass / leek seed raw medicinal material mass;

[0067] The extraction rate of allicin in garlic and leek seeds = allicin mass / (total mass of garlic raw medicinal materials + leek seed raw medicinal materials).

[0068] 1.2 Polysaccharide content detection

[0069] 1.2.1 Materials and Equipment

[0070] Methanol (AR), sulfuric acid (AR), distilled water, phenol (AR), sodium hydroxide, iodine, sodium thiosulfate, soluble starch, anhydrous sodium carbonate, potassium iodide, reference substance (glucose)

[0071] Electronic balance (1 / 100000), glass instruments: round-bottom flask, volumetric flask, graduated cylinder, graduated pipette (pipette or electric pipette), ultrasonic cleaner, water bath, refrigerator, UV spectrophotometer, cuvette (1 cm)

[0072] 1.2.2 Methods

[0073] ① Preparation of glucose standard solution: Dry the standard glucose in an oven at 105℃ to constant weight. Accurately weigh 10 mg of glucose into a 100 ml volumetric flask, add appropriate amount of distilled water to dissolve, then dilute to 100 ml with distilled water, shake well, and set aside.

[0074] ②Preparation of 5% phenol solution:

[0075] 80% phenol: Weigh 80.00 g of phenol (analytical grade redistilled), dissolve it in 20.00 g of water, and store it in a brown bottle in the refrigerator away from light.

[0076] 5% phenol: Prepare with 80% phenol before use and should be prepared immediately before use.

[0077] ③Determination:

[0078] To prepare a standard curve: Pipette 0, 0.2, 0.4, 0.6, 0.8, and 1.0 mL of each solution, respectively, and make up to 2.0 mL with distilled water. Then add 1.0 mL of 5% phenol and 5.0 mL of concentrated sulfuric acid, shake well, heat in boiling water for 10 minutes, shake well again, and let stand at room temperature for 20 minutes. Measure the absorbance at 490 nm and regress the absorbance against the glucose concentration to create a standard curve. A = kc + b (A is absorbance, c is solution concentration, and k and b are constants).

[0079] Determination of Total Polysaccharides: Accurately weigh 100 mg of the extract powder (0-50%) and place it in a 500 mL round-bottom flask. Add 100 mL of 80% methanol and heat under reflux for 1 hour. Filter while hot. Wash the filter with 30 mL of hot 80% methanol three times. Place the filter residue and filter paper in a 500 mL round-bottom flask. Add 100 mL of water and heat under reflux for 1 hour. Filter the mixture while hot into a 250 mL volumetric flask. Wash the residue and filter with a small amount of water. After cooling, dilute to the mark with water.

[0080] 1.3 Detection of terpenoid content

[0081] 1.3.1 Materials and Equipment

[0082] Methanol (AR), glacial acetic acid (AR), perchloric acid (AR), ethyl acetate (AR), vanillin, distilled water, reference substance (oleanolic acid)

[0083] Electronic balance (1 / 100000), glass instruments: volumetric flask, graduated cylinder, graduated pipette (pipette or electric pipette), ultrasonic cleaner, water bath, refrigerator, UV spectrophotometer, cuvette (1 cm)

[0084] 1.3.2 Methods

[0085] Vanillin glacial acetic acid solution: Accurately weigh 0.5g of vanillin and add glacial acetic acid to dissolve it into 10ml.

[0086] 1.3.3 Determination

[0087] Preparation of reference solution: Take an appropriate amount of oleanolic acid reference substance, weigh accurately, and add methanol to make a solution containing 0.2 mg per 1 ml.

[0088] Preparation of standard curve: Accurately measure 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml and 0.5 ml of reference solution, respectively, and place them in 15 ml stoppered test tubes, evaporate to dryness, let cool, accurately add 0.2 ml of freshly prepared vanillin glacial acetic acid solution and 0.8 ml of perchloric acid, shake well, heat in a 70°C water bath for 15 minutes, immediately cool in an ice bath for 5 minutes, take out, accurately add 4 ml of ethyl acetate, shake well, use the corresponding reagent as blank, measure the absorbance at a wavelength of 546 nm according to the UV-visible spectrophotometry, and draw a standard curve with absorbance as the ordinate and concentration as the abscissa.

[0089] Preparation of test solution: Take about 2 g of the powder of this product, weigh it accurately, place it in a stoppered conical flask, add 50 ml of ethanol, and ultrasonically treat it (power 140 W, frequency 42 kHz) for 45 minutes. Filter it, and place the filtrate in a 100 ml volumetric flask. Use appropriate amount of ethanol to wash the filter and filter residue in several times. Combine the washing liquid into the same volumetric flask, add ethanol to the scale, shake well, and obtain the solution.

[0090] Determination method: Accurately measure 0.2 ml of the test solution and place it in a 15 ml stoppered test tube. According to the method under the standard curve preparation, from "evaporation to dryness", operate in the same way to determine the absorbance. Read the content of oleanolic acid in the test solution from the standard curve and calculate it.

[0091] Example 1

[0092] Preparation of plant compound extracts for inhibiting Helicobacter pylori

[0093] 1. Ingredients: 200g minced garlic, 30g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0094] 2. Preparation steps:

[0095] (1) Garlic and leek seeds were mixed and crushed, 0.23 kg of 50° C. water was added, stirred for 1 h, 2.3 kg of n-hexane was added and soaked for 2 h, and then filtered to obtain a first filtrate and a first filter residue. The solvent was recovered from the first filtrate to obtain 4.064 g of garlic / leek seed volatile oil;

[0096] To the garlic / leek seed volatile oil, 40.64 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 0.5 h to emulsify the mixture. The emulsion was freeze-dried and pulverized to obtain 44 g of sulfide volatile oil.

[0097] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0098] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0099] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 169 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0100] (5) The powders obtained in step (1) and step (4) were combined to obtain 213 g of a plant composite extract.

[0101] According to tests, the extraction rate of allicin in garlic and leek seeds in this embodiment was 1.97 mg / g. In the plant composite extract, the proportion of sulfide volatile oil was 20.66%, the content of mushroom terpenoids was 3.37%, and the content of complex polysaccharides was 20.92%.

[0102] Example 2

[0103] Preparation of plant compound extracts for inhibiting Helicobacter pylori

[0104] 1. Ingredients: 200g minced garlic, 100g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0105] 2. Preparation steps:

[0106] (1) Garlic and leek seeds were mixed and crushed, 0.3 kg of 50° C. water was added, stirred for 1 h, 3 kg of n-hexane was added and soaked for 2 h, and filtered after soaking to obtain a first filtrate and a first filter residue. The solvent was recovered from the first filtrate to obtain 11.13 g of garlic / leek seed volatile oil;

[0107] To the garlic / leek seed volatile oil, 111.3 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 0.5 h to emulsify the obtained emulsion, which was freeze-dried and pulverized to obtain 122 g of sulfide volatile oil powder.

[0108] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0109] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0110] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 182 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0111] (5) The powders obtained in step (1) and step (4) were combined to obtain 304 g of a plant composite extract.

[0112] According to tests, the extraction rate of allicin in garlic and leek seeds in this embodiment was 1.72 mg / g. In the plant composite extract, the proportion of sulfide volatile oil was 40.13%, the content of mushroom terpenoids was 2.41%, and the content of complex polysaccharides was 11.37%.

[0113] Example 3

[0114] Preparation of capsules for inhibiting Helicobacter pylori

[0115] 1. Raw materials: 2kg crushed garlic, 0.3kg crushed leek seeds, 6kg crushed Ganoderma lucidum, 1kg crushed Hericium erinaceus;

[0116] 2. Preparation steps:

[0117] (1) Garlic and leek seeds were mixed and crushed, 2.3 kg of 50° C. water was added, stirred for 1 h, 23 kg of n-hexane was added and soaked for 2 h, and filtered after soaking to obtain a first filtrate and a first filter residue. The solvent was recovered from the first filtrate to obtain 40.94 g of garlic / leek seed volatile oil;

[0118] To the garlic / leek seed volatile oil, 409.4 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 2 h to emulsify the obtained emulsion, which was freeze-dried and pulverized to obtain 450 g of sulfide volatile oil powder.

[0119] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 56 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0120] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0121] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 1.69 kg of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0122] (5) combining the powders obtained in step (1) and step (4) to obtain 2.14 kg of the plant composite extract;

[0123] (6) The obtained plant compound extract was passed through an 80-mesh sieve, and then the sieved plant compound extract, pregelatinized starch, talc and magnesium stearate were mixed uniformly in a mass ratio of 70:25:3:2, and filled into capsules to prepare capsules.

[0124] Example 4

[0125] Preparation of tablets for inhibiting Helicobacter pylori

[0126] 1. Raw materials: 2kg crushed garlic, 0.3kg crushed leek seeds, 6kg crushed Ganoderma lucidum, 1kg crushed Hericium erinaceus;

[0127] 2. Preparation steps:

[0128] (1) Mix crushed garlic and crushed leek seeds, add 2.3 kg of 50° C. water, stir for 1 hour, add 23 kg of n-hexane and soak for 2 hours, filter after soaking to obtain a first filtrate and a first filter residue, recover the solvent from the first filtrate, and obtain 40.61 g of garlic / leek seed volatile oil;

[0129] To the garlic / leek seed volatile oil, 406.1 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 2 h to emulsify the solution. The emulsion was freeze-dried and pulverized to obtain 446 g of sulfide volatile oil powder.

[0130] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 56 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0131] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0132] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 1.69 kg of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0133] (5) combining the powders obtained in step (1) and step (4) to obtain 2.136 kg of the plant composite extract;

[0134] (6) The obtained plant compound extract was passed through an 80-mesh sieve, and then the sieved plant compound extract, pregelatinized starch, talc and magnesium stearate were mixed uniformly in a mass ratio of 70:26:3:1, pressed into tablets, and film-coated to prepare tablets.

[0135] Comparative Example 1

[0136] 1. Ingredients: 200g minced garlic, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0137] 2. Preparation steps:

[0138] (1) Add 0.2 kg of 50°C water to the minced garlic, stir for 1 hour, add 2 kg of n-hexane and soak for 2 hours. After extraction, filter to obtain a first filtrate and a first filter residue. Recover the solvent from the first filtrate to obtain 1.0 g of volatile oil;

[0139] 10 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added to the volatile oil and stirred for 0.5 h for emulsification to obtain an emulsion, which was freeze-dried and pulverized to obtain 11 g of sulfide volatile oil powder;

[0140] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0141] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0142] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 163 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0143] (5) The powders obtained in step (1) and step (4) were combined to obtain 174 g of a plant composite extract.

[0144] The results showed that the extraction rate of allicin in garlic in this comparative example was 1.08 mg / g. The plant compound extract contained 6.32% sulfide volatile oil, 4.06% mushroom terpenoids, and 29.82% complex polysaccharides.

[0145] Comparative Example 2

[0146] 1. Ingredients: 30g crushed leek seeds, 600g crushed Ganoderma lucidum, 100g crushed Hericium erinaceus;

[0147] 2. Preparation steps:

[0148] (1) Add 0.03 kg of 50° C. water to the crushed leek seeds, stir for 1 h, add 0.3 kg of n-hexane and soak for 2 h, extract and filter to obtain a first filtrate and a first filter residue, recover the solvent from the first filtrate, and obtain 2.961 g of volatile oil;

[0149] 29.61 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added to the volatile oil and stirred for 0.5 h for emulsification to obtain an emulsion, which was freeze-dried and pulverized to obtain 32 g of sulfide volatile oil powder;

[0150] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0151] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0152] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 84 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0153] (5) The powders obtained in step (1) and step (4) were combined to obtain 116 g of a plant composite extract.

[0154] The results showed that the extraction rate of allicin from leek seeds in this comparative example was 0.90 mg / g. The plant compound extract contained 27.59% sulfide volatile oil, 6.79% mushroom terpenoids, and 6.40% complex polysaccharides.

[0155] Comparative Example 3

[0156] 1. Ingredients: 200g minced garlic, 30g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0157] 2. Preparation steps:

[0158] (1) Garlic and leek seeds were mixed and crushed, 0.23 kg of 50° C. water was added, stirred for 1 h, 2.3 kg of n-hexane was added and soaked for 2 h, and then filtered to obtain a first filtrate and a first filter residue. The solvent was recovered from the first filtrate to obtain 4.056 g of garlic / leek seed volatile oil;

[0159] To the garlic / leek seed volatile oil, 40.56 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 0.5 h to emulsify the mixture. The emulsion was freeze-dried and pulverized to obtain 44 g of sulfide volatile oil powder.

[0160] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0161] (3) mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue, heating and refluxing for 1 hour, and filtering after extraction to obtain a second extract;

[0162] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 156 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0163] (5) The powders obtained in step (1) and step (4) were combined to obtain 200 g of the plant composite extract.

[0164] The results showed that the extraction rate of allicin in garlic and leek seeds in this comparative example was 1.93 mg / g. The plant compound extracts contained 22% sulfide volatile oil, 2.37% mushroom terpenoids, and 20.59% complex polysaccharides.

[0165] Comparative Example 4

[0166] 1. Ingredients: 200g minced garlic, 30g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0167] 2. Preparation steps:

[0168] (1) Garlic and leek seeds were mixed and crushed, 0.23 kg of 50° C. water was added, stirred for 1 h, 2.3 kg of n-hexane was added and soaked for 2 h, and then filtered to obtain a first filtrate and a first filter residue. The solvent was recovered from the first filtrate to obtain 4.151 g of garlic / leek seed volatile oil;

[0169] To the garlic / leek seed volatile oil, 41.51 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added and stirred for 0.5 h to emulsify the obtained emulsion, which was freeze-dried and pulverized to obtain 45 g of sulfide volatile oil powder.

[0170] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0171] (3) mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue, heating and refluxing for 3 hours, and filtering after extraction to obtain a second extract;

[0172] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 159 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0173] (5) The powders obtained in step (1) and step (4) were combined to obtain 204 g of a plant composite extract.

[0174] The results showed that the extraction rate of allicin in garlic and leek seeds in this comparative example was 1.96 mg / g. The plant compound extracts contained 22.06% sulfide volatile oil, 2.54% mushroom terpenoids, and 20.92% complex polysaccharides.

[0175] Comparative Example 5

[0176] 1. Raw materials: 200g minced garlic;

[0177] 2. Preparation steps:

[0178] (1) Add 0.2 kg of 50°C water to the minced garlic, stir for 1 h, add 2 kg of n-hexane and soak for 2 h. After soaking, filter to obtain a filtrate and a filter residue. Recover the solvent from the filtrate to obtain 1.02 g of volatile oil;

[0179] 5 mL of a 10.2 g / mL aqueous solution of sodium starch octenylsuccinate was added to the volatile oil and stirred for 0.5 h for emulsification to obtain an emulsion, which was freeze-dried and pulverized to obtain 11 g of sulfide volatile oil powder;

[0180] (2) Add 6 times the weight of water to the filter residue and perform microwave extraction for 1 h at a microwave power of 500 W. After extraction, filter to obtain the extract, concentrate until the extract has a Baume of 35°, dry under reduced pressure, and then grind to obtain 98 g of powder;

[0181] (3) The powder obtained in step (2) and the volatile oil powder were combined to obtain 109 g of plant extract.

[0182] The results showed that the extraction rate of allicin in garlic and leek seeds in this comparative example was 1.11 mg / g. The proportion of sulfide volatile oil in the plant composite extract was 10.09%, and the content of composite polysaccharides was 40.60%.

[0183] Comparative Example 6:

[0184] 1. Raw materials: 200g crushed leek seeds;

[0185] 2. Preparation steps:

[0186] (1) Add 0.2 kg of 50°C water to the crushed leek seeds, stir for 1 hour, add 2 kg of n-hexane and soak for 2 hours. After extraction, filter to obtain a filtrate and a filter residue. Recover the solvent from the filtrate to obtain 20 g of volatile oil;

[0187] 200 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added to the volatile oil and stirred for 0.5 h for emulsification to obtain an emulsion, which was freeze-dried and pulverized to obtain 220 g of sulfide volatile oil powder;

[0188] (2) Add 6 times the weight of water to the filter residue and perform microwave extraction for 1 h at a microwave power of 500 W. After extraction, filter to obtain the extract, concentrate until the extract has a Baume of 35°, dry under reduced pressure, and then grind to obtain 36 g of powder;

[0189] (3) The powder obtained in step (2) and the volatile oil powder were combined to obtain 256 g of plant extract.

[0190] After testing, in this comparative example, the proportion of sulfide volatile oil in the plant composite extract was 85.94%, and the content of composite polysaccharide was 0.06%.

[0191] Comparative Example 7

[0192] 1. Raw materials: 600g crushed Ganoderma lucidum;

[0193] 2. Preparation steps:

[0194] (1) adding 4.8 kg of 70% by volume ethanol to the Ganoderma lucidum shreds, heating and refluxing, and extracting for 2 hours. After extraction, filtering to obtain a filtrate and a filter residue; recovering the solvent from the filtrate to obtain a first extract;

[0195] (2) adding 6 times the weight of water to the residue mixture and performing microwave extraction for 1 h at a microwave power of 500 W. After extraction, filtering was performed to obtain a second extract;

[0196] (3) The first extract and the second extract were combined, concentrated until the extract Baume reached 35°, dried under reduced pressure, and then crushed to obtain 34 g of plant extract.

[0197] According to the test, in this comparative example, the content of mushroom terpenoid compounds in the plant composite extract is 20.00%, and the content of composite polysaccharides is 5.00%.

[0198] Comparative Example 8

[0199] 1. Raw materials: 100g minced Hericium erinaceus;

[0200] 2. Preparation steps:

[0201] (1) adding 0.8 kg of 70% by volume ethanol to the Hericium erinaceus powder, heating and refluxing, and extracting for 2 hours. After extraction, filtering to obtain a filtrate and a filter residue; recovering the solvent from the filtrate to obtain a first extract;

[0202] (3) adding 6 times the mass of water to the mixed residue and performing microwave extraction for 1 h at a microwave power of 500 W. After extraction, filtering was performed to obtain a second extract;

[0203] (4) The first extract and the second extract were combined, concentrated until the extract Baume reached 35°, dried under reduced pressure, and then crushed to obtain 32 g of plant extract.

[0204] According to the test, in this comparative example, the content of mushroom terpenoid compounds in the plant composite extract is 0.80%, and the content of composite polysaccharides is 15.00%.

[0205] Comparative Example 9

[0206] 1. Ingredients: 600g chopped Ganoderma lucidum, 100g chopped Hericium erinaceus;

[0207] 2. Preparation steps:

[0208] (1) Ganoderma lucidum and Hericium erinaceus were mixed, 5.6 kg of 70% ethanol by volume was added, and the mixture was heated under reflux for extraction for 2 h. After extraction, the mixture was filtered to obtain a filtrate and a filter residue; the solvent was recovered from the filtrate to obtain a first extract;

[0209] (2) adding 6 times the mass of water to the mixed residue and performing microwave extraction for 1 h at a microwave power of 500 W. After extraction, filtering was performed to obtain a second extract;

[0210] (3) The first extract and the second extract were combined, concentrated until the extract Baume reached 35°, dried under reduced pressure, and then crushed to obtain 66 g of the plant composite extract.

[0211] According to the test, in this comparative example, the content of mushroom terpenoid compounds in the plant composite extract is 10.71%, and the content of composite polysaccharides is 9.84%.

[0212] Comparative Example 10

[0213] 1. Ingredients: 200g minced garlic, 30g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0214] 2. Preparation steps:

[0215] (1) Add 0.2 kg of 50°C water to the minced garlic, stir for 1 hour, add 2 kg of n-hexane and soak for 2 hours. After soaking, filter to obtain a first filtrate and a first filter residue. Recover the solvent from the first filtrate to obtain 1.02 g of garlic volatile oil;

[0216] Add 0.03 kg of 50°C water to the crushed leek seeds, stir for 1 hour, add 0.3 kg of n-hexane and soak for 2 hours. After soaking, filter to obtain a first filtrate and a first filter residue. Recover the solvent from the first filtrate to obtain 3.0 g of leek seed volatile oil;

[0217] Garlic volatile oil and leek seed volatile oil were mixed to obtain a mixed volatile oil. 40.2 mL of a 1 g / mL aqueous solution of sodium starch octenylsuccinate was added to the mixed volatile oil, and the mixture was stirred for 0.5 h for emulsification to obtain an emulsion, which was freeze-dried and pulverized to obtain 44 g of sulfide volatile oil powder.

[0218] (2) Mixing the crushed Ganoderma lucidum and Hericium erinaceus, adding 5.6 kg of 70% by volume ethanol, heating under reflux and extracting for 2 hours, filtering after extraction to obtain a second filtrate and a second filter residue; recovering the solvent from the second filtrate to obtain a first extract;

[0219] (3) Mixing the first filter residue and the second filter residue to obtain a mixed filter residue, adding water 6 times the mass of the mixed filter residue and performing microwave extraction for 1 hour at a microwave power of 500W, and filtering after extraction to obtain a second extract;

[0220] (4) combining the first extract and the second extract, concentrating until the extract has a Baume of 35°, drying under reduced pressure, and then pulverizing to obtain 169 g of powder containing mushroom terpenoid compounds and complex polysaccharides;

[0221] (5) The powders obtained in step (1) and step (4) were combined to obtain 213 g of a plant composite extract.

[0222] Testing showed that the extraction rate of allicin in garlic and leek seeds in this comparative example was 1.10 mg / g. The plant composite extracts contained 20.66% sulfide volatile oil, 3.26% mushroom terpenoids, and 20.96% complex polysaccharides.

[0223] Comparative Example 11

[0224] 1. Ingredients: 200g minced garlic, 30g minced leek seeds, 600g minced Ganoderma lucidum, 100g minced Hericium erinaceus;

[0225] 2. Preparation steps:

[0226] Preparation steps: Add chopped garlic, chopped leek seeds, chopped Ganoderma lucidum and chopped Hericium erinaceus to 9.3 kg of purified water, heat and reflux extract for 2 hours, and recover the extract; add 9.3 kg of water to the medicinal residue and heat and reflux extract for a second time for 2 hours; combine the two filtrates, concentrate under reduced pressure, and dry under reduced pressure to obtain 203 g of plant composite extract.

[0227] According to the test, the extraction rate of allicin in garlic and leek seeds in this comparative example is 1.29 mg / g. The content of terpenoid compounds in the plant composite extract is 0.24%, and the content of composite polysaccharides is 26.45%.

[0228] Test Example 1:

[0229] The in vitro antibacterial experiment on Helicobacter pylori (H. pylori) was used to evaluate the synergistic effects between the extract formulas based on the in vitro antibacterial results of the extracts against Helicobacter pylori (H. pylori).

[0230] 1. Test methods

[0231] 1.1 Instruments and Equipment

[0232] 0.9% normal saline, distilled water, bacterial urease biochemical identification tubes, hydrogen peroxide solution, H. pylori liquid culture medium, solid culture medium, fresh defibrinated sheep blood, microaerophilic culture bags, microaerophilic gas release bags. Instruments: pipettes and tips, sterile pipettes, sterile Oxford cups, glass Petri dishes, L-shaped glass rods, electronic balance, glass rods, flasks, sterile test tubes, droppers, electric thermostatic water tank, inoculation loops, slides, electron microscope, ultra-clean workbench, vertical electric high-pressure steam sterilizer, and water-tight thermostatic incubator.

[0233] 1.2 Experimental steps

[0234] The agar diffusion method was used to measure the sizes of the inhibition zones of the extracts in the examples and comparative examples against Helicobacter pylori (H. pylori); the liquid continuous dilution method was used to observe the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extracts in the examples and comparative examples against Helicobacter pylori (H. pylori).

[0235] Preparation of bacterial suspension: Remove a standard strain of Helicobacter pylori (H. pylori) from a -80°C freezer. Dip a sterile inoculating loop into the bacterial suspension and inoculate the strain onto the surface of an H. pylori liquid culture plate using the streak method. Cover the plate with a lid and place the plate flat in a microaerobic culture bag. Also, insert a microaerobic gas release packet. Place the bag in a constant-temperature incubator and incubate at 37°C for 72 hours. Observe the morphology and characteristics of the colonies. H. pylori colonies are Gram-negative under a microscope, and their arrangement is S-shaped, curved, or seagull-shaped.

[0236] The cultured colonies were confirmed as H. pylori after Gram staining, positive catalase test, and positive urease test. After these identifications, the H. pylori culture was enriched in liquid medium and incubated in a microaerophilic environment for 72 h. The culture suspension was then removed and the concentration of the bacterial suspension was adjusted to 1 × 10 based on the A625 mm value on a UV spectrophotometer. 9 CFU / mL.

[0237] Liquid continuous dilution method The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the examples and comparative examples for H. pylori were determined by liquid continuous dilution method. Each group of plant extracts was prepared into a 1 g / ml solution with H. pylori liquid culture medium, and then diluted with liquid culture medium in a multiple ratio to 8 dilutions of 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256, and 1:512, that is, the concentrations of the Chinese medicine contained in the test tubes were 250 mg / ml, 125 mg / ml, 62.5 mg / ml, 31.25 mg / ml, 15.63 mg / ml, 7.81 mg / ml, 3.91 mg / ml,

[0238] 1.95mg / ml, take 1ml of each concentration of drug-containing culture medium, place it in a 2ml test tube, and add 0.05ml of bacterial solution to each tube. Simultaneously make a bacterial solution control tube and a culture medium control tube. Place the above culture solution in a culture bag, microaerobic environment, and culture in a 37℃ constant temperature incubator for 72 hours to observe the results. The experimental results use the lowest concentration level at which the liquid does not become turbid as the MIC of the test drug. Use an inoculation loop to dip the liquid culture medium that does not become turbid, streak it on the H. pylori plate culture medium, place it in a 37℃ incubator, microaerobic environment, and culture at a constant temperature for 72 hours. Observe the results, and use the lowest drug concentration at which no bacteria grow as the MBC of the test drug.

[0239] Agar diffusion method: Use a sterile pipette to pipette 0.1 ml of H. pylori bacterial solution onto the surface of an H. pylori culture plate. Spread the solution evenly with an L-shaped glass rod. Place four sterile Oxford cups equidistantly on the surface of the culture plate and remove the agar from the wells to create 6 mm diameter wells. Use a sterile pipette to pipette 0.1 ml of the 10 mg / ml solution from the Example group and the control group into the Oxford cups. Place the culture plate and a microaerobic gas generating bag into a sealed culture bag. Incubate in a constant temperature incubator at 37°C for 72 hours, then measure the diameter of the inhibition zone.

[0240] 2. Experimental Results

[0241] 2.1 Formulation dosage screening

[0242] Table 1 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of H. pylori

[0243]

[0244] The MIC and MBC values of garlic, leek seed, Ganoderma lucidum, and Hericium erinaceus extracts obtained from the above extractions were used to determine the screening dose range of 3.91-125 mg / ml. Garlic and leek seed extracts showed superior antibacterial effects, with dosages designed to be 1 / 4 MBC, 1 / 2 MBC, and MBC, respectively. Ganoderma lucidum and Hericium erinaceus extracts had lower antibacterial effects than garlic and leek seed extracts, and their dosages were designed to be 1 / 4 MIC, 1 / 2 MIC, and MIC, respectively.

[0245] Taking the size of the inhibition zone as the investigation factor, an orthogonal test was designed and composite extraction was performed. The results are as follows:

[0246] Table 2

[0247]

[0248] Table 3

[0249]

[0250]

[0251] Table 4

[0252]

[0253] According to the results of orthogonal experiment, the optimal concentration of allicin is 15.63 mg / ml, leek seeds, hericium erinaceus and Ganoderma lucidum are 7.81 mg / ml, and the dosage of extracts converted into raw medicinal materials is 2:0.33:1:6.

[0254] Table 5 Comparison of antibacterial effects of formulas

[0255]

[0256]

[0257] The inhibition zone values for H. pylori measured in this experiment for Example Group 1 and Example Group 2 were 22.87±0.43 and 20.37±0.35, respectively. Compared with Comparative Examples 1, 2, and 9, the antibacterial effect on H. pylori was greater than that of the formula without leek seeds, the formula without garlic, and the formula without leek seeds and garlic. Compared with Comparative Example Groups 5, 6, 7, and 8, the antibacterial effect was greater than that of the individual components of garlic extract, leek seed extract, Ganoderma lucidum extract, and Hericium erinaceus extract. Compared with Comparative Example Groups 3, 4, 10, and 11, the antibacterial effect on H. pylori was greater than that of the heating reflux extraction process, the separate extraction process of garlic and leek seeds, and the traditional water extraction process.

[0258] According to the degree of inhibition zone measured this time, the antibacterial strength of the whole formula increased, indicating that the simultaneous use of each group of drugs has a synergistic effect.

Claims

1. A plant compound extract for inhibiting Helicobacter pylori, characterized in that: The plant composite extract comprises 7%-45% of sulfide volatile oil, 1%-9% of mushroom terpenoid compounds and 10%-33% of complex polysaccharides; The plant compound extract is obtained from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum; Among them, sulfide volatile oil is extracted from garlic and leek seeds, and sulfide volatile oil includes allicin; Mushroom terpenoids are extracted from Hericium erinaceus and Ganoderma lucidum; The complex polysaccharide is extracted from garlic, leek seeds, Hericium erinaceus and Ganoderma lucidum.

2. The plant composite extract according to claim 1, characterized in that The plant compound extract is extracted from 1-3 parts of garlic, 0.1-1 parts of leek seeds, 1-3 parts of hericium erinaceus and 6-12 parts of ganoderma lucidum.

3. The plant composite extract according to claim 1, characterized in that The plant compound extract is extracted from 2 parts of garlic, 0.3 parts of leek seeds, 1 part of Hericium erinaceus and 6 parts of Ganoderma lucidum.

4. The method for preparing the plant composite extract according to any one of claims 1 to 3, characterized in that: The following steps are involved: Mixing garlic and leek seeds and crushing them, adding water and keeping warm for 1-2 hours, adding n-hexane for extraction, and filtering after extraction to obtain a first filtrate and a first filter residue; The solvent is recovered from the first filtrate to obtain garlic / leek seed volatile oil, and an aqueous solution of sodium starch octenylsuccinate is added and stirred for 0.5-2 hours for emulsification to obtain an emulsion, which is freeze-dried and pulverized to obtain sulfide volatile oil.

5. The preparation method according to claim 4, characterized in that The following steps are also included: The crushed Ganoderma lucidum and Hericium erinaceus are mixed, ethanol is added, and the mixture is heated under reflux for extraction. After extraction, the mixture is filtered to obtain a second filtrate and a second filter residue. The solvent is recovered from the second filtrate, and the mixture is concentrated and dried to obtain mushroom terpenoid compounds.

6. The preparation method according to claim 5, characterized in that The following steps are also included: The first filter residue and the second filter residue are mixed to obtain a mixed filter residue, water is added to perform microwave extraction, and after extraction, an extract is obtained by filtering, and the extract is concentrated and dried to obtain a complex polysaccharide.

7. The preparation method according to claim 4, characterized in that The mass ratio of garlic, leek seeds and n-hexane is (1-3):(0.1-1):(11-80); The ratio of the added amount of garlic / leek seed volatile oil to the sodium starch octenylsuccinate aqueous solution is 1g:6-10mL, and the concentration of the sodium starch octenylsuccinate aqueous solution is 0.5-1.5g / mL.

8. The preparation method according to claim 5, characterized in that The mass ratio of Ganoderma lucidum crushed, Hericium erinaceus crushed and ethanol is (6-12):(1-3):(42-150); The volume fraction of ethanol is 70%-80%, and the heating reflux extraction time is 1-2 hours.

9. The preparation method according to claim 6, characterized in that The mass ratio of the mixed filter residue to water is 1:(6-10), the power of the microwave extraction is 500-1000W, and the time is 0.5-1h.

10. Use of the plant composite extract according to any one of claims 1 to 3 in the preparation of a product for inhibiting Helicobacter pylori, characterized in that: The dosage form of the product is selected from at least one of powder, tablet, granule and capsule.

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