Preparation method of astragalus mongholicus fermented ointment for improving immunity

Abstract

The invention relates to the technical field of preparation of fermented ointment, and particularly discloses a preparation method of astragalus membranaceus fermented ointment for improving immunity. The method comprises the following steps: cutting astragalus membranaceus into sections, cleaning at low temperature, performing enzymolysis, adding saccharomycetes, performing anaerobic fermentation, drying and crushing to obtain astragalus membranaceus ferment powder; cleaning endothelium corneum gigeriae galli, drying and crushing at low temperature, extracting a stomach phase and an intestine phase, and merging filtrate to obtain an endothelium Steaming glutinous rice and Chinese yam, cooling, adding saccharomyces cerevisiae and the like, fermenting for 12-24 hours, adding enzyme, treating, filtering and centrifuging to obtain Chinese yam fermentation liquor; the three components are mixed, homogenized, concentrated, sterilized under ultrahigh pressure, packaged and then subjected to irradiation sterilization, and the finished ointment is obtained. The finished ointment prepared by the method realizes a multi-component synergistic effect, improves the availability of active components, retains thermosensitive components, gives consideration to the safety and stability of the product through double sterilization, and has an obvious benefit for improving the immunity.

Description

Technical Field

[0001] This invention belongs to the field of fermented paste preparation technology, specifically relating to a method for preparing Astragalus fermented paste to enhance immunity. Background Technology

[0002] With the fast pace of modern life and increasing environmental pressures, weakened immunity has become a global health problem. World Health Organization data shows that approximately 70% of the global population is in a sub-healthy state, with immune dysfunction being a core contributing factor. Traditional Chinese medicine theory holds that weakened immunity is closely related to "insufficient qi and blood" and "weak spleen and stomach." The combination of astragalus, chicken gizzard lining, yam, and glutinous rice has long been used for immune regulation due to its effects of tonifying the middle energizer, replenishing qi, strengthening the spleen, and harmonizing the stomach.

[0003] However, clinical practice shows that directly consuming the above four-ingredient formula through decoctions or medicinal diets generally has limited immune-boosting effects, especially for middle-aged and elderly people, patients with chronic diseases, and other groups with weakened immune function, where the effectiveness rate is significantly lower. The main reason is that conventional water extraction can only extract polar components and the extraction rate is low, resulting in insufficient release of effective components. At the same time, traditional processing methods require high-temperature treatment, which also causes a significant degree of damage to the active ingredients, leading to poor results. Summary of the Invention

[0004] In view of the shortcomings and problems of low utilization rate of effective ingredients and easy destruction of activity by high temperature processing in the traditional consumption method of the four-ingredient formula, the present invention provides a method for preparing Astragalus fermented paste to improve immunity.

[0005] A method for preparing Astragalus fermented extract to enhance immunity includes the following steps: S1. Weigh out 400-600 parts of Astragalus membranaceus, 600-800 parts of fresh chicken gizzard lining, 500-800 parts of fresh yam, and 100-150 parts of glutinous rice by weight. S2. Preparation of Astragalus Fermentation Powder: Astragalus is cut into sections, washed at low temperature, and then enzymatically hydrolyzed. After enzymatic hydrolysis, yeast is added for anaerobic fermentation. The fermented Astragalus sections are dried and then pulverized to obtain Astragalus Fermentation Powder. S3. Preparation of chicken gizzard lining extract: After cleaning the inner wall of fresh chicken gizzard, dry it at low temperature and crush it. After crushing, extract the gastric phase to obtain gastric phase extract and residue. Extract the residue into the intestinal phase to obtain intestinal phase extract. Combine the gastric phase extract and intestinal phase extract and filter to obtain chicken gizzard lining extract. S4. Preparation of yam fermentation liquid: Glutinous rice and yam are steamed and cooled separately. Then, brewer's yeast, Lactobacillus plantarum and glucoamylase are added to the pre-treated glutinous rice and yam for fermentation for 12-24 hours. After fermentation, heat-resistant α-amylase is added to the fermented solution and boiled. When the temperature drops to 60℃, β-glucosidase is added and the reaction continues for 2 hours. The solution is then filtered to obtain the treatment liquid. The treatment liquid is then filtered and centrifuged to obtain yam fermentation liquid. S5. Ointment preparation: Astragalus fermented powder, chicken gizzard extract and yam fermented liquid are mixed in proportion, homogenized and concentrated to obtain concentrated liquid. The concentrated liquid is sterilized by ultra-high pressure and packaged. After packaging, it is sterilized by irradiation to obtain the finished ointment.

[0006] The above-mentioned method for preparing Astragalus fermented paste to enhance immunity includes the following steps in step S2: (1) Cut Astragalus membranaceus into sections, clean it with ultrasonic cleaning at low temperature, add 1.5% cellulase and 2.0% pectinase, and enzymatically hydrolyze it for 2 hours at 45℃ and pH 5.0. (2) Mix the enzymatically hydrolyzed Astragalus membranaceus segments with yeast at a ratio of 10:1 and add glucose solution. Ferment at 37°C under anaerobic conditions for 48 hours. (3) After fermentation, take out the Astragalus segments and dry them in a low-temperature dryer. Place the dried Astragalus segments in an air jet mill and grind them to 300-500 mesh under low-temperature conditions to obtain Astragalus fermentation powder.

[0007] In the above-mentioned method for preparing Astragalus fermented paste to enhance immunity, the cellulase used in step (1) has an enzyme activity of 2000 U / g, and the pectinase has an enzyme activity of 1000 U / g.

[0008] The above-mentioned method for preparing Astragalus fermented paste to enhance immunity includes the following steps in step S3: preparation of chicken gizzard extract: (1) Take the inner wall of fresh chicken gizzard, wash it repeatedly with deionized water, dry it in a low temperature dryer, and then pulverize it to 300-500 mesh in an air jet mill under low temperature conditions. (2) Gastric phase extraction: Mix 0.1 mol / L hydrochloric acid solution and 0.2 mol / L potassium chloride solution at a volume ratio of 1:2, adjust the pH to 1.5~2.0, add chicken gizzard powder and 2% pepsin, and extract with magnetic stirring at 37℃ for 2h. After extraction, centrifuge at 4℃ and 8000rpm for 15min, collect the supernatant, and keep the residue for intestinal phase extraction; (3) Intestinal phase extraction: Mix 0.1 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution at a volume ratio of 3:1, adjust the pH to 7.5~8.0, add gastric phase extraction residue, 1.5% trypsin and 1.0% α-amylase, and extract at 37℃ with magnetic stirring for 3h. After extraction, centrifuge at 4℃ and 8000rpm for 15min, collect the supernatant and combine it with the gastric phase extract; (4) The combined extract was pre-filtered through a 0.45 μm microfiltration membrane to remove solid particles, and then purified by ultrafiltration to obtain chicken gizzard extract.

[0009] In the above-mentioned method for preparing Astragalus fermented paste to enhance immunity, the enzyme activity of the pepsin used in the gastric phase extraction in step (2) is ≥3000U / mg.

[0010] The above-mentioned method for preparing Astragalus fermented paste to enhance immunity includes step (3) in which the activity of trypsin used in intestinal extraction is ≥2000U / mg and the activity of α-amylase used is ≥5000U / mg.

[0011] In the above-mentioned method for preparing Astragalus fermented paste to enhance immunity, the molecular weight cutoff of the ultrafiltration membrane in step (4) is 3000 Da.

[0012] The above-mentioned method for preparing Astragalus fermented extract to enhance immunity includes the following steps in step S4: (1) Soak glutinous rice in warm water at 37℃ for 8 hours. After the water absorption rate reaches 40%, steam it at 100℃ for 15 minutes to gelatinize the outer layer. Then, cool it down to 85℃ and steam it for 20 minutes. Let it cool and set aside. (2) Peel and cut the yam into pieces, then vacuum steam it at -0.06MPa and 90℃ for 30 minutes, mash it, and let it cool before use; (3) Add 5.0% brewer's yeast, 2.0% Lactobacillus plantarum and 2.0% glucoamylase to the pretreated glutinous rice and yam, mix well, seal and incubate in a 25-30℃ incubator for 18h; (4) Add 1.5% heat-resistant α-amylase to the fermented solution and boil it. When the temperature drops to 60℃, add 1.0% β-glucosidase and continue to act for 2 hours. Then filter to obtain the treatment solution. (5) After filtering the treatment liquid, centrifuge it at 8000 r / min and 20℃ for 15 min to obtain a preliminary clarified liquid. Then, centrifuge the preliminary clarified liquid a second time at high speed to obtain yam fermentation liquid.

[0013] The above-mentioned method for preparing Astragalus fermented paste to enhance immunity involves using Lactobacillus plantarum, which is Lactobacillus plantarum that has been domesticated in yam juice for three generations. The yam juice is obtained by mixing yam chunks and distilled water in a ratio of 1:1.1-1.2, then pulping the mixture using a tissue homogenizer, and finally removing the residue by passing it through an 80-100 mesh sieve.

[0014] The above-mentioned method for preparing Astragalus fermented paste to enhance immunity includes the following steps in step S5: (1) Mix Astragalus fermented powder, chicken gizzard extract and yam fermented liquid according to the proportion, and then circulate them 3-5 times at 150-200MPa through a homogenizing valve to obtain a fermented powder suspension; (2) The yeast suspension was concentrated to one-fifth to one-quarter using a hydrophobic polytetrafluoroethylene membrane with a pore size of 0.2 μm under operating conditions of -0.09 MPa and 40 °C. (3) After the concentrate is sterilized by ultra-high pressure for 3 minutes, it is packaged. After packaging, the finished product is sterilized by cobalt-60 γ-ray irradiation at ≤10℃ with a dose of 2-5kGy to obtain the finished ointment.

[0015] Compared with the prior art, the beneficial effects of the present invention are: This invention achieves a synergistic effect of multiple components working together to exert a compound effect of "immune regulation, digestive aid, and intestinal health maintenance" through differentiated and precise processing of Astragalus membranaceus, chicken gizzard membrane, and Dioscorea opposita. For Astragalus membranaceus, a combined approach of probiotic fermentation and enzymatic hydrolysis is used to synergistically process the product, converting poorly absorbed primary saponins into easily absorbed secondary glycosides. Low-temperature pulverization further enhances the release efficiency of the active ingredients, significantly improving the bioavailability of Astragalus membranaceus and strengthening its core activities of immune regulation and antioxidation. For chicken gizzard lining, an innovative biomimetic enzymatic hydrolysis process is employed, mimicking the human gastrointestinal environment. Pepsin is used in the gastric phase, while trypsin and α-amylase are used for stepwise extraction in the intestinal phase. Purification is achieved using an ultrafiltration membrane with a molecular weight cutoff of ≤3000 Da, specifically retaining digestive active ingredients such as small-molecule peptides, significantly improving the extraction rate and purity of proteins and peptides. For yam, a combined process of microbial fermentation and dual enzymatic hydrolysis is used. Saccharin yeast and Lactobacillus plantarum domesticated from yam juice are used for co-fermentation, combined with dual enzymatic hydrolysis using glucosyl amylase and β-glucosidase. This converts large-molecule polysaccharides in yam into small-molecule polysaccharides and functional oligosaccharides. Simultaneously, organic acids and prebiotics are produced through probiotic metabolism, enhancing intestinal regulation and nutritional supplementation activities.

[0016] This invention also introduces nano-homogenization technology for mixed powders, processing the mixed fermentation powder suspension to the nanoscale, significantly improving the dissolution and absorption efficiency of components; combined with a low-temperature process throughout, it maximizes the retention of heat-sensitive active ingredients; at the same time, it adopts a dual aseptic protection system of ultra-high pressure sterilization and cobalt-60 gamma irradiation, combined with membrane filtration purification to ensure efficacy, so as to ensure product sterility while avoiding the destruction of active ingredients by high-temperature sterilization, thus taking into account both safety and stability. Detailed Implementation

[0017] To address the shortcomings and problems of low utilization rate of effective ingredients and easy destruction of activity during high-temperature processing in traditional four-ingredient formulas, this invention provides a method for preparing Astragalus fermented paste to enhance immunity. The invention is further illustrated below with specific embodiments.

[0018] The materials and equipment used in this invention are as follows: Material: Cellulase: Nanning Pangbo Biotechnology Co., Ltd.; Pectinase: Shandong Longket Enzyme Preparation Co., Ltd.; Yeast: Angel Yeast Co., Ltd.; Pepsin: Nanning Pangbo Biotechnology Co., Ltd.; Trypsin: Nanning Pangbo Biotechnology Co., Ltd.; α-Amylase: Shandong Longket Enzyme Preparation Co., Ltd.; Brewing yeast: Angel Yeast Co., Ltd.; Lactobacillus plantarum: Angel Yeast Co., Ltd.; Glucoamylase: Hunan Xinhongying Biotechnology Co., Ltd.; High-temperature resistant α-amylase: Baiweixiang Biotechnology Co., Ltd.; The astragalus, chicken gizzard lining, yam, and glutinous rice used are all commercially available.

[0019] equipment: Electric heating forced-air drying oven: DHG-9140A, Shanghai Yiheng Scientific Instruments Co., Ltd.; Vacuum drying oven: DZF-1, Beijing Yongguangming Medical Instrument Factory; Ultrasonic Cleaner: KQ-100V, Kunshan Ultrasonic Instrument Co., Ltd. Electric thermostatic incubator: BXP-280, Shanghai Boxun Medical Bio-instrument Co., Ltd.; Magnetic Stirred Water Bath: DK-10S, Ningguo Shaying Science; High-speed refrigerated centrifuge: SIGMA3K15, SIGMA GmbH, Germany.

[0020] Example 1: This example provides a method for preparing Astragalus fermented extract to enhance immunity. Step 1: Weigh out 600 parts of Astragalus membranaceus, 800 parts of fresh chicken gizzard lining, 800 parts of fresh yam, and 150 parts of glutinous rice according to their weight proportions.

[0021] Step 2: Preparation of Astragalus Fermentation Powder: (1) Cut the astragalus into sections and use a low-temperature ultrasonic cleaning device with the temperature set at 5-10℃ to remove the mud, microorganisms and impurities attached to the surface of the astragalus by high-frequency vibration.

[0022] (2) Add a compound enzyme solution containing 1.5% (by weight of Astragalus membranaceus) of cellulase with an activity of 2000 U / g and 2.0% (by weight of Astragalus membranaceus) of pectinase with an activity of 1000 U / g to treat Astragalus membranaceus segments. Enzymatic hydrolysis was carried out at 45℃ and pH 5.0 for 2 h to destroy the cell wall fiber structure.

[0023] (3) Mix the enzymatically hydrolyzed Astragalus membranaceus segments with yeast at a ratio of 10:1, add an appropriate amount of glucose solution, and ferment at 37°C under anaerobic conditions for 48 h.

[0024] (4) After fermentation, remove the Astragalus membranaceus segments and dry them in a low-temperature dryer at a temperature of 50-60℃ until dry.

[0025] (5) Place the dried Astragalus segments in an air jet mill and grind them to 300-500 mesh at a low temperature of ≤20℃ to obtain Astragalus fermentation powder for later use.

[0026] Step 3: Preparation of chicken gizzard extract (1) Select fresh chicken gizzard lining, rinse repeatedly with deionized water until no impurities remain, and dry in a low temperature dryer at a temperature of 50-60℃ until dry.

[0027] (2) Place the dried chicken gizzard in an air jet mill and grind it to 300-500 mesh at a low temperature of ≤20℃ to achieve a cell wall breakage rate of over 90%.

[0028] (3) Extraction of the gastric phase in a simulated human gastrointestinal digestive environment: Mix 0.1 mol / L hydrochloric acid solution and 0.2 mol / L potassium chloride solution at a volume ratio of 1:2, adjust the pH to 1.5~2.0, add chicken gizzard powder and 2% pepsin with enzyme activity ≥3000U / mg, and extract at 37℃ with magnetic stirring for 2 h. After extraction, centrifuge at 4℃ and 8000rpm for 15 min, collect the supernatant, and retain the residue for intestinal phase extraction. The supernatant mainly contains protein, polypeptide and other components.

[0029] (4) Intestinal phase extraction simulating the human gastrointestinal digestive environment: Mix 0.1 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution at a volume ratio of 3:1, adjust the pH to 7.5~8.0, add the residue after gastric phase extraction, 1.5% trypsin with enzyme activity ≥2000U / mg and 1.0% α-amylase with enzyme activity ≥5000U / mg, and extract at 37℃ with magnetic stirring for 3h. After extraction, centrifuge at 4℃ and 8000rpm for 15 min, collect the supernatant and combine it with the gastric phase extract.

[0030] (5) The combined extract was pre-filtered through a 0.45 μm microfiltration membrane to remove solid particles, and then purified by ultrafiltration membrane with a molecular weight cutoff of 3000 Da to obtain chicken gizzard extract for later use.

[0031] Step 4: Preparation of Yam Fermentation Broth: (1) Soak glutinous rice in warm water at 37℃ for 8 hours. After the water absorption rate reaches 40%, use a steam jacketed kettle to steam at 100℃ for 15 minutes to gelatinize the outer layer, then cool down to 85℃ and steam for 20 minutes. Let it cool before use.

[0032] (2) Peel and cut the yam into pieces, then vacuum steam it at -0.06MPa and 90℃ for 30 minutes, crush it, let it cool, and set it aside.

[0033] (3) Add 5.0% brewer's yeast, 2.0% Lactobacillus plantarum (pre-cultured in yam juice for three generations) and 2.0% glucoamylase to the pre-treated glutinous rice and yam, mix well, seal and incubate in a 25-30℃ incubator for 18 h.

[0034] (4) Add 1.5% heat-resistant α-amylase to the fermented solution and boil it. When the temperature drops to 60℃, add 1.0% β-glucosidase and continue to act for 2 h to obtain the treatment solution.

[0035] (5) Filter the treatment liquid obtained in (4) and centrifuge it at 8000 r / min and 20℃ for 15 min to obtain a preliminary clarified liquid.

[0036] (6) The preliminary clarified liquid was centrifuged twice at 11000 r / min and 4℃ for 30 min to obtain yam fermentation liquid for later use.

[0037] Step 5: Ointment Preparation (1) The processed Astragalus fermented powder, chicken gizzard extract and yam fermented liquid are mixed according to the ratio and circulated through a homogenizer at 150-200MPa for 3-5 times to prepare the fermented powder suspension into nanoscale.

[0038] (2) The yeast suspension was concentrated to one-fifth to one-quarter using a hydrophobic polytetrafluoroethylene membrane with a pore size of 0.2 μm at an operating pressure of -0.09 MPa and a temperature of 40°C.

[0039] (3) The concentrate is sterilized under ultra-high pressure at 600MPa for 3 minutes and then packaged.

[0040] (4) After packaging, the product is sterilized by irradiation with cobalt-60 γ rays at a low temperature of ≤10℃ with a dose of 2-5kGy to obtain the finished ointment.

[0041] The prepared ointment is brownish-black to dark brown in color, with a uniform and delicate appearance, no obvious insoluble matter, and when picked up with a glass rod, it slowly falls off in flakes, exhibiting a flag-hanging phenomenon and having a certain degree of fluidity.

[0042] Example 2: The similarities between this example and Example 1 will not be repeated here. The difference is that the raw materials used are: 400 parts of Astragalus membranaceus, 600 parts of fresh chicken gizzard lining, 600 parts of fresh yam, and 100 parts of glutinous rice.

[0043] Example 3: The similarities between this example and Example 1 will not be repeated here. The difference is that the raw materials used are: 500 parts of Astragalus membranaceus, 700 parts of fresh chicken gizzard lining, 500 parts of fresh yam, and 120 parts of glutinous rice.

[0044] Comparative Example 1: This comparative example provides an astragalus paste, the similarities with Example 1 will not be repeated, the difference being: in this example, astragalus, chicken gizzard lining, yam and glutinous rice are directly boiled, specifically: Soak astragalus, yam and glutinous rice in 5 times the amount of water for 1 hour. Bring to a boil over high heat, then simmer over low heat for 1 hour. Filter out the liquid, add 5 times the amount of water and decoct again. Combine the two liquids. After drying and pulverizing the chicken gizzard lining, add it to the medicinal liquid and concentrate it to one-fifth to one-quarter of the medicinal liquid to obtain Astragalus ointment.

[0045] Comparative Example 2: This comparative example provides a fermented Astragalus extract, which differs from Example 1 in that Astragalus, chicken gizzard lining, yam, and glutinous rice are fermented as a whole. The preparation method is as follows: (1) Preparation of chicken gizzard extract: Place the dried chicken gizzard in an air jet mill and grind it at low temperature to 300-500 mesh. Add 5 times the amount of water to the chicken gizzard and boil for 2 hours. Filter and collect the filtrate. Add 5 times the amount of water to the residue and continue to boil for 2 hours. Filter and collect the filtrate. Combine the two filtrates to obtain chicken gizzard extract for later use.

[0046] (2) Pulverize the dried Astragalus membranaceus and pass it through a 300-500 mesh sieve; (3) Steam the glutinous rice and yam separately until cooked, then let them cool and set aside. (4) Mix Astragalus powder, chicken gizzard extract, steamed glutinous rice and yam in proportion and put them into a fermentation tank. Add Lactobacillus plantarum and Saccharomyces cerevisiae to the fermentation tank and ferment for 18 hours. After filtration, concentrate and sterilize the fermentation liquid to obtain fermentation paste.

[0047] Comparative Example 3: This comparative example provides an Astragalus fermented extract. The similarities to Example 1 will not be repeated here. The difference is that the Astragalus fermented powder used in this comparative example only undergoes fermentation treatment without enzymatic hydrolysis. Astragalus segments and yeast are directly mixed at a ratio of 10:1 and then added to a glucose solution. Fermentation is carried out at 37°C under anaerobic conditions for 48 hours. After fermentation, the Astragalus segments are removed and dried in a low-temperature dryer (50-60°C) until dry. The dried Astragalus segments are then placed in an air jet mill and pulverized to 300-500 mesh at a low temperature of ≤20°C to obtain Astragalus fermented powder for later use.

[0048] Comparative Example 4: This comparative example provides an Astragalus fermented paste. The similarities with the examples will not be repeated. The difference is that when processing chicken gizzard lining in this comparative example, the chicken gizzard lining is pulverized by airflow, then 5 times the amount of water is added to the chicken gizzard lining and boiled for 2 hours. The filtrate is collected by filtration, and 5 times the amount of water is added to the residue and boiled for another 2 hours. The filtrate is collected by filtration, and the two filtrates are combined to obtain chicken gizzard lining extract for later use.

[0049] Experimental Example: In this experimental example, multiple batches of fermented paste were prepared according to the method of Example 1, and the relative density and insoluble matter of different batches of paste were measured to verify the stability of the paste prepared by the present invention.

[0050] 1. Relative density measurement According to Item 0601 of General Chapter IV of the 2020 Chinese Pharmacopoeia, five batches of fermentation paste were weighed and their relative densities were calculated using the hydrostatic bottle method. The results are shown in Table 1 below.

[0051]

[0052] As can be seen from Table 1, the density of multiple batches of samples is stable, all not lower than 1.18, and the RSD value is 0.13%, which meets the requirements.

[0053] 2. Insoluble matter detection Five batches of fermented paste samples were taken. Following the instructions in Section 0183 of Part IV of the 2020 Chinese Pharmacopoeia, 5g of each sample was weighed and placed in a beaker. 300mL of hot water was poured in, and the mixture was stirred with a glass rod. After the sample was completely dissolved, it was left to stand for 5 minutes, and the insoluble matter was observed. The results are shown in Table 2 below.

[0054]

[0055] As can be seen from Table 2, none of the above samples contained obvious foreign matter.

[0056] Experimental Example 2: In this experimental example, a rat experiment was conducted to compare the effects of ointments obtained by different preparation methods, as follows.

[0057] (1) Experimental animals Thirty male SD rats at 25 weeks of age with a body weight of 450 - 550 g were randomly selected, and thirty female SD rats at 25 weeks of age with a body weight of 300 - 400 g were randomly selected. They were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (License No.: SYXK(Beijing)2022 - 0052), and the experiment was carried out in accordance with the regulations of the "Regulations on the Administration of Laboratory Animals". The male rats and female rats were each randomly divided into 6 groups, with 1 group in 1 cage and 5 rats in each group. All rats were raised under the conditions of an environmental temperature of 20 - 24°C, a humidity of 40% - 60%, 12 - hour daylight and night alternation lighting, and during this period, they had free access to water and food, and the air was regularly ventilated and exhausted.

[0058] (2) Experimental grouping The male rats and female rats were respectively divided into a blank control group, Experimental Group 1, Experimental Group 2, Experimental Group 3, Experimental Group 4, and Experimental Group 5 according to the grouping. The initial body weight of each rat in each group was weighed and recorded, and the average body weight was used as the initial body weight of the group. Among them, the blank control group was intragastrically administered 5 ml of drinking water; Experimental Group 1 was intragastrically administered 5 ml of the 1:10 diluted liquid medicine of the fermented ointment prepared in Example 1; Experimental Group 2 was intragastrically administered 5 ml of the 1:10 diluted liquid medicine of the ointment prepared in Comparative Example 1; Experimental Group 3 was intragastrically administered 5 ml of the 1:10 diluted liquid medicine of the ointment prepared in Comparative Example 2; Experimental Group 4 was intragastrically administered 5 ml of the 1:10 diluted liquid medicine of the ointment prepared in Comparative Example 3; Experimental Group 5 was intragastrically administered 5 ml of the 1:10 diluted liquid medicine of the ointment prepared in Comparative Example 4; Intragastric administration was carried out continuously for 8 weeks. During the intragastric administration period, the rats in each group had normal diet and water intake, and the signs and behavioral activities of the rats in each group were observed and recorded for any abnormalities.

[0059] After 8 weeks, the body weights of the rats in each group were weighed and recorded, and blood was collected from the tail vein to detect the blood glucose concentration, serum IgG concentration, serum TNF-α concentration, and hemoglobin (Hb) concentration of the rats. The results are shown in Table 3 and Table 4 below.

[0060]

[0061]

[0062] It can be seen from the results of Table 3 and Table 4 that taking the ointment made of Astragalus membranaceus, Gallus gallus domesticus Gmelin, Dioscorea opposita Thunb, and glutinous rice has a certain promoting effect on the weight gain, immune improvement, and hemoglobin increase of rats. The fermented ointment prepared by the method of the present invention has a more obvious effect on the weight gain and immune improvement of rats through the dual effects of enzymatic hydrolysis and fermentation, indicating that the ointment prepared by the process of the present invention can effectively retain the active ingredients of the raw materials and is beneficial to the exertion of the efficacy of the active ingredients.

[0063] The above description is only a preferred embodiment of the present invention and does not limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing Astragalus fermented extract to enhance immunity, characterized in that: Includes the following steps: S1. Weigh out 400-600 parts of Astragalus membranaceus, 600-800 parts of fresh chicken gizzard lining, 500-800 parts of fresh yam, and 100-150 parts of glutinous rice by weight. S2. Preparation of Astragalus Fermentation Powder: Astragalus is cut into sections, washed at low temperature, and then enzymatically hydrolyzed. After enzymatic hydrolysis, yeast is added for anaerobic fermentation. The fermented Astragalus sections are dried and then pulverized to obtain Astragalus Fermentation Powder. S3. Preparation of chicken gizzard lining extract: After cleaning the inner wall of fresh chicken gizzard, dry it at low temperature and crush it. After crushing, extract the gastric phase to obtain gastric phase extract and residue. Extract the residue into the intestinal phase to obtain intestinal phase extract. Combine the gastric phase extract and intestinal phase extract and filter to obtain chicken gizzard lining extract. S4. Preparation of yam fermentation liquid: Glutinous rice and yam are steamed and cooled separately. Then, brewer's yeast, Lactobacillus plantarum and glucoamylase are added to the pre-treated glutinous rice and yam for fermentation for 12-24 hours. After fermentation, heat-resistant α-amylase is added to the fermented solution and boiled. When the temperature drops to 60℃, β-glucosidase is added and the reaction continues for 2 hours. The solution is then filtered to obtain the treatment liquid. The treatment liquid is then filtered and centrifuged to obtain yam fermentation liquid. S5. Ointment preparation: The prepared Astragalus fermented powder, chicken gizzard extract and yam fermented liquid are mixed, homogenized and concentrated to obtain a concentrated liquid. The concentrated liquid is sterilized under ultra-high pressure and then packaged. After packaging, it is sterilized by irradiation to obtain the finished ointment.

2. The method for preparing Astragalus fermented extract to enhance immunity according to claim 1, characterized in that: The preparation of Astragalus fermented powder in step S2 includes the following steps: (1) Cut Astragalus membranaceus into sections, clean it with ultrasonic cleaning at low temperature, add 1.5% cellulase and 2.0% pectinase, and enzymatically hydrolyze it for 2 hours at 45℃ and pH 5.

0. (2) Mix the enzymatically hydrolyzed Astragalus membranaceus segments with yeast at a ratio of 10:1 and add glucose solution. Ferment at 37°C under anaerobic conditions for 48 hours. (3) After fermentation, take out the Astragalus segments and dry them in a low-temperature dryer. Place the dried Astragalus segments in an air jet mill and grind them to 300-500 mesh under low-temperature conditions to obtain Astragalus fermentation powder.

3. The method for preparing Astragalus fermented extract to enhance immunity according to claim 2, characterized in that: The cellulase used in step (1) has an enzyme activity of 2000 U / g, and the pectinase has an enzyme activity of 1000 U / g.

4. The method for preparing Astragalus fermented extract to enhance immunity according to claim 1, characterized in that: Step S3, the preparation of chicken gizzard extract, includes the following steps: (1) Take the inner wall of fresh chicken gizzard, wash it repeatedly with deionized water, dry it in a low temperature dryer, and then pulverize it to 300-500 mesh in an air jet mill under low temperature conditions. (2) Gastric phase extraction: Mix 0.1 mol / L hydrochloric acid solution and 0.2 mol / L potassium chloride solution at a volume ratio of 1:2, adjust the pH to 1.5~2.0, add chicken gizzard powder and 2% pepsin, and extract with magnetic stirring at 37℃ for 2h. After extraction, centrifuge at 4℃ and 8000rpm for 15min, collect the supernatant, and keep the residue for intestinal phase extraction; (3) Intestinal phase extraction: Mix 0.1 mol / L disodium hydrogen phosphate solution and 0.1 mol / L citric acid solution at a volume ratio of 3:1, adjust the pH to 7.5~8.0, add gastric phase extraction residue, 1.5% trypsin and 1.0% α-amylase, and extract at 37℃ with magnetic stirring for 3h. After extraction, centrifuge at 4℃ and 8000rpm for 15min, collect the supernatant and combine it with the gastric phase extract; (4) The combined extract was pre-filtered through a 0.45 μm microfiltration membrane to remove solid particles, and then purified by ultrafiltration to obtain chicken gizzard extract.

5. The method for preparing Astragalus fermented extract to enhance immunity according to claim 4, characterized in that: In step (2), the pepsin used for gastric extraction has an enzyme activity ≥3000U / mg.

6. The method for preparing Astragalus fermented extract to enhance immunity according to claim 4, characterized in that: Step (3) The trypsin activity used for intestinal phase extraction is ≥2000U / mg, and the α-amylase activity used is ≥5000U / mg.

7. The method for preparing Astragalus fermented extract to enhance immunity according to claim 4, characterized in that: The ultrafiltration membrane described in step (4) has a molecular weight cutoff of 3000 Da.

8. The method for preparing Astragalus fermented extract to enhance immunity according to claim 1, characterized in that: The preparation method of yam fermentation broth in step S4 includes the following steps: (1) Soak glutinous rice in warm water at 37℃ for 8 hours. After the water absorption rate reaches 40%, steam it at 100℃ for 15 minutes to gelatinize the outer layer. Then, cool it down to 85℃ and steam it for 20 minutes. Let it cool and set aside. (2) Peel and cut the yam into pieces, then vacuum steam it at -0.06MPa and 90℃ for 30 minutes, mash it, and let it cool before use; (3) Add 5.0% brewer's yeast, 2.0% Lactobacillus plantarum and 2.0% glucoamylase to the pretreated glutinous rice and yam, mix well, seal and incubate in a 25-30℃ incubator for 18h; (4) Add 1.5% heat-resistant α-amylase to the fermented solution and boil it. When the temperature drops to 60℃, add 1.0% β-glucosidase and continue to act for 2 hours. Then filter to obtain the treatment solution. (5) After filtering the treatment liquid, centrifuge it at 8000 r / min and 20℃ for 15 min to obtain a preliminary clarified liquid. Then, centrifuge the preliminary clarified liquid a second time at high speed to obtain yam fermentation liquid.

9. The method for preparing Astragalus fermented extract to enhance immunity according to claim 8, characterized in that: The plant lactobacillus is a plant lactobacillus that has been domesticated in yam juice for three generations. The yam juice is obtained by mixing yam chunks and distilled water in a ratio of 1:1.1-1.2, then pulping the mixture with a tissue homogenizer, and finally removing the residue by passing it through an 80-100 mesh sieve.

10. The method for preparing Astragalus fermented extract to enhance immunity according to claim 1, characterized in that: Step S5, the preparation of the ointment, includes the following steps: (1) Mix Astragalus fermented powder, chicken gizzard extract and yam fermented liquid according to the proportion, and then circulate them 3-5 times at 150-200MPa through a homogenizing valve to obtain a fermented powder suspension; (2) The yeast suspension was concentrated to one-fifth to one-quarter using a hydrophobic polytetrafluoroethylene membrane with a pore size of 0.2 μm under operating conditions of -0.09 MPa and 40 °C. (3) After the concentrate is sterilized by ultra-high pressure for 3 minutes, it is packaged. After packaging, the finished product is sterilized by cobalt-60 γ-ray irradiation at ≤10℃ with a dose of 2-5kGy to obtain the finished ointment.

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