Probiotic culture medium and method for preparing probiotic preparation

Abstract

The invention provides a probiotic culture medium and a method for preparing a probiotic preparation, and belongs to the technical field of microbial culture. According to the preparation method, the probiotic culture medium is firstly prepared, the probiotic compound is obtained by utilizing the probiotic culture medium and bifidobacteria, and then the dietary fiber compound, the quercetin compound and the liquorice-rhizoma cyperi compound are used together, so that the probiotic preparation is obtained, and the preparation can effectively improve related symptoms of intestinal tracts, stomachs and respiratory tracts.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial culture, and in particular relates to a probiotic culture medium and a method for preparing a probiotic preparation. Background Art

[0002] Probiotics are a type of living microorganisms that can secrete substances that are beneficial to other microorganisms in a suitable living environment, thereby having a positive effect on the host, so they are called probiotics. Preparations rich in beneficial substances obtained by cultivating one or more probiotics through biological methods or intervening through physical means are called probiotic preparations. Probiotics or probiotic preparations can colonize in the host's body and produce a large amount of beneficial substances through continuous reproduction, such as digestive enzymes, vitamins, short-chain fatty acids, etc. These substances can regulate the composition of bacterial communities in certain parts of the host, enhance immunity and antioxidant capacity, and improve the conversion and absorption capacity of nutrients, thereby having a positive effect on the host.

[0003] A Chinese patent (publication number CN117965368A) discloses a method for preparing a probiotic culture medium and a probiotic preparation, which can improve the problems of a small number of microorganisms and low activity in the probiotic preparations in the soil, thereby increasing the number of microorganisms in the soil and improving the activity of microorganisms; improving the crop yield of saline-alkali soil and the pollution problem of metal cadmium. The probiotic preparation has a significant effect on improving the structure of the soil and has a good effect on increasing land production. However, the main purpose of the patented technology is to improve the soil, and the existing probiotic preparations, when applied to the human body, have no significant help for the intestines, stomach and respiratory tract, and cannot effectively improve related symptoms. Related problems have yet to be solved.

[0004] Therefore, how to obtain probiotic culture medium and probiotic preparations through the ratio of multiple components and Chinese medicine components as important components to effectively improve related symptoms of the intestinal tract, stomach and respiratory tract has become a direction that needs to be focused on. Summary of the invention

[0005] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a probiotic culture medium and a method for preparing a probiotic preparation, aiming to solve the problem that existing probiotic preparations, when applied to the human body, have no significant help to the intestines, stomach and respiratory tract and cannot effectively improve related symptoms.

[0006] The present invention first prepares a probiotic culture medium, and uses the probiotic culture medium and bifidobacteria to obtain a probiotic complex, which is then used in combination with a dietary fiber complex, a quercetin complex and a liquorice-cyperus complex to obtain a probiotic preparation, which can effectively improve related symptoms of the intestinal tract, stomach and respiratory tract.

[0007] The technical solution of the present invention is as follows:

[0008] In one aspect, the present invention provides a probiotic culture medium, wherein the preparation method thereof comprises the following steps:

[0009] In parts by weight, 5 to 8 parts of glucose, 3 to 6 parts of lactose, 8 to 10 parts of skim milk powder, 1 to 2 parts of soy peptone, 0.5 to 0.8 parts of betaine and 0.5 to 1 part of Tween 80 are dissolved in 15 to 20 parts of deionized water, and after the dissolution is completed, a sodium hydroxide solution with a mass concentration of 10% is added dropwise to adjust the pH value to 6.5 to 7.0, so as to prepare a probiotic culture medium;

[0010] The glucose is purified glucose; the preparation method of the purified glucose comprises: adding 100 to 150 parts of glucose by weight to 400 to 500 parts of deionized water to fully dissolve, then adding 7 to 10 parts of nickel composite material and stirring for 1 to 2 hours, adjusting the pH to 6.5 to 6.7, the temperature to 85 to 95°C, the pressure to 8.5 to 9.5 MPa, and performing purification treatment for 2 to 3 hours; after the purification treatment is completed, cooling, sedimentation, filtration, concentration and crystallization are performed in sequence to obtain purified glucose.

[0011] As a preferred technical solution of the present invention, the preparation method of the nickel composite material comprises: adding 10 to 15 parts of graphene oxide to 100 to 120 parts of deionized water by weight, stirring evenly, adding 3 to 5 parts of 10% mass concentration sodium hydroxide solution and 5 to 8 parts of nickel catalyst, and then ball milling to obtain a nickel-graphene oxide mixed solution; freeze-drying the nickel-graphene oxide mixed solution to obtain a nickel composite material.

[0012] Purified glucose is prepared using nickel composite materials. After freeze-drying, the nickel composite material can form a porous structure. The micropores allow small molecule glucose to pass through smoothly, while blocking large molecular impurities outside the micropores, thereby achieving glucose purification.

[0013] Another aspect of the present invention provides a method for preparing a probiotic preparation using the probiotic culture medium, comprising the following steps:

[0014] Step S1: sterilizing and disinfecting 10 parts of the probiotic culture medium in step S1, adding 2 to 4 parts of bifidobacteria for oxygen-isolated constant temperature culture, and then performing vacuum freeze drying to obtain a probiotic complex;

[0015] Step S2: In parts by weight, 40 to 70 parts of the probiotic complex described in step S2, 20 to 30 parts of the dietary fiber complex, 10 to 20 parts of the quercetin complex and 10 to 20 parts of the licorice-cyperus complex are mixed evenly, shielded from light, and allowed to ferment. After the fermentation is completed, 0.5 to 1 part of a thickener and 0.3 to 0.5 part of a preservative are added, and the mixture is mixed evenly to obtain a probiotic preparation.

[0016] Preferably, the constant temperature culture in step S1 is at a temperature of 25 to 30° C. and a time of 24 to 48 hours.

[0017] As a preferred technical solution of the present invention, the preparation method of the dietary fiber complex in step S2 includes: soaking 30 to 50 parts of astragalus granules and 20 to 30 parts of psyllium powder in 100 to 150 parts of vinegar by weight, heating to 70 to 75°C and maintaining the constant temperature for 0.5 to 1 hour, washing with deionized water and draining; then adding 40 to 50 parts of propylene glycol, reflux extraction at 80 to 85°C for 3 to 4 hours, and filtering to obtain an extract; adding 20 to 30 parts of the extract and 15 to 20 parts of water-soluble dietary fiber to 100 to 120 parts of deionized water, stirring and mixing for 2 to 4 hours to obtain a dietary fiber complex liquid; removing impurities from the dietary fiber complex liquid through a de-impurity membrane, and then freeze-drying to obtain a dietary fiber complex.

[0018] Water-soluble dietary fiber can be fermented by beneficial bacteria in the intestine to produce short-chain fatty acids, which can provide energy for intestinal cells and promote intestinal health. In addition, short-chain fatty acids also help maintain the integrity of the intestinal mucosal barrier and reduce the absorption of harmful substances. Astragalus can adjust the proportion of intestinal flora through multiple targets and multiple pathways, significantly increase the abundance of intestinal flora, improve the intestinal microenvironment, and repair the diversity of intestinal bacteria. The active substances in Plantago can form a gel-like substance to help absorb excess water and reduce the fluidity of intestinal contents. Through the synergistic effect of dietary fiber, astragalus, and rhein, intestinal symptoms can be effectively improved.

[0019] As a preferred technical solution of the present invention, the preparation method of the impurity removal membrane includes: adding 10 to 20 parts of polyvinyl alcohol to 100 to 120 parts of deionized water by weight, stirring evenly, adding 2 to 4 parts of mesoporous material, stirring for 1 to 2 hours to obtain a suspension; electrospinning the suspension to obtain an electrospun fiber membrane, drying it at 50 to 60°C for 6 to 8 hours to obtain an impurity removal membrane; the preparation method of the mesoporous material includes: mixing 100 to 120 parts of anhydrous ethanol and 20 to 30 parts of deionized water by weight to obtain a mixed solution, weighing 0.2 to 0.4 parts of ferrosoferric oxide and adding it to the mixed solution, ultrasonically dispersing it for 1 to 2 hours, and then dripping 2 to 4 parts of tetraethyl silicate and 3 to 5 parts of ammonia water into the suspension, reacting for 8 to 10 hours under stirring at room temperature, and then washing it with deionized water 3 times to obtain composite particles, and drying the composite particles in a vacuum oven at 50 to 60°C for 10 to 12 hours to obtain a mesoporous material.

[0020] The impurity removal membrane is composed of an electrospun fiber membrane, using polyvinyl alcohol with good biocompatibility as the main material, and adding mesoporous materials for modification. The silicon dioxide as the surface layer of the mesoporous material has a high specific surface area and good adsorption properties, which can effectively adsorb impurities; and the core layer of the mesoporous material is magnetic ferrosoferric oxide, which can effectively adsorb trace elements contained in the raw materials themselves or introduced during the processing, thereby reducing the impurity content.

[0021] As a preferred technical solution of the present invention, the preparation method of the quercetin complex described in step S2 includes: mixing 10 to 15 parts of quercetin and 2 to 4 parts of zein by weight, stirring at 1200 to 1500 r / min for 0.5 to 1 h, then adding 2 to 4 parts of Panax notoginseng total saponins and 1 to 3 parts of polyethylene glycol, performing ultrasonic treatment, and obtaining the quercetin complex after drying.

[0022] The quercetin in the complex can enhance the defense ability of the gastric mucosa and reduce the gastric mucosal damage caused by alcohol, non-steroidal anti-inflammatory drugs and other substances; while Panax notoginseng total saponins can activate signal pathways, induce apoptosis of gastric mucosal epithelial cells, control inflammatory responses, and delay the malignant progression of gastric mucosal tissue, thereby effectively improving adverse gastric symptoms.

[0023] As a preferred technical solution of the present invention, the preparation method of the licorice-cyperus complex described in step S2 includes: mixing 50 to 60 parts of licorice extract, 10 to 20 parts of cyperus extract and 5 to 10 parts of chitosan by weight, adding 100 to 120 parts of deionized water, adjusting the pH to 7.4 to 7.6, heating to 60 to 65° C. to react for 2 to 4 hours, filtering, and naturally drying and crushing the filter cake to obtain the licorice-cyperus complex.

[0024] The active ingredients of licorice in the licorice-cyperus complex can cover the inflamed throat mucosa, relieve cough by alleviating inflammatory stimulation, and promote bronchial mucosal secretion to cough up phlegm; the cyperus in the complex can regulate the function of airway smooth muscle, has a certain anti-asthmatic effect, and can help relieve bronchospasm. At the same time, the volatile oils and other ingredients contained in cyperus have anti-inflammatory effects, which help to reduce respiratory inflammation and are beneficial to diseases such as chronic bronchitis; through the combined effects of the licorice-cyperus complex, respiratory symptoms are improved.

[0025] Preferably, the temperature of the static fermentation in step S2 is 20-35° C. and the time is 2-4 days.

[0026] Preferably, in step S2, the thickener is xanthan gum, and the preservative is potassium sorbate.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] (1) The present invention uses a dietary fiber complex to improve intestinal symptoms. The water-soluble dietary fiber in the complex can be fermented by beneficial bacteria in the intestine to produce short-chain fatty acids. These short-chain fatty acids can provide energy for intestinal cells and promote intestinal health. In addition, short-chain fatty acids can help maintain the integrity of the intestinal mucosal barrier and reduce the absorption of harmful substances. Astragalus can use multiple targets and multiple pathways to adjust the proportion of intestinal flora, significantly increase the abundance of intestinal flora, improve the intestinal microenvironment, and repair the diversity of intestinal bacteria. The active substances in Plantago can form a gel-like substance to help absorb excess water and reduce the fluidity of intestinal contents.

[0029] (2) The present invention uses a quercetin complex to improve gastric symptoms. The quercetin in the quercetin complex can enhance the defense ability of the gastric mucosa and reduce gastric mucosal damage caused by substances such as alcohol and non-steroidal anti-inflammatory drugs; and Panax notoginseng total saponins can activate signal pathways, induce apoptosis of gastric mucosal epithelial cells, control inflammatory reactions, and delay the malignant progression of gastric mucosal tissue, thereby effectively improving gastric adverse symptoms. The combined effect effectively improves gastric symptoms.

[0030] (3) The present invention uses a licorice-cyperus rotundus complex to improve respiratory symptoms. The effective ingredients of licorice in the complex can cover the inflamed throat mucosa, relieve cough by alleviating inflammatory stimulation, and promote bronchial mucosal secretion to cough up phlegm; the cyperus rotundus in the complex can regulate the function of airway smooth muscle, has a certain anti-asthmatic effect, and can help relieve bronchospasm. At the same time, the volatile oil and other ingredients contained in the cyperus rotundus have anti-inflammatory effects, which help to reduce respiratory inflammation and are beneficial to diseases such as chronic bronchitis. DETAILED DESCRIPTION

[0031] For the convenience of understanding the present invention, the present invention lists the following embodiments. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0032] The sources of some components in the embodiments and comparative examples are as follows:

[0033] Commercially available glucose, catalog number PA30754, was purchased from Shanghai Chuangsai Technology Co., Ltd.;

[0034] Lactose, product number PA38891, was purchased from Shanghai Chuangsai Technology Co., Ltd.;

[0035] Skim milk powder, product number CM0681B, purchased from Thermo Fisher Scientific, USA;

[0036] Soybean peptone, product number 050220, was purchased from Guangdong Huankai Microbiological Technology Co., Ltd.;

[0037] Betaine, product number S18046, was purchased from Shanghai Yuanye Biotechnology Co., Ltd.;

[0038] Tween 80, catalog number MB2675, was purchased from Dalian Meilun Biotechnology Co., Ltd.;

[0039] Sodium hydroxide, CAS No. 1310-73-2, was purchased from Sinopharm Chemical Reagent Co., Ltd.;

[0040] Bifidobacterium, catalog number Bio-81738, was purchased from Beijing Bio-Bowei Biotechnology Co., Ltd.;

[0041] Xanthan gum, product number 174931, purchased from Wilder (Beijing) Flavor Co., Ltd.;

[0042] Potassium sorbate, product number 56567, was purchased from Suzhou Zhuoxin Biotechnology Co., Ltd.;

[0043] Astragalus granules, product number 2111024101, were purchased from Jiangyin Tianjiang Pharmaceutical Co., Ltd.;

[0044] Plantago seed powder, product number 20201125003, was purchased from Shaanxi New Horizon Biotechnology Co., Ltd.;

[0045] Vinegar was purchased from Beijing Ershang Longhe Food Co., Ltd.;

[0046] Water-soluble dietary fiber, product number TB-0181, was purchased from Zhengzhou Taber Trading Co., Ltd.;

[0047] Propylene glycol, CAS No. 57-55-6, food grade, purchased from Shandong Aicai Biotechnology Co., Ltd.;

[0048] Quercetin, catalog number Q817162, was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.;

[0049] Zein, product number Z3625, was purchased from Shanghai Sigma-Aldrich Trading Co., Ltd.;

[0050] Panax notoginseng total saponins, product number SS9450, were purchased from Beijing Solaibao Technology Co., Ltd.;

[0051] Polyethylene glycol, product number 20240201, was purchased from Jiangxi Yipsheng Pharmaceutical Co., Ltd.;

[0052] Licorice extract, product number BH0432, was purchased from Xi'an Best Biotechnology Co., Ltd.;

[0053] Cyperus rotundus extract, product number CY230625, was purchased from Shaanxi Ciyuan Biotechnology Co., Ltd.;

[0054] Chitosan, product number PA20151, was purchased from Perfemiker, USA;

[0055] Graphene oxide, model 102740, was purchased from Jiangsu Xianfeng Nanomaterials Technology;

[0056] Nickel catalyst, model RCZ-80-2, purchased from Jiangsu Rayni Metal Technology Co., Ltd.;

[0057] Polyvinyl alcohol, product number P815725-500g, was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.;

[0058] Anhydrous ethanol, CAS No. 64-17-5, was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.;

[0059] Ferroferric oxide, CAS No. 12227-89-3; purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.;

[0060] Tetraethyl silicate, product number T819505, was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.;

[0061] Ammonia, CAS No. 1336-21-6; purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0062] Preparation of purified glucose: (1) By weight, add 15 parts of graphene oxide to 120 parts of deionized water, stir evenly, add 5 parts of 10% mass concentration sodium hydroxide solution and 8 parts of nickel catalyst, then ball mill and mix to obtain a nickel-graphene oxide mixed solution; freeze-dry the nickel-graphene oxide mixed solution to obtain a nickel composite material. (2) By weight, add 150 parts of commercially available glucose to 500 parts of deionized water and fully dissolve, then add 10 parts of nickel composite material and stir for 2 hours, adjust the pH to 6.7, the temperature to 85°C, the pressure to 9.5MPa, and purify for 3 hours; after the purification treatment is completed, cool down, settle, filter, concentrate and crystallize in sequence to obtain purified glucose.

[0063] Preparation of impurity removal membrane: (1) In parts by weight, 120 parts of anhydrous ethanol and 30 parts of deionized water were mixed to obtain a mixed solution, 0.4 parts of ferrosoferric oxide was weighed and added to the mixed solution, ultrasonically dispersed for 2 hours, and then 4 parts of tetraethyl silicate and 5 parts of ammonia water were dropped into the suspension, stirred at room temperature for 10 hours, and then washed with deionized water 3 times to obtain composite particles, and the composite particles were dried in a vacuum oven at 60°C for 12 hours to obtain a mesoporous material. (2) In parts by weight, 20 parts of polyvinyl alcohol were added to 120 parts of deionized water, stirred evenly, and then 4 parts of mesoporous material were added, stirred for 2 hours to obtain a suspension; the suspension was electrospun to obtain an electrospun fiber membrane, and dried at 60°C for 8 hours to obtain an impurity removal membrane.

[0064] Preparation of dietary fiber complex: by weight, 50 parts of astragalus granules and 30 parts of psyllium powder are soaked in 150 parts of vinegar, heated to 75°C and kept at a constant temperature for 1 hour, then washed with deionized water and drained; then 50 parts of propylene glycol are added, refluxed and extracted at 85°C for 4 hours, and an extract is obtained by filtration; 30 parts of the extract and 20 parts of water-soluble dietary fiber are added to 120 parts of deionized water, and stirred and mixed for 4 hours to obtain a dietary fiber complex liquid; the dietary fiber complex liquid is subjected to impurity removal treatment through an impurity removal membrane, and then freeze-dried to obtain a dietary fiber complex.

[0065] Preparation of quercetin complex: 15 parts of quercetin and 4 parts of zein were mixed by weight, stirred at 1500 r / min for 1 hour, then 4 parts of notoginseng total saponins and 3 parts of polyethylene glycol were added, ultrasonically treated, and dried to obtain the quercetin complex.

[0066] Preparation of licorice-cyperus complex: In parts by weight, 60 parts of licorice extract, 20 parts of cyperus extract and 10 parts of chitin are mixed, 120 parts of deionized water are added, the pH is adjusted to 7.6, and the mixture is heated to 65°C for 2 hours, filtered, and the filter cake is naturally dried and crushed to obtain a licorice-cyperus complex.

[0067] Example 1

[0068] This embodiment provides a probiotic culture medium and a method for preparing a probiotic preparation, comprising the following steps:

[0069] Step (1): dissolving 8 parts of purified glucose, 6 parts of lactose, 10 parts of skim milk powder, 2 parts of soy peptone, 0.8 parts of betaine and 1 part of Tween 80 in 20 parts of deionized water by weight, and after the dissolution is completed, adding a 10% sodium hydroxide solution by weight concentration to adjust the pH value to 7.0, thereby preparing a probiotic culture medium;

[0070] Step (2): sterilize and disinfect 10 parts of the probiotic culture medium in step (1) by weight, then add 4 parts of bifidobacteria for oxygen-free constant temperature culture (temperature is 30° C., time is 24 h), and then vacuum freeze-dry to obtain a probiotic complex;

[0071] Step (3): In parts by weight, 70 parts of the probiotic complex described in step (2), 30 parts of the dietary fiber complex, 20 parts of the quercetin complex and 20 parts of the licorice-cyperus complex are mixed evenly, shielded from light, and allowed to ferment (temperature is 35° C., time is 2 days). After the fermentation is completed, 1 part of xanthan gum and 0.5 part of potassium sorbate are added, and the mixture is mixed to obtain a probiotic preparation.

[0072] Example 2

[0073] This embodiment provides a probiotic culture medium and a method for preparing a probiotic preparation, comprising the following steps:

[0074] Step (1): dissolving 5 parts of purified glucose, 3 parts of lactose, 8 parts of skim milk powder, 1 part of soy peptone, 0.5 parts of betaine and 0.5 parts of Tween 80 in 15 parts of deionized water by weight, and after the dissolution is completed, adding a 10% sodium hydroxide solution dropwise to adjust the pH value to 6.5, thereby preparing a probiotic culture medium;

[0075] Step (2): sterilize and disinfect 10 parts of the probiotic culture medium in step (1) by weight, then add 2 parts of bifidobacteria for oxygen-free constant temperature culture (temperature is 25° C., time is 48 hours), and then perform vacuum freeze drying to obtain a probiotic complex;

[0076] Step (3): In parts by weight, 40 parts of the probiotic complex described in step (2), 20 parts of the dietary fiber complex, 10 parts of the quercetin complex and 10 parts of the licorice-cyperus complex are mixed evenly, shielded from light, and allowed to ferment (temperature is 20° C., time is 4 days). After the fermentation is completed, 0.5 parts of xanthan gum and 0.3 parts of potassium sorbate are added, and the mixture is mixed to obtain a probiotic preparation.

[0077] Example 3

[0078] This embodiment provides a probiotic culture medium and a method for preparing a probiotic preparation, comprising the following steps:

[0079] Step (1): dissolving 6 parts of purified glucose, 4 parts of lactose, 9 parts of skim milk powder, 2 parts of soy peptone, 0.6 parts of betaine and 0.8 parts of Tween 80 in 18 parts of deionized water by weight, and after the dissolution is completed, adding a 10% sodium hydroxide solution dropwise to adjust the pH value to 6.8, thereby preparing a probiotic culture medium;

[0080] Step (2): sterilize and disinfect 10 parts of the probiotic culture medium in step (1) by weight, then add 3 parts of bifidobacteria for oxygen-free constant temperature culture (temperature is 28° C., time is 36 hours), and then vacuum freeze-dry to obtain a probiotic complex;

[0081] Step (3): In parts by weight, 60 parts of the probiotic complex described in step (2), 25 parts of the dietary fiber complex, 15 parts of the quercetin complex and 15 parts of the licorice-cyperus complex are mixed evenly, shielded from light, and allowed to ferment (temperature is 28° C., time is 3 days). After the fermentation is completed, 0.8 parts of xanthan gum and 0.4 parts of potassium sorbate are added, and the mixture is mixed to obtain a probiotic preparation.

[0082] Comparative Example 1

[0083] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that no dietary fiber complex is added in step (3), and other conditions are the same as those in Example 1.

[0084] Comparative Example 2

[0085] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that no quercetin complex is added in step (3), and other conditions are the same as those in Example 1.

[0086] Comparative Example 3

[0087] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that no licorice-cyperus complex is added in step (3), and other conditions are the same as those in Example 1.

[0088] Comparative Example 4

[0089] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that no quercetin complex and licorice-cyperus complex are added in step (3), and other conditions are the same as those in Example 1.

[0090] Comparative Example 5

[0091] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which is different from Example 1 in that no dietary fiber complex and licorice-cyperus complex are added in step (3), and other conditions are the same as those in Example 1.

[0092] Comparative Example 6

[0093] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that dietary fiber complex and quercetin complex are not added in step (3), and other conditions are the same as those in Example 1.

[0094] Comparative Example 7

[0095] This comparative example provides a probiotic culture medium and a method for preparing a probiotic preparation, which differs from Example 1 in that commercially available glucose is used instead of purified glucose in step (1), and other conditions are the same as those in Example 1.

[0096] The performance of the probiotic preparations prepared by the methods provided in the above embodiments and comparative examples was tested, and the testing method was as follows:

[0097] The subjects of this experiment were selected from office workers who often had high work pressure and abnormal diet, mainly those who were prone to colds, diarrhea, low immunity and frequent gastrointestinal discomfort; the subjects were randomly divided into groups, with 100 people in each group, and all male and female subjects were between 20 and 45 years old, and took 2g of the probiotic preparation prepared in the embodiment and the comparative example respectively; the subjects in different groups took the probiotic preparation half an hour before breakfast and dinner every day, and each group of subjects took it continuously for 1 month, and the subjects did not change their daily life and eating habits during the test period.

[0098] This experiment uses the subject's own control method to evaluate the effect. The subjects subjectively evaluate the changes in their own intestinal, gastric and respiratory symptoms before and after the test. The evaluation method is that if the probiotic preparation of the present invention has obvious improvement compared with before taking it and it lasts for more than 3 days (including 3 days), it is recorded as effective; if it is more than 5 days, it is recorded as significantly effective; if it is less than 3 days, it is recorded as ineffective. The total effective rate is the percentage of the effective number and the significantly effective number of people in the total number of people.

[0099] The above performance test data is shown in Table 1.

[0100] Table 1 Performance test results

[0101]

[0102] From the above content, it can be seen that the present invention first prepares a probiotic culture medium, and uses the probiotic culture medium and bifidobacteria to obtain a probiotic complex, and then uses a dietary fiber complex, a quercetin complex and a licorice-cyperus complex together to obtain a probiotic preparation (Examples 1 to 3), which has a total effective rate of 90 to 92% in improving intestinal symptoms, a total effective rate of 87 to 89% in improving gastric symptoms, and a total effective rate of 86 to 88% in improving respiratory symptoms.

[0103] Compared with Example 1, if the dietary fiber complex is not added, the total effective rate of improving intestinal symptoms decreases (Comparative Example 1); compared with Example 1, if the quercetin complex is not added, the total effective rate of improving gastric symptoms decreases (Comparative Example 2); compared with Example 1, if the licorice-cyperus complex is not added, the total effective rate of improving respiratory symptoms decreases (Comparative Example 3); compared with Example 1, if the quercetin complex and the licorice-cyperus complex are not added, the total effective rate of improving gastric symptoms decreases, and the total effective rate of improving respiratory symptoms decreases (Comparative Example 4); compared with Example 1, Compared with Example 1, if the dietary fiber complex and the liquorice-cyperus complex are not added, the total effective rate of improving intestinal symptoms decreases, and the total effective rate of improving respiratory symptoms decreases (Comparative Example 5); compared with Example 1, if the dietary fiber complex and the quercetin complex are not added, the total effective rate of improving intestinal symptoms decreases, and the total effective rate of improving gastric symptoms decreases (Comparative Example 6); compared with Example 1, if commercially available glucose is used instead of purified glucose, the total effective rate of improving intestinal symptoms decreases, the total effective rate of improving gastric symptoms decreases, and the total effective rate of improving respiratory symptoms decreases (Comparative Example 7).

[0104] In summary, the present invention prepares a probiotic culture medium first, obtains a probiotic complex using the probiotic culture medium and bifidobacteria, and then uses the probiotic complex, the quercetin complex and the liquorice-cyperus complex together to obtain a probiotic preparation, which can effectively improve the related symptoms of the intestines, stomach and respiratory tract.

[0105] The applicant declares that the present invention illustrates the detailed process flow of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned detailed process flow, that is, it does not mean that the present invention must rely on the above-mentioned detailed process flow to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A probiotic culture medium, characterized in that The preparation method comprises the following steps: In parts by weight, 5 to 8 parts of glucose, 3 to 6 parts of lactose, 8 to 10 parts of skim milk powder, 1 to 2 parts of soy peptone, 0.5 to 0.8 parts of betaine and 0.5 to 1 part of Tween 80 are dissolved in 15 to 20 parts of deionized water, and after the dissolution is completed, a sodium hydroxide solution with a mass concentration of 10% is added dropwise to adjust the pH value to 6.5 to 7.0, so as to prepare a probiotic culture medium; The glucose is purified glucose; the preparation method of the purified glucose comprises: adding 100 to 150 parts of glucose by weight to 400 to 500 parts of deionized water to fully dissolve, then adding 7 to 10 parts of nickel composite material and stirring for 1 to 2 hours, adjusting the pH to 6.5 to 6.7, the temperature to 85 to 95°C, the pressure to 8.5 to 9.5 MPa, and performing purification treatment for 2 to 3 hours; after the purification treatment is completed, cooling, sedimentation, filtration, concentration and crystallization are performed in sequence to obtain purified glucose.

2. The probiotic culture medium according to claim 1, characterized in that The preparation method of the nickel composite material comprises: adding 10 to 15 parts of graphene oxide to 100 to 120 parts of deionized water by weight, stirring evenly, adding 3 to 5 parts of 10% mass concentration sodium hydroxide solution and 5 to 8 parts of nickel catalyst, and then ball milling and mixing to obtain a nickel-graphene oxide mixed solution; and freeze-drying the nickel-graphene oxide mixed solution to obtain the nickel composite material.

3. A method for preparing a probiotic preparation using the probiotic culture medium according to claim 1, characterized in that: The following steps are involved: Step S1: sterilizing and disinfecting 10 parts of the probiotic culture medium in step S1, adding 2 to 4 parts of bifidobacteria for oxygen-isolated constant temperature culture, and then performing vacuum freeze drying to obtain a probiotic complex; Step S2: In parts by weight, 40 to 70 parts of the probiotic complex described in step S2, 20 to 30 parts of the dietary fiber complex, 10 to 20 parts of the quercetin complex and 10 to 20 parts of the licorice-cyperus complex are mixed evenly, shielded from light, and allowed to ferment. After the fermentation is completed, 0.5 to 1 part of a thickener and 0.3 to 0.5 part of a preservative are added, and the mixture is mixed evenly to obtain a probiotic preparation.

4. The method for preparing a probiotic preparation according to claim 3, characterized in that: The constant temperature culture in step S1 is 25-30° C. and the time is 24-48 hours.

5. The method for preparing a probiotic preparation according to claim 3, characterized in that: The preparation method of the dietary fiber complex in step S2 comprises: soaking 30 to 50 parts of astragalus granules and 20 to 30 parts of psyllium powder in 100 to 150 parts of vinegar by weight, heating to 70 to 75° C. and maintaining the constant temperature for 0.5 to 1 hour, washing with deionized water and draining; then adding 40 to 50 parts of propylene glycol, refluxing and extracting at 80 to 85° C. for 3 to 4 hours, and filtering to obtain an extract; adding 20 to 30 parts of the extract and 15 to 20 parts of water-soluble dietary fiber to 100 to 120 parts of deionized water, stirring and mixing for 2 to 4 hours to obtain a dietary fiber complex liquid; removing impurities from the dietary fiber complex liquid through a de-impurity membrane, and then freeze-drying to obtain a dietary fiber complex.

6. The method for preparing a probiotic preparation according to claim 5, characterized in that: The preparation method of the impurity removal membrane comprises: adding 10 to 20 parts of polyvinyl alcohol to 100 to 120 parts of deionized water by weight, stirring evenly, adding 2 to 4 parts of mesoporous material, stirring for 1 to 2 hours to obtain a suspension; electrostatically spinning the suspension to obtain an electrostatically spun fiber membrane, and drying at 50 to 60° C. for 6 to 8 hours to obtain an impurity removal membrane; The preparation method of the mesoporous material comprises: in parts by weight, mixing 100 to 120 parts of anhydrous ethanol and 20 to 30 parts of deionized water to obtain a mixed solution, weighing 0.2 to 0.4 parts of ferrosoferric oxide and adding it to the mixed solution, ultrasonically dispersing for 1 to 2 hours, then dropping 2 to 4 parts of tetraethyl silicate and 3 to 5 parts of ammonia water into the suspension, reacting for 8 to 10 hours under stirring at room temperature, then washing with deionized water for 3 times to obtain composite particles, and drying the composite particles in a vacuum oven at 50 to 60° C. for 10 to 12 hours to obtain the mesoporous material.

7. The method for preparing a probiotic preparation according to claim 3, characterized in that: The preparation method of the quercetin complex described in step S2 includes: mixing 10 to 15 parts of quercetin and 2 to 4 parts of zein by weight, stirring at 1200 to 1500 r / min for 0.5 to 1 hour, then adding 2 to 4 parts of Panax notoginseng total saponins and 1 to 3 parts of polyethylene glycol, performing ultrasonic treatment, and obtaining the quercetin complex after drying.

8. The method for preparing a probiotic preparation according to claim 3, characterized in that: The preparation method of the licorice-cyperus complex described in step S2 includes: mixing 50 to 60 parts of licorice extract, 10 to 20 parts of cyperus extract and 5 to 10 parts of chitosan by weight, adding 100 to 120 parts of deionized water, adjusting the pH to 7.4 to 7.6, heating to 60 to 65° C. to react for 2 to 4 hours, filtering, and naturally drying and crushing the filter cake to obtain the licorice-cyperus complex.

9. The method for preparing a probiotic preparation according to claim 3, characterized in that: The temperature of the static fermentation in step S2 is 20-35° C. and the time is 2-4 days.

10. The method for preparing a probiotic preparation according to claim 3, characterized in that: In step S2, the thickener is xanthan gum, and the preservative is potassium sorbate.

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