CN114504623A - Traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation and preparation method thereof - Google Patents
Traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation, which is at least formed by compounding traditional Chinese medicine leavening and composite probiotics; the traditional Chinese medicine fermented product is prepared by fermenting traditional Chinese medicine extracting solution obtained by extracting a traditional Chinese medicine formula by saccharomycetes, wherein the traditional Chinese medicine formula comprises 10-30 parts of astragalus membranaceus, 10-30 parts of cistanche, 10-25 parts of semen cassiae, 10-25 parts of fructus cannabis, 10-25 parts of rhizoma polygonati, 5-20 parts of dark plum and 5-20 parts of fingered citron; the composite probiotics comprise bifidobacterium animalis bifidobacterium lactis subspecies, lactobacillus paracasei and pediococcus pentosaceus. The food is suitable for long-term use, can safely and stably solve the problem of intestinal health with definite curative effect and regulate the intestinal tract, is particularly suitable for improving qi deficiency type constipation of middle-aged and elderly people, has good market prospect, is convenient to prepare, is easy to operate in quality control, and is beneficial to realizing industrial production.
Description
Technical Field
The invention relates to the technical field of intestinal health, in particular to a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation and a preparation method thereof.
Background
Constipation is a common clinical digestive system disease, is usually chronic, namely, defecate is obstructed, the time of defecating is prolonged, one time is carried out for 2 to 3 days, one time is carried out for 4 to 5 days, and in severe cases, one defecate can be released for a week or even a longer time. According to survey, the incidence rate of constipation in people is 3-4%, and the elderly can reach more than 3.
Constipation is recorded in the literature of traditional Chinese medicine, for example, the "defecate difficulty" in the "internal classic", and the therapeutic principle of "five cereals nourishing, five fruits assisting, five animals benefiting and five vegetables supplementing" is mentioned. The syndrome differentiation from Shanghai treatise on Cold-induced diseases is manifested as yang nodulation, yin nodulation or spleen constraint, there are Canren Wan in Jing Fang to nourish yin and moisten the lower energizer, and Mi Jiang dao Fang (suppository). Constipation is called constipation as "stool failure" in the treatise on the source of various diseases, and constipation is called stuffiness in the body from the formula of Qian jin Yao Fang. The traditional Chinese medicine considers that the basic pathogenesis of constipation is that pathogenic factors stagnate in the large intestine, the qi of the fu organs is blocked or the intestine is not warm and moist, the promotion is weak, the conduction function of the large intestine is abnormal, and the constipation of the middle-aged and the elderly people, especially the chronic constipation with slow disease course, is the main cause.
Traditional Chinese medicine cognition for normal stool: the normal defecation of people is once a day, the defecation is early (5-7 points in the morning, the Yangming large intestine meridian is right), the defecation is thick, long and yellow, the defecation is not sticky and greasy, and no special odor is generated, so the normal defecation is normal. Thus, more than one stool or less than one stool per day is not normal; the stool time is not correct and abnormal; the stool has different colors, is green/black and the like, is sticky and greasy, is sticky to the closestool and is abnormal particularly when smelly; the powder is scattered when being touched with water, and more undigested food residues are not normal; the feces are dry and egg-shaped, which is not normal. Constipation is dry stool with egg-shaped stool (qi and yin deficiency), or green and black greasy stool (spleen and stomach deficiency with severe damp qi) or other constipation, but most of them are accompanied by qi deficiency, while constipation of middle-aged and elderly people is accompanied by qi deficiency with excessive yin deficiency.
At present, the problem of constipation is mainly solved by drug therapy, which can be divided into purgative, intestinal peristalsis promoting, secretion promoting and enema. The medicines have quick response but side effects, have dependence after long-term administration, cause more serious constipation after no administration, possibly cause intestinal function reduction after long-term administration, and are not suitable for long-term administration. Therefore, the development of a product which is particularly suitable for middle-aged and elderly people and mainly aims at improving qi deficiency type constipation, particularly a safe and effective food product, is suitable for long-term, safe and effective use, is very necessary, and has very good market prospect.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a traditional Chinese medicine probiotic compound product for improving qi-deficiency constipation, which is a food product suitable for long-term use, can safely and stably solve the problem of intestinal health with definite curative effect, regulates the intestinal tract, is particularly suitable for improving qi-deficiency constipation of middle-aged and elderly people, and has good market prospect.
The second purpose of the invention is to provide a preparation method of the traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation, which has the advantages of convenient flow, easy quality control and operation and contribution to realizing industrial production.
One of the purposes of the invention is realized by adopting the following technical scheme:
the traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is at least compounded by traditional Chinese medicine leavening and compound probiotics; the traditional Chinese medicine fermented product is prepared by fermenting traditional Chinese medicine extracting solution obtained by extracting a traditional Chinese medicine formula by saccharomycetes, wherein the traditional Chinese medicine formula comprises 10-30 parts of astragalus membranaceus, 10-30 parts of cistanche, 10-25 parts of semen cassiae, 10-25 parts of fructus cannabis, 10-25 parts of rhizoma polygonati, 5-20 parts of dark plum and 5-20 parts of fingered citron; the composite probiotics comprise bifidobacterium animalis bifidobacterium lactis subspecies bifidus, lactobacillus paracasei and pediococcus pentosaceus.
The medicinal materials in the traditional Chinese medicine formula provided by the invention can be used for food (part of the medicinal materials are used in test points) in latest food related regulations. The middle-aged and old people, especially the elderly, have the most obvious phenomena of unsmooth defecation and constipation due to the decline of the body functions of the old people, the decline of the digestive capacity of the gastrointestinal tract and the weakened intestinal peristalsis after the deficiency of yang qi. From the traditional Chinese medicine, body fluid is insufficient to cause dry stool, and qi deficiency cannot push the stool to creep forward and discharge the stool out of the body. Therefore, treating constipation of middle-aged and elderly people, regulating intestinal tract, invigorating qi, producing body fluid, invigorating spleen and dredging intestinal tract are all considered at the same time. In the traditional Chinese medicine formula, the astragalus root tonifies qi and strengthens the exterior, the cistanche deserticola tonifies kidney yang, replenishes essence and blood, and loosens the bowel to relieve constipation, the astragalus root tonifies acquired spleen qi and lung qi, the cistanche deserticola tonifies yang qi of the acquired kidney, and loosens the bowel to relieve constipation, the first and the latter tonics are simultaneously performed, so that the yang qi is generated, the spleen and the stomach are dissolved, the lung qi is fully gained, and the cistanche deserticola falls in the large intestine to push the excrement to be discharged out of the body; rhizoma Polygonati has effects of invigorating qi, nourishing yin, invigorating spleen, moistening lung, invigorating kidney, nourishing yin, promoting fluid production, resolving constipation problem, and simultaneously assisting radix astragali in invigorating spleen and lung qi and herba cistanches in invigorating kidney qi; semen cassiae has the effects of clearing heat and improving eyesight and relaxing bowel, fructus cannabis has the effect of relaxing bowel, and is a purgative medicament which plays a role of purgation and is just suitable for middle-aged and elderly people (the old people with deficiency of vital energy and physical weakness cannot bear the medicament when rheum officinale and folium sennae are completed); the smoked plums astringe lung and promote fluid production, so that body fluids distributed by lung qi can win the intestinal tract, the problem of constipation of the intestinal tract can be solved, the large intestine is facilitated (the small intestine is obstructed in stool and the qi is going to die, ten smoked plums are placed in soup, the small intestine is denucleated, the pestle is a pill such as jujube, the small intestine is accommodated in the lower part, and the small intestine is communicated), the fingered citron can reduce the qi of the lung and the stomach, the gastrointestinal tract power can be increased, the excrement discharge can be facilitated, and the problem of the fullness and pain of the spleen and the stomach caused by constipation can be solved. The medicines are combined, so that the effects of tonifying qi and spleen, tonifying lung and producing body fluid and relaxing bowel are achieved, the intestinal health of middle-aged and old people can be commonly conditioned, and dendrobium can be used for treating constipation.
Further, the yeast is Kluyveromyces marxianus SMN-S7-LBK with the preservation number of CCTCC M2017643; the yeast is used for partial fermentation of traditional Chinese medicines, and the Kluyveromyces marxianus SMN-S7-LBK is separated from self-made fermented yogurt (mare milk fermentation) of Xinjiang shepherd and belongs to aroma-producing yeast. Research shows that after the yoghourt is fermented by saccharomycetes, the content of cysteine Cys and tryptophan in whey is increased, and the yoghourt is beneficial to reducing blood fat; lactase produced by Kluyveromyces lactis during fermentation is beneficial to relieving lactose intolerance. In the invention, the saccharomycetes also have the functions of helping digestion, promoting metabolism, enhancing nutrition, supplementing B vitamins, and also contain rich dietary fibers, thereby being beneficial to improving constipation, and the saccharomycetes and the metabolites thereof also have the functions of improving the flavor of the traditional Chinese medicine in the formula and covering the bitter taste and the bad taste of the traditional Chinese medicine.
In the research process, the invention discovers that after the traditional Chinese medicine is fermented by Kluyveromyces marxianus SMN-S7-LBK, (1) bitter and astringent tastes of the traditional Chinese medicine are reduced, the bitter taste of the traditional Chinese medicine is reduced under the fermentation action of yeast, and after prebiotics are added, the sweet taste is increased, and the taste and the astringent taste are covered; (2) the bad smell of the traditional Chinese medicine is improved, the Kluyveromyces marxianus SMN-S7-LBK belongs to aroma-producing yeast, and a special mellow smell is generated after the fermentation, so that the smell of the traditional Chinese medicine is more easily accepted; (3) the polysaccharide component in the traditional Chinese medicine components in the formula is increased after fermentation.
Further, the compound probiotics comprise bifidobacterium animalis bifidobacterium lactis subspecies Y6, lactobacillus paracasei K9 and pediococcus pentosaceus LN-PT16, and are named as 'compound probiotics Y30'. The composite probiotics are products of vitamin benefiting technology limited company, are embedded technically and have good stability when stored at normal temperature (non-water environment). The bifidobacterium animalis bifidobacterium lactis subspecies is bifidobacterium animalis subspecies Y6 with the preservation number of CGMCC No. 15026; the lactobacillus paracasei is lactobacillus paracasei K9 with the preservation number of CGMCC No. 15025; the Pediococcus pentosaceus is Pediococcus pentosaceus LN-PT16 with the preservation number of GDMCC NO. 62044.
Bifidobacterium animalis bifidobacterium lactis subspecies Y6: the composition has high acid resistance, maintains the intestinal mucosa barrier, reduces inflammation and effectively takes care of intestinal health; acid resistance, bile salt resistance and promotion of forward balance of intestinal microecology.
Lactobacillus paracasei K9: acid resistance, cholate resistance and high colonization ability, and promotes the forward balance of intestinal microecology; high yield of lactic acid, repair of intestinal barrier and maintain intestinal wall integrity.
Pediococcus pentosaceus LN-PT 16: acid resistance and bile salt resistance, excellent intestinal tract colonization ability, high extracellular polysaccharide yield, excellent oxidation resistance and strong pathogenic bacteria inhibition ability.
Further, the addition amount of the composite probiotic Y30 is 100-300 hundred million cfu. The composite probiotics comprise the following components in percentage by weight: lactobacillus paracasei: pediococcus pentosaceus = 25-45: 25-45: 20-40, preferably, the ratio of the components is = 30-40: 30-40: 25-35.
Further, the traditional Chinese medicine fermentation product-probiotic compound product also comprises one or any combination of prebiotics, dietary fibers and auxiliary materials; the prebiotics are one or any combination of fructo-oligosaccharide, isomaltooligosaccharide, galacto-oligosaccharide and stachyose; the dietary fiber is one or any combination of psyllium seed husk, chia seed and polydextrose; the auxiliary material is maltodextrin and/or silicon dioxide.
Constipation patients often have intestinal flora changes, such that the number of harmful bacteria such as enterobacteria, enterococci and clostridium in intestinal tracts is increased, and the number of probiotics such as lactic acid bacteria, bifidobacteria and bacteroides is reduced. The composite product provided by the invention is supplemented with suitable probiotics, and matched with prebiotics such as oligosaccharide and the like and dietary fiber, so that the distribution of intestinal flora can be improved, the abundance of intestinal probiotics is improved, and the composite product is helpful for improving the intestinal constipation condition. The prebiotics such as fructo-oligosaccharide, stachyose and the like have sweet taste, and can cover the bitter taste and bad taste of the traditional Chinese medicine; the dietary fiber (prebiotics is also one of the dietary fibers) absorbs water, swells and maintains water, increases the volume of excrement, stimulates intestinal tract peristalsis, accelerates defecation frequency, and has the effects of relaxing bowel and preventing constipation.
The traditional Chinese medicine formula, the prebiotics, the dietary fibers and the probiotics of the invention act together to play a role in conditioning the gastrointestinal tract, so that the constipation gastrointestinal tract is unobstructed, the gastrointestinal tract environment is improved, and the recovery of body functions is promoted.
Further, the traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is formed by compounding traditional Chinese medicine leavening, compound probiotics, prebiotics, dietary fibers and auxiliary materials; the traditional Chinese medicine fermented product is prepared by fermenting traditional Chinese medicine extracting solution obtained by extracting a traditional Chinese medicine formula by saccharomycetes, wherein the traditional Chinese medicine formula comprises 15-25 parts of astragalus membranaceus, 15-25 parts of cistanche, 10-20 parts of semen cassiae, 10-20 parts of fructus cannabis, 10-20 parts of rhizoma polygonati, 5-15 parts of dark plum and 5-15 parts of fingered citron; the addition amount of the composite probiotics is 100-300 hundred million cfu, and the composite probiotics comprise the following components in parts by weight: 30-40: 25-35 of Bifidobacterium animalis, Bifidobacterium lactis, and Pediococcus pentosaceus; the prebiotics comprise 0-5 parts of fructo-oligosaccharide, 0-15 parts of isomaltooligosaccharide, 0-5 parts of galacto-oligosaccharide and 0-2 parts of stachyose; the dietary fiber comprises 0-3 parts of psyllium husk, 0-3 parts of chia seed and 0-3 parts of polydextrose; the auxiliary materials comprise 5-10 parts of maltodextrin and 0.05-0.2 part of silicon dioxide.
The second purpose of the invention is realized by adopting the following technical scheme:
a preparation method of a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation comprises the following steps:
the traditional Chinese medicine extraction step: extracting traditional Chinese medicinal materials in a traditional Chinese medicine formula to obtain a traditional Chinese medicine extracting solution;
a sterilization step: adjusting pH of the Chinese medicinal extractive solution to 6.0-6.5, sterilizing, and transferring sterilized solution to cleaned and sterilized fermentation tank;
a fermentation step: adding the yeast live bacteria powder (after activation) into the fermentation tank, fermenting, and finishing fermentation (the pH of the fermentation liquid is 4.0-4.5, and the viable bacteria density is more than 1 multiplied by 106cfu/ml) to obtain a traditional Chinese medicine fermentation extract;
and (3) filtering: filtering the traditional Chinese medicine fermentation extract to obtain traditional Chinese medicine fermentation liquor;
a concentration and drying step: concentrating and drying the traditional Chinese medicine fermentation liquor to obtain dry paste powder;
mixing: and mixing the dry paste powder with composite probiotics, and if the dry paste powder also comprises prebiotics, dietary fibers and auxiliary materials, uniformly mixing the prebiotics, the dietary fibers and the auxiliary materials to obtain a powder product.
Further, in the traditional Chinese medicine extraction step, the traditional Chinese medicines are firstly crushed, then water is added for extraction, the crushing mesh number of the traditional Chinese medicines is controlled to be 8-24 meshes, and the total multiple of the added water is 7-15 times of the weight of the traditional Chinese medicines (more preferably, the total multiple of the added water is 10-15 times of the weight of the traditional Chinese medicines, if the total multiple of the added water is more than the weight of the traditional Chinese medicines, the water is appropriately concentrated and adjusted to be 10-15 times); during extraction, the traditional Chinese medicinal materials are extracted by a heating reflux extraction method, an ultrasonic extraction method or an ultrahigh pressure extraction method; when a heating reflux extraction method is adopted, the extraction time is 1-2 hours, and the extraction times are 2-3 times; when an ultrasonic extraction method is adopted, the extraction time is 0.5-1 hour, and the extraction conditions are as follows: temperature 25-45 ℃, ultrasonic power: stopping the ultrasonic treatment for 1-3 seconds at the speed of 1100-1300W/single ultrasonic head for 2-5 seconds; when the ultrahigh pressure extraction method is adopted, the pressure is 100-600MPa, the pressure is maintained for 1-6 minutes, the temperature is 25-60 ℃, and the cycle period is 8-12 times; after the traditional Chinese medicine extracting solution is obtained, adding sucrose (used for yeast fermentation and providing nutrients), wherein the adding amount of the sucrose is 1.5-2.5% of the weight of the traditional Chinese medicine extracting solution.
Further, in the fermentation step, the adding amount of the live yeast powder during fermentation is 0.025-0.04% of the weight of the traditional Chinese medicine extracting solution, the yeast is activated before being added into the fermentation tank, the dry yeast powder is added into 1-2.5% of cane sugar water in an amount which is 8-12 times of the weight of the yeast powder, the mixture is uniformly mixed, the mixture is added into the fermentation tank after being activated for 1.2-1.8 hours, and the mixture is shaken every 8-12 minutes during the activation process.
Further, in the fermentation step, the fermentation process comprises the following operations:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation;
secondly, adjusting the oxygen introduction in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing oxygen in a proper amount, and introducing clean air for 1.5-2.5 hours every day (yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield, the main purpose of the process is to produce the yeast, and alcohol does not need to be produced);
thirdly, index control, sampling for 1 time every 8 hours after fermenting for 24 hours, detecting the pH value, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5;
fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth speed reaching 1 multiplied by 10 according to normal fermentation6More than cfu/ml; and fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
Further, in the filtering step, coarse powder of the medicinal materials is firstly removed by a 100-mesh filter screen, and then the coarse powder is filtered by a 400-mesh filter cloth to obtain the Chinese medicinal fermentation liquor. The filtering method can adopt plate-and-frame filter pressing, flat centrifuge centrifugal filtration and other methods.
Further, in the concentration and drying step, a concentration method is a reduced pressure concentration method; the drying method is spray drying method or one-step drying granulation method.
Further, the method also comprises a granulating or tabletting step, wherein the powder preparation product obtained in the mixing step is pressed into granules to obtain a granular product, or is further pressed into other non-water type preparations, such as tablets and the like.
Compared with the prior art, the invention has the beneficial effects that:
(1) the traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation provided by the invention is a food product suitable for long-term use, can safely and stably solve the problem of intestinal health with a definite curative effect and regulate intestinal tracts, is particularly suitable for improving qi deficiency type constipation of middle-aged and elderly people, and has a very good market prospect. The traditional Chinese medicine formula is fermented by saccharomycetes after being extracted, and then forms a compound with probiotics, and all components in the formula act together to condition the gastrointestinal tract, so that the constipation gastrointestinal tract is unobstructed, the gastrointestinal tract environment is improved, and the recovery of body functions is promoted.
(2) The preparation method of the traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation provided by the invention is convenient in process, easy to operate in quality control and beneficial to realization of industrial production.
Biological material preservation information:
the strain provided by the invention is Kluyveromyces marxianus SMN-S7-LBK with the preservation number of CCTCC NO: m2017643, classified and named Kluyveromyces marxianus SMN-S7-LBK, which has been preserved in the China center for type culture Collection (address: Wuhan university, Wuhan, Wuchang Lodoya mountain postal code: 430072) in 2017, 10 months and 30 days, the preservation unit is abbreviated as CCTCC.
Bifidobacterium animalis Bifidobacterium lactis subspecies Y6 with the preservation number of CGMCC NO.15026, and is classified and named as: bifidobacterium animalis subsp lactis, which has been deposited in the general microbiological center of China Committee for culture Collection of microorganisms (address: Ministry No. 3 of West Lu 1 of Beijing, Inward area of the republic of China, microbiological research institute of Chinese academy of sciences, postal code: 100101) 12.7.12.7.2017.A short name of the unit for deposition is CGMCC;
the lactobacillus paracasei K9 has a preservation number of CGMCC No.15025 and is classified and named as: lactobacillus paracasei, which has been deposited in the China general microbiological culture Collection center (address: No. 3, institute of microbiology, China academy of sciences, and postal code: 100101) of China Committee for culture Collection of microorganisms in 12.01.12.2017.A., Lactobacillus paracasei, which has been deposited in CGMCC (China general microbiological culture Collection center) (address: No. 3, Ministry of sciences, China, Ministry of Japan);
pediococcus pentosaceus LN-PT16 with the collection number GDMCC NO.62044, and classified and named as: pediococcus pentosaceus LN-PT16, which was deposited at 08.11.2021 at Guangdong microbial cultures Collection center (address: Shiu 5, Middleyao 100, Md., Guangdong province scientific microbiology research institute, postal code: 510070), was abbreviated as GDMCC.
Detailed Description
The present invention is further described below with reference to specific embodiments, and it should be noted that, without conflict, any combination between the embodiments or technical features described below may form a new embodiment.
Experiment system one: screening of probiotic bacteria
When the traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation is researched and developed, the screening of probiotics is very critical, the properties and the effects of the probiotics need to be considered, and the matching of the probiotics and the traditional Chinese medicine formula also needs to be considered. Specific screening procedures included the following experiments.
Research shows that constipation of middle-aged and elderly people is related to intestinal flora imbalance, the quantity of enterobacteria, enterococci, clostridium and the like in the intestinal tract is increased, and the quantity of lactic acid bacteria, bifidobacterium, bacteroides and the like is reduced, so that probiotics is supplemented, the abundance of beneficial bacteria in the intestinal tract is improved, the growth of harmful bacteria is inhibited, and the method is a powerful way for improving the intestinal health of constipation people.
In the research process, strains developed by yikoku vitamin technology limited are selected for screening, and the strains are as follows:
bifidobacterium longum W68: has high acid resistance, cholate resistance, strong intestinal tract colonization ability, better oxidation resistance and capability of reducing oxidative stress injury.
Bifidobacterium animalis bifidobacterium lactis subspecies Y6: the composition has high acid resistance, maintains the intestinal mucosa barrier, reduces inflammation and effectively takes care of intestinal health; acid resistance, bile salt resistance and promotion of forward balance of intestinal microecology.
Lactobacillus paracasei K9: acid resistance, cholate resistance and high colonization ability, and promotes the forward balance of intestinal microecology; high yield of lactic acid, repair of intestinal barrier and maintain intestinal wall integrity.
Lactobacillus rhamnosus YGG: the intestinal tract colonization ability is strong, the number of harmful bacteria in the intestinal tract is reduced, and the positive balance of intestinal tract flora is promoted.
Lactobacillus reuteri K07: acid resistance and bile salt resistance, can generate short-chain fatty acid and antibacterial substances, inhibit the growth of harmful bacteria, and promote the positive balance of intestinal flora.
Lactobacillus plantarum fed 8: acid resistance, cholate resistance and strong intestinal tract colonization ability, and promotes the positive balance of intestinal tract flora; has excellent antioxidant capacity and can reduce the absorption of harmful factors by organisms during constipation.
Pediococcus pentosaceus LN-PT 16: acid resistance and bile salt resistance, excellent intestinal tract colonization ability, high extracellular polysaccharide yield, excellent oxidation resistance and strong pathogenic bacteria inhibition ability.
The inhibition capacity of the above strains on common pathogenic bacteria is used as an index for optimization. The common pathogenic bacteria are selected from Escherichia coli, Salmonella thuringiensis and Bacillus cereus as indicator bacteria.
The test method adopts an Oxford cup method to prepare a flat plate with punched holes, probiotic solution is respectively added into the holes, anaerobic culture is carried out at 37 ℃, the diameter of a transparent ring around the holes is observed, data is recorded, the diameter represents the strength of inhibiting pathogenic bacteria, and the larger the diameter is, the stronger the ability of inhibiting pathogenic bacteria is.
Colony counts and absorbance (OD) were previously established according to GB4789.2600) The corresponding relationship of (1). Respectively inoculating Escherichia coli, Salmonella thuringiensis and Bacillus cereus to Nutrient Broth (NB) liquid culture medium for activation, taking third generation activated culture solution as indicator bacteria solution, adjusting to appropriate light absorption value to make its suspension concentration be 1 × 108 cfu/ml~5×108cfu/ml, adding 1% of the cfu/ml into a Nutrient Agar (NA) culture medium cooled to 46-50 ℃, uniformly mixing, measuring 20ml, adding into a plane culture dish in which 5-6 Oxford cups are placed at equal intervals in advance, taking out the Oxford cups after solidification, and placing for later use.
Firstly, animal bifidobacterium and bifidobacterium lactis subspecies Y6 are selected for preliminary experiments, and proper addition amount is selected. Activating the strain, taking the third generation activated culture solution as experimental bacteria solution, adjusting to appropriate light absorption value to make the concentration of the suspension of the strain be 1 × 108The probiotic liquid of cfu/ml is added into the holes according to the amount of 0.1ml, 0.05ml, 0.03ml and 0.01ml respectively, the anaerobic culture is carried out for 48 +/-2 hours at the temperature of 37 ℃, until the inhibition zone is clear, a vernier caliper is used for measuring the diameter of the transparent zone by taking the hole as the center, each inhibition zone is measured for 3 times along three different directions, the average value is taken, 3 parallel experiments are carried out, and the recorded data are as follows:
TABLE 1 Bifidobacterium animalis Bifidobacterium lactis Y6 preliminary experiment bacteriostatic circle diameter data
The diameter of the bacteriostatic zone is about 20mm, which is proper and too large, the number of holes punched in the plate is small, too small and the error is large, therefore, in the screening process of the above 7 probiotics, the activated bacteria content is 1 multiplied by 1080.03ml of probiotic solution was added per well.
Respectively activating the remaining 6 strains, namely bifidobacterium longum W68, lactobacillus paracasei K9, lactobacillus rhamnosus YGG, lactobacillus reuteri K07, lactobacillus plantarum FEED8 and pediococcus pentosaceus LN-PT16, taking the third generation activated culture solution as an experimental bacterial solution, adjusting the absorption value to a proper value to ensure that the concentration of the bacterial suspension is 1 × 108 Adding the probiotic liquid cfu/ml into a perforated NA plate, adding 0.03ml into each hole, preparing 3 parallel samples, carrying out anaerobic culture at 37 ℃ for 48 +/-2 hours until the inhibition zone is clear, measuring the diameter of a transparent ring by using a vernier caliper by taking the hole as the center, and measuring the diameter of each inhibition zone along three different directionsMeasure 3 times, measure NA plate zone diameter of restraining, the data is as follows:
TABLE 27 zone of inhibition data of Lactobacillus on E.coli NA plates
TABLE 37 inhibition zone data of Lactobacillus on Salmonella thuringiensis NA plates
TABLE 47 zone of inhibition data of Lactobacillus on Bacillus cereus NA plates
From the results, the diameters of inhibition zones of bifidobacterium animalis bifidobacterium lactis subspecies Y6, pediococcus pentosaceus LN-PT16 and lactobacillus paracasei K9 on the NA plates of the three indicator bacteria are larger, which indicates that the capability of inhibiting pathogenic bacteria is stronger, so that the three bacteria are selected as compound probiotic bacteria for regulating gastrointestinal tracts and improving constipation formulas.
Experiment system two: traditional Chinese medicine probiotic composite product for improving qi deficiency type constipation
The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation provided by the embodiment of the invention is a traditional Chinese medicine fermentation product capable of adjusting intestinal tracts and improving constipation (qi deficiency type) of middle-aged and elderly people, and is prepared by fermenting astragalus membranaceus, cistanche deserticola, semen cassiae, fructus cannabis, rhizoma polygonati, dark plum, fingered citron and yeast, and then adding compound probiotics, prebiotics, dietary fibers and auxiliary materials.
In the following examples, the yeast is Kluyveromyces marxianus SMN-S7-LBK with the preservation number of CCTCC M2017643; the Bifidobacterium animalis Bifidobacterium lactis subspecies is Bifidobacterium animalis subspecies Y6 with the preservation number of CGMCC No. 15026; the lactobacillus paracasei is lactobacillus paracasei K9 with the preservation number of CGMCC No. 15025; the Pediococcus pentosaceus is Pediococcus pentosaceus LN-PT16 with the collection number of GDMCC NO. 62044. The Plantago ovata husk and chia seed used are all in powder form.
In the process of product development, the inventor tries to use Kluyveromyces marxianus SMN-S7-LBK and composite probiotic Y30 for co-fermentation to obtain fermentation liquor. The final product is either liquid viable bacteria liquid preparation or lyophilized powder. When the finished product is a viable bacteria liquid preparation, the product cannot be stored for a long time, and the shelf life of the product is only slightly longer than that of the yoghourt (18-21 days), so that the finished product is not sold; if the freeze-dried powder is prepared, a large amount of traditional Chinese medicine extract components and live bacteria are required to be freeze-dried, and 2 problems also exist, namely, a large amount of materials are freeze-dried and are not beneficial to the production process, and secondly, because a large amount of traditional Chinese medicine extracts are arranged in the freeze-drying process, the embedding of the live bacteria is not beneficial, and the long-term storage is also not beneficial. The process which is most suitable for the production is therefore considered at the end of the invention: the traditional Chinese medicine liquid is added with yeast for fermentation, the fermentation liquid is concentrated and dried, and then prebiotics, dietary fiber and compound probiotics Y30 are added to prepare the solid beverage, so that the preparation stability is ensured on the premise of ensuring the efficacy, and the shelf life of the product is ensured. In the process of researching the fermentation of the kluyveromyces marxianus SMN-S7-LBK on the traditional Chinese medicine extracting solution (without centrifugation, with the traditional Chinese medicine residues in the extracting solution), the growth speed of the yeast is found to be slower than that of the yeast growing on the YPD agar medium, the kluyveromyces marxianus SMN-S7-LBK can reach the stationary phase 24 hours on the YPD agar medium, the kluyveromyces marxianus SMN-S7-LBK can grow in the traditional Chinese medicine liquid in an inoculated mode, except the nutrients in the traditional Chinese medicine, only sucrose is added, the growth speed is slower, and the fermentation time basically reaches 48 hours or even longer.
Example 1
A traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 10 parts of astragalus membranaceus, 30 parts of cistanche, 20 parts of rhizoma polygonati, 20 parts of semen cassiae, 15 parts of fructus cannabis, 5 parts of dark plum, 10 parts of fingered citron, 3 parts of fructo-oligosaccharide, 10 parts of isomalto-oligosaccharide, 1.5 parts of chia seed, 1.5 parts of polydextrose, 100 hundred million cfu of composite probiotics Y30, 10 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 35: 35: 30".
The preparation method of the product comprises the following steps:
(1) pulverizing the Chinese medicinal materials, sieving with 8 mesh sieve to obtain coarse powder, adding 8 times of water, heating and reflux-extracting for 1.5 hr, filtering, collecting filtrate, adding 7 times of water into residue, heating and reflux-extracting for 1.5 hr, mixing with the first extractive solution, adding 2.0% sucrose (by weight), adjusting pH to 6.0-6.5, boiling and sterilizing for 30 min.
(2) And transferring the inactivation solution to a cleaned and sterilized fermentation tank for later use.
(3) Adding kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.03%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank.
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing oxygen in proper amount, and introducing clean air for 2 hours every day (yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth speed reaching 1 multiplied by 10 according to normal fermentation6More than cfu/ml.
And fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
(4) Completing the fermentation (pH value is 4.0-4.5, viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at 70 deg.C and-0.085 Mpa to obtain concentrated solution (relative density of 1.08-1.10 and 50 deg.C), slowly adding 5 parts of fructus Hordei Germinatus paste into the concentrated solution, stirring, and spray drying to obtain spray dried powder. And (3) drying: air inlet temperature 175-: 75-80 ℃, atomization pressure: 0.24Mpa, and keeping the temperature of the thick paste at 70-75 ℃.
(6) And (3) adding 3 parts of fructo-oligosaccharide, 10 parts of isomaltooligosaccharide, 1.5 parts of chia seed, 1.5 parts of polydextrose, the rest 5 parts of maltodextrin, 0.1 part of silicon dioxide and the compound probiotics Y30 in the formula amount into the dry paste powder obtained in the step (5), and uniformly mixing in a multi-directional motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. Uniformly mixing materials except the compound probiotics Y30 and 0.05 part of silicon dioxide to obtain a mixed material, uniformly mixing the compound probiotics Y30, 0.05 part of silicon dioxide and 10 times of the mixed material, adding 90 times of the mixed material, uniformly mixing, and uniformly mixing with the rest mixed material to obtain the solid beverage (powder).
(7) Or further adding the mixed powder obtained in step (6) into a dry granulating machine, pressing, and grading to obtain granules, thus obtaining the solid beverage (granule type).
Example 2:
a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 20 parts of astragalus membranaceus, 15 parts of cistanche, 15 parts of rhizoma polygonati, 10 parts of semen cassiae, 20 parts of fructus cannabis, 10 parts of dark plum, 5 parts of fingered citron, 15 parts of isomaltooligosaccharide, 1 part of stachyose, 1.5 parts of Plantago ovata seed husk, 1 part of polydextrose, 250 hundred million cfu of composite probiotics Y30, 5 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 45: 45: 40".
The preparation method of the product comprises the following steps:
(1) crushing the traditional Chinese medicinal materials, sieving the crushed traditional Chinese medicinal materials by a 24-mesh sieve to obtain coarse powder, putting the coarse powder into an ultrasonic extractor, adding 12 times of water, and carrying out ultrasonic extraction for 40 minutes under the extraction conditions: temperature 35 ℃, ultrasonic power: 1200 watts per single head, 2 seconds on ultrasound stop for 1 second. Ultrasonic treating, adding 1.5% sucrose (weight ratio of extractive solution), adjusting pH to 6.0-6.5, boiling, and sterilizing for 30 min.
(2) And transferring the inactivation solution to a cleaned and sterilized fermentation tank for later use.
(3) Adding Kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.025%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank.
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing oxygen in proper amount, and introducing clean air for 2 hours every day (yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth speed reaching 1 multiplied by 10 according to normal fermentation6More than cfu/ml.
And fifthly, keeping the workshops of the fermentation tank and the fermentation liquor storage tank dry and sanitary to avoid breeding of mixed bacteria.
(4) The fermentation is finished (the pH value is 4.0-4.5, the viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at 70 deg.C under-0.085 Mpa to obtain concentrated solution (relative density of 1.08-1.10, 50 deg.C), and granulating with other materials (except probiotic embedded powder). Putting 1 part of stachyose, 15 parts of isomaltooligosaccharide, 1.5 parts of Plantago ovata forsk, 1 part of polydextrose and 5 parts of maltodextrin in a one-step granulator, spraying the extraction concentrated solution, and granulating, wherein the granulating conditions are as follows: inlet air temperature 102 ℃, material temperature 60 ℃, outlet air temperature: 45-50 ℃, atomization pressure: 0.22 MPa.
(6) And (3) uniformly mixing the granules prepared in the step (5) with 0.1 part of silicon dioxide and the compound probiotics Y30 in a formula amount in a multi-direction motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. And (3) adding 0.1 part of silicon dioxide and 10 times of the particles obtained in the step (5) into the composite probiotics Y30, uniformly mixing, adding 90 times of the particles obtained in the step (5), uniformly mixing, and uniformly mixing with the rest particles obtained in the step (5) to obtain the solid beverage (particle dosage form).
Example 3:
a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 15 parts of astragalus membranaceus, 25 parts of cistanche, 25 parts of rhizoma polygonati, 15 parts of semen cassiae, 10 parts of fructus cannabis, 20 parts of dark plum, 15 parts of fingered citron, 5 parts of fructo-oligosaccharide, 5 parts of galacto-oligosaccharide, 3 parts of Plantago ovata seed husk, 1 part of chia seed, 150 hundred million cfu of composite probiotics Y30, 10 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 25: 25: 20".
The preparation method of the product comprises the following steps:
(1) crushing the traditional Chinese medicinal materials, sieving the crushed traditional Chinese medicinal materials by a 16-mesh sieve to obtain coarse powder, putting the coarse powder into an ultrahigh pressure extractor, adding 10 times of water, and extracting under the extraction conditions: soaking for 30 minutes under the pressure of 400MPa, maintaining the pressure for 4 minutes, and circulating for 10 times at normal temperature. After extraction, 2.5% sucrose (by weight) is added, the pH value is adjusted to 6.0-6.5, and the mixture is boiled and sterilized for 30 minutes.
(2) And transferring the inactivation solution to a cleaned and sterilized fermentation tank for later use.
(3) Adding Kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.035%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank.
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in the fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen supply in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing proper amount of oxygen, and supplying clean air for 2 hours every day (the yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield, the process mainly aims at producing the yeast and does not need to produce alcohol).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control is carried out, fermentation is finished, the density of live bacteria is detected, and the density can reach 1 multiplied by 10 according to the growth speed of probiotics in normal fermentation6More than cfu/ml.
And fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
(4) Completing the fermentation (pH value is 4.0-4.5, viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at 70 deg.C under-0.085 Mpa to obtain concentrated solution (relative density of 1.08-1.10, 50 deg.C), slowly adding 5 parts of fructus Hordei Germinatus paste into the concentrated solution, stirring, and spray drying to obtain spray dried powder. And (3) drying: air inlet temperature 175-: 75-80 ℃, atomization pressure: 0.24Mpa, and keeping the temperature of the thick paste at 70-75 ℃.
(6) And (3) adding 5 parts of fructo-oligosaccharide, 5 parts of galacto-oligosaccharide, 3 parts of Plantago ovata seed husk, 1 part of chia seed and the remaining 5 parts of maltodextrin, 0.1 part of silicon dioxide and the compound probiotics Y30 in formula amount into the dry paste powder obtained in the step (5), and uniformly mixing in a multi-direction motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. Uniformly mixing materials except the compound probiotics Y30 and 0.05 part of silicon dioxide to obtain a mixed material, uniformly mixing the compound probiotics Y30, 0.05 part of silicon dioxide and 10 times of the mixed material, adding 90 times of the mixed material, uniformly mixing, and uniformly mixing with the rest mixed material to obtain the solid beverage (powder).
(7) Or further adding the mixed powder obtained in step (6) into a dry granulating machine, pressing, grading to obtain granules, and making into solid beverage (granule type)
Example 4:
a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 25 parts of astragalus membranaceus, 15 parts of cistanche, 10 parts of rhizoma polygonati, 10 parts of semen cassiae, 25 parts of fructus cannabis, 15 parts of dark plum, 20 parts of fingered citron, 10 parts of isomaltooligosaccharide, 2 parts of galactooligosaccharide, 1 part of chia seed, 3 parts of polydextrose, 250 hundred million cfu of composite probiotics Y30, 5 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 40: 30: 25".
The preparation method of the product comprises the following steps:
(1) pulverizing the Chinese medicinal materials, sieving with 8 mesh sieve to obtain coarse powder, placing in an extraction tank, adding 8 times, 6 times and 6 times of water, respectively, heating and reflux-extracting for 3 times (1.5 hr each time), concentrating the extractive solution of the first 2 times, mixing with the last time, adjusting to 15 times of the medicinal material amount of the extractive solution of 3 times, adding 1.5% sucrose (by weight), adjusting pH to 6.0-6.5, boiling and sterilizing for 30 min.
(2) And transferring the inactivation solution to a fermentation tank which is cleaned and disinfected for standby.
(3) Adding Kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.04%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank. .
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing oxygen in proper amount, and introducing clean air for 2 hours every day (yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth according to normal fermentationLong speed up to 1 × 106More than cfu/ml.
And fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
(4) The fermentation is finished (the pH value is 4.0-4.5, the viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at 70 deg.C under-0.085 Mpa to obtain concentrated solution (relative density of 1.08-1.10, 50 deg.C), and granulating with other materials (except probiotic embedded powder). Placing 2 parts of galacto-oligosaccharide, 10 parts of isomaltooligosaccharide, 1 part of chia seed, 3 parts of polydextrose and 5 parts of maltodextrin in a one-step granulator, spraying the extraction concentrated solution, and granulating under the following granulating conditions: inlet air temperature 102 ℃, material temperature 60 ℃, outlet air temperature: 45-50 ℃, atomization pressure: 0.22 MPa.
(6) And (3) uniformly mixing the granules prepared in the step (5) with 0.1 part of silicon dioxide and the compound probiotics Y30 in a formula amount in a multi-direction motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. And (3) adding 0.1 part of silicon dioxide and 10 times of the particles obtained in the step (5) into the composite probiotics Y30, uniformly mixing, adding 90 times of the particles obtained in the step (5), uniformly mixing, and uniformly mixing with the rest particles obtained in the step (5) to obtain the solid beverage (particle dosage form).
Example 5:
a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 30 parts of astragalus membranaceus, 10 parts of cistanche deserticola, 15 parts of polygonatum sibiricum, 25 parts of semen cassiae, 10 parts of fructus cannabis, 10 parts of dark plum fruit, 5 parts of fingered citron, 2 parts of galactooligosaccharide, 2 parts of stachyose, 1 part of psyllium husk, 3 parts of chia seed, 300 hundred million cfu of composite probiotics Y30, 10 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 30: 40: 35".
The preparation method of the product comprises the following steps:
(1) crushing the traditional Chinese medicinal materials, sieving the crushed traditional Chinese medicinal materials by a 24-mesh sieve to obtain coarse powder, putting the coarse powder into an ultrahigh pressure extractor, adding 12 times of water, and extracting under the extraction conditions: soaking for 30 minutes under the pressure of 400MPa, maintaining the pressure for 4 minutes, and circulating for 10 times at normal temperature. After extraction, 2.5% sucrose (by weight) is added, the pH value is adjusted to 6.0-6.5, and the mixture is boiled and sterilized for 30 minutes.
(2) And transferring the inactivation solution to a cleaned and sterilized fermentation tank for later use.
(3) Adding Kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.035%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank.
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing oxygen in proper amount, and introducing clean air for 2 hours every day (yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth speed reaching 1 multiplied by 10 according to normal fermentation6More than cfu/ml.
And fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
(4) The fermentation is finished (the pH value is 4.0-4.5, the viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at 70 deg.C under-0.085 Mpa to obtain concentrated solution (relative density of 1.08-1.10, 50 deg.C), slowly adding 5 parts of fructus Hordei Germinatus paste into the concentrated solution, stirring, and spray drying to obtain spray dried powder. And (3) drying: air inlet temperature 175-: 75-80 ℃, atomization pressure: 0.24Mpa, and keeping the temperature of the thick paste at 70-75 ℃.
(6) Adding 2 parts of galacto-oligosaccharide, 2 parts of stachyose, 1 part of Plantago ovata seed husk, 3 parts of chia seed and the rest 5 parts of maltodextrin, 0.1 part of silicon dioxide and the compound probiotics Y30 in formula amount into the dry paste powder obtained in the step (5), and uniformly mixing in a multi-direction motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. Uniformly mixing materials except the compound probiotics Y30 and 0.05 part of silicon dioxide to obtain a mixed material, uniformly mixing the compound probiotics Y30, 0.05 part of silicon dioxide and 10 times of the mixed material, adding 90 times of the mixed material, uniformly mixing, and uniformly mixing with the rest mixed material to obtain the solid beverage (powder).
(7) Or further adding the mixed powder obtained in step (6) into a dry granulating machine, pressing, and grading to obtain granules, thus obtaining the solid beverage (granule type).
Example 6:
a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is prepared from the following materials in parts by weight: 20 parts of astragalus membranaceus, 20 parts of cistanche, 15 parts of rhizoma polygonati, 15 parts of semen cassiae, 15 parts of fructus cannabis, 10 parts of dark plum, 10 parts of fingered citron, 3 parts of fructo-oligosaccharide, 0.5 part of stachyose, 1.5 parts of Plantago ovata seed husk, 1.5 parts of chia seed, 200 hundred million cfu of composite probiotics Y30, 10 parts of maltodextrin and 0.1 part of silicon dioxide. Wherein the composition of the composite probiotics Y30 is as follows, "Bifidobacterium animalis Bifidobacterium lactis subspecies Y6: lactobacillus paracasei K9: pediococcus pentosaceus LN-PT16= 35: 35: 30".
The preparation method of the product comprises the following steps:
(1) crushing the traditional Chinese medicinal materials, sieving the crushed traditional Chinese medicinal materials by a 16-mesh sieve to obtain coarse powder, putting the coarse powder into an ultrasonic extractor, adding 10 times of water, and extracting the coarse powder for 40 minutes by an ultrasonic extraction method under the extraction conditions: temperature 35 ℃, ultrasonic power: 1200 watts per single head, 2 seconds on ultrasound stop for 1 second. Ultrasonic treating, adding 2.0% sucrose (weight ratio of extractive solution), adjusting pH to 6.0-6.5, boiling, and sterilizing for 30 min.
(2) And transferring the inactivation solution to a cleaned and sterilized fermentation tank for later use.
(3) Adding Kluyveromyces marxianus SMN-S7-LBK live bacteria powder into the sterilized solution obtained in the step (2), and culturing, wherein the adding amount of the yeast live bacteria powder is 0.040%. Before yeast is added into a fermentation tank, activation is carried out, dried yeast powder is added into 10 times of 2% sucrose water, the mixture is uniformly mixed, and after 1.5 hours of activation (in the activation process, the yeast powder is shaken every 10 minutes) the mixture is added into the fermentation tank.
After the inoculation of the traditional Chinese medicine liquid is finished, fermentation management is carried out according to the following modes:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation.
Secondly, adjusting the oxygen supply in the tank, fully increasing oxygen in the liquid in the fermentation tank before inoculation, subsequently supplementing proper amount of oxygen, and supplying clean air for 2 hours every day (the yeast is facultative/anaerobic bacteria, aerobic fermentation, good bacterial growth, anaerobic fermentation, low yeast activity, but high alcohol yield, the process mainly aims at producing the yeast and does not need to produce alcohol).
Thirdly, index control, pH value detection, sampling for 1 time every 8 hours after the fermentation period reaches 24 hours, pH value detection, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5.
Fourthly, index control is carried out, fermentation is finished, the density of live bacteria is detected, and the density can reach 1 multiplied by 10 according to the growth speed of probiotics in normal fermentation6More than cfu/ml.
And fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
(4) The fermentation is finished (the pH value is 4.0-4.5, the viable bacteria density is more than 1 multiplied by 106cfu/ml), filtering, removing coarse powder with 100 mesh filter screen, and filtering with 400 mesh filter cloth to obtain Chinese medicinal fermentation liquid.
(5) Concentrating the Chinese medicinal fermentation liquid under reduced pressure at-0.085 Mpa at 70 deg.C to obtain concentrated solution (1.08-1.10, 50 deg.C), and granulating with other materials (except probiotic embedded powder). Putting 3 parts of fructo-oligosaccharide, 0.5 part of stachyose, 1.5 parts of Plantago ovata forsk, 1.5 parts of chia seed and 10 parts of maltodextrin in the formula into a one-step granulator, spraying the extraction concentrated solution, and granulating under the following granulating conditions: inlet air temperature 102 ℃, material temperature 60 ℃, outlet air temperature: 45-50 ℃, atomization pressure: 0.22 MPa.
(6) And (3) uniformly mixing the granules prepared in the step (5) with 0.1 part of silicon dioxide and the compound probiotics Y30 in a formula amount in a multi-direction motion mixer.
And (4) mixing the materials in the step (6) in order to fully and uniformly mix the probiotic mixed powder in the formula with other materials, and adopting an equivalent incremental mixing method. And (3) adding 0.1 part of silicon dioxide and 10 times of the particles obtained in the step (5) into the composite probiotics Y30, uniformly mixing, adding 90 times of the particles obtained in the step (5), uniformly mixing, and uniformly mixing with the rest particles obtained in the step (5) to obtain the solid beverage (particle dosage form).
Comparative example 1
Comparative example 1 differs from example 6 in that: the composition and preparation method were the same as in example 6 except that Kluyveromyces marxianus SMN-S7-LBK was not added to ferment the Chinese medicinal extract.
Comparative example 2
Comparative example 2 differs from example 6 in that: the preparation material lacks the composite probiotics Y30, prebiotics and dietary fibers, and the rest components and the preparation method are the same as those in the example 6.
Comparative example 3
Comparative example 3 differs from example 6 in that: in the case of fermenting the extract solution of the traditional Chinese medicine, the composition and preparation method were the same as those of example 6 except that the yeast Kluyveromyces marxianus SMN-S7-LBK was not fermented, but "Saccharomyces cerevisiae (selected from Saccharomyces cerevisiae manufactured by Angel Yeast Co., Ltd.) was fermented".
Experiment system three: study on Change of substances before and after fermentation
First, the comparison study of the total nitrogen content
The kluyveromyces marxianus SMN-S7-LBK is one kind of yeast, contains almost no fat and starch, contains high-quality protein, complete B vitamins, various biological minerals and high-quality dietary fibers, has the characteristics of low fat, low sugar and low calorie (no cholesterol), and is very balanced in nutrition. The nitrogen content is used as an index to research the change of protein components before and after fermentation, and the quality of the fermentation process can be reflected. The specific research process is as follows:
in example 6 and comparative example 3, 3 parts of each of the pre-fermentation extract and the post-fermentation extract was measured by the first method under the item of the nitrogen measurement method 0704, which is the fourth guideline of the "chinese pharmacopoeia", and the measurement results are shown in table 5 below.
TABLE 5 table of nitrogen content measurement results
The nitrogen content increase ratio: kluyveromyces marxianus SMN-S7-LBK = (2.317-0.525)/0.525 × 100= 341.3%; brewers yeast = (2.033-0.528)/0.528 × 100= 285.0%;
in the above table, the nitrogen content is greatly increased after the fermentation in example 6, which shows that the protein, amino acid and nitrogen-containing vitamin in the fermentation liquid are greatly increased after the fermentation of Kluyveromyces marxianus SMN-S7-LBK, and the efficacy of the extract is also greatly improved. Comparative example 3, the nitrogen content was also greatly increased by fermentation using Saccharomyces cerevisiae, but the increase rate was not as high as that of Kluyveromyces marxianus SMN-S7-LBK.
Second, the content comparison research of total polysaccharide
The polysaccharide produced by the traditional Chinese medicine and the probiotics has excellent physiological activity, such as improving immunity, helping to repair skin and mucosa, forming a skin and mucosa protective barrier, resisting tumor, resisting virus, reducing blood sugar, resisting aging and the like. In the formula of the product, the astragalus, the rhizoma polygonati, the cistanche salsa, the dark plum, the cassia seed and other medicinal materials all have a large amount of polysaccharide, and the yeast can also generate the polysaccharide in the fermentation process; in addition, during the fermentation process, part of starch substances in the traditional Chinese medicine can be converted into polysaccharide under the action of bacteria and part of enzyme. Therefore, it is necessary to study the change of polysaccharide in the extract before and after fermentation, and the specific implementation process is as follows.
In example 6 and comparative example 3, 3 parts of each of the pre-fermentation extract and the post-fermentation extract were taken and measured.
The determination method comprises the following steps: referring to the determination of the wolfberry polysaccharide under the term of wolfberry in 'Chinese pharmacopoeia 2020 edition', the determination method of the total polysaccharide in the sample is formulated as follows:
preparing reference solution by accurately weighing 25mg of anhydrous glucose reference, placing in a 250ml measuring flask, adding appropriate amount of water to dissolve, diluting to scale, and shaking to obtain the final product (each 1ml contains 0.1mg of anhydrous glucose).
Preparation of standard curve reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0ml are precisely measured, respectively placed in test tubes with plugs, respectively added with water to supplement to 2.0ml, respectively and precisely added with 5% phenol solution 1ml, shaken, rapidly and precisely added with sulfuric acid 5ml, shaken, placed for 10 minutes, placed in a water bath at 40 ℃ for heat preservation for 15 minutes, taken out, rapidly cooled to room temperature, and measured at 490nm wavelength absorbance with absorbance as ordinate and concentration as abscissa by using corresponding reagent as blank and ultraviolet-visible spectrophotometry (general rule 0401).
Measuring a sample by accurately weighing 12g, slowly adding 15ml of ethanol, uniformly shaking while adding, centrifuging at 5000 revolutions, discarding a supernatant, adding 60% of ethanol into residues, uniformly dispersing by shaking, centrifuging again, discarding a supernatant, adding 100ml of water into residues to dissolve, transferring into a beaker, heating to boil, filtering while hot, washing a filter by using a small amount of hot water, combining a filtrate and a washing solution, cooling, transferring into a 250ml measuring flask, diluting with water to a scale, uniformly shaking, accurately measuring 1ml, placing into a test tube with a plug, adding 1.0ml of water, measuring absorbance by the method from the point that 1ml of 5% phenol solution is accurately added, reading the weight (mg) of glucose in a sample solution to be measured from a standard curve, and calculating to obtain the glucose concentration test solution. The specific measurement results are shown in table 6 below.
TABLE 6 polysaccharide test results Table
The increasing ratio of polysaccharide: kluyveromyces marxianus SMN-S7-LBK = (1.32-1.14)/1.14 × 100= 15.8%; brewer's yeast = (1.30-1.13)/1.13 × 100= 14.2%;
from the results shown in the above table, the amounts of polysaccharides before and after fermentation in example 6 and comparative example 3 were increased, but they were not particularly high and were not greatly different.
And (4) experiment system IV: sensory testing
Firstly, the method comprises the following steps: comparing the color, smell and taste of the Chinese medicinal liquid before and after fermentation
To compare the fermentation effect of yeast, example 6 and comparative example 3 were used for comparison, and 1 part of each of the pre-fermentation sample and the post-fermentation sample (the samples were centrifuged and the supernatant was collected) was taken, and the color, smell and taste of the solution were visually checked by non-testers, and the color, smell and taste of the solution before and after fermentation were compared, and the results are as follows:
TABLE 7 comparison table of the properties of Chinese medicinal liquid before and after fermentation
Remarking: the more "+" in the color column, the darker the color is.
From the above table, after yeast fermentation, the traditional Chinese medicine has lighter color, increased fragrance, reduced traditional Chinese medicine taste, and reduced bitter taste and light astringent taste. In contrast, kluyveromyces marxianus fermented product has stronger mellow fragrance, smells better and has lighter bitter and astringent taste. In general, the fermentation of Kluyveromyces marxianus has better improvement on the color, smell and taste of traditional Chinese medicines than the fermentation of beer yeast.
Experiment system five: efficacy verification
Mouse experiment of example 6, comparative example 1 and comparative example 2
The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation (traditional Chinese medicine fermentation product for regulating intestinal tract and improving constipation (qi deficiency type) of middle-aged and elderly people) is prepared by extracting a traditional Chinese medicine formula of raw astragalus, cistanche deserticola, rhizoma polygonati, cassia seed, fructus cannabis, dark plum and fingered citron, fermenting in YPD through Kluyveromyces marxianus SMN-S7-LBK, and mixing with prebiotics, dietary fibers, bifidobacterium animalis, lactobacillus lactis strain Y6, lactobacillus paracasei L9S and pediococcus pentosaceus LN-PT16, and has the effects of regulating gastrointestinal tract and improving constipation. To verify product efficacy, animal experimental studies were performed.
Meanwhile, in order to verify the efficacy of Kluyveromyces marxianus SMN-S7-LBK in YPD fermentation, the same process as that of example 6 was carried out except that Kluyveromyces marxianus SMN-S7-LBK was not added in YPD fermentation, and comparative example 1 was obtained; in order to verify the efficacies of the probiotics, the prebiotics and the diet, the preparation method is the same as the preparation method in the example 6 except that the probiotics Y30, the prebiotics and the dietary fibers are not added, the comparative example 2 is obtained, and meanwhile, the efficacy comparison research is carried out.
Construction of mouse model
Materials: senna leaf, raw rice, activated charcoal, animal: kunming white mouse is from southern medical university animal experiment center, has a weight of 18-22g, half male and half female, and is 6-8 weeks old.
In the qi deficiency model, the spleen and stomach deficiency is caused by purgative (senna leaf), and then the spleen deficiency and qi deficiency are further caused by limited diet. On the basis of the qi-deficiency model, drinking water is further limited, diet is further controlled, and a constipation model (qi-deficiency type) is caused. The molding process is as follows:
preparing a folium sennae extracting solution: adding 8 times of water into senna leaves, heating, refluxing and extracting for 2 times, filtering, and concentrating the filtrate under reduced pressure to obtain the following medicinal materials: the concentrated solution is 1: 1, and refrigerating at low temperature for later use.
Modeling a mouse: 132 mice, half of each male and female, were raised in cages. After three days, the molding was started, and 12 (half each of the male and female) groups were kept as normal control groups (normal diet with the same amount of distilled water as the molding group in the molding process). Constructing 120 models, and irrigating with folium sennae extract at a ratio of 0.8 g/day for 1-7 days to obtain spleen deficiency model when diet and drinking water are normal; feeding raw rice with low fiber feed at a ratio of 1.6 g/day from 8 days, continuously drinking water for 5 days, and adding a qi deficiency model after spleen deficiency; 5 g of raw rice fed with the low-fiber feed every other day after 13 days, and the raw rice is freely drunk for 1 time, 0.5 hour each time and fed for 6 days. Under the condition of controlling diet with interval of limiting drinking water and hunger, the constipation model is continuously caused, and the molding time is 18 days.
And (3) molding results: mice show shriveled appearance, weight loss, stiff and fluffy arch back, reduced activity, dry feces, reduced quantity and successful model building.
Grouping mice: the model group 108 was divided into 9 groups, and the samples of example 6 were gavaged in groups 1 to 3, and were gavaged in groups of low, medium and high at 0.05 g/day, 0.1 g/day and 0.2 g/day, respectively. The samples of comparative example 1 (fermented without Kluyveromyces marxianus SMN-S7-LBK) were divided into low, medium and high groups, and gavage was performed at 0.05 g/day, 0.1 g/day and 0.2 g/day, respectively. The samples of comparative example 2 (without probiotic Y30, prebiotics and dietary fibers) were gavaged in groups 7, 8 and 9 at 0.05 g/day, 0.1 g/day and 0.2 g/day for the low, medium and high groups, respectively. Group 10 was a blank model control group fed 0.1 grams of distilled water. Group 11 was a normal mouse control group and was fed normally.
The treatment method comprises the following steps: on days 19-26, except for the normal control group, the mice of each group were fed with 5 g of raw rice every day and freely drunk water 1 time for 0.5 hour each time except for the gavage treatment. On day 26, the animal experiment was completed and the index was measured.
Observation indexes are as follows: during the modeling and medication period, the eating condition, body weight, feces and the like of each group of mice are observed and recorded every day.
Detecting the intestinal propulsion function: after the treatment group had taken off the drug, fasted and deprived of water for 24 hours, mice were gavaged with 0.3ml of activated carbon suspension (concentration 0.1 g/ml), 25 minutes later, sacrificed, dissected immediately, and the entire intestine from the pylorus to the end of the rectum was removed and measured under tension-free conditions: 1 intestinal tract full length, 2 active carbon suspension in intestinal tract propulsion length, and calculating the percentage of active carbon propulsion length to the intestinal tract full length (black tube length/total tube length).
As a result:
general conditions of mice in each group
(1) Normal control group (group 11): eyes have spirit, the activity is rapid, the back hair is dense and glossy, the diet is normal, and the people can feel normal.
(2) Model group (pre-treatment): on the 3 rd day of molding, the mice begin to have hair uprising, lackluster body hair, arch back and reduced activity, and then slowly feel withered, tired, lie, sleepy, loose stool, emaciation and shrivelled, and after the 13 th day, constipation symptoms appear, the stool quantity is reduced, and the particles become small and hard.
(3) Treatment groups (groups 1-9) following treatment: from the 4 th day of administration, mice in each treatment group slowly improved, had increased food intake, were active, had increased stool numbers, had large granules, and had increased water content.
(4) Model control group (gavage distilled water, group 10): the mice are shriveled and emaciated, the body hair is lackluster, the food intake is less, the activity is less, the stool is dry and hard, the quantity is less, the grain is small, and compared with the symptoms after the model is made, the symptoms are basically not improved.
The detection results of the mouse indexes of each group are as follows:
according to the comparison before and after the administration, the weight and the average stool amount (granules) of each group of mice are detected, and the intestinal propulsion rate is calculated. Before administration, after successful model building, the average weight of 1-10 groups is 16.84 + -0.61 g, and the average stool amount is 12.27 + -2.11 granules. The results after administration are shown in table 8 below.
Table 8 statistical table of mouse test results
Regarding the water content of the excrement, the defecation time of the mouse is uncertain, the mouse is possible in the daytime and at night, if the excrement is not discharged in time, the water content is slowly reduced, and the mouse can not be continuously observed for 24 hours. Therefore, the water content measurement cannot be accurately carried out, but the defecation condition of the mice is observed in time, after the products of the embodiment group and the control group are used, the defecation particles become relatively large, the water content is increased visually, and the mice are not dried before being used; the stool particles in the comparative example 2 were slightly smaller than those in the examples 6 and 1, and the water was slightly dry.
From the above table, the weight of the mice increased after the administration, the average number of particles per day increased, and the intestinal propulsion rate of the mice was tested after the administration, which was significantly changed from that before the administration. Example 6 compared with comparative example 1 (no fermentation, and other processes consistent with the example 6) and comparative example 2 (no addition of probiotics, prebiotics and dietary fibers), the sample of example 6 has better effect, and the yeast fermentation and the fermented product have very good effect on gastrointestinal tract; the probiotics, prebiotics and dietary fibers have good effects on the health of the gastrointestinal tract.
Overall, the sample of example 6 has very good effects of conditioning the gastrointestinal tract and improving constipation.
Second, the trial of the crowd
In the product development process, trial tests of healthy people, people with poor gastrointestinal tract conditions and people with constipation are performed, and the trial sample is the sample of embodiment 6.
As a result, after the health people use the traditional Chinese medicine composition, the appetite becomes better, the defecation is more regular, the defecation generally starts early, the stool character is thick and long, yellow, and the defecation is easier. The defecation condition of the crowd with bad gastrointestinal tract condition is improved, the people turns to regular and normal defecation, and the appetite slowly gets better. The defecation interval of constipation people is shortened, after the constipation people use the composition for 1 week, the constipation people basically recover to 1 day for 1 time or 3 days for 2 times, the conditions of abdominal distension and mild abdominal pain are improved, and the intestinal defecation pain feeling is not caused after the composition is finished; the oral odor is improved for some people with halitosis. Some trial examples are as follows:
test example 1: female, 58 years old, bad gastrointestinal tract condition, constipation, general under the condition of no medicine, defecating for 1 time in 3-4 days, defecating for no more than 2 times in 1 week, less meal, dare not to eat more, eat more abdominal distension, especially in evening, eat more abdominal distension, pain, be difficult to fall asleep, go to toilet and defecate difficult to discharge, hard and dry defecate, yellow face, visible color and pigmentation, and breath. The patients with qi deficiency may get up the stairs and have dyspnea. Other constipation-regulating products such as frequently-moistened tea are eaten before, the tea is effective when being drunk at first, the tea is ineffective when being drunk later, the tea is only required to be stopped, and the constipation is more serious after the tea is stopped; but also solves the fundamental problems of lactulose oral liquid, cisapride and the like prescribed by doctors. Usually, the enema should be used for many days without stool.
Test example 1 the product of example 6 was used after meal, 1 bag 1 time, with warm water. As a result, after taking for 1 week, 3 times of defecation, and after 20 days, 1 time of defecation basically takes 1-1.5 days, the defecation is yellow and thick, and basically returns to normal, the abdominal pain feeling caused by the prior medication is not generated in the whole process, the appetite is greatly improved, the peculiar smell in the mouth is also greatly improved, the people do not feel hard to go upstairs, and the face is ruddy.
Test case 2: for men, in age 29, they like to eat peppery in home and country, and they often eat peppers after leaving hometown. After a long time, the face has acne, the people eat more spicy and more acne, the face, particularly the nose and the periphery are greasy, the breath is heavier, the abdomen is large, the weight is increased more, the stool is cleared 1 day 2-3, the stool is hard first and then soft, the head is nearly black and dry first, the back is green, soft and sticky, the toilet is stuck, the tongue coating is greasy, the tongue body is fat, and the edge has tooth marks.
After the product of the example 6 is tried, the pox on the back part of 1 week gradually becomes less and lighter, and the stool is not sticky any more; after 2 weeks, there was a reduction in body weight from about 76kg to 74.6kg, with substantial improvement in pox and yellowing of stool. After 3 weeks, the stool is adjusted to 1 time for 1 day, the stool is yellow and not sticky, no acne grows on the face, the tongue is observed, the coating is recovered to be white, the edge of the tongue body has no tooth marks, the body weight is not obviously reduced, the weight is weighed to be 73.9kg, but after 3 weeks, the total abdominal circumference is reduced by about 3-5 cm, the belt contracts 2 holes, the person is fresh and cool, and the oil on the face is reduced.
From the trial example 2, even for people who are non-qi-deficiency, after the laxative traditional Chinese medicine and the probiotics are added and the prebiotics and the dietary fibers are matched, the intestinal environment is integrally improved, and the improvement effect on related problems caused by the gastrointestinal tract is very good.
Test case 3: for a male, the male is 62 years old, the body is originally good, after last-year gallstone operation, the stomach is not good, poor appetite and dyspepsia are caused, if people eat greasy food, the people feel nausea and vomiting immediately, and the people have slight abdominal distension and pain feeling and are not serious, the constipation is relieved after defecating, the people feel relieved after 3-4 days, the people feel dry and hard, the time for going to the toilet is long, the face is slightly dull, the physical strength self-sensation is much worse than that before the operation, and the people feel dry and alive and tired.
The product of example 6 was used and taken after meal, 1 bag 1 time, with warm water.
After 1 week, the condition of going to the toilet is improved, the toilet is easy to be disassembled, and the excrement is required to be disassembled by force originally; after 3 weeks, the appetite is greatly improved, no nausea is caused when people eat greasy food for a little, 3 times of defecating are carried out for 2 days, the stomachache is greatly improved, no abdominal pain symptom exists, the complexion is ruddy, people feel energetic, and the symptoms are greatly improved.
The trial product (example 6) is used by healthy people and some people with gastrointestinal tract problems (such as constipation), and has no report on the use safety problem.
The above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are within the protection scope of the present invention.
Claims (10)
1. The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation is characterized by being at least formed by compounding traditional Chinese medicine leavening and compound probiotics; the traditional Chinese medicine fermented product is prepared by fermenting traditional Chinese medicine extracting solution obtained by extracting a traditional Chinese medicine formula by saccharomycetes, wherein the traditional Chinese medicine formula comprises 10-30 parts of astragalus membranaceus, 10-30 parts of cistanche, 10-25 parts of semen cassiae, 10-25 parts of fructus cannabis, 10-25 parts of rhizoma polygonati, 5-20 parts of dark plum and 5-20 parts of fingered citron; the composite probiotics comprise bifidobacterium animalis bifidobacterium lactis subspecies bifidus, lactobacillus paracasei and pediococcus pentosaceus.
2. The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 1, wherein the yeast is kluyveromyces marxianus SMN-S7-LBK with the preservation number of CCTCC M2017643; the bifidobacterium animalis bifidobacterium lactis subspecies is bifidobacterium animalis subspecies Y6 with the preservation number of CGMCC No. 15026; the lactobacillus paracasei is lactobacillus paracasei K9 with the preservation number of CGMCC No. 15025; the Pediococcus pentosaceus is Pediococcus pentosaceus LN-PT16 with the preservation number of GDMCC NO. 62044.
3. The traditional Chinese medicine probiotic compound product for improving qi-deficiency constipation according to claim 1, wherein the compound probiotics comprises the following components in parts by weight: lactobacillus paracasei: pediococcus pentosaceus = 25-45: 25-45: 20-40.
4. The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 1, wherein the traditional Chinese medicine fermentation product-probiotic compound product further comprises one or any combination of prebiotics, dietary fibers and auxiliary materials; the prebiotics are one or any combination of fructo-oligosaccharide, isomaltooligosaccharide, galacto-oligosaccharide and stachyose; the dietary fiber is one or any combination of psyllium seed husk, chia seed and polydextrose; the auxiliary material is maltodextrin and/or silicon dioxide.
5. The traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 1, which is prepared by compounding traditional Chinese medicine fermentation products, compound probiotics, prebiotics, dietary fibers and auxiliary materials; the traditional Chinese medicine fermented product is prepared by fermenting traditional Chinese medicine extracting solution obtained by extracting a traditional Chinese medicine formula by saccharomycetes, wherein the traditional Chinese medicine formula comprises 15-25 parts of astragalus membranaceus, 15-25 parts of cistanche, 10-20 parts of semen cassiae, 10-20 parts of fructus cannabis, 10-20 parts of rhizoma polygonati, 5-15 parts of dark plum and 5-15 parts of fingered citron; the addition amount of the composite probiotics is 100-300 hundred million cfu, and the composite probiotics comprise the following components in parts by weight: 30-40: 25-35 of bifidobacterium animalis bifidobacterium lactis subspecies, lactobacillus paracasei and pediococcus pentosaceus; the prebiotics comprise 0-5 parts of fructo-oligosaccharide, 0-15 parts of isomaltooligosaccharide, 0-5 parts of galacto-oligosaccharide and 0-2 parts of stachyose; the dietary fiber comprises 0-3 parts of psyllium husk, 0-3 parts of chia seed and 0-3 parts of polydextrose; the auxiliary materials comprise 5-10 parts of maltodextrin and 0.05-0.2 part of silicon dioxide.
6. The preparation method of the traditional Chinese medicine probiotic compound product for improving qi-deficiency constipation according to any one of claims 1 to 5, which is characterized by comprising the following steps:
the traditional Chinese medicine extraction step: extracting traditional Chinese medicinal materials in a traditional Chinese medicine formula to obtain a traditional Chinese medicine extracting solution;
a sterilization step: adjusting the pH of the Chinese medicinal extract to 6.0-6.5, sterilizing, and transferring the sterilized solution to a fermentation tank for later use;
a fermentation step: activating yeast, adding into the fermentation tank, fermenting to obtain Chinese medicinal fermented extract;
and (3) filtering: filtering the traditional Chinese medicine fermentation extract to obtain traditional Chinese medicine fermentation liquor;
a concentration and drying step: concentrating and drying the traditional Chinese medicine fermentation liquor to obtain dry paste powder;
mixing: mixing the dry extract powder with compound probiotic, and mixing well if prebiotics, dietary fiber and adjuvants are included.
7. The method for preparing a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 6, wherein in the traditional Chinese medicine extraction step, the traditional Chinese medicine is firstly crushed, and then water is added for extraction, the crushing mesh number of the traditional Chinese medicine is controlled to be 8-24 meshes, and the total multiple of the added water is 7-15 times of the weight of the traditional Chinese medicine; during extraction, the traditional Chinese medicinal materials are extracted by a heating reflux extraction method, an ultrasonic extraction method or an ultrahigh pressure extraction method; when a heating reflux extraction method is adopted, the extraction time is 1-2 hours, and the extraction times are 2-3 times; when an ultrasonic extraction method is adopted, the extraction time is 0.5-1 hour, and the extraction conditions are as follows: temperature 25-45 ℃, ultrasonic power: stopping the ultrasonic treatment for 1-3 seconds at the speed of 1100-1300W/single ultrasonic head for 2-5 seconds; when the ultrahigh pressure extraction method is adopted, the pressure is 100-600MPa, the pressure is maintained for 1-6 minutes, the temperature is 25-60 ℃, and the cycle period is 8-12 times; after the traditional Chinese medicine extracting solution is obtained, adding sucrose, wherein the adding amount of the sucrose is 1.5-2.5% of the weight of the traditional Chinese medicine extracting solution.
8. The method for preparing a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 6, wherein in the fermentation step, the addition amount of yeast live bacteria powder during fermentation is 0.025-0.04% of the weight of the traditional Chinese medicine extract, the yeast is activated before being added into a fermentation tank, the dry yeast powder is added into 1-2.5% of sucrose water in an amount of 8-12 times of the weight of the yeast live bacteria powder, the mixture is uniformly mixed, the mixture is added into the fermentation tank after being activated for 1.2-1.8 hours, and the mixture is shaken every 8-12 minutes during the activation process.
9. The method for preparing a traditional Chinese medicine probiotic compound product for improving qi-deficiency constipation according to claim 6, wherein in the fermentation step, the fermentation process comprises the following operations:
firstly, controlling the temperature, and after inoculation, controlling the temperature in a fermentation tank to be 28-30 ℃ to perform yeast fermentation;
adjusting oxygen to be introduced into the fermentation tank, fully increasing oxygen to the liquid in the fermentation tank before inoculation, and subsequently supplementing oxygen in a proper amount, wherein clean air is introduced for 1.5-2.5 hours every day;
thirdly, index control, sampling for 1 time every 8 hours after fermenting for 24 hours, detecting the pH value, wherein the normal required value is less than 4.5, and if the normal required value is more than 4.5, the fermentation time is prolonged until the pH value is less than 4.5;
fourthly, index control, fermentation completion, viable bacteria density detection and probiotic growth speed reaching 1 multiplied by 10 according to normal fermentation6The above;
and fifthly, the workshops of the fermentation tank and the fermentation liquor storage tank are kept dry and sanitary, so that the breeding of mixed bacteria is avoided.
10. The method for preparing a traditional Chinese medicine probiotic compound product for improving qi deficiency type constipation according to claim 6, wherein in the concentration and drying step, the concentration method is a reduced pressure concentration method; the drying method is spray drying method or one-step drying granulation method.
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