CN106727731A - A kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating and preparation method thereof - Google Patents

A kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating and preparation method thereof Download PDF

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Publication number
CN106727731A
CN106727731A CN201710021903.1A CN201710021903A CN106727731A CN 106727731 A CN106727731 A CN 106727731A CN 201710021903 A CN201710021903 A CN 201710021903A CN 106727731 A CN106727731 A CN 106727731A
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preparation
stomach invigorating
freeze drying
tiny ecosystem
whole
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储卫华
周淑鑫
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China Pharmaceutical University
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China Pharmaceutical University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/064—Saccharomycetales, e.g. baker's yeast
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions

Abstract

The invention discloses a kind of whole Enteral formulationses of compound microecological stomach invigorating and preparation method thereof.The compound micro-ecological preparation is individually fermented in suitable culture medium by the lactobacillus acidophilus with prebiotic effect, bacillus licheniformis and saccharomyces cerevisiae; according to certain ratio after fermentation; freeze drying protectant is added, and is equipped with oligosaccharide, then carried out vacuum freeze drying and be obtained.Gained Tiny ecosystem lyophilized formulations have frankincense taste, and bacterial activity is strong, the features such as preserving for a long time.Directly supplement this kind of compound micro-ecological preparation, coordinative role is played by multi-cultur es, gastrointestinal bacterial flora can effectively be adjusted, promote the propagation of beneficial bacterium, enhance immunity, help digest, suppress harmful bacteria breeding etc. effect, whole intestines effect of stomach invigorating of composite bacteria preparation is played, and composite bacteria preparation obtained by this preparation method can more effectively ensure activity and the preservation time of beneficial bacterium.

Description

A kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating and preparation method thereof
Technical field
Patent of the present invention belongs to microbial technology field.It is specifically related to a kind of with the compound micro- of the whole intestines prebiotic effect of stomach invigorating Ecological agent and preparation method thereof.
Background technology
With the progress in epoch, the development speed of society is more and more faster.Facing to various live and work pressure, people are not The trend of era development must be not suitable with, immediately following the paces of social development.The body and mind that so allegro life style gives people is good for Health brings strong influence, and especially diet and the irregular of daily life cause modern generally with serious gastrointestinal disease, While oneself healthy is compromised, the normal live and work of people is also have impact on.Gastrointestinal bacterial flora is to maintain people The necessary factor of body health, physiological condition lower gastrointestinal tract flora may also provide vitamin in addition to certain " colonization resistance ", Synthesis digestive ferment, participates in the metabolism of protein, and cholesterol degradation forms bile acid, and short of money with immunostimulation and biology Anti- effect, is internal environment balance of humanbody whether mark.If body internal and external environment changes, human gastrointestinal tract will occur Disorder, causes digestion and absorption function to weaken, and enterogastric diseases occur, and shows gastritis, enteritis, diarrhoea and constipation etc., or even Immunity degradation etc. can be caused, such as adjusted not in time, the probability of intestines and stomach canceration can be increased.In order to treat the stomaches such as diarrhoea, constipation Intestinal disorder, clinically commonly uses some antibiotic or charthartic to improve symptom, but because these medicine long-term uses can be produced Raw tolerance, therefore be not suitable for persistently taking for a long time.It has recently been demonstrated that flora is adjusted by absorbing some probiotics putting down Weighing apparatus, environment in adjustment intestines and stomach, can effectively improve gastrointestinal disturbance.Therefore, probiotics turns into the intestines and stomach such as diarrhoea, constipation The auxiliary replenishers of disease are paid attention to and are applied.
It is liquid or pulvis, tablet that probiotics currently on the market is most of.Liquid micro-ecological preparations are due to thin The growth of bacterium, the death of viable count bacterium quickly will be reduced largely；The probiotics of pulvis or tablet removes gemma mushroom In addition, because, with the presence of moisture, bacterium also can be dead in air or auxiliary material, thus activity can also decline.The present invention is based on intestines The one kind researched and developed in road colony balance theoretical foundation is aided with prebiotics to adjust gastrointestinal bacterial flora by multiple beneficial bacterium, reaches extensive The purpose of multiple normal flora, carrys out regulating gastointestinal tract disease.The present invention will also be answered for the Vacuum Freezing ＆ Drying Technology of culture presevation Used in probiotics industrialization production so that obtained compound probiotic is in a kind of resting state, can for a long time keep micro- Biological activity.
The content of the invention
It is an object of the invention to provide a kind of probiotics of the whole intestines of compound stomach invigorating, while it is corresponding to provide its Preparation method is another object of the present invention.
Based on above-mentioned purpose, the present invention is adopted the following technical scheme that：A kind of compound whole Enteral formulationses of Tiny ecosystem stomach invigorating, per 1g Contain lactobacillus acidophilus >=5 × 10 in preparation7CFU, bacillus licheniformis >=5 × 107CFU, saccharomyces cerevisiae >=5 × 107CFU, freezes Protective agent 0.1-0.2g.
The mass ratio of the freeze drying protectant is：10-20% skimmed milk powers, 5-10% sucrose, 5-10% oligosaccharide.It is described Oligosaccharide includes FOS, oligomeric maltose, xylo-oligosaccharide.
A kind of preparation method of mixed type enteral microecological formulation, comprises the following steps：
A () prepares compound bacteria bacterium solution：
By lactobacillus acidophilus, bacillus licheniformis and saccharomyces cerevisiae be inoculated into respectively MRS culture mediums, LB culture mediums and Fermented 18-36 hours in YPD culture mediums, 30-40 DEG C of fermentation temperature, each bacterial content >=5 × 10 of gained8CFU/ml, is then carried out Mix to obtain mixed bacteria liquid in proportion；
B bacterium solution that () is obtained (a) adds freeze drying protectant and oligosaccharide, ready bacterium solution is dispensed into lyophilized In bottle, precooling is then carried out；
C () vacuum freeze drying, obtains finished product.
A kind of preparation method of described whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that bacteria used thereby in the step (a) Strain is lactobacillus acidophilus CGMCC1.1854, bacillus licheniformis CGMCC1.105, saccharomyces cerevisiae CICC1249；It is each in mixed liquor Bacterium accounts for cumulative volume ratio for 20-50%V/V.
The preparation method of described a kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that in the step (b), with matter Amount percentages, the formula of the freeze drying protectant includes：10-20% skimmed milk powers, 5-10% sucrose, 5-10% oligosaccharide.
The preparation method of described a kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that in the step (b), -70 Pre-freeze 2-4 hours at~-80 DEG C.
The preparation method of described a kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that in the step (c), vacuum Freeze-drying 36 hours.
The preparation method of described a kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that oligomeric in the step (b) Sugar includes FOS, oligomeric maltose, xylo-oligosaccharide.
It is an advantage of the invention that：Probiotics with stomach invigorating, whole gut function is carried out into external individually fermentation, then according to one Fixed ratio is mixed, and is equipped with the prebioticses such as freeze drying protectant and oligosaccharide, then carries out vacuum freeze drying, can improve thin Bacterium survival rate in its natural state.The compound micro-ecological preparation can enter into intestines and stomach with the thalline of resting state, in stomach and intestine Oxygen is customized and consumed in road, and creating the prebioticses such as anaerobic environment, oligosaccharide can promote the growth of beneficial bacterium, the compound microecological Preparation can effectively suppress the field planting of harmful bacteria, reduce the generation of harmful substance, and the balance of normal flora, improves in regulation intestines and stomach The immunity of body, it is ensured that health.
Specific embodiment
Following examples are only to further illustrate the present invention, and the invention is not limited in any way, involved in the present embodiment And strains lactobacillus acidophilus CGMCC1.1854, bacillus licheniformis CGMCC1.105 purchased from China General Microbiological strain protect Tibetan center, saccharomyces cerevisiae CICC1249 is purchased from Chinese industrial Culture Collection.
Embodiment 1
By lactobacillus acidophilus CGMCC1.1854, tri- kinds of bacterium of bacillus licheniformis CGMCC1.105, saccharomyces cerevisiae CICC1249 Strain is cultivated according to the order of inclined-plane, liquid seeds, liquid fermentation, and inclined-plane and liquid fermentation medium are respectively：Acidophilus breast Baccilus medium is MRS culture mediums, consisting of (W/V)：Peptone 1%, powdered beef 0.5%, dusty yeast 0.4%, glucose 2%, Tween 80 0.1%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, Triammonium citrate 0.2%, magnesium sulfate 0.02%, sulphur Sour manganese 0.005%, solid medium adds agar powder 1.5-2%, pH 6.2 or so；Saccharomyces cerevisiae culture medium is YPD culture mediums, Consisting of (W/V)：Glucose 2%, peptone 1%, yeast extract 0.5%, solid medium adds agar powder 1.5-2%, pH It is natural；Bacillus licheniformis culture medium is LB culture mediums, consisting of：Tryptone 1%, yeast extract 0.5%, sodium chloride 1%, solid medium adds agar powder 1.5-2%, pH7.4.Seed liquor is inoculated into fermentation medium according to 5% inoculum concentration In, at 30 DEG C, at 37 DEG C, 24-48h is cultivated in ventilation to saccharomycete so that the bacterium of every kind of bacterium for lactobacillus acidophilus and bacillus licheniformis Content >=109cfu/ml。
Three kinds of fermented liquids are pressed into percent by volume 20%:30%:50% mixing, obtains composite bacteria liquid, adds frozen-dried protective The formula of the agent freeze drying protectant includes：Skimmed milk, sucrose and FOS, wherein, in freeze drying protectant, skimmed milk is 10%, next sucrose 5%, FOS 10%, packing carry out pre-freeze 4 hours, last freeze-drying 36 hours with -80 DEG C .
Embodiment 2
By lactobacillus acidophilus CGMCC1.1854, tri- kinds of bacterium of bacillus licheniformis CGMCC1.105, saccharomyces cerevisiae CICC1249 Strain is cultivated according to the order of inclined-plane, liquid seeds, liquid fermentation, and inclined-plane and liquid fermentation medium are respectively：Acidophilus breast Baccilus medium is MRS culture mediums, consisting of (W/V)：Peptone 1%, powdered beef 0.5%, dusty yeast 0.4%, glucose 2%, Tween 80 0.1%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, Triammonium citrate 0.2%, magnesium sulfate 0.02%, sulphur Sour manganese 0.005%, solid medium adds agar powder 1.5-2%, pH 6.2 or so；Saccharomyces cerevisiae culture medium is YPD culture mediums, Consisting of (W/V)：Glucose 2%, peptone 1%, yeast extract 0.5%, solid medium adds agar powder 1.5-2%, pH It is natural；Bacillus licheniformis culture medium is LB culture mediums, consisting of：Tryptone 1%, yeast extract 0.5%, sodium chloride 1%, solid medium adds agar powder 1.5-2%, pH7.4.Seed liquor is inoculated into fermentation medium according to 5% inoculum concentration In, at 30 DEG C, at 37 DEG C, 24-48h is cultivated in ventilation to saccharomycete so that the bacterium of every kind of bacterium for lactobacillus acidophilus and bacillus licheniformis Content >=109cfu/ml。
Three kinds of fermented liquids are pressed into percent by volume 30%:30%:40% mixing, obtains composite bacteria liquid, adds frozen-dried protective The formula of the agent freeze drying protectant includes：Skimmed milk, sucrose and FOS, wherein, in freeze drying protectant, skimmed milk is 10%, next sucrose 5%, FOS 10%, packing carry out pre-freeze 4 hours, last freeze-drying 36 hours with -80 DEG C .
Embodiment 3
By lactobacillus acidophilus CGMCC1.1854, tri- kinds of bacterium of bacillus licheniformis CGMCC1.105, saccharomyces cerevisiae CICC1249 Strain is cultivated according to the order of inclined-plane, liquid seeds, liquid fermentation, and inclined-plane and liquid fermentation medium are respectively：Acidophilus breast Baccilus medium is MRS culture mediums, consisting of (W/V)：Peptone 1%, powdered beef 0.5%, dusty yeast 0.4%, glucose 2%, Tween 80 0.1%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, Triammonium citrate 0.2%, magnesium sulfate 0.02%, sulphur Sour manganese 0.005%, solid medium adds agar powder 1.5-2%, pH 6.2 or so；Saccharomyces cerevisiae culture medium is YPD culture mediums, Consisting of (W/V)：Glucose 2%, peptone 1%, yeast extract 0.5%, solid medium adds agar powder 1.5-2%, pH It is natural；Bacillus licheniformis culture medium is LB culture mediums, consisting of：Tryptone 1%, yeast extract 0.5%, sodium chloride 1%, solid medium adds agar powder 1.5-2%, pH7.4.Seed liquor is inoculated into fermentation medium according to 5% inoculum concentration In, at 30 DEG C, at 37 DEG C, 24-48h is cultivated in ventilation to saccharomycete so that the bacterium of every kind of bacterium for lactobacillus acidophilus and bacillus licheniformis Content >=109cfu/ml。
Three kinds of fermented liquids are pressed into percent by volume 50%:30%:20% mixing, obtains composite bacteria liquid, adds frozen-dried protective The formula of the agent freeze drying protectant includes：Skimmed milk, sucrose and FOS, wherein, in freeze drying protectant, skimmed milk is 10%, next sucrose 5%, FOS 10%, packing carry out pre-freeze 4 hours, last freeze-drying 36 hours with -80 DEG C .
Embodiment 4 is sold probiotic products and preserves stabilization according to composite probiotics preparations prepared by embodiment 1 with the market Property compares.Market product selects Sanzhu oral liquid, commercially available highly enriched probiotics bacterium powder respectively.Routinely accelerated test, if will produce Product are put 37~40 DEG C, are placed under the conditions of relative humidity 75% 3 months, after 12 months, determine bacterial content.Result shows institute of the present invention The compound probiotic of obtained vacuum freeze drying is by after long-term preservation, viable bacteria content is still very high, the results are shown in Table 1.
The probiotics keeping quality of table 1 is tested
Embodiment 5
Treatment and the recovery effects to gut flora that composite probiotics preparations are suffered from diarrhoea to Clinical Acute Infections.The experiment Completed in Zhejiang Province Lishui the People's Hospital, the acute infectious diarrhea for choosing in March, -2016 in June, 2015 in hospital therapy is suffered from Person 60, inclusive criteria：(1) there are the clinical symptoms such as diarrhoea, stomachache, the infectious diarrhea diagnosis basis People's Republic of China (PRC) defends What life portion promulgated《Infectious diarrhea diagnostic criteria》(WS271-2007) (2) go to a doctor within 48h after illness.(3) research is included It is preceding not take antibacterials and microecologic regulator is treated.(4) age ＞ 18 years old, ＜ 60 years old.Exclusion standard：(1) it is chronic Diarrhoea, noninfectious diarrhea.(2) other infectious diseases are merged.(3) disease of immune system, malignant tumour, serious conscience are merged The other organs functional disorder disease such as kidney.Patient is randomly divided into compound probiotic group (invention formulation), commercially available Infantile diarrhea Piece (Bifidobacterium lactobacillus live triple piece) is for control group 1 and does not use probiotic group for control group 2, every group 20, as a result It was found that diarrhoea status (table 2) can be effectively improved using compound probiotic group, and can fast quick-recovery gut flora (table 3).
The use probiotics of table 2 three groups of diarrhoea situations of patient after a day
Influence of the compound probiotic of table 3 to the main bacterial content of enteron aisle (1g CFU/g are wet just, x ± s, n=5)

Claims (7)

1. a kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating, are primarily characterized in that：Said preparation is by compound bacteria, oligosaccharide and freeze drying protectant Composition, lactobacillus acidophilus >=5 × 10 are contained in every 1g preparations7CFU, bacillus licheniformis >=5 × 107CFU, saccharomyces cerevisiae >=5 ×107CFU, freeze drying protectant 0.1-0.2g.
2. it is a kind of as claimed in claim 1 a kind of preparation method of the whole Enteral formulationses of Tiny ecosystem stomach invigorating, it is characterised in that including as follows Step：
Prepare compound bacteria bacterium solution：
Lactobacillus acidophilus, bacillus licheniformis and saccharomyces cerevisiae are inoculated into MRS culture mediums, LB culture mediums and YPD training respectively Fermented 24 hours in foster base, 35-40 DEG C of fermentation temperature, gained bacterial content >=5 × 108CFU/ml, is then mixed in proportion Close to obtain mixed bacteria liquid；
Will（a）The bacterium solution for being obtained adds freeze drying protectant, and ready bacterium solution is dispensed into lyophilized bottle, then carries out pre- It is cold；
Vacuum freeze drying, obtains finished product.
3. a kind of preparation method of whole Enteral formulationses of Tiny ecosystem stomach invigorating according to claim 2, it is characterised in that the step （a）Middle bacterial strain uses therefor is lactobacillus acidophilus CGMCC1.1854, bacillus licheniformis CGMCC1.105, saccharomyces cerevisiae CICC1249； Each bacterium accounts for cumulative volume ratio for 20-50% in mixed liquor.
4. a kind of preparation method of the whole Enteral formulationses of Tiny ecosystem stomach invigorating as described in claim 2, it is characterised in that the step （b）In, by percentage to the quality, the formula of the freeze drying protectant includes：10-20 % skimmed milk powers, 5-10% sucrose, 5-10% Oligosaccharide.
5. a kind of preparation method of the whole Enteral formulationses of Tiny ecosystem stomach invigorating as described in claim 2, it is characterised in that the step （b）In, pre-freeze 2-4 hours at -70~-80 DEG C.
6. a kind of preparation method of the whole Enteral formulationses of Tiny ecosystem stomach invigorating as claimed in claim 2, it is characterised in that the step （c）In, vacuum freeze drying 36 hours.
7. a kind of preparation method of the whole Enteral formulationses of Tiny ecosystem stomach invigorating as described in claim 4, it is characterised in that the step （b）In, oligosaccharide includes FOS, oligomeric maltose, xylo-oligosaccharide.
CN201710021903.1A 2017-01-12 2017-01-12 A kind of whole Enteral formulationses of Tiny ecosystem stomach invigorating and preparation method thereof Pending CN106727731A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107468721A (en) * 2017-09-07 2017-12-15 鄂尔多斯康医院 It is a kind of that there is raising immunity, the probiotics compound for adjusting stomach, weight losing function and its preparation method and application
CN107714738A (en) * 2017-09-07 2018-02-23 鄂尔多斯康医院 It is a kind of that there is probiotics compound for improving immunity, reducing blood lipid, the function that enhances metabolism and its preparation method and application

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101496822A (en) * 2009-02-18 2009-08-05 上海谱莱生物技术有限公司 Composite probiotics micro-ecological formulation and preparation method

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101496822A (en) * 2009-02-18 2009-08-05 上海谱莱生物技术有限公司 Composite probiotics micro-ecological formulation and preparation method

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107468721A (en) * 2017-09-07 2017-12-15 鄂尔多斯康医院 It is a kind of that there is raising immunity, the probiotics compound for adjusting stomach, weight losing function and its preparation method and application
CN107714738A (en) * 2017-09-07 2018-02-23 鄂尔多斯康医院 It is a kind of that there is probiotics compound for improving immunity, reducing blood lipid, the function that enhances metabolism and its preparation method and application

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