CN104642870A - Prebiotics composition - Google Patents

Abstract

The invention provides a composition for adjusting intestinal microecology of a human body. The composition comprises stachyose and galactooligosaccharides as active ingredients. The composition provided by the invention can be used for effectively adjusting the intestinal bacteria community function.

Description

A kind of prebiotic compositions

Technical field

The present invention relates to the composition for regulating human body intestinal microecology, particularly relating to and comprising stachyose and the galactooligosaccharide composition as effective active composition.

Background technology

Microbial cell number entrained by human body reaches 100,000,000,000,000 more than, and the symbiotic microorganism colony of these substantial amounts is mainly distributed in the skin of human body, oral cavity and intestines and stomach, and wherein enteron aisle is the position that microorganism is concentrated the most.Experiment confirms, gut flora serves vital effect to the nutrition of host, metabolism and immunologic function, very important to maintenance health, is realize the indispensable part of host's normal physiological function.

In human body microecosystem, intestinal microecology is main, most active, has significant impact to health.Intestinal microecology system refers to by the biotic components such as gut flora and food source property abiotic component (not digested food) and the ecosystem being positioned at the local of enteron aisle that jointly forms from the secretion (as hormone, enzyme, mucus and cholate etc.) of stomach, intestines, pancreas and liver.

Research shows, after neonate leaves parent, skin, tracheae and alimentary canal etc. just start to grow a large amount of bacteriums on the surface.The meconium of neonate's first time excretion is aseptic, just finds that there is bacterium, bacterial number sharp increase after lactation after 3 ~ 4 hours.After neonate is born in 3 ~ 4 days, in ight soil, just there will be Bifidobacterium, become dominant bacteria to when the 5th day.Along with growing of baby, a kind of metastable state can be reached to the composition of gut flora during ablactation.Under normal circumstances, this metastable gut flora forms the growth of meeting along with human body, until the manhood.And after the senescence phase, gut flora composition can have greatly changed, the bifidobacteria in ight soil greatly reduces, and bacteroid and clostridium quantity then significantly increase.

Gut flora and human body establish close relationship in very long evolution, are therebetween not only to have opposed but also unified unity of contradictions body.Through long-term natural selection process, gut flora and human body reach the metastable dynamic balance state that a kind of mutual reciprocity and mutual benefit restrict again mutually.Human body provides living space and energy source for flora, and flora then helps human body to synthesize some nutrients and directly or the physiological function of remote-effects human body.

Great many of experiments confirms, gut flora is formed the immune response of the normal metabolic activity of human body, nutrient absorption, epithelial Growth and Differentiation, normal bowel mucous membrane and the aspect such as maintenance plays very important effect, simultaneously they and some disease also have and contact closely, comprise the lienteric diseases such as abdominal distension stomachache, IBD, cancer, Multiple Organ's disorder, allergy etc.

Gut flora not only affects the absorption and digestion of food, also can have influence on absorption and the metabolism of oral drugs.Therefore, gut flora all can have an impact to the metabolism of medicine and result for the treatment of, may be also that drug therapy produces one of reason of individual difference.

Human body intestinal canal flora can be divided into according to effect classification: probio, conditioned pathogen and pathogenic bacteria.(1) namely probio, as the term suggests be the bacterium useful to health.Probio is indispensable part in human body intestinal canal micro-ecological environment, is enteron aisle dominant microflora, has the effect such as nutrition and immunological regulation in healthy human body.The main probio of human body comprises Bacillus acidi lactici, Bifidobacterium etc.; (2) conditioned pathogen, refers to when intestinal microecology balances harmless, but has again an invasive class bacterium under given conditions, is the non-dominant microflora of enteron aisle, as enterococcus, enterobacteria etc. in healthy human body; (3) pathogenic bacteria, refer to harmful, can cause the bacterium that human pathology reacts.Pathogenic bacteria are mostly the bacterium that passes by one's way, and the chance being colonizated in enteron aisle is for a long time fewer, in host internal memory residual hour, a couple of days or several weeks, as proteus, pseudomonad and C.perfringens etc.

Human body intestinal canal flora can be divided into according to aerobic situation: anaerobic bacteria, facultative anaerobe and aerobic bacteria.Wherein obligate anaerobe accounts for more than 99%, and only bacteroid and Bifidobacterium just account for more than 90% of total number of bacteria.Because hydrochloric acid in gastric juice, bile effect and intestinal fluid flow are large, it is fast to wriggle, thus the kind of Stomach duodenum, jejunum bacterium and quantity few, be mainly gram-positive aerobic bacteria.Terminal ileum is because intestinal juice flow is few, wriggling slows down, and bacterial population increases gradually, mainly containing Bacillus acidi lactici, EHEC, bacteroid and clostridium etc.To colon, bacterial population obviously increases, and defines the bacterium colony group that microbe density is the highest, and wherein great majority are anaerobic bacteria, are mainly Bifidobacterium, bacteroid and Bacillus acidi lactici etc.

Some scholars think, the effect of probio may be summarized to be: control diarrhoea, the lactose intolerance symptom alleviated, prevention vagina infection, strengthen body immunity, alleviate irritated, reduce serum cholesterol, pre-anti-cancer and Tumor suppression growth.Prevent the effect of bone loss and osteoporosis in addition in addition.

Probio has been widely known by the people for the health promotion effect of human body.Therefore corresponding product also slowly more gets up.Product in the market for human body intestinal canal Tiny ecosystem mainly contains three classes: probio series products, prebiotics series products and the product being called " symphysis unit " class combined by the former two.

" probio series products " and before described this concept of human body intestinal canal probio are distinguishing.Human body intestinal canal probio belongs to ancestral home bacterium, is through human body and the long-term natural selection process of microbial body is finally recognized each other and is colonizated in the bacterium general name in human body intestinal canal for a long time.And " probio series products " belongs to foreign nationality bacterium, be people be the bacterial classification similar with human body intestinal canal probio joined in body.Only have those can be survived by the test of gastrointestinal tract environment and the bacterial strain of successful field planting could really be converted into " human body intestinal canal probio ".This wherein also will through the immune system cleaning of human body and the Reverse transcriptase of enteron aisle ancestral home bacterium own, and therefore, success rate is very low.

At present, Food and Drug Administration (FDA) thinks that safe probio has reached 40 kinds more than, along with the development of science and technology, the going deep into of research, also can get more and more.According to report, human body intestinal canal bacterium comprises more than 200 kind of Pseudomonas, more than totally 800 bacterial classifications, and bacterial strain kind then reaches more than 7000 and plants.There is following worry for additional probio in academia: in the face of so various flora system always, merely supplement the effect that several probio can play and the allergic reaction that whether can cause human body, and external source probio can no because production accumulating condition change and morph and become pathogenic bacteria, etc.With regard to security standpoint, probiotics preparation is directly used to have certain risk.

Prebiotics (Prebiotics) refers to the material that optionally can promote original beneficial bacteria (probio) growth and breeding in host intestine.Increasing by beneficial bacterium, suppresses harmful bacterial growth, thus reaches adjustment gut flora, promotes the object of body health.It has sharp in vivo proliferation of probiotics, suppresses harmful bacteria, the health benefit of activated cell vigor, enhancing human immunity vigor.

The current research to prebiotics and use mainly concentrate on functional oligose.Functional oligose generally has following characteristics: 1, be by 2 ~ 10 identical or different monose, (can be straight chain, also can be side chain) that be polymerized with glycosidic bond; 2, there is the characteristic of carbohydrate, can directly as food ingredient, but not by human body alimentary canal enzyme and gastric acid degradation, not by (or difficult quilt) intestinal absorption; 3, there is physiological functions such as promoting human body Bifidobacterium propagation.Functional oligose comprises FOS, galactooligosaccharide, oligoisomaltose, xylo-oligosaccharide, soyabean oligosaccharides, Oligomeric manna sugar, stachyose, raffinose etc.

Functional oligose is fermented by intestinal beneficial bacterium, produces SCFA, and wherein acetic acid, butyric acid account for 95% of total product acid amount.The generation of SCFA can reduce intestinal pH, suppresses the growth and breeding of pathogenic entero becteria and conditioned pathogen further; Regulate body water-electrolyte balance, promote calcium uptake; Suppress the propagation of enterocyte, differentiation, the synthesis of transfer and DNA and methylation level, play antineoplastic action.

" symphysis unit " series products is that " probio " series products combines with " prebiotics " series products, because it is containing profitable probliotics, so there is no the safety issue worry solving " probio " series products.

" prebiotics " series products compares that " probio " series products character is more stable, quality controllability is stronger, and play " prebiotic " effect by propagation ancestral home bacterium, do not worry activity problems and the variation problem of artificial foreign nationality bacterium of adding, therefore obtain the trust of more scholars and even consumer.On January 16th, 2007, the Public nutrition that OLIGO (compound sugar) project is put into China improves project, becomes the another Public nutrition improvement project that China starts after implementing the measure such as " supplementation of iodine salt ", " enriched nutritive flour ".This describes the extensive market of " prebiotics " series products on the one hand, also illustrate that the accreditation of industry to " prebiotics " series products on the other hand.

In recent years, the various composition comprising prebiotics is proposed.

Such as, patent document 1 (CN102488100A) discloses a kind of feed addictive containing composite prebiotics, and its raw material components is cooked corn skin, corn steep liquor, oligoisomaltose, Oligomeric manna sugar, galactooligosaccharide, saccharomyces boulardii liquid, Bacillus licheniformis liquid and Bacillus acidi lactici bacterium liquid.In described feed addictive, the survival bacterium amount of probio is high, has the advantages that safety, environmentally degradable and cost are low.

Patent document 2 (CN101263841A) discloses a kind of functional health care dairy products, it is characterized in that with the addition of function compound sugar, comprise Oligomeric manna sugar, xylo-oligosaccharide, FOS, galactooligosaccharide, stachyose, raffinose, oligoisomaltose, oligomeric maltose, soyabean oligosaccharides, chitooligosaccharide-, lactosucrose, oligomeric dragon gallbladder sugar, particularly Oligomeric manna sugar, xylo-oligosaccharide, FOS can be wherein a kind of or more than one mixture.

Patent document 3 (CN1731938A) discloses a kind of nutrition or the pharmaceutical composition that contain fructose oligosaccharides (FOS) and GOS (GOS).

Patent document 4 (TWI337079B) discloses the composition (claim 1) of numerous compound sugar such as methyl α manna oligosacchride, galactooligosacchari(es, FOS, chitosan oligosaccharide and OPC.

In above-mentioned patent document 1, because feed addictive is for animal, different with the range of application of the food of people; In addition, the object of the technical scheme of document 1 is to make the survival bacterium amount of probio in product high, and human internal environment and external environment have the difference of essence, and the probability that the bacterium of Motility also survives in vivo is not high; In addition, in composition of the present invention, do not add external source bacterium, do not add the material such as maize peel, corn steep liquor yet.In addition, above-mentioned document 1 ~ 4 does not all disclose the specific composition of the present invention.

Therefore, although have a lot about the research of various compound sugar, most of research direction is all confined to single compound sugar functionally.Few people carry out deep research for compound sugar combination.Due to the system that human body intestinal canal Tiny ecosystem is complicated, wherein comprised bacterial species is various, and various bacterium is have specific for the utilization of different compound sugar, and its result causes single compound sugar often can only to part proliferation of probiotics.And this unbalanced propagation is actually the poised state changing the intestinal microecology of human body own.The change of this poised state does not have risk also not have enough evidences to show at present.According to bibliographical information, the functional oligose realizing the application of suitability for industrialized production at present more has FOS, oligoisomaltose, xylo-oligosaccharide, galactooligosaccharide, soyabean oligosaccharides, stachyose, raffinose, Oligomeric manna sugar etc.Wherein the consumption of FOS, oligoisomaltose, galactooligosaccharide is bigger than normal; The consumption of xylo-oligosaccharide is little, but its to probios such as such as bifidobacterium bifidum, bifidobacterium breve, lactobacillus acidophilus, Lactobacillus caseis without proliferation function; Soyabean oligosaccharides, stachyose, raffinose have aerogenesis effect according to report; Oligomeric manna sugar has its propagation probiotic effect of bibliographical information imprecise.Generally speaking, each compound sugar in use has respective feature, also all there is certain limitation.

The research of prior art to stachyose is less, and about the combination research of stachyose and galactooligosaccharide, there is not yet relevant report at home and abroad at present.

Inventor herein by conducting in-depth research the combination of stachyose and galactooligosaccharide, and then achieves the experimental result of making us unexpected, surpassing the expectation.

Summary of the invention

The present inventor is surprisingly found out that: by combinationally using galactooligosaccharide and stachyose, there is provided extensively breed probio, suppress pathogenic bacteria, stable condition pathogenic bacteria, and then balanced human's intestinal microecology and consumption is little, the composition safely and effectively of taking convenience.The combination of galactooligosaccharide and stachyose significantly improves prebiotics characteristic, demonstrates cooperative effect.Further preferably, the combination of galactooligosaccharide, stachyose and Oligomeric manna sugar demonstrates the prebiotics characteristic raising effect improved further.

Instant invention overcomes the deficiencies in the prior art, provide and extensively can breed probio, suppression/eliminating pathogenic bacteria, stable condition pathogenic bacteria, correct flora imbalance, balanced human's intestine microenvironment, and then eliminate the sub-health state such as such as abdominal distension, diarrhoea, indigestion and associated conditions that cause because of human body intestinal canal Dysbiosis and consumption is little, taking convenience, safely and effectively composition.

Technical solution of the present invention comprises:

1, a kind ofly to it is characterized in that for regulating the composition of human body intestinal microecology, comprising stachyose and galactooligosaccharide as effective active composition.

2, the composition as described in item 1, it also comprises Oligomeric manna sugar and/or xylo-oligosaccharide.

3, the composition as described in item 1, wherein, the weight ratio of described stachyose and galactooligosaccharide is 1:10 ~ 10:1, is preferably 1:1 ~ 5:1, or is preferably 1:5 ~ 1:1.

4, the composition according to any one of item 1 ~ 3, wherein, described galactooligosaccharide in the oligomeric disaccharides of gala to oligomeric eight sugar content >=57% (w/w) of gala, preferably >=70% (w/w), more preferably >=90% (w/w), further preferably >=99% (w/w), and lactose Han Liang≤23% (w/w), You Xuan≤20% (w/w), Geng You Xuan≤10% (w/w), and glucose Han Liang≤22% (w/w), You Xuan≤10% (w/w), Geng You Xuan≤5% (w/w).

5, the composition as described in item 2, wherein, in described Oligomeric manna sugar, mannobiose is to content >=85% (w/w), preferably >=90% (w/w) of sweet dew ten sugar, and mannobiose is to content >=50% (w/w) of sweet dew six sugar.

6, the composition as described in item 2, wherein, described xylo-oligosaccharide is in xylobiose extremely wooden seven sugar content >=70% (w/w), preferably >=80% (w/w), more preferably >=90% (w/w), and xylobiose is to content >=50% (w/w), preferably >=60% (w/w) of Xylotetrose.

7, the composition according to any one of item 1 ~ 6, wherein, content of stachyose >=60% (w/w), preferably >=70% (w/w), more preferably >=80% (w/w).

8, the composition according to any one of item 1 ~ 7, wherein, also comprises carbohydrate, such as chitosan oligosaccharide, oligoisomaltose, yeast beta-dextran, inulin, resistant dextrin, cottonseed compound sugar.

9, the composition according to any one of item 1 ~ 7, wherein, also comprises auxiliary material and/or food materials.

Item 10, a kind of food, health food or pharmaceutical preparation, containing, for example the composition according to any one of item 1 ~ 9, described food, health food or pharmaceutical preparation are preferably solid, semisolid or liquid form.

11, the composition according to any one of item 1 ~ 9 is for the preparation of the purposes regulated in the food of human body intestinal microecology, health food or pharmaceutical preparation.

The invention has the beneficial effects as follows: (1) present composition can make prebiotic index (Prebiotic Index in given scope, PI) be significantly improved, demonstrate obvious synergy, regulating intestinal canal flora function is apparently higher than single compound sugar, consumption is little, can realize better probiotic effects when taking in heat and being low; (2) gas-forming reaction of the present composition significantly reduces compared with the stachyose single dose of Isodose; (3) bacterial classification that covers of present composition propagation probio more complete, suppress/get rid of the bacterial classification of pathogenic bacteria also more; (4) present composition all demonstrates obvious effect for the symptom improvement of abdominal distension and diarrhea group, shows that the present composition has dual regulation to gastrointestinal function.

Detailed description of the invention

Namely " regulate human body intestinal microecology " described in this description is by regulating intestinal canal flora, makes the poised state that human body intestinal canal micro-ecological environment reaches healthy, and then reveals various health benefits to body.

Described in this description, " stachyose " is: have the implication that it is general in the art, and it can be synthesis, also can obtain from natural origin processing.The stachyose that preferred natural origin processing obtains.It represents the tetrose being combined two α-galactolipin formation by the glucosyl group side of sucrose with α-1,6 glycosidic bond." stachyose " herein also comprises the mixture that principal component is stachyose.Preferably, principal component is in the mixture of stachyose, content of stachyose >=60% (w/w), preferably >=70% (w/w), more preferably >=80% (w/w).

" galactooligosaccharide " described in this description has its implication general in the art, and it can be synthesis, also can obtain from natural origin processing.The galactooligosaccharide that preferred natural origin processing obtains.It represents based on lactose molecule in conjunction with the galactooligosaccharide containing 2 ~ 10 (preferably 2 ~ 8) monosaccharide molecule that galactose molecule generates." galactooligosaccharide " herein also comprises the mixture that principal component is galactooligosaccharide.Preferably, principal component is in the mixture of galactooligosaccharide, the oligomeric disaccharides of gala is to oligomeric eight sugared content >=57% (w/w) of gala, preferably >=70% (w/w), more preferably >=90% (w/w), further preferably >=99% (w/w), and lactose Han Liang≤23% (w/w), You Xuan≤20% (w/w), Geng You Xuan≤10% (w/w), and glucose Han Liang≤22% (w/w), You Xuan≤10% (w/w), Geng You Xuan≤5% (w/w).

" Oligomeric manna sugar " described in this description has its implication general in the art, and it can be synthesis, also can obtain from natural origin processing.The Oligomeric manna sugar that preferred natural origin processing obtains.It represents the polymer containing 2 ~ 10 (preferably 2 ~ 6) mannose molecules." Oligomeric manna sugar " herein also comprises the mixture that principal component is Oligomeric manna sugar.Preferably, principal component is in the mixture of Oligomeric manna sugar, and mannobiose is to content >=85% (w/w), preferably >=90% (w/w) of sweet dew ten sugar, and mannobiose is to content >=50% (w/w) of sweet dew six sugar.)

" xylo-oligosaccharide " described in this description has its implication general in the art, and it can be synthesis, also can obtain from natural origin processing.The xylo-oligosaccharide that preferred natural origin processing obtains.It represents the compound sugar be combined into by 2 ~ 10 wood sugar molecules.Preferably connected with β-Isosorbide-5-Nitrae-glycosidic bond by 2 ~ 7 wood sugar molecules, and with the xylo-oligosaccharide mixture that xylobiose, xylotriose, Xylotetrose are main component." xylo-oligosaccharide " herein also comprises the mixture that principal component is xylo-oligosaccharide.Preferably, principal component is in the mixture of xylo-oligosaccharide, xylobiose is to wood seven sugared content >=70% (w/w), preferably >=80% (w/w), more preferably >=90% (w/w), and xylobiose is to content >=50% (w/w), preferably >=60% (w/w) of Xylotetrose.

In composition of the present invention, the weight ratio of stachyose and galactooligosaccharide is preferably 1:10 ~ 10:1, is more preferably 1:5 ~ 5:1.In composition of the present invention, except stachyose and galactooligosaccharide are as except effective active composition, preferably, also comprise Oligomeric manna sugar and/or xylo-oligosaccharide.Preferably, when composition of the present invention comprises stachyose, galactooligosaccharide, Oligomeric manna sugar, the weight ratio of Oligomeric manna sugar/stachyose preferably in the scope of 1:5 ~ 5:1, more preferably in the scope of 1:5 ~ 1:1.

The auxiliary material that described composition of the present invention comprises and food materials are conventional those in food and drug world, such as: glidant, pH adjusting agent, disintegrant, starch, resistant starch, dextrin, resistant dextrin, the Chinese medicine belonging to integration of drinking and medicinal herbs, flavoring essence, flavoring.

Embodiment

Below in conjunction with embodiment, the present invention is illustrated further, but the present invention is not limited thereto.

Product embodiments 1:

Stachyose and galactooligosaccharide cross 100 mesh sieves respectively, take 1000 grams of stachyoses, 3000 grams of galactooligosaccharides, are distributed into 1000 bags after mixing, preferably aluminum-plastic packaged.

Product embodiments 2:

Take 3000 grams of stachyoses, 1000 grams of galactooligosaccharides, after mixing with appropriate dextrin, to be packaged into 1000 bags after 85% alcohol granulation, oven dry, preferably aluminum-plastic packaged.

Product embodiments 3:

Take 1 gram of stachyose, 10 grams of galactooligosaccharides, join after dissolving with a small amount of distilled water in the taste of various cream.

Product embodiments 4:

Take 5000 grams of stachyoses, 1000 grams of galactooligosaccharides, respectively with distilled water 10000 milliliters, 5000 milliliters be mixed with solution after mix and be mixed with 15000 ml solns with distilled water after adjusted volume to 15000 milliliter or first mixing, to filter, filling one-tenth 1000 oral liquids.

Product embodiments 5:

Take 1000 grams of stachyoses, 5000 grams of galactooligosaccharides, 1000 grams of Oligomeric manna sugars, after dissolving with 15000 ml distilled waters, be concentrated into syrup form, be distributed into 1000 parts, preferably aluminum-plastic packaged.

Product embodiments 6:

1000 grams of stachyoses, 600 grams of galactooligosaccharides, 500 grams of Oligomeric manna sugars, anhydrous citric acid 680 grams, sodium acid carbonate 500 grams, microcrystalline cellulose 100 grams, PEG6000110 gram, dolomol 10 grams is taken after raw material drying, mix, direct powder compression, makes 1000.

Testing example:

One, materials and methods

(1) in the present invention, compound sugar source is respectively:

Stachyose: commercially available prod, content of stachyose 80%.

Galactooligosaccharide: commercially available prod, galactooligosaccharide in the oligomeric disaccharides of gala to the oligomeric eight sugar content 90% of gala.

Oligomeric manna sugar: commercially available prod, Oligomeric manna sugar in mannobiose to sweet dew ten sugar content 90%.

Xylo-oligosaccharide: commercially available prod, xylo-oligosaccharide is in xylobiose extremely wooden seven sugar content 95%.

FOS: commercially available prod, the percentage compositions that FOS accounts for dry with ketose, fruit fruit trisaccharide, Nystose, fruit fruit tetrose, GF4, fruit fruit pentasaccharides, sugarcane fruit six sugar, fruit fruit six sugared sums count 96%.

Above-mentioned compound sugar obtains by domestic market buying, is powdery product, meets state food, new resource food/new raw-food material requirement.

(2) analytical standard

1, the qualitative judgement standard for regulating intestinal canal flora function positive findings in " health food inspection and assessment technical specification " (version in 2003) is:

A, zoopery

The situation of change of Bifidobacterium, lactobacillus, enterococcus, enterobacteria, C.perfringens between self and group before and after comparative experiments, before and after experimental group experiment, self comparing difference has conspicuousness, or experiment after between experimental group and control group group comparing difference have conspicuousness and experimental group test front and back self comparing difference have conspicuousness, meet following any one, can judge that this given the test agent results of animal is positive.

1) in ight soil, Bifidobacterium and/or lactobacillus obviously increase, and C.perfringens reduces or do not increase, and enterobacteria, enterococcus are without significant change.

2) in ight soil, Bifidobacterium and/or lactobacillus obviously increase, and C.perfringens reduces or do not increase, and enterobacteria and/or enterococcus obviously increase, but the amplitude that the amplitude increased increases lower than Bifidobacterium/lactobacillus.

B, human experiment are tested

Meet following any one, and self to compare before and after test group test-meal and after test-meal, test-meal group compares with control group, difference all has conspicuousness, can judge that this given the test agent has the effect of regulating intestinal canal flora function.

1) in ight soil, Bifidobacterium and/or lactobacillus obviously increase, and C.perfringens reduces or do not increase, and enterobacteria, enterococcus, bacteroid are without significant change.

2) in ight soil, Bifidobacterium and/or lactobacillus obviously increase, and C.perfringens reduces or do not increase, and enterobacteria and/or enterococcus, bacteroid obviously increase, but the amplitude that the amplitude increased increases lower than Bifidobacterium/lactobacillus.

2, people (the Development of a quantitative tool for thecomparison of the prebiotic effect of dietary oligosaccharides.Letters inApplied Microbiology 2003 such as R.Palframan, 37,281-284) propose the index of quantitative analysis prebiotic function: prebiotic index (Prebiotic Index, PI).The present inventor, according to the existing standard judged for regulating intestinal canal flora function of China, under the prerequisite additionally not increasing experiment burden, devises zoopery PI computational methods and human experiment experiment PI computational methods that applicable China uses.Be respectively:

Animal PI=(two/total)+(breast/total)-(shuttle/total)-(bar/total)-(ball/total)

Bifidobacterium number, lactobacillus number, C.perfringens number, enterobacteria number and enterococcus number after wherein " two ", " breast ", " shuttle ", " bar ", " ball " represent experiment respectively test with its each corresponding bacterium before the ratio of bacterium number; " always " represents the ratio of the front bacterium number sum of the above-mentioned 5 kinds of bacterium bacterium number sums after experiment and experiment.

People PI=(two/total)+(breast/total)-(shuttle/total)-(bar/total)-(ball/total)-(intend/total)

Bifidobacterium number after wherein " two ", " breast ", " shuttle ", " bar ", " ball ", " plan " represent experiment respectively, lactobacillus number, C.perfringens number, enterobacteria number, enterococcus number and bacteroid test with its each corresponding bacterium before the ratio of bacterium number; " always " represents the ratio of the front bacterium number sum of the above-mentioned 6 kinds of bacterium bacterium number sums after experiment and experiment.

Two, zoopery

1, animal used as test

SPF inbred strais BALB/c male mice, 18-22g, often organizes 10.

2, grouping and given the test agent give the time

Test group and blank group are established in experiment, and given the test agent gives 14 days time.

3, experimental procedure

Before giving given the test agent, asepticly take stool in mice 0.1g, 10 times of serial dilutions, select suitable dilution factor to be seeded in respectively on each culture medium.After cultivation, with qualification counting bacterium colonies such as colonial morphology, gram stain microscopy, biochemical reactions, calculate every gram of bacterium number wet just, after taking the logarithm, carry out statistical disposition.24h after giving given the test agent for the last time, with the same manner cut-off intestines ight soil before experiment, detect gut flora, method is the same.

4, data processing:

Data variance analysis, first carries out homogeneity test of variance by the program of variance analysis, and variance is neat, calculates F value, F value <0.05, conclusion: no significant difference between each group mean; F value >=0.05, P≤0.05, adds up with the comparative approach between two of mean between multiple experimental group and a control group; Suitable variable transitions is carried out to the data of abnormal or heterogeneity of variance, after meeting normal state or variance and requiring together, adds up by the data after conversion; If do not reach normal state or the neat object of variance after variable transitions yet, use rank test instead and add up.

5, experimental result and analysis

5.1 experiments (1)

It is that 0.1g/kg, 0.1g/kg, 0.4g/kg give mouse that stachyose, Oligomeric manna sugar amount to into mouse daily dose with document for minimum day effective dose (0.5g, 0.5g and 2g) that people reports with galactooligosaccharide.A+B group daily dose is stachyose 0.1g/kg+ galactooligosaccharide 0.4g/kg; B+C group daily dose is galactooligosaccharide 0.4g/kg+ Oligomeric manna sugar 0.1g/kg; A+B+C group daily dose is stachyose 0.1g/kg+ galactooligosaccharide 0.4g/kg+ Oligomeric manna sugar 0.1g/kg.Each tested material is airtight stored refrigerated in 4 DEG C after dissolving preparation with sterile distilled water, and the holding time is no more than 5 days, takes out and be placed to room temperature before gavage prerequisite.Mouse stomach volume is 0.1ml/10g.Each group of mouse respectively continuously gavage give measurement result after 14 days.Blank group gavage gives the SPSS of respective volume.

[table 1]

(self compare and test before and after experimental group experiment and compare with control group afterwards, "-" represents without significant change; "+" representative has significant difference, P<0.05; " ++ " representative has pole significant difference, P<0.01; )

Experimental result shows: when stachyose, galactooligosaccharide and Oligomeric manna sugar individually use, and does not demonstrate the proliferation function to probio under low dosage, and regulating intestinal canal flora function is judged to be feminine gender; And the composition of same stachyose+galactooligosaccharide under same dose demonstrates significant prebiotics function, regulating intestinal canal flora function is judged to be the positive, A+B demonstrates significant difference (P<0.05), and the composition of Oligomeric manna sugar+galactooligosaccharide under same dose fails to make a significant impact on prebiotic index.Imply that the composition of stachyose+galactooligosaccharide creates cooperative effect.The composition cooperative effect of stachyose+galactooligosaccharide+Oligomeric manna sugar is stronger.

5.2 experiments (2)

All mouse is given with the dosage of 0.6g/kg every day (amounting to adult body dosage 3g/d) with experiment (1) described method by stachyose, Oligomeric manna sugar, galactooligosaccharide and their combination (composition proportioning is with experiment (1)).Give measurement result after 14 days continuously.

[table 2]

(self compare and test before and after experimental group experiment and compare with control group afterwards, "-" represents without significant change; "+" representative has significant difference, P<0.05; " ++ " representative has pole significant difference, P<0.01; " ± " representative has effect tendency, but there was no significant difference).

Experimental result shows: although demonstrate certain prebiotics function when each component is used alone under the dosage of 0.6g/kg/d, unbalanced to the propagation of Bifidobacterium, lactobacillus and the inhibitory action of C.perfringens.And the composition of stachyose+Oligomeric manna sugar+galactooligosaccharide is balanced and strong to the effect that Bifidobacterium, lactobacillus and C.perfringens produce.And the more each component of animal PI value of the composition group animal PI value that is used alone group all has and significantly improves.This result further demonstrates the synergy of combinations thereof.

We test the composition (3:1 of stachyose and galactooligosaccharide different ratio under the dosage of 0.6g/kg every day in addition, 1:1,1:3,1:5,1:7) impact on prebiotic index, method of testing is with experiment (1) described method.It is positive that result shows that each composition all shows as regulating intestinal canal flora function, wherein animal PI value is the highest in the composition group of 1:3 and 1:5 two ratios, but between statistical analysis is respectively organized, compare there was no significant difference (P>0.05).

For investigating the synergy of said composition further, we test the impact of each independent component when increasing dosage on prebiotic index in addition.Suitable for animal PI value during 0.6g/kg/d with the stachyose+Oligomeric manna sugar shown in table 1+galactooligosaccharide composition (0.1+0.1+0.4g/kg/d) when the dosage that result shows when stachyose and galactooligosaccharide are used alone brings up to 0.8g/kg/d and 1.2g/kg/d respectively, and same effect will be reached when Oligomeric manna sugar is used alone need consumption more.Result illustrate by combination obviously can reduce taking dose, from economy or practicality aspect all significant.

5.3 experiments (3)

All mouse is given continuously 14 days with the dosage of 0.6g/kg every day (wherein A+B assembly ratio is for 1:4, A+B+C assembly ratio is for 1:4:1) with testing (1) described method by stachyose, Oligomeric manna sugar, galactooligosaccharide and their composition.Often organize mouse quantity and be increased to 20, pour into salmonella typhimurium in addition more respectively giving within 7 days, respectively to organize mouse afterwards continuously, bacterial concentration is 1 × 10 ⁸cFU/ml, 0.2ml/ only, continuous 2 days, continue observation 5 days, record dead mouse situation and diarrhoea situation.

[table 3]

Experimental result shows, diarrhoea all appears in blank group, wherein has 13 to occur dead; The Incidence of Diarrhea of each compound sugar group and the death rate all have and reduce in various degree, and stachyose+galactooligosaccharide composition group reduces amplitude and is obviously greater than each independent compound sugar group, and stachyose+galactooligosaccharide+Oligomeric manna sugar composition group effect is optimum.Imply that said composition have suppression/removing pathogenic bacteria, prevention bacterial diarrhea and mitigation symptoms, minimizing death effect.

Three, human feeding trial

1, experimenter's inclusive criteria

Gastrointestinal disease person was not suffered from one month; Antibiotic person was not taken in one month.

2, experimental design and grouping requirement

Adopt two kinds of control design between self and group.Be divided into test-meal and control group at random by the flora situation of experimenter, consider that the principal element affecting result is as age, sex, dietary factor etc. as far as possible, carry out harmony inspection, to ensure the comparativity between group.Often organize experimenter 50 example.

3, the dosage of given the test agent and using method

Everyone daily this composition 3g of test-meal group, is dissolved in this product when taking in 100 milliliters of warm water, a clothes.Control group adopts blank.Given the test agent gives 14 days time.Duration of test does not change original eating habit, normal diet.

4, observation index

Bifidobacterium, lactobacillus, enterococcus, enterobacteria, bacteroid, C.perfringens, aerogenesis situation etc.

5, test procedure

Before giving given the test agent, asepticly take experimenter's ight soil 1.0g, 10 times of serial dilutions, select suitable dilution factor to be seeded in respectively on each culture medium.After cultivation, with qualification counting bacterium colonies such as colonial morphology, gram stain microscopy, biochemical reactions, calculate every gram of bacterium number wet just, after taking the logarithm, carry out statistical disposition.24hr after giving given the test agent for the last time, uses the same method and again detects.120hr after giving given the test agent for the last time, uses the same method and again detects.

6, data processing

Test data is measurement data, analyzes with t inspection.Own control data adopts paired t-test, two groups of means compare employing independent samples t-test, and the latter carries out homogeneity test of variance, carry out suitable variable transitions to the data of Non-Gaussian Distribution or heterogeneity of variance, after meeting normal state variance and be neat, carry out t inspection by the data of conversion; If translation data still can not meet the neat requirement of normal state variance, use t ' inspection or rank test instead; To the logging data application rank test of the coefficient of variation too large (as CV>50%).

7, experimental result and analysis

The composition of the composition of stachyose and stachyose+galactooligosaccharide+Oligomeric manna sugar and FOS+galactooligosaccharide is all carried out human experiment experiment with the dosage of 3g every day (composition is equal proportion mixing) according to " the regulating intestinal canal flora functional check method " in " health food inspection and assessment technical specification " (version in 2003) function assessment evaluation test method.Give measurement result after 14 days continuously.

[table 4] human experiment tests measurement result after 14 days

(self compare before and after test group test-meal and compare with control group after test-meal, "-" represents without significant change; "+" representative has significant difference, P<0.05; " ++ " representative has pole significant difference, P<0.01; " ± " representative has effect tendency, but there was no significant difference)

Experimental result shows: the composition of stachyose and stachyose+galactooligosaccharide+Oligomeric manna sugar three all has regulating intestinal canal flora function.And the effect that stachyose+galactooligosaccharide+Oligomeric manna sugar composition shows is stronger.And be obviously better than the effect of composition under the dosage of 3g/d of FOS+galactooligosaccharide.In addition, the person of participating in the experiment of stachyose group 92% reports that aerogenesis increases, and reports in composition experimental group that the number that aerogenesis increases only has 28%, considerably reduces the side effect of stachyose aerogenesis.

[table 5] human experiment tests measurement result after 5 days

(self compare before and after test group test-meal and compare with control group after test-meal, "-" represents without significant change; "+" representative has significant difference, P<0.05; " ± " representative has effect tendency, but there was no significant difference).

Experimental result shows: after stachyose withdraws 5 days (120hr), probio quantity falls after rise fast, and stachyose+galactooligosaccharide+Oligomeric manna sugar composition group also falls after rise to some extent, but decrease speed is considerably slower than stachyose group.The composition group of FOS+galactooligosaccharide has got back to original state after (120hr) in 5 days withdrawing.In addition, stachyose group is withdrawn and within 5 days, is still had the person of participating in the experiment of 32% to report that aerogenesis increases after (120hr), reports that the number that aerogenesis increases is 22% in stachyose+galactooligosaccharide+Oligomeric manna sugar composition experimental group.

Other human experiment experiment uses separately this composition 3g/d (stachyose+galactooligosaccharide 1:1) continuously to 20 routine acute diarrhea crowds, and wherein 15 examples are at the same day and rapid relief of symptoms, and defecation frequency obviously reduces.To 30 routine constipation old men use continuously this composition 3g/d (stachyose+galactooligosaccharide 1:2) after 3 days defecation frequency and difficult defecation degree all obviously improve.Show that the present composition has dual regulation to gastrointestinal function.

Four, experiment in vitro

By in vitro test qualitative test under same dose (2% (w/v)) stachyose and the composition of stachyose+galactooligosaccharide+Oligomeric manna sugar three and the composition of stachyose+both FOSs for the impact of the probios such as bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium infantis, bifidobacterium breve, bifidobacterium bifidum, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus plantarum.Assay method is the method under the qualification item of lactobacillus inspection and GB 4789.34-2012 Bifidobacterium in GB 4789.35-2010.

Experimental result (table 6) shows that the different proportion composition of stachyose (A)+galactooligosaccharide (B)+Oligomeric manna sugar (C) three all has proliferation function to 10 kinds of probios, covers bacterial classification comprehensive time more alone than stachyose.The bacterial classification covered than the composition of stachyose+both FOSs is also comprehensive.

[table 6]

(compare with control group, "-" represents without proliferation function; "+" representative has proliferation function).

Comprehensive above-mentioned experimental result, the compositions table of stachyose+galactooligosaccharide reveals good synergy.Especially the composition of stachyose+galactooligosaccharide+Oligomeric manna sugar overcome well stachyose alone time to the incomplete deficiency of proliferation of probiotics, substantially improve the bad body sense of stachyose aerogenesis simultaneously.Said composition also shows unexpected synergy, has been issued to significant effect and its effect can also maintain the long period after cutting out at very low dosage.This products application for the later stage has very positive effect.

Claims (11)

1. for regulating a composition for human body intestinal microecology, it is characterized in that, comprising stachyose and galactooligosaccharide as effective active composition.

2. composition as claimed in claim 1, it also comprises Oligomeric manna sugar and/or xylo-oligosaccharide.

3. composition as claimed in claim 1, wherein, the weight ratio of described stachyose and galactooligosaccharide is 1:10 ~ 10:1, is preferably 1:1 ~ 5:1, or is preferably 1:5 ~ 1:1.

4. the composition according to any one of claims 1 to 3, wherein, described galactooligosaccharide in the oligomeric disaccharides of gala to oligomeric eight sugar content >=57% (w/w) of gala, preferably >=70% (w/w), more preferably >=90% (w/w), further preferably >=99% (w/w), and lactose Han Liang≤23% (w/w), You Xuan≤20% (w/w), Geng You Xuan≤10% (w/w), and glucose Han Liang≤22% (w/w), You Xuan≤10% (w/w), Geng You Xuan≤5% (w/w).

5. composition as claimed in claim 2, wherein, in described Oligomeric manna sugar, mannobiose is to content >=85% (w/w), preferably >=90% (w/w) of sweet dew ten sugar, and mannobiose is to content >=50% (w/w) of sweet dew six sugar.

6. composition as claimed in claim 2, wherein, described xylo-oligosaccharide is in xylobiose extremely wooden seven sugar content >=70% (w/w), preferably >=80% (w/w), more preferably >=90% (w/w), and xylobiose is to content >=50% (w/w), preferably >=60% (w/w) of Xylotetrose.

7. the composition according to any one of claim 1 ~ 6, wherein, content of stachyose >=60% (w/w), preferably >=70% (w/w), more preferably >=80% (w/w).

8. the composition according to any one of claim 1 ~ 7, wherein, also comprises carbohydrate, such as chitosan oligosaccharide, oligoisomaltose, yeast beta-dextran, inulin, resistant dextrin, cottonseed compound sugar.

9. the composition according to any one of claim 1 ~ 7, wherein, also comprises auxiliary material and/or food materials.

10. food, health food or a pharmaceutical preparation, containing, for example the composition according to any one of claim 1 ~ 9, described food, health food or pharmaceutical preparation are preferably solid, semisolid or liquid form.

11. compositions according to any one of claim 1 ~ 9 are for the preparation of the purposes regulated in the food of human body intestinal microecology, health food or pharmaceutical preparation.