CN104642870B - A kind of prebiotic compositions - Google Patents

Abstract

The present invention provides a kind of composition for being used to adjust human body intestinal canal Tiny ecosystem, and it includes stachyose and galactooligosaccharide as effective active composition.The present composition can effectively adjust gut flora function.

Description

A kind of prebiotic compositions

Technical field

The present invention relates to the composition for adjusting human body intestinal canal Tiny ecosystem, more particularly to including stachyose and oligomeric half Composition of the lactose as effective active composition.

Background technology

For microbial cell number entrained by human body up to as many as 100,000,000,000,000, the symbiotic microorganism colony of these substantial amounts is main Skin, oral cavity and the intestines and stomach of human body are distributed in, wherein enteron aisle is the position that microorganism is concentrated the most.It is experimentally confirmed that enteron aisle Nutrition, metabolism and immunologic function of the flora to host serve vital effect, to maintaining health particularly significant, It is the part for realizing that host's normal physiological function is indispensable.

In human body microecosystem, intestinal microecology be it is main, it is most active, there is significant shadow to health Ring.Intestinal microecology system refer to by the biotic components such as gut flora and food-borne abiotic component (undigested food) with And it is located at the local of enteron aisle from what stomach, intestines, pancreas and the secretion of liver (such as hormone, enzyme, mucus and cholate) collectively formed The ecosystem.

Research shows, after neonate leaves parent, it is substantial amounts of thin that growth is begun on the surface such as skin, tracheae and alimentary canal Bacterium.The meconium of neonate's excretion for the first time is sterile, bacterium has been found that after 3~4 hours, bacterial number drastically increases after lactation It is long.After neonate's birth in 3~4 days, Bifidobacterium just occurs in excrement, turns into dominant bacteria during by the 5th day.With baby's Grow, the composition to gut flora during ablactation can reach a kind of relatively steady state.Under normal circumstances, it is this relative Stable gut flora composition can be along with the growth of human body, until the manhood.And to after the senescence phase, gut flora composition It can have greatly changed, the bifidobacteria in excrement is greatly reduced, and bacteroid and clostridium quantity are then big Width increase.

Gut flora establishes close relationship with human body in very long evolution, is not only to have opposed but also unified therebetween Unity of contradictions body.By long-term natural selection process, gut flora and human body have reached a kind of mutual reciprocity and mutual benefit and mutually made again Metastable dynamic balance state about.Human body provides living space for flora and energy source, flora then help human body Synthesize some nutrients and directly or indirectly influence the physiological function of human body.

Many experiments confirm that growth of the gut flora to the normal metabolic activity of human body, nutrient absorption, epithelial cell divides Very important effect, while they and some diseases are played in change, the immune response formation of normal bowel mucous membrane and maintenance etc. Disease, which also has, closely to be contacted, including lienteric disease, IBD, cancer, the Multiple Organ such as abdominal distension stomachache are disorderly Disorderly, allergy etc..

Gut flora not only influences the digestion and absorption of food, also affects the absorption and metabolism of oral drugs.Therefore, Metabolism and therapeutic effect of the gut flora to medicine can all have an impact, it is also possible to which drug therapy produces the original of individual difference Therefore one.

Human body intestinal canal flora can be divided into according to effect classification：Probiotics, conditioned pathogen and pathogenic bacteria.(1) probiotics, As the term suggests it is the bacterium beneficial to health.Probiotics is indispensable composition portion in human body intestinal canal micro-ecological environment Point, it is enteron aisle dominant microflora in healthy human body, there is the effect such as nutrition and immunological regulation.The main probiotics of human body includes lactic acid Bacillus, Bifidobacterium etc.；(2) conditioned pathogen, refer to it is harmless when intestinal microecology balances, but under given conditions again have There is a kind of bacterium of invasion, be the non-advantage flora of enteron aisle in healthy human body, such as enterococcus, enterobacteria；(3) pathogenic bacteria, it is Refer to the bacterium harmful, human pathology can be caused to react.Pathogenic bacteria are mostly the bacterium that passes by one's way, and are colonized in the chance ratio of enteron aisle for a long time It is less, in host internal memory residual hour, a couple of days or several weeks, such as proteus, pseudomonad and C.perfringens.

Human body intestinal canal flora can be divided into according to aerobic situation：Anaerobic bacteria, facultative anaerobic bacteria and aerobic bacteria.It is wherein obligate to detest Oxygen bacterium accounts for more than 99%, and only bacteroid and Bifidobacterium just account for more than the 90% of total number of bacteria.Due to hydrochloric acid in gastric juice, bile effect and Small intestine flow quantity is big, it is fast to wriggle, thus Stomach duodenum, the species of jejunum bacterium and quantity are few, predominantly Gram-positive Aerobic bacteria.Because intestinal juice flow is few, wriggling slows down, bacterial population gradually increases terminal ileum, mainly uncommon containing Bacillus acidi lactici, large intestine angstrom Bacterium, bacteroid and clostridium etc..To colon, bacterial population substantially increases, and forms microbe density highest bacterium colony group, And wherein most of is anaerobic bacteria, predominantly Bifidobacterium, bacteroid and Bacillus acidi lactici etc..

Some scholars think that the effect of probiotics may be summarized to be：Preventing and treating diarrhoea, lactose intolerance symptom of alleviating, prevention Vagina infection, enhancing body immunity, Reduce allergy, reduction serum cholesterol, pre- anti-cancer and suppression tumour growth.In addition also Prevent bone loss and osteoporosis.

Health promotion effect of the probiotics for human body is widely known.Therefore corresponding product is also slowly more Come.Mainly there are three classes for the product of human body intestinal canal Tiny ecosystem in the market：Prebiotic becteriums product, prebiotic metaclass product and by The product for entitled " symphysis unit " class that the former two combines.

" prebiotic becteriums product " this concept is distinguishing with foregoing description human body intestinal canal probiotics.Human body intestinal canal is prebiotic Pseudomonas is finally to be recognized each other by human body and the long-term natural selection process of microbial body and be colonized in human body intestines for a long time in ancestral home bacterium Bacterium general name in road.And " prebiotic becteriums product " belongs to foreign nationality bacterium, be artificially be added to it is internal with human body intestinal canal probiotics Similar strain.The bacterial strain that only those can be survived by the test of gastrointestinal tract environment and success is colonized could be true Just it is converted into " human body intestinal canal probiotics ".Also to pass through immune system cleaning and the enteron aisle ancestral home bacterium itself of human body among these Reverse transcriptase, therefore, success rate are very low.

At present, Food and Drug Administration (FDA) thinks safe probiotics up to as many as 40 kinds, with the hair of science and technology Exhibition, research go deep into, can also be more and more.Root is it is reported that human body intestinal canal bacterium includes more than 200 kinds of Pseudomonas, more than totally 800 strain, Bacterial strain species is then planted up to more than 7000.Following worries always be present for additional probiotics in academia：In face of so various flora System, merely supplement the allergic reaction that several probiotics can play a part of and whether can cause human body, and external source benefit Raw bacterium no can morph as pathogenic bacteria, etc. because of the change of production accumulating condition.For security standpoint, directly make There is certain risk with probiotics preparation.

Prebiotics (Prebiotics) is that to refer to selectively promote original beneficial bacteria in host intestine (prebiotic Bacterium) growth and breeding material.By increasing for beneficial bacterium, suppress harmful bacterial growth, so as to reach adjustment gut flora, promote The purpose of body health.It, which has, activates internal proliferation of probiotics, suppresses harmful bacteria, activated cell vigor, enhancing human immunity The health benefit of vigor.

The research to prebiotics and use are concentrated mainly on functional oligose at present.Functional oligose typically has Following characteristics：1st, be by 2~10 same or different monose, with glycosidic bond be polymerized (can be straight chain, can also It is side chain)；2nd, there is the characteristic of carbohydrate, can be directly as food ingredient, but not by human body alimentary canal enzyme and gastric acid degradation, no By (or difficult quilt) intestinal absorption；3rd, have and promote the physiological functions such as human body Bifidobacterium propagation.Functional oligose includes oligomeric Fructose, galactooligosaccharide, oligoisomaltose, xylo-oligosaccharide, soyabean oligosaccharides, Oligomeric manna sugar, stachyose, raffinose etc..

Functional oligose is fermented by intestinal beneficial bacterium, produces short chain fatty acids, and wherein acetic acid, butyric acid accounts for total yield acid amount 95%.The generation of short chain fatty acids can reduce intestinal pH, further suppress the growth of pathogenic entero becteria and conditioned pathogen Breeding；Adjust body water-electrolyte balance, promote calcium uptake；Suppress the propagation of enterocyte, differentiation, transfer and DNA Synthesis and methylation level, play antineoplastic action.

" symphysis unit " class product is that " probiotics " class product forms with " prebiotics " class product mix because its contain it is prebiotic Bacterium, therefore worry without the safety issue for solving " probiotics " class product.

" prebiotics " class product is more stable compared to " probiotics " class product property, quality controllability is stronger, and passes through Breed ancestral home bacterium and play " prebiotic " effect, activity problems and the problem that makes a variation without the foreign nationality bacterium for worrying artificially to add, because This obtains the trust of more scholars or even consumer.On January 16th, 2007, OLIGO (oligosaccharide) project are put into the public affairs in China Many nutrition improvement projects, turn into the another public affairs that China starts after the measures such as " supplementation of iodine salt ", " enriched nutritive flour " are implemented Many nutrition improvement projects.This aspect illustrates the extensive market of " prebiotics " class product, on the other hand also illustrate that industry pair The accreditation of " prebiotics " class product.

In recent years it has been proposed that the various compositions for including prebiotics.

For example, patent document 1 (CN102488100A) discloses a kind of feed addictive containing composite prebiotics, it is former Material component be cooked corn skin, corn steep liquor, oligoisomaltose, Oligomeric manna sugar, galactooligosaccharide, Bradley yeast bacterium solution, Clothing bacillus bacterium solution and Bacillus acidi lactici bacterium solution.The survival bacterium amount of probiotics is high in the feed addictive, has safety, environment The characteristics of degradable and cost is low.

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

Patent document 3 (CN1731938A) discloses one kind and contains fructose oligosaccharides (FOS) and GOS (GOS) Nutrition or pharmaceutical composition.

It is numerous that patent document 4 (TWI337079B) discloses methyl α manna oligosacchrides, galactooligosacchari(es, FOS, chitosan oligosaccharide etc. The composition (claim 1) of oligosaccharide and OPC.

In above-mentioned patent document 1, because feed addictive is for animal, differ with the application of the food of people Sample；In addition, the purpose of the technical scheme of document 1 is in order that the survival bacterium amount of probiotics is high in product, and human internal environment with Difference of the external environment with essence, the probability that the bacterium of Motility also survives in vivo be not high；In addition, composition of the present invention In do not add external source bacterium, also without adding maize peel, the material such as corn steep liquor.In addition, above-mentioned document 1~4 is no to disclose this hair Bright specific composition.

Therefore, although the research about various oligosaccharide has a lot, it is low that most of research directions are all confined to single Glycan is functionally.Few people combine for oligosaccharide carries out in-depth study.Because human body intestinal canal Tiny ecosystem is a complexity System, the bacterial species included in it are various, and utilization of the various bacterium for different oligosaccharide be have it is specific, its As a result cause single oligosaccharide often can only be to part proliferation of probiotics.And this unbalanced propagation is actually to change The poised state of human body intestinal microecology itself.Show that the change of this poised state is that do not have there is presently no enough evidences Risk.According to the literature, that realizes industrialized production at present has FOS, oligomeric using more functional oligose Isomaltose, xylo-oligosaccharide, galactooligosaccharide, soyabean oligosaccharides, stachyose, raffinose, Oligomeric manna sugar etc..Wherein oligomeric fruit Sugar, oligoisomaltose, the dosage of galactooligosaccharide are bigger than normal；The dosage of xylo-oligosaccharide is small, but its to such as bifidobacterium bifidum, The probiotics such as bifidobacterium breve, lactobacillus acidophilus, Lactobacillus casei are no proliferation functions；Soyabean oligosaccharides, stachyose, cottonseed Sugar has aerogenesis effect according to report；Oligomeric manna sugar has its propagation probiotic effect of document report imprecise.It is sum it up, every A kind of oligosaccharide has the characteristics of respective in use, and also all there is certain limitation.

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

Inventor herein is had made intensive studies by the combination to stachyose and galactooligosaccharide, and then achieves order People is unexpected, beyond expected experimental result.

The content of the invention

The present inventor is surprisingly found out that：By the way that galactooligosaccharide and stachyose is applied in combination, there is provided extensively Breed probiotics, suppress pathogenic bacteria, stable condition pathogenic bacteria, and then balanced human's intestinal microecology and dosage is small, takes Convenient safely and effectively composition.The combination of galactooligosaccharide and stachyose significantly improves prebiotics characteristic, shows association Same effect.It is further preferred that galactooligosaccharide, the combination of stachyose and Oligomeric manna sugar show further improve it is prebiotic First characteristic improves effect.

Instant invention overcomes the deficiencies in the prior art, there is provided can breed extensively probiotics, suppression/exclusion pathogenic bacteria, Stable condition pathogenic bacteria, flora imbalance, balanced human's intestine microenvironment are corrected, and then eliminated because human body intestinal canal Tiny ecosystem loses Sub-health state and the associated conditions such as abdominal distension, diarrhoea, indigestion caused by weighing apparatus and dosage is small, convenient to take, safe Effective composition.

Technical solution of the present invention includes：

Item 1, a kind of composition for being used to adjust human body intestinal canal Tiny ecosystem, it is characterised in that including stachyose and oligomeric half Lactose is as effective active composition.

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

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

4, the composition as any one of item 1~3, wherein, the galactooligosaccharide with the oligomeric disaccharides of gala extremely Gala it is oligomeric eight sugar meter content >=57% (w/w), preferably >=70% (w/w), more preferably >=90% (w/w), further preferably >= 99% (w/w), and newborn sugared content≤23% (w/w) ,≤20% (w/w) of excellent choosing ,≤10% (w/w) of more excellent choosing, and glucose contains 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 the Oligomeric manna sugar mannobiose to the sugar of sweet dew ten content >= 85% (w/w), preferably >=90% (w/w), and mannobiose is to content >=50% (w/w) of the sugar of sweet dew six.

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

7, the composition as 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 as any one of item 1~7, wherein, in addition to carbohydrate, for example, it is chitosan oligosaccharide, low IMO, yeast beta-dextran, inulin, resistant dextrin, raffinose-oligosaccharide.

Item 9, the composition as any one of item 1~7, wherein, also comprising auxiliary material and/or food materials.

Item 10, a kind of food, health food or pharmaceutical preparation, contain the composition as any one of item 1~9, institute It is preferably solid, semisolid or liquid form to state food, health food or pharmaceutical preparation.

11, the composition as any one of item 1~9 prepare be used to adjusting human body intestinal canal Tiny ecosystem food, Purposes in health food or pharmaceutical preparation.

The beneficial effects of the invention are as follows：(1) present composition can be so that prebiotic index in given scope (Prebiotic Index, PI) is significantly improved, and shows obvious synergy, and it is obvious to adjust gut flora function Higher than single oligosaccharide, dosage is small, can realize more preferable probiotic effects in the case where intake heat is low；(2) of the present invention group The gas-forming reaction of compound significantly reduces compared with the stachyose single dose of Isodose；(3) present composition propagation probiotics is covered Strain is more complete, the strain of suppression/exclusion pathogenic bacteria is also more；(4) present composition is for abdominal distension and the disease of diarrhea group Shape, which improves, shows obvious effect, shows that the present composition has dual regulation to gastrointestinal function.

Embodiment

" regulation human body intestinal canal Tiny ecosystem " is by adjusting gut flora, making the micro- life of human body intestinal canal described in this specification State environment reaches the poised state of health, and then various health benefits are embodied to human body.

" stachyose " is described in this specification：With its general implication in the art, it can be synthesis, It can process and obtain from natural origin.It is preferred that the stachyose that natural origin processing obtains.It represents the glucose by sucrose With α -1,6 glycosidic bonds combine the tetrose that two α-galactolipin is formed for base side." stachyose " herein also includes principal component For the mixture of stachyose.Preferably, principal component is in the mixture of stachyose, and content of stachyose >=60% (w/w), preferably >= 70% (w/w), more preferably >=80% (w/w).

" galactooligosaccharide " described in this specification has its general implication in the art, and it can be synthesis, It can also process and obtain from natural origin.It is preferred that the galactooligosaccharide that natural origin processing obtains.It is represented with lactose point The galactooligosaccharide containing 2~10 (preferably 2~8) monosaccharide molecules of galactose molecule generation is combined based on son.Herein In " galactooligosaccharide " also include principal component be galactooligosaccharide mixture.Preferably, principal component is galactooligosaccharide In mixture, the oligomeric disaccharides of gala 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 newborn sugared content≤23% (w/w) ,≤20% (w/w) of excellent choosing, more preferably ≤ 10% (w/w), and grape sugared content≤22% (w/w) ,≤10% (w/w) of excellent choosing ,≤5% (w/w) of more excellent choosing.

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

" xylo-oligosaccharide " described in this specification has its general implication in the art, and it can be synthesis, It can process and obtain from natural origin.It is preferred that the xylo-oligosaccharide that natural origin processing obtains.It is represented by 2~10 xyloses The oligosaccharide that molecule is combined into.It is preferred that by 2~7 xylose molecules with β-Isosorbide-5-Nitrae-glucosides key connection, and with xylobiose, wood three Sugar, the xylo-oligosaccharide mixture that Xylotetrose is main component." xylo-oligosaccharide " herein is also xylo-oligosaccharide including principal component Mixture.Preferably, principal component is in the mixture of xylo-oligosaccharide, xylobiose to wooden seven sugared content >=70% (w/w), preferably >=80% (w/w), more preferably >=90% (w/w), and xylobiose is to content >=50% (w/w) of Xylotetrose, preferably >=60% (w/ w)。

In the composition of the present invention, the weight ratio of stachyose and galactooligosaccharide is preferably 1:10~10:1, more preferably 1:5~5:1.In the composition of the present invention, in addition to stachyose and galactooligosaccharide are as effective active composition, it is preferable that and also Include Oligomeric manna sugar and/or xylo-oligosaccharide.Preferably, when the composition of the present invention includes stachyose, oligomeric gala When sugar, Oligomeric manna sugar, the weight ratio of Oligomeric manna sugar/stachyose is preferably 1:5~5:In the range of 1, more preferably 1:5~ 1:In the range of 1.

The auxiliary material and food materials that the composition of the present invention includes are those conventional, example in food and drug field Such as：Glidant, pH adjusting agent, disintegrant, starch, resistant starch, dextrin, resistant dextrin, the Chinese medicine for belonging to integration of drinking and medicinal herbs, food With essence, flavoring.

Embodiment

The present invention is further illustrated with reference to embodiment, but the invention is not limited in this.

Product embodiments 1：

Stachyose and galactooligosaccharide cross 100 mesh sieves respectively, weigh 1000 grams of stachyoses, 3000 grams of galactooligosaccharides, mix 1000 bags are distributed into after closing uniformly, it is preferably aluminum-plastic packaged.

Product embodiments 2：

3000 grams of stachyoses, 1000 grams of galactooligosaccharides are weighed, after being mixed with appropriate dextrin, with 85% alcohol granulation, are dried It is packaged into 1000 bags after dry, it is preferably aluminum-plastic packaged.

Product embodiments 3：

1 gram of stachyose, 10 grams of galactooligosaccharides are weighed, are added to after being dissolved with a small amount of distilled water in various cream tastes.

Product embodiments 4：

5000 grams of stachyoses, 1000 grams of galactooligosaccharides are weighed, respectively with 10000 milliliters, 5000 milliliters preparations of distilled water It is well mixed and adjusts after into solution and 15000 milliliters of solution is configured to distilled water after volume mixes to 15000 milliliters or first, Filter, be filling into 1000 oral liquids.

Product embodiments 5：

1000 grams of stachyoses, 5000 grams of galactooligosaccharides, 1000 grams of Oligomeric manna sugars are weighed, with 15000 milliliters of distilled water After dissolving, syrup form is concentrated into, be distributed into 1000 parts, it is preferably aluminum-plastic packaged.

Product embodiments 6：

1000 grams of stachyoses, 600 grams of galactooligosaccharides, 500 grams of Oligomeric manna sugars, anhydrous lemon are weighed after raw material drying 680 grams of acid, 500 grams of sodium acid carbonate, 100 grams of microcrystalline cellulose, PEG6000110 grams, 10 grams of magnesium stearate, it is well mixed, powder Direct tablet compressing, it is made 1000.

Testing example：

First, materials and methods

(1) oligosaccharide source is respectively in the present invention：

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

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

Oligomeric manna sugar：Commercially available prod, Oligomeric manna sugar content 90% in terms of mannobiose to the sugar of sweet dew ten.

Xylo-oligosaccharide：Commercially available prod, xylo-oligosaccharide content 95% in terms of xylobiose to wooden seven sugar.

FOS：Commercially available prod, FOS is with ketose, fruit fruit trisaccharide, Nystose, fruit fruit tetrose, sugarcane fruit The percentage composition that pentasaccharides, fruit fruit pentasaccharides, the sugar of sugarcane fruit six, six sugared sum of fruit fruit account for dry is calculated as 96%.

Above-mentioned oligosaccharide is purchased and obtained by domestic market, is powdery product, meets state food, new resources food Product/new raw-food material requirement.

(2) analytical standard

1、《Health food is examined and assessment technique specification》For the positive knot of regulation gut flora function in (version in 2003) The qualitative judgement standard of fruit is：

A, zoopery

The change of Bifidobacterium, lactobacillus, enterococcus, enterobacteria, C.perfringens between comparative experiments front and rear itself and group Change situation, itself front and rear comparing difference of experimental group experiment have after conspicuousness, or experiment comparing difference between experimental group and control group group There is conspicuousness and itself front and rear comparing difference of experimental group experiment has conspicuousness, meet any one of following, it is possible to determine that the test sample Product results of animal is positive.

1) Bifidobacterium and/or lactobacillus substantially increase in excrement, and C.perfringens is reduced or do not increased, enterobacteria, Enterococcus is without significant change.

2) Bifidobacterium and/or lactobacillus substantially increase in excrement, and C.perfringens is reduced or do not increased, enterobacteria And/or enterococcus substantially increases, but increased amplitude is less than the increased amplitude of Bifidobacterium/lactobacillus.

B, human experiment is tested

Meet it is any one of following, and test group test-meal it is front and rear itself compare and test-meal after test-meal group compared with control group, it is poor It is different to have conspicuousness, it is possible to determine that the given the test agent has the function that to adjust gut flora function.

1) Bifidobacterium and/or lactobacillus substantially increase in excrement, and C.perfringens is reduced or do not increased, enterobacteria, Enterococcus, bacteroid are without significant change.

2) Bifidobacterium and/or lactobacillus substantially increase in excrement, and C.perfringens is reduced or do not increased, enterobacteria And/or enterococcus, bacteroid substantially increase, but increased amplitude is less than the increased amplitude of Bifidobacterium/lactobacillus.

2nd, R.Palframan et al. (Development of a quantitative tool for the comparison of the prebiotic effect of dietary oligosaccharides.Letters in Applied Microbiology 2003,37,281-284) propose the index of quantitative analysis prebiotic function：Prebiotics refers to Number (Prebiotic Index, PI).The present inventor according to China it is existing for regulation gut flora function judge standard, On the premise of additionally burden is not tested in increase, zoopery PI computational methods and human experiment that suitable China uses are devised Test PI computational methods.Respectively：

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

Wherein " double ", " breast ", " shuttle ", " bar ", " ball " represent Bifidobacterium number, lactobacillus number, aerogenesis after experiment respectively The ratio between capsular clostridium number, enterobacteria number and enterococcus number and bacterium number before its each corresponding bacterium experiment；It is " total " to represent after testing The ratio of above-mentioned 5 kinds of bacterium bacterium number sums and bacterium number sum before experiment.

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

Wherein " double ", " breast ", " shuttle ", " bar ", " ball ", " plan " represent respectively experiment after Bifidobacterium number, lactobacillus number, The ratio between C.perfringens number, enterobacteria number, enterococcus number and bacteroid and bacterium number before its each corresponding bacterium experiment；" total " table Show the ratio of above-mentioned 6 kinds of bacterium bacterium number sums after experiment and bacterium number sum before experiment.

2nd, zoopery

1st, experimental animal

SPF inbred strais BALB/c male mices, 18-22g, every group 10.

2nd, packet and given the test agent give the time

Experiment sets test group and blank control group, and given the test agent gives the time 14 days.

3rd, experimental procedure

Before given the test agent is given, sterile to take stool in mice 0.1g, 10 times are serially diluted, and select suitable dilution factor It is seeded in respectively on each culture medium.After culture, bacterium colony is counted with identifications such as colonial morphology, gram stain microscopy, biochemical reactions, Calculate every gram it is wet just in bacterium number, carry out statistical disposition after taking the logarithm.Last time gives 24h after given the test agent, with reality The same manner cut-off intestines excrement before testing, detects gut flora, and method is same as above.

4th, data processing：

Data variance analysis, homogeneity test of variance is first carried out by the program of variance analysis, variance is neat, calculates F values, F values< 0.05, conclusion：No significant difference between each group mean；F value >=0.05, P≤0.05, between multiple experimental groups and a control group The comparative approach two-by-two of mean is counted；Appropriate variable is carried out to the data of abnormal or heterogeneity of variance to change, and waits to meet After normal state or variance require together, counted with the data after conversion；It is neat that if normal state or variance are still not up to after variable conversion Purpose, use rank test instead and counted.

5th, experimental result and analysis

5.1 experiments (1)

Stachyose, Oligomeric manna sugar and galactooligosaccharide with document for people report minimum day effective dose (0.5g, 0.5g and 2g) it is converted into mouse daily dose and gives mouse for 0.1g/kg, 0.1g/kg, 0.4g/kg.A+B groups daily dose is wood Sugared 0.1g/kg+ galactooligosaccharides 0.4g/kg；B+C groups daily dose is galactooligosaccharide 0.4g/kg+ Oligomeric manna sugars 0.1g/kg； A+B+C groups daily dose is stachyose 0.1g/kg+ galactooligosaccharide 0.4g/kg+ Oligomeric manna sugars 0.1g/kg.Each tested material is with nothing The dissolving preparation of bacterium distilled water is closed stored refrigerated after 4 DEG C, and the holding time is no more than 5 days, takes out and places to room in advance before gavage Temperature.Mouse stomach volume is 0.1ml/10g.Continuous gavage gives measurement result after 14 days to each group mouse respectively.Blank control group Gavage gives the sterile saline of respective volume.

[table 1]

(after itself comparing and test before and after experimental group experiment compared with control group, "-" is represented without significant change；“+” Representative has significant difference, P<0.05；" ++ ", which represents, pole significant difference, P<0.01；)

Test result indicates that：Stachyose, galactooligosaccharide and Oligomeric manna sugar individually in use, under low dosage not The proliferation function to probiotics is shown, regulation gut flora function is determined as feminine gender；And the wood equally under same dose The composition of sugar+galactooligosaccharide but shows significant prebiotic meta function, and regulation gut flora function is determined as the positive, A+B Show significant difference (P<0.05) composition of Oligomeric manna sugar+galactooligosaccharide, and under same dose is to prebiotic First index fails to make a significant impact.It imply that the composition of stachyose+galactooligosaccharide generates cooperative effect.Stachyose+ The composition cooperative effect of galactooligosaccharide+Oligomeric manna sugar is stronger.

5.2 experiments (2)

By stachyose, Oligomeric manna sugar, galactooligosaccharide and combinations thereof (composition proportioning is with experiment (1)) with every The dosage (being converted into human dose 3g/d) of day 0.6g/kg gives mouse with experiment (1) methods described.Surveyed after continuously giving 14 days Determine result.

[table 2]

(after itself comparing and test before and after experimental group experiment compared with control group, "-" is represented without significant change；“+” Representative has significant difference, P<0.05；" ++ ", which represents, pole significant difference, P<0.01；" ± " representative has an impact trend, but nothing Significant difference).

Test result indicates that：Although shown when each component is used alone under 0.6g/kg/d dosage certain prebiotic Meta function, but to Bifidobacterium, the inhibitory action of the propagation of lactobacillus and C.perfringens and unbalanced.And stachyose+ The composition of Oligomeric manna sugar+galactooligosaccharide is balanced to effect caused by Bifidobacterium, lactobacillus and C.perfringens And strong.And the animal PI values of composition group compare the animal PI values of each component exclusive use group and are respectively provided with and significantly improve. This result further demonstrates the synergy of combinations thereof.

We test the composition of stachyose and galactooligosaccharide different ratio under daily 0.6g/kg dosage in addition (3：1,1：1、1：3、1：5、1：7) to the influence of prebiotic index, method of testing is the same as experiment (1) methods described.As a result show each Composition shows as adjusting the gut flora function positive, and wherein animal PI values are 1：3 and 1：The composition group of 5 two ratios is most Height, but through the (P that compares that there was no significant difference between statistical analysis each group>0.05).

Further to investigate the synergy of said composition, we test feelings of each independent component in incremental dose in addition To the influence of prebiotic index under condition.As a result show that the dosage when stachyose and galactooligosaccharide exclusive use is respectively increased Stachyose+Oligomeric manna sugar+galactooligosaccharide composition (0.1+0.1+ during 0.8g/kg/d and 1.2g/kg/d and shown in table 1 0.4g/kg/d) animal PI values when being 0.6g/kg/d are suitable, and to reach same effect when Oligomeric manna sugar is used alone Need dosage more.As a result explanation can significantly reduce taking dose by combination, no matter in terms of economy or practicality It is all significant.

5.3 experiments (3)

By stachyose, Oligomeric manna sugar, galactooligosaccharide and their compositions with daily 0.6g/kg dosage (its Middle A+B is assembled than for 1：4th, A+B+C is assembled than for 1：4：1) mouse is continuously given 14 days with experiment (1) methods described.It is every group small Mouse quantity increases to 20, and each group mouse still further irrigates salmonella typhimurium, bacterial concentration respectively after continuous give 7 days For 1 × 10⁸CFU/ml, 0.2ml/ only, continuous 2 days, continue observation 5 days, record dead mouse situation and diarrhoea situation.

[table 3]

Test result indicates that blank control group is all suffered from diarrhoea, wherein there is 13 death occur；The abdomen of each oligosaccharide group Rushing down incidence and the death rate has different degrees of reduction, and it is significantly greater than each that stachyose+galactooligosaccharide composition group reduces amplitude Independent oligosaccharide group, stachyose+galactooligosaccharide+Oligomeric manna sugar composition group effect are optimal.It imply that said composition has Suppression/removing pathogenic bacteria, prevention bacterial diarrhea and mitigation symptom, reduce dead effect.

3rd, human feeding trial

1st, subject's inclusive criteria

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

2nd, experimental design and packet require

Using itself between group two kinds of control designs.Test-meal and control group are randomly divided into by the flora situation of subject, to the greatest extent Principal element such as age, sex, dietary factor etc. of influence result may be considered, harmonious inspection is carried out, between guarantee group Comparativity.Every group of subject 50.

3rd, the dosage and application method of given the test agent

This product, is dissolved in 100 milliliters of warm water by everyone daily this composition 3g of test-meal group when taking, a clothes.Control Group uses blank control.Given the test agent gives the time 14 days.Original eating habit, normal diet are not changed during experiment.

4th, observation index

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

5th, test procedure

Before given the test agent is given, sterile to take subject's excrement 1.0g, 10 times are serially diluted, and select suitable dilution Degree is seeded on each culture medium respectively.After culture, bacterium is counted with identifications such as colonial morphology, gram stain microscopy, biochemical reactions Fall, calculate every gram it is wet just in bacterium number, carry out statistical disposition after taking the logarithm.Last time gives 24hr after given the test agent, Detected again with same method.Last time gives 120hr after given the test agent, is detected again with same method.

6th, data processing

Test data is measurement data, is examined and is analyzed with t.Own control data uses paired t-test, two groups of means Compare and use independent samples t-test, the latter carries out homogeneity test of variance, and Non-Gaussian Distribution or the data of heterogeneity of variance are carried out suitably Variable is changed, and after meeting that normal state variance is neat, t inspections are carried out with the data of conversion；If change data can not still meet normal state side Poor neat requirement, uses t ' inspections or rank test instead；(such as CV too big to the coefficient of variation>50%) logging data application rank test.

7th, experimental result and analysis

By stachyose and the composition and FOS+galactooligosaccharide of stachyose+galactooligosaccharide+Oligomeric manna sugar Composition with daily 3g (composition be equal proportion mixing) dosage according to《Health food is examined advises with assessment technique Model》It is real that " regulation gut flora functional check method " in (version in 2003) function assessment evaluation test method carries out human experiment Test.Continuously give measurement result after 14 days.

Measurement result after [table 4] human experiment is tested 14 days

(test group test-meal is front and rear itself compare and test-meal after compared with control group, "-" is represented without significant change；"+" generation Table has significant difference, P<0.05；" ++ ", which represents, pole significant difference, P<0.01；" ± ", which represents, has an impact trend, but without aobvious Write sex differernce)

Test result indicates that：Stachyose and the composition of stachyose+galactooligosaccharide+Oligomeric manna sugar three are respectively provided with Adjust gut flora function.And the effect that stachyose+galactooligosaccharide+Oligomeric manna sugar composition shows is stronger.And It is substantially better than effect of the composition of FOS+galactooligosaccharide under 3g/d dosage.In addition, the ginseng of stachyose group 92% Examination person reports that aerogenesis increases, and reports that the number that aerogenesis increases only has 28% in composition experimental group, hence it is evident that reduces stachyose The side effect of aerogenesis.

Measurement result after [table 5] human experiment is tested 5 days

(test group test-meal is front and rear itself compare and test-meal after compared with control group, "-" is represented without significant change；"+" generation Table has significant difference, P<0.05；" ± " representative has an impact trend, but there was no significant difference).

Test result indicates that：After stachyose withdraws 5 days (120hr), probiotics quantity quickly falls after rise, stachyose+oligomeric half Lactose+Oligomeric manna sugar composition group has also fallen after rise, but decrease speed is considerably slower than stachyose group.FOS+oligomeric half The composition group of lactose has had returned to original state withdrawing 5 days after (120hr).In addition, stachyose group is withdrawn 5 days The person of participating in the experiment for still having 32% after (120hr) reports that aerogenesis increases, stachyose+galactooligosaccharide+Oligomeric manna sugar composition experiment Report that the number that aerogenesis increases is 22% in group.

Other human experiment experiment 20 acute diarrhea crowds are individually used continuously this composition 3g/d (stachyose+ Galactooligosaccharide 1：1), wherein 15 are rapid relief of symptoms on the day of, defecation frequency substantially reduces.30 constipation old men are connected It is continuous to use this composition 3g/d (stachyose+galactooligosaccharides 1：2) defecation frequency and difficult defecation degree are obviously improved after 3 days. Show that the present composition has dual regulation to gastrointestinal function.

4th, experiment in vitro

By in vitro test qualitative test stachyose and stachyose+galactooligosaccharide under same dose (2% (w/v))+ Composition and stachyose+FOS combination thing of Oligomeric manna sugar three is for bifidobacterium adolescentis, long bifid Bacillus, bifidobacterium infantis, bifidobacterium breve, bifidobacterium bifidum, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus salivarius, mouse The influence of the probiotics such as Lee's sugar lactobacillus, Lactobacillus plantarum.Assay method is lactobacillus inspection and GB in GB 4789.35-2010 4789.34-2012 the method under the identification item of Bifidobacterium.

Experimental result (table 6) shows the different proportion of stachyose (A)+galactooligosaccharide (B)+Oligomeric manna sugar (C) three Composition, all with proliferation function, it is comprehensive to cover strain to 10 kinds of probiotics when more alone than stachyose.Than stachyose+oligomeric The strain of fructose combination thing covering is also comprehensive.

[table 6]

(compared with control group, "-" is represented without proliferation function；"+", which represents, proliferation function).

Summary experimental result, the composition of stachyose+galactooligosaccharide show good synergy.Especially The composition of stachyose+galactooligosaccharide+Oligomeric manna sugar overcome well stachyose it is alone when it is incomplete to proliferation of probiotics The deficiency in face, while substantially improve the bad body-sensing of stachyose aerogenesis.Said composition also shows unexpected association Same-action, when very low dosage has been issued to significant effect and its effect can also remain longer after cutting out Between.This has the function that very positive for the products application in later stage.

Claims (15)

1. a kind of be used to adjust the composition of human body intestinal canal Tiny ecosystem, it is characterised in that including stachyose, galactooligosaccharide and low For poly- mannose as effective active composition, the weight ratio of the stachyose and the galactooligosaccharide is 1：10~10：1, and institute The weight ratio for stating stachyose and the Oligomeric manna sugar is 1：5~5：1.

2. composition as claimed in claim 1, wherein, the oligomeric disaccharides of gala is sugared to gala oligomeric eight in the galactooligosaccharide Content >=57% (w/w), and content≤23% (w/w) of lactose, and content≤22% (w/w) of glucose.

3. composition as claimed in claim 1, wherein, the oligomeric disaccharides of gala is sugared to gala oligomeric eight in the galactooligosaccharide Content >=70% (w/w), and content≤20% (w/w) of lactose, and content≤10% (w/w) of glucose.

4. composition as claimed in claim 1, wherein, the oligomeric disaccharides of gala is sugared to gala oligomeric eight in the galactooligosaccharide Content >=90% (w/w), and content≤10% (w/w) of lactose, and content≤5% (w/w) of glucose.

5. composition as claimed in claim 1, wherein, in the Oligomeric manna sugar mannobiose to the sugar of sweet dew ten content >= 85% (w/w), and mannobiose is to content >=50% (w/w) of the sugar of sweet dew six.

6. composition as claimed in claim 1, wherein, in the Oligomeric manna sugar mannobiose to the sugar of sweet dew ten content >= 90% (w/w), and mannobiose is to content >=50% (w/w) of the sugar of sweet dew six.

7. composition as claimed in claim 1, wherein, purity >=60% (w/w) of the stachyose.

8. composition as claimed in claim 1, wherein, purity >=70% (w/w) of the stachyose.

9. composition as claimed in claim 1, wherein, purity >=80% (w/w) of the stachyose.

10. such as composition according to any one of claims 1 to 9, wherein, in addition to other carbohydrate.

11. composition as claimed in claim 10, wherein, other described carbohydrate be chitosan oligosaccharide, oligoisomaltose, Yeast beta-dextran, inulin, resistant dextrin, raffinose-oligosaccharide, xylo-oligosaccharide.

12. such as composition according to any one of claims 1 to 9, wherein, also comprising auxiliary material and/or food materials.

13. a kind of food, health food or pharmaceutical preparation, it contains the composition any one of claim 1~12.

14. food as claimed in claim 13, health food or pharmaceutical preparation, wherein, the food, health food or medicine Preparation is solid, semisolid or liquid form.

15. the composition any one of claim 1~12 is preparing the food for being used to adjust human body intestinal canal Tiny ecosystem, protected Purposes in health food or pharmaceutical preparation.