CN110172106A - A kind of peach gum polysaccharide iron and its application - Google Patents
A kind of peach gum polysaccharide iron and its application Download PDFInfo
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- CN110172106A CN110172106A CN201910502603.4A CN201910502603A CN110172106A CN 110172106 A CN110172106 A CN 110172106A CN 201910502603 A CN201910502603 A CN 201910502603A CN 110172106 A CN110172106 A CN 110172106A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/26—Iron; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/02—Nutrients, e.g. vitamins, minerals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/06—Antianaemics
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
Abstract
The present invention provides a kind of peach gum polysaccharide iron, it is by chelating peach gum polysaccharide and iron, form a kind of peach gum polysaccharide iron complexes at iron core center with albuminoid iron, the beneficial effect is that, the obtained peach gum polysaccharide iron iron-content of the present invention is high, also there is complex stability, no free Fe in product2+, nonirritant to gastrointestinal tract, benefit iron effect is good, which does not have the use of noxious material, and Product safety is good, and additive amount is low；After iron is released in peach gum polysaccharide iron, peach gum polysaccharide itself has probiotic supplemented nutrition, to have the biological action for promoting probiotics in intestinal colonisation, simultaneously, peach gum polysaccharide has the function of natural absorption harmful substance, and in addition it is also a kind of good water-soluble dietary fiber, good water-retaining property, the moisture and loose degree that will increase excrement are conducive to improve function of intestinal canal.
Description
Technical field
The invention belongs to food pharmaceutical technology fields, more particularly, to a kind of peach gum polysaccharide iron and its application.
Background technique
Iron is microelement necessary to human body, if human body is insufficient to the intake of iron, can make the synthesis of hemoglobin
It reduces, and then reduces the content of hemoglobin in red blood cell so that erythrocyte number is reduced, result then will lead in human body
Each histoorgan for hypoxgia.Hypoferric anemia is one of highest nutrient deficiency disease of current disease incidence, seriously threatens people
Body health.Global disease burden mechanisms (GBD) in 2013 are research shows that anemia is to damage the disease of life-span TOP V.It is poor
The function of blood injury human internal organ tissue hinders the growth and development in people's each stage, is easy the functional developmental disorder of initiation, human body is exempted from
The decline of epidemic disease ability, digestion function of stomach and intestine decline, children's intelligence development is slow, pregnant woman anemia can cause fetus premature labor and neonate
The reaction for the Disadvantage pregnancies such as weight is relatively low, constitution is poor.
Currently used two kinds of relatively broad iron supplementaries have inorganic iron and Organic Iron iron supplementary.Ferrous sulfate is as inorganic iron
The representative of iron supplementary, although mending, iron effect is good, it is easy stimulating gastrointestinal road, generates the poison pair such as oral peculiar smell and iron poisoning and makees
With.Although common ferrous pyrophosphate effect is preferable, it is as Fe2+There are many side effects, such as to GI irritation compared with
Greatly, oral peculiar smell is serious and there are risks of iron poisoning etc., and most of patient is made to be difficult to bear.Amino acid iron as albumen iron with
The same PORPHYRIN IRON is all Organic Iron, although effect is preferable, produces difficult complicated, and synthesis cost is high, allow many patients without
Method is assumed charge.
The very big factor of infant's iron deficiency is caused by itself congenital Blood lipids deficiency.Pregnant woman during pregnancy, take the photograph by iron
Enter amount deficiency, makes infant that can not obtain enough ferro elements in parent.Pregnant woman itself takes in that irony is less or external condition
Influence, the problems such as will lead to fetus premature labor or fetus blood loss.After baby due, the middle and later periods of nursing period is baby from parent
The important stage for obtaining nutriment can make iron content in lotion lower if maternal nutrition is insufficient.In addition, nourishing twins
Or the pregnant woman of multiparity, child's demand is larger, and iron content deficiency can also increase anaemic probability in breast milk.These are all made
The reason of at congenital under-reserve.The reason of another baby's iron deficiency is malabsorption: iron content is high in pregnant woman's milk, is born
Six months childs are preferably fed by the iron supplementary of " natural ".But for can not normal lactation pregnant woman, milk powder generation can be selected
For milk, protein and calcium content are higher in milk powder, will lead to the generation that RBC acceptor garland rate lacks phenomenon.Baby is (big for a long time
About six months) by milk powder nursing to will lead to irony content in infants low, while iron absorbs and utilization rate is lower, and this will cause
Iron malabsorption.Some babies there is also allergic reaction, lead to the generation of gastrointestinal bleeding phenomenon, and then cause anemia.
It is larger to nutritional need during infant growth is developed, while blood volume can increased, and ferro element
Demand also will increase.But the demand of iron deficiency infant is not able to satisfy by tonic merely, the digestive system of infant is also not
It is perfect, it is higher to safety, the validity requirement of iron-supplementing preparation, currently, common baby's iron supplementary has: ferrous gluconate,
Ferrous lactate, lactoferrin etc..Wherein, lactoferrin is a kind of important nonheme iron combination glycoprotein in milk, at the beginning of people
Lactoferrin is up to 6-14mg/ml in cream, only 1-3.2mg/ml in normal breast milk, content only 0.02-0.35mg/ml in cow's milk,
So real lactoferrin is at high price.
Summary of the invention
The present invention is directed to problem above, provides a kind of peach gum polysaccharide iron.The obtained peach gum polysaccharide iron iron content of the present invention
Amount is high, and complexing is stablized, nonirritant to gastrointestinal tract, and benefit iron effect is good, is conducive to improve function of intestinal canal.
The present invention, which adopts the following technical scheme that, realizes above-mentioned technical purpose:
A kind of peach gum polysaccharide iron, the peach gum polysaccharide iron the preparation method comprises the following steps:
S1: 1:1 ~ 3 weigh peach gum polysaccharide extract and trisodium citrate and dissolve in the water of 40 ~ 80 times of volumes in mass ratio, delay
Slowly the liquor ferri trichloridi and sodium hydroxide solution for stirring and being added dropwise 2 ~ 5mol/L are heavy until occurring rufous in reaction liquid
It forms sediment, and precipitates and stop being added dropwise when not being redissolved, then be placed in 1 ~ 2h of reaction in 70 ~ 90 DEG C of water-baths, obtain end reaction liquid；
S2: being centrifuged off precipitating for 500 ~ 1000rpm/min of end reaction liquid in step S1, takes liquid that the nothing of 1 ~ 5 times of volume is added
Water-ethanol is precipitated precipitating and is washed repeatedly 2 ~ 3 times with 70 ~ 80% ethanol solutions, dry peach gum polysaccharide iron.
Former peach gum (crude peach gum, CPG) is a kind of resourceful natural biomass, is a kind of natural suction
Attached dose, CPG contains a large amount of negatively charged carboxylic acid groups, can pass through strong electrostatic interaction Liquidity limit type dye.
CPG main component is peach gum polysaccharide, contains a small amount of protein and impurity etc..Peach gum polysaccharide is a kind of acidic polysaccharose, by gala
Sugared (42%), arabinose (36-37%), uronic acid (7-13%), xylose (7%) and mannose (2%) pass through glucosides key connection group
At.
Peach gum polysaccharide contains a large amount of negatively charged carboxyl functional groups, and opposite electricity can be adsorbed by strong electrostatic interaction
The Fe ion of lotus forms the iron core center of an albuminoid iron, and do not dissociate Fe stimulating intestine and stomach, and peach gum polysaccharide is with hydrophilic
The characteristic of colloid and film forming can form one of natural preventive barrier in alimentary canal, more mild easily to absorb.
Further, in the step S1 in peach gum polysaccharide extract peach gum polysaccharide content >=80%.
Further, the peach gum polysaccharide extract is prepared by the following method:
S11: former peach gum being cleaned and is cleaned, and 10 ~ 30 times of amount water are added and impregnate 8 ~ 12h, 50 ~ 70 DEG C of holdings are heated to after homogenate is broken
1 ~ 2h, 2000 ~ 4000rpm/min are centrifuged 3 ~ 5min, take supernatant, obtain raw sugar liquid；
S12: dehydrated alcohol is added in raw sugar liquid described in step S11 by 1 ~ 5:1 times, sufficiently oscillation after in 0 ~ 4 DEG C stand 10 ~
Pellet frozen dry, pulverize after taking precipitating to repeat sedimentation 2 ~ 3 times, obtain peach gum polysaccharide by 12h, 4000 ~ 6000rpm/min centrifugation
Crude extract；
S13: add water to be configured to the polysaccharide solution of 2 ~ 8mol/mL the peach gum polysaccharide crude extract in step S12, it is fine to cross DEAE-52
Plain column is tieed up, the NaCl solution elution of 0.1 ~ 2mol/mL is collected elution samples, refiltered, and peach gum polysaccharide extraction is concentrated and dried to obtain
Object.
Former peach gum dissolubility is poor, needs to impregnate removing protein and impurity again after swelling, obtains crude extract and chromatograph again to be separated by filtration
Purification, using extract obtained by this method, peach gum polysaccharide yield is high, and purity is good, and preparation process is easily waved using moderate notoxic
The solvent of hair, product are safer.
Further, 0.45 μm of miillpore filter or bag filter filtering are filtered into the step S13.
The present invention protects the peach gum polysaccharide iron preparing the application in iron-supplementing preparation simultaneously.Peach gum provided by the invention is more
Sugared iron iron-content is high, also has complex stability, nonirritant to gastrointestinal tract, and it is good to mend iron effect.
Above-mentioned preparation can be used in food simultaneously, especially prepare enhancing infant immunity food.
Peach gum polysaccharide iron is non-stimulated to gastrointestinal tract, bioavilability is high, is the benefit iron of the hypoferric anemia of good validity
Agent, while growth and development and the enhancing hematopoiesis function of baby can also be promoted, first to grow up healthy and sound has been constructed for baby have been prevented
Line can remove interior free yl, and infant is helped to resist the harmful microorganisms such as bacterium, virus, breathing caused by pre- anti-virus
Road infection and babies' common disease such as diarrhea, play regulatory role the generation of body antibody, erect one of antibacterial, disease-resistant for human body
The defence line that poison is invaded and harassed, can help them excite, establish, repairing, improve immune system, enhance disease resistance.
The present invention also provides a kind of for enhancing the preparation of infant immunity, including following components: peach gum polysaccharide iron 30 ~
50 parts, 20 ~ 40 parts of active microbial inoculum, wherein the active microbial inoculum includes animal bifidobacteria, bifidobacterium lactis, rhamnose cream bar
One of bacterium is a variety of.
After iron is released in peach gum polysaccharide iron, peach gum polysaccharide belongs to nutrition physiology active material, on the top of gastrointestinal tract, no
By the digestion enzymic digestion of humans and animals gastro-intestinal secretion, it cannot be hydrolyzed absorption, it can only be selectively (such as double to enteral probiotics
Discrimination bacillus) stimulating growth breeding or activation metabolism are carried out, increase probiotics absolute quantity, improves probiotics institute in intestinal flora
The ratio accounted for reduces pH, improves intestinal environment, fosters normal the gut flora, inhibits pathogenic bacteria, and then cause flora imbalance
Constipation symptom corrected.In addition peach gum polysaccharide liquid is a kind of good water-soluble dietary fiber, will increase the moisture of excrement
With loose degree, be conducive to improve function of intestinal canal.
It further, further include vitamin C in the component of the preparation for enhancing infant immunity.Vitamin C can
Promote Fe after discharging3+Absorption.
Further, said preparation can be used as nutrition enhancer.
The deficiency of element iron content will lead to the decline of body's immunity, the normal operation of function of human body, such as breathe
When the processes such as oxygen transportation, DNA reparation, cell division have the participation of trace iron, the activity of a variety of enzymes and immunogenic compound
Inseparable with ferro element, and during brain development, iron content is insufficient easily to cause mental retardation, said preparation colorless and odorless,
It is good with the interaction of milk powder, the nutritive value of milk powder can be enhanced, meet the needs of infant is to various nutrition.
Further, adding proportion of the nutrition enhancer in nutriment is 0.1 ~ 1%.
Compared with the existing technology, the present invention has the advantage that and effect:
The obtained peach gum polysaccharide iron iron-content of the present invention is high, also has complex stability, nonirritant to gastrointestinal tract, mends iron effect
Fruit is good, which does not have the use of noxious material, and Product safety is good, and additive amount is low.Iron is released in peach gum polysaccharide iron
Afterwards, peach gum polysaccharide itself has probiotic supplemented nutrition, thus promote probiotics in the bioactivity of intestinal colonisation, meanwhile, peach
Glue polysaccharide has the function of natural absorption harmful substance, and in addition it is also a kind of good water-soluble dietary fiber, water-retaining property
It is good, it will increase the moisture and loose degree of excrement, be conducive to improve function of intestinal canal.
Specific embodiment
Further illustrate the present invention below in conjunction with specific embodiment, but embodiment the present invention is not done it is any type of
It limits.Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, method and apparatus.
Unless stated otherwise, agents useful for same and material of the present invention are commercially available.
Embodiment 1:
The preparation of peach gum polysaccharide extract:
S11: 50g original peach gum being cleaned and is cleaned, and 1000ml water is added and impregnates 8h, is heated to 100 DEG C of holding 1h after homogenate is broken,
2000rpm/min is centrifuged 3min, takes supernatant, obtains raw sugar liquid；
S12: dehydrated alcohol is added in raw sugar liquid described in step S11 by 1:1 times, sufficiently in 4 DEG C of standing 12h after oscillation,
4000rpm/min centrifugation dry, pulverize pellet frozen, obtaining peach gum polysaccharide crude extract is after taking precipitating to repeat S12 step 2 time
42g；
S13: the peach gum polysaccharide crude extract in step S12 is added water to be configured to the polysaccharide solution of 2mol/mL, pre-processed fine by purifying
Column is tieed up, then uses the DEAE-52 cellulose column of deionized water balance, polysaccharide solution is slow to cross sample, finally with the NaCl of 0.2mol/mL
The solution being collected into Phenol-sulphate acid method is finally detected sugared content therein, according to its suction at 490 nm by solution elution
Luminosity draws elution curve.It selects optimal crest segment to collect sample, merges the sample of gradient crest segment, then at 10000 u bag filters
Filtering, in 50 DEG C of rotary evaporations, be concentrated and dried peach gum polysaccharide extract be 36g.
The preparation of peach gum polysaccharide iron:
S1: weigh peach gum polysaccharide extract 3g and trisodium citrate 3g dissolved in 300ml water be respectively configured to 10mg/mL(with
Na+ meter) aqueous solution, the liquor ferri trichloridi and 2mol/L sodium hydroxide solution of 2mol/L are slowly stirred and are added dropwise, until anti-
It answers and occurs red-brown precipitation in liquid, and precipitate and stop being added dropwise when not being redissolved, then be placed in 70 DEG C of water-baths and react 2h, obtain anti-eventually
Answer liquid；
S2: being centrifuged off precipitating for end reaction liquid 1000rpm/min in step S1, takes liquid that the dehydrated alcohol of 1 times of volume is added,
Precipitating is precipitated to be washed repeatedly 2 times with 70% ethanol solution, dry peach gum polysaccharide iron.
It is a kind of for enhancing the preparation of infant immunity, including following components:
30 parts of peach gum polysaccharide iron, 40 parts of active microbial inoculum (wherein, 30 parts of bifidobacterium lactis (bacterial strain HN019), rhamnose cream bar
10 parts of bacterium (bacterial strain LGG)), 0. 1 parts of vitamin C.
Embodiment 2:
The preparation of peach gum polysaccharide extract:
S11: 50g original peach gum being cleaned and is cleaned, and 500ml water is added and impregnates 10h, is heated to 100 DEG C of holding 2h after homogenate is broken,
4000rpm/min is centrifuged 3min, takes supernatant, obtains raw sugar liquid；
S12: dehydrated alcohol is added in raw sugar liquid described in step S11 by 2 times of amounts of raw sugar liquid, sufficiently quiet in 0 DEG C after oscillation
12h is set, pellet frozen dry, pulverize after taking precipitating to repeat S12 step 3 time, it is thick to obtain peach gum polysaccharide by 4000rpm/min centrifugation
Extract；
S13: add water to be configured to the polysaccharide solution of 2.5mol/mL the peach gum polysaccharide crude extract in step S12, it is fine to cross DEAE-52
Plain column is tieed up, the NaCl solution elution of 0.5mol/mL is collected elution samples and is concentrated and dried then at 0.45 μm of filtering with microporous membrane
Peach gum polysaccharide extract 30g.
The preparation of peach gum polysaccharide iron:
S1: peach gum polysaccharide extract 1g and trisodium citrate 3g dissolves in 150mL water is respectively configured to 20mg/mL(with Na+
Meter) aqueous solution, be slowly stirred and be added dropwise the liquor ferri trichloridi and sodium hydroxide solution of 2mol/L, until in reaction liquid go out
Existing red-brown precipitation, and precipitate and stop being added dropwise when not being redissolved, then be placed in 60 DEG C of water-baths and react 1h, obtain end reaction liquid；
S2: being centrifuged off precipitating for end reaction liquid 500rpm/min in step S1, takes liquid that the dehydrated alcohol of 2 times of volumes is added,
Precipitating is precipitated to be washed repeatedly 2 times with 75% ethanol solution, dry peach gum polysaccharide iron.
It is a kind of for enhancing the preparation of infant immunity, including following components:
50 parts of peach gum polysaccharide iron, 20 parts of active microbial inoculum, wherein including 10 parts of animal bifidobacteria (bacterial strain Bb-12), newborn bifid
10 parts of bacillus (bacterial strain Bi-07).
Embodiment 3:
The preparation of peach gum polysaccharide extract:
S11: 50g original peach gum being cleaned and is cleaned, and 800ml water is added and impregnates 10h, is heated to 100 DEG C of holding 2h after homogenate is broken,
4000rpm/min is centrifuged 3min, takes supernatant, obtains raw sugar liquid；
S12: it by the dehydrated alcohol of 5 times of volumes by being added in raw sugar liquid described in step S11, is sufficiently stood after oscillation in 4 DEG C
Pellet frozen dry, pulverize after taking precipitating to repeat S12 step 3 time, obtain peach gum polysaccharide and slightly mention by 12h, 6000rpm/min centrifugation
Object；
S13: add water to be configured to the polysaccharide solution of 5mol/mL the peach gum polysaccharide crude extract in step S12, cross DEAE-52 fiber
Plain column, the NaCl solution elution of 1mol/mL, collects elution samples, filters then at bag filter, is concentrated and dried to obtain peach gum polysaccharide extraction
Object 37g.
The preparation of peach gum polysaccharide iron:
S1: peach gum polysaccharide extract 1g and trisodium citrate 2g dissolves in 200mL water is respectively configured to 10mg/mL(with Na+
Meter) aqueous solution, the liquor ferri trichloridi and sodium hydroxide solution of 2.5mol/L are slowly stirred and are added dropwise, until in reaction liquid
There is red-brown precipitation, and precipitate and stop being added dropwise when not being redissolved, then be placed in 50 DEG C of water-baths and react 2h, obtains end reaction liquid；
S2: being centrifuged off precipitating for end reaction liquid 1000rpm/min in step S1, takes liquid that the dehydrated alcohol of 2 times of volumes is added,
Precipitating is precipitated to be washed repeatedly 3 times with 75% ethanol solution, dry peach gum polysaccharide iron.
It is a kind of for enhancing the preparation of infant immunity, including following components:
50 parts of peach gum polysaccharide iron, 40 parts of Lactobacillus rhamnosus (bacterial strain LGG), 0.2 part of vitamin C.
Embodiment 4:
The preparation of peach gum polysaccharide extract:
S11: 50g original peach gum being cleaned and is cleaned, and 1000ml water is added and impregnates 10h, is heated to 100 DEG C of holding 2h after homogenate is broken,
2000rpm/min is centrifuged 4min, takes supernatant, obtains raw sugar liquid；
S12: dehydrated alcohol is added in raw sugar liquid described in step S11 by 1:1 times, sufficiently in 0 DEG C of standing 10h after oscillation,
Pellet frozen dry, pulverize after taking precipitating to repeat S12 step 2 time, obtain peach gum polysaccharide crude extract by 6000rpm/min centrifugation；
S13: the peach gum polysaccharide crude extract in step S12 is added water to be configured to the polysaccharide solution of 2mol/mL, pre-processed fine by purifying
Column is tieed up, then uses the DEAE-52 cellulose column of deionized water balance, polysaccharide solution is slow to cross sample, finally with the NaCl of 0.2mol/mL
The solution being collected into Phenol-sulphate acid method is finally detected sugared content therein, according to its suction at 490 nm by solution elution
Luminosity draws elution curve.It selects optimal crest segment to collect sample, merges the sample of gradient crest segment, then at 10000 u bag filters
Filtering, in 50 DEG C of rotary evaporations, be concentrated and dried peach gum polysaccharide extract be 31g.
The preparation of peach gum polysaccharide iron:
S1: weigh peach gum polysaccharide extract 3g and trisodium citrate 6g dissolved in 300ml water be respectively configured to 20mg/mL(with
Na+ meter) aqueous solution, the liquor ferri trichloridi and 3mol/L sodium hydroxide solution of 3mol/L are slowly stirred and are added dropwise, until anti-
It answers and occurs red-brown precipitation in liquid, and precipitate and stop being added dropwise when not being redissolved, then be placed in 70 DEG C of water-baths and react 2h, obtain anti-eventually
Answer liquid；
S2: being centrifuged off precipitating for end reaction liquid 1000rpm/min in step S1, takes liquid that the dehydrated alcohol of 1 times of volume is added,
Precipitating is precipitated to be washed repeatedly 2 times with 75% ethanol solution, dry peach gum polysaccharide iron.
It is a kind of for enhancing the preparation of infant immunity, including following components:
35 parts of peach gum polysaccharide iron, 35 parts of active microbial inoculum (wherein, 10 parts of bifidobacterium lactis (bacterial strain HN019), bifidobacterium lactis
15 parts of (bacterial strain Bi-07), 10 parts of Lactobacillus rhamnosus (bacterial strain LGG)), 0.5 part of vitamin C.
Embodiment 5:
The preparation of peach gum polysaccharide extract:
S11: 50g original peach gum being cleaned and is cleaned, and 1000ml water is added and impregnates 10h, is heated to 90 DEG C of holding 2h after homogenate is broken,
4000rpm/min is centrifuged 5min, takes supernatant, obtains raw sugar liquid；
S12: it by the dehydrated alcohol of 3 times of volumes by being added in raw sugar liquid described in step S11, is sufficiently stood after oscillation in 4 DEG C
Pellet frozen dry, pulverize after taking precipitating to repeat S12 step 3 time, obtain peach gum polysaccharide and slightly mention by 10h, 4000rpm/min centrifugation
Object；
S13: add water to be configured to the polysaccharide solution of 3mol/mL the peach gum polysaccharide crude extract in step S12, cross DEAE-52 fiber
Plain column, the NaCl solution elution of 1mol/mL, collects elution samples, filters then at bag filter, is concentrated and dried to obtain peach gum polysaccharide extraction
Object 35g.
The preparation of peach gum polysaccharide iron:
S1: peach gum polysaccharide extract 1g and trisodium citrate 2g dissolves in 200mL water is respectively configured to 10mg/mL(with Na+
Meter) aqueous solution, be slowly stirred and be added dropwise the liquor ferri trichloridi and sodium hydroxide solution of 2mol/L, until in reaction liquid go out
Existing red-brown precipitation, and precipitate and stop being added dropwise when not being redissolved, then be placed in 50 DEG C of water-baths and react 2h, obtain end reaction liquid；
S2: being centrifuged off precipitating for end reaction liquid 1000rpm/min in step S1, takes liquid that the dehydrated alcohol of 2 times of volumes is added,
Precipitating is precipitated to be washed repeatedly 3 times with 80% ethanol solution, dry peach gum polysaccharide iron.
It is a kind of for enhancing the preparation of infant immunity, including following components:
50 parts of peach gum polysaccharide iron, 20 parts of bifidobacterium lactis (bacterial strain HN019), 20 parts of Lactobacillus rhamnosus (bacterial strain LGG), dimension
Raw 0.2 part of C of element.
Experimental example 1:
Proliferation function of the peach gum polysaccharide to Bifidobacterium
It is 1,2,3(peach gum polysaccharide concentration of test group that peach gum polysaccharide extract in embodiment 1 is added in Bifidobacterium basic culture solution
It is respectively set to 10mg/mL, 20mg/mL, 30mg/mL), basic culture solution is control group, and each culture solution is respectively charged into test tube,
10 mL/ branch loading amounts are inoculated with the initial bacteria concentration of bifidobacterium lactis strain (bacterial strain HN019) 1 mL(after 121 DEG C of 15 min of sterilizing
It is 102, 37 DEG C of Anaerobic culturel 1h ~ 4h.
1 mL Spawn incubation sample liquid is taken to measure bifidobacterium cells quantity respectively.With bifidobacterium cells in basic culture solution
Quantity is reference, measures each concentration peach gum polysaccharide to the proliferation function of Bifidobacterium.
As the result is shown: addition peach gum polysaccharide can promote the rapid growth of Bifidobacterium, and with the increase of peach gum polysaccharide content,
The increased speed of thallus significantly increases.
Experimental example 2:
The physicochemical property of peach gum polysaccharide iron
Smell color: colorless and odorless.
Stability Determination, room temperature, which dries in the shade, to be located to place, and no property changes within one month, and only water imbibition is strong.
Experimental example 3
Mend iron effect measuring
By gained peach gum polysaccharide iron in embodiment 3, to be deployed with iron deficiency milk powder, preparation additive amount is respectively 0.1%, 0.3%, 0.5%,
0.7%, 1% is made nutritional agents.
The foundation of Anemia model carries out 2 % phenylhydrazines of subcutaneous injection by 0.04 ml/kg body weight dose, establishes mouse anaemia
Model.The OD380nm value of mouse hemoglobin is measured after injection 2 d of phenylhydrazine.
Anemia model mouse is grouped by the mouse for choosing the congenital hypoferric anemia of 7 ages in days at random, and every group of male and female are half-and-half.Hello
The above nutritional agents is supported, iron deficiency milk powder, 4h/ times every, each 10mL, experimental group and control group milk powder concentration phase are only fed in blank control
Deng taking a blood sample within every 5 days primary.Mouse tail takes blood, measures the OD380 nm value of its hemoglobin.
As the result is shown: peach gum polysaccharide iron is obvious to the benefit iron effect of anemia mice, and in peach gum polysaccharide iron in milk powder
Content increasing with its content in 0.1 ~ 1% range, it is faster to mend iron effect.
Experimental example 4
Immunity effect measuring:
Influenza virus is provided by Zhejiang Center For Disease Control and Prevention's virus, in -80 DEG C of refrigerator cryopreservations.It is connect before use
Kind 9 d instar chicken embryo allantoic cavities inoculation passage, 72 h of routine culture carry out conventional hemagglutination test, after measured, virus titer 1:
512。
(half male and half female, 4 ~ 6 week old (18 ~ 22g of weight), is provided mouse by Zhejiang Province's Experimental Animal Center.
Positive control medicine is Oseltamivir phosphate capsule.
Experimental group is the preparation for enhancing infant immunity.
Mouse is divided into 4 groups by weight stratified random, Normal group, virus model group, positive group, experimental group is (except just
The isodose physiological saline of normal control group collunarium is outer, and ((this experiment is using 20 μ L's under ether light anesthesia for remaining each group mouse
Influenza virus collunarium infecting mouse.
Starting gastric infusion treatment after influenza infection mouse 2d, (concrete condition is as follows: Normal group, viral mould
Type group gives the physiological saline of equivalent, and 21.63mg/kg is given in positive group stomach-filling, and 10mg/kg is given in experimental group stomach-filling), dosage ginseng
It (presses body surface area to be calculated as ,-HA weight adult's clinical equivalent agent, successive administration according to zoopery early period of this seminar
4 days, until experiment terminates.
Each group mouse is weighed the 5th day after infection, is plucked eyeball sacrificed by exsanguination mouse, is taken lung, spleen and thymic tissue,
It is cleaned with physiological saline, filter paper blots surface moisture, claims its weight respectively, calculates Lung Exponent, spleen index and thymus index.
Each group mouse Lung Exponent and immune organ index compare (table 3), compared with Normal group, virus model group mouse
Lung Exponent increases, and spleen index, thymus index reduces, compared with virus model group, positive group and experimental group, and Lung Exponent and immune
Organ index has different degrees of improvement.
Normal group mouse alveolar is positive Chang great little, and lung tissue structure is clear, and alveolar wall is without thickening, bronchiole mucous membrane
It is perfect, inflammatory cell infiltration, virus model group mouse visible many places pulmonary consolidation area, part lung are had no in interstitial lung and alveolar
Existing fusion is bubbled out, lung tissue structure's damage is serious, and Diffuse alveolar wall thickening is congested, ties hoof hyperblastosis, and bronchiole is small
Visible a large amount of active chronic inflammation cellular infiltrations around vascular wall are partially diffused into alveolar compared with model group, each dosing group lung
Lesion tissue range, degree is substantially reduced, and pulmonary morphology is substantially complete, and alveolar septum slightly thickens, and lesion area is reduced.
The acute and chronic inflammatory cell infiltrated in alveolar substantially reduces, congested, and the pathological changes such as alveolar consolidation obtain different degrees of change
It is kind.
Within the scope of cell maximal non-toxic mass concentration, the inhibiting rate of preparation infected by influenza of the invention is with mass concentration
Increase and be incremented by, i.e. antiviral activity gradually increases, and significant dose-effect relationship is presented.Show that the present invention is prepared
Preparation for enhancing infant immunity has resisiting influenza virus effect.
Experimental example 5
The studies on acute toxicity of peach gum polysaccharide iron
Mouse 20 of health, are randomly divided into 2 groups, every group 10, half male and half female is standard according to the body of mouse using 10 g/kg
It weighs a stomach-filling and takes peach gum polysaccharide iron orally, 14 d are observed continuously.Observed content includes: dead mouse number and respective death time；
Mouse weight；Whether poisoning and its reaction symptom, observation skin, coat, eyes and the case where mucous membrane are breathing, circulation, autonomous
With central nervous system and four limbs activity and the variation of behavior etc., pay special attention to have atremia, convulsions, diarrhea, drowsiness
Phenomena such as；
When mouse takes 10000 mg/kg of peach gum polysaccharide iron orally, within the observation period, drinking-water and normal, body weight increase each group of searching for food
Unanimously, do not occur being poisoned and dead.By toxicological evaluation standard, it may not be necessary to measure median lethal dose (LD50).According to externalization
The classification of object acute toxicity is learned, peach gum polysaccharide iron is nontoxic or toxicity is minimum.
Under the premise of without departing substantially from the spirit and scope of the present invention (being more specifically set forth in appended claims), ability
Domain those of ordinary skill can be to present invention implementation these or other modifications and changes.It is further understood that the side of different embodiments
Face can exchange in whole or in part.Moreover, it will be appreciated by the skilled addressee that the description of front is exemplary only and nothing
Meaning is limited in the invention further described in these appended claims in this way.Therefore, spirit and scope of the appended claims
It should not be limited by description form wherein included.
Claims (9)
1. a kind of peach gum polysaccharide iron, which is characterized in that the peach gum polysaccharide iron the preparation method comprises the following steps:
S1: 1:1 ~ 3 weigh peach gum polysaccharide extract and trisodium citrate and dissolve in the water of 40 ~ 80 times of volumes in mass ratio, stir
Liquor ferri trichloridi and sodium hydroxide solution that concentration is 2 ~ 5mol/L are mixed and are added dropwise to, it is reddish brown until occurring in reaction liquid
Color precipitating, and precipitate and stop being added dropwise when not being redissolved, then be placed in 1 ~ 2h of reaction in 70 ~ 90 DEG C of water-baths, obtain end reaction liquid；
S2: being centrifuged off precipitating for 500 ~ 1000rpm/min of end reaction liquid in step S1, takes liquid that the nothing of 1 ~ 5 times of volume is added
Water-ethanol is precipitated precipitating and is washed repeatedly 2 ~ 3 times with 70 ~ 80% ethanol solutions, dry peach gum polysaccharide iron；
In the step S1 in peach gum polysaccharide extract peach gum polysaccharide content >=80%.
2. peach gum polysaccharide iron according to claim 1, which is characterized in that in step S1, the peach gum polysaccharide extract passes through
Following methods preparation:
S11: former peach gum being cleaned and is cleaned, and 10 ~ 30 times of amount water are added and impregnate 8 ~ 12h, 50 ~ 70 DEG C of holdings are heated to after homogenate is broken
1 ~ 2h, 2000 ~ 4000rpm/min are centrifuged 3 ~ 5min, take supernatant, obtain raw sugar liquid；
S12: dehydrated alcohol is added in raw sugar liquid described in step S11 by 1 ~ 5:1 times, sufficiently oscillation after in 0 ~ 4 DEG C stand 10 ~
Pellet frozen dry, pulverize after taking precipitating to repeat step 2 ~ 3 time S12, obtain peach gum by 12h, 4000 ~ 6000rpm/min centrifugation
Polysaccharide crude extract；
S13: add water to be configured to the polysaccharide solution of 2 ~ 8mol/mL the peach gum polysaccharide crude extract in step S12, it is fine to cross DEAE-52
Tie up plain column, then eluted using the NaCl solution of 0.1 ~ 2mol/mL, collect elution samples, refilter, be concentrated and dried peach gum is more
Sugar extract.
3. peach gum polysaccharide extract according to claim 2, which is characterized in that be filtered into the step S13 0.45 μm it is micro-
Hole filter membrane or bag filter filtering.
4. peach gum polysaccharide iron described in a kind of claim 1 ~ 3 any one is preparing the application in iron-supplementing preparation.
5. a kind of the answering in preparation enhancing infant immunity food of peach gum polysaccharide iron described in claim 1 ~ 3 any one
With.
6. a kind of preparation for being used to enhance infant immunity comprising peach gum polysaccharide iron described in claim 1, which is characterized in that
According to parts by weight, the preparation includes following components: 30 ~ 50 parts of peach gum polysaccharide iron, 20 ~ 40 parts of active microbial inoculum, wherein described
Active bacteria is one of animal bifidobacteria, bifidobacterium lactis, Lactobacillus rhamnosus or a variety of in active microbial inoculum.
7. according to claim 6 for enhancing the preparation of infant immunity, which is characterized in that further include in the component
Vitamin C.
8. a kind of preparation for enhancing infant immunity of claim 6 or 7 is as nutrition enhancer in nutriment
Application.
9. preparing the application in nutrition enhancer according to claim 8, which is characterized in that the nutrition enhancer is in nutrition
Adding proportion in product is 0.1 ~ 1%.
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Application publication date: 20190827 |