CN101491329B - Immunity adjustment function of corn extract and use thereof in food - Google Patents
Immunity adjustment function of corn extract and use thereof in food Download PDFInfo
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Abstract
The invention discloses a corn extract for adjusting the immunologic function of organisms and application thereof in foods. The invention particularly discloses a corn extract, a method for preparing the extract, a composition containing the extract, an acceptable carrier and/or an auxiliary material and applications of corns, the corn extract and the composition containing the corn extract in preparing products for adjusting the immunologic function, wherein the products comprise foods, health products and medicines, and the method comprises the following steps: extracting the corns for multiple times with a solvent; combining the extracting solutions and reclaiming the solvent; and drying the combined extracting solution to obtain the corn extract.
Description
Technical field
The present invention relates to corn extract, preparation method of extract contains the composition of this type extract, and this type extract is regulated the application in the body's immunity product in preparation.
Background technology
Corn is one of China three generalized grain crops, has another name called maize, big chinese sorghum, ear of maize, maize, maize, beautiful water chestnut, maize, maize, reed broomcorn millet, maize, maize, corn etc.; Be grass family Gramineae Zea annual herb plant, Latin is called Zea mays Linn..Corn originates in Central and South America, mainly is distributed between 30 °-50 ° the latitude; Cultivation is all arranged now all over the world, and what cultivated area was maximum is the U.S., China, Brazil, Mexico, South Africa, India and Romania.The corn belt of China is northeast, North China and southwestern mountain area.Corn not only can be used as food and feed, and still a kind of important reproducible raw material of industry occupies an important position in national food safety.According to National Development and Reform Commission's " about instruction that promotes that corn deep processing already develops in a healthy way " statistics, Tenth Five-Year Plan Period China's corn consumption figure from 1.27 hundred million tons of rising to 2005 of 1.12 hundred million tons in 2000, increase by 2.5% every year.2006 domestic corn consumption figure (not containing outlet) be 1.34 hundred million tons, increased by 5.5% than 2005; Wherein, 8,400 ten thousand tons of feeding consumption account for 64.2% of domestic corn total quantity consumed, and proportion is on a declining curve; Deep processing consumes 3,589 ten thousand tons of corns, accounts for 26.8% of total quantity consumed, and proportion shows a rising trend; Kind usefulness and edible consumption are relatively stable.It should be noted that especially over the past two years along with fossil energy supply in the world is becoming tight, is that the corn deep processing of representative already develops rapidly with cornstarch, ethanol and derived product thereof, becomes one of industry with fastest developing speed in the farming industry.
In order to make full use of the resource of corn, improve human dietary structure, the applicant of this patent studies the chemistry and the physiologically active thereof of corn extract.In addition, the bibliographical information corn contains micro-phosphorus, calcium, iron, selenium, magnesium etc.Also contain vitamin E, B1, B2, B6 and carrotene, nicotinic acid, lysine etc.
Summary of the invention
The object of the present invention is to provide a kind of corn extract.
Another object of the present invention is to provide a kind of method for preparing corn extract.
A purpose more of the present invention is to provide a kind of composition that contains corn extract.
Another purpose of the present invention is to provide a kind of corn extract to regulate the application in the body's immunity product in preparation.
In order to accomplish the present invention's purpose, can adopt following production corn extract technical scheme:
(a) take by weighing corn, clean with flushing with clean water, pulverize, put in the extraction element and preserve moisture infiltration;
(b) use alcohol extracting, filter, merge extract, recovered under reduced pressure alcohol gets ethanol extract to rare medicinal extract shape;
(c) the residue water after the said extracted is extracted, filter, merge extract, recovered under reduced pressure water causes rare medicinal extract shape, gets water extract.
(d) alcohol extract and water extract are merged mixing, drying obtains corn extract of the present invention.
The preferred corn that meets GB GB1353-1999 that uses in the step (a); Pulverizing the preferred fashion of extrusion that uses pulverizes; Time of soaking into of preserving moisture is 0.5-4 hour, preferred 1-2 hour.
The alcohol that step (b) is used can use various lower alcohols, ethanol preferably, and the concentration of preferred alcohol is 30%-95%; Extraction can be carried out under the condition of solvent refluxing in room temperature, preferably the temperature of solvent refluxing; The number of times that extracts is 1-4 time, preferably 2 times; The amount of solvent is that the ratio of solvent and corn is 2-6: 1 (L of unit: kg), 4-5 preferably: the 1 (L of unit: kg);
During recovered under reduced pressure alcohol to rare medicinal extract shape, the 0.3-0.4 that is equivalent to raw material when its volume doubly (extract/raw material=L/kg), density are about at 1.10 o'clock stop the corn alcohol extract.
The mode that step (c) water extracts is that extraction can be carried out under 20-100 ℃ condition; Preferably reflux or decoction,, then must note supplementing solvent water if decoct; The number of times that extracts is that the number of times that extracts is 1-4 time, preferably 2 times; The amount of water, the ratio of water and corn is 2-6: 1 (L of unit: kg), 4-5 preferably: the 1 (L of unit: kg);
Recovered under reduced pressure water causes rare medicinal extract shape, the 0.3-0.4 that its volume is equivalent to raw material doubly (stop during extract/raw material=L/kg), the corn water extract.
All conventional drying modes of step (d) can, preferably adopt dry, the freeze drying of spray pattern.
The corn extract preferred manufacturing procedure:
(a) take by weighing the corn that meets GB GB1353-1999, clean with flushing with clean water, adopt fashion of extrusion to pulverize, put in the refluxing extraction device and preserve moisture and soaked into 1-2 hour;
(b) extract 2 times with the 50%-95% alcohol reflux, (quantity of solvent: 5 times, 4 times); Filter, merge extract, decompression recycling ethanol to rare medicinal extract shape; The 0.3-0.4 that its volume is equivalent to raw material doubly (extract/raw material=L/kg), density is about 1.10, ethanol extract;
(c) the residue water after will extracting reflux or decoct extract 2 times (quantity of solvent: 5 times, 4 times, if decoct; Then must note supplementing solvent water); Filter, merge extract, recovered under reduced pressure water causes rare medicinal extract shape; Its volume is equivalent to 0.3-0.4 times of (extract/raw material=L/kg), get water extract of raw material;
(d) ethanol extract and water extract are merged mixing, adopt spray pattern dry, obtain this patent corn extract.
Corn extract of the present invention can also be used following method preparation:
Corn is decocted extraction 3 times (quantity of solvent: 3-6 times, 3-6 times, 3-6 is doubly) with pure water under the condition of pressurization 0.01-0.1MPa, each time is 30-60min, filtration, and the merging extract, concentrate drying gets the corn water extract.
The remnants corn also adds 50% ethanol atmospheric pressure reflux again and extracts 2 times, and each 1 hour, filter, merge extract, reclaim solvent, concentrate drying gets extract.If pressurised extraction was changed into atmospheric pressure reflux 3 hours, should be also near effect same.
The present invention sees through the chemical identification reaction and confirms that corn extract of the present invention mainly contains corn oligosaccharides and corn polysaccharide.
Corn extract of the present invention can be used for food, health products, medicine.
Described food comprises functional food, functional food additives.
Corn extract can be used as food additives to be added in the present varieties of food items, makes it become functional food.
For example can add in the various types of beverages.
For example can add in all kinds of dairy produces, for example various milk, comprise respectively to children, middle age, old milk and to patient's milk.
For example can add in flour of the prior art, the rice, prepare all kinds of staple foods, non-staple foodstuff such as functional bread, biscuit.
For the convenience of using, food that corn extract is processed or health products can be prepared into various forms such as powder, tablet, capsule, oral liquid.
Corn extract of the present invention forms pharmaceutical composition as acceptable carrier on the active ingredient of medicine and the pharmaceutics.This pharmaceutical composition can be according to method preparation well known in the art.Can be through the pharmaceutically acceptable solid of extract of the present invention and one or more or liquid excipient and/or assistant agent being combined, process to be suitable for any formulation of human or animal's use.The content of extract of the present invention in its pharmaceutical composition is generally 0.1-95 weight %.
Extract of the present invention or contain its pharmaceutical composition can the unit dosage form administration; Method of administration can be enteron aisle or non-enteron aisle, like oral, intravenous injection, intramuscular injection, hypodermic injection, nasal cavity, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum etc.
Form of administration can be liquid dosage form, solid dosage forms or semisolid dosage form.Liquid dosage form can be solution (comprising true solution and colloidal solution), emulsion (comprising o/w type, w/o type and emulsion), supensoid agent, injection (comprising liquid drugs injection, powder-injection and transfusion), eye drops, nasal drop, lotion and liniment etc.; Solid dosage forms can be tablet (comprising ordinary tablet, enteric coatel tablets, lozenge, dispersing tablet, chewable tablets, effervescent tablet, oral disnitegration tablet), capsule (comprising hard shell capsules, soft capsule, capsulae enterosolubilis), granule, powder, micropill, dripping pill, suppository, film, paster, the agent of gas (powder) mist, spray etc.; Semisolid dosage form can be ointment, gel, paste etc.
Extract of the present invention can be processed ordinary preparation, also process is sustained release preparation, controlled release preparation, targeting preparation and various particulate delivery system.
For extract of the present invention is processed tablet, the various excipient well known in the art that can be widely used comprises diluent, binder, wetting agent, disintegrant, lubricant, glidant.Diluent can be starch, dextrin, sucrose, glucose, lactose, sweet mellow wine, sorbierite, xylitol, microcrystalline cellulose, calcium sulfate, calcium monohydrogen phosphate, calcium carbonate etc.; Wetting agent can be water, ethanol, isopropyl alcohol etc.; Adhesive can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, mucialga of arabic gummy, gelatine size, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol etc.; Disintegrant can be dried starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, PVPP, Ac-Di-Sol, sodium carboxymethyl starch, sodium acid carbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, dodecyl sodium sulfate etc.; Lubricant and glidant can be talcum powder, silica, stearate, tartaric acid, atoleine, polyethylene glycol etc.
Can also tablet further be processed coating tablet, for example sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet.
For capsule is processed in the administration unit, can active ingredient extract of the present invention be mixed with diluent, glidant, mixture is directly placed hard shell capsules or soft capsule.Also can active ingredient extract of the present invention be processed particle or micropill with diluent, binder, disintegrant earlier, place hard shell capsules or soft capsule again.Each diluent, binder, wetting agent, disintegrant, the glidant kind that are used to prepare extract tablet of the present invention also can be used for preparing the capsule of extract of the present invention.
For extract of the present invention is processed injection, can water, ethanol, isopropyl alcohol, propane diols or their mixture as solvent and add an amount of this area solubilizer commonly used, cosolvent, pH and adjust agent, osmotic pressure regulator.Solubilizer or cosolvent can be poloxamer, lecithin, HP-etc.; PH adjustment agent can be phosphate, acetate, hydrochloric acid, NaOH etc.; Osmotic pressure regulator can be sodium chloride, sweet mellow wine, glucose, phosphate, acetate etc.As prepare freeze drying powder injection, also can add sweet mellow wine, glucose etc. as proppant.
In addition, like needs, also can in pharmaceutical preparation, add colouring agent, anticorrisive agent, spices, flavouring or other additive.
For reaching the medication purpose, enhancing treatment effect, medicine of the present invention or pharmaceutical composition can be used any known medication administration.
Proved with oligosaccharides and polysaccharide to be that the corn extract of main component has the adjusting body's immunity first.
The effect of corn extract is mainly estimated in this research through B cell and T cell proliferation.Visible by the result; Corn extract induces the normal mouse spleen lymphocyte proliferation of generation that significant inhibitory effect is all arranged for ConA and LPS when 100 μ g/ml concentration; And the SPL (cell proliferation minimizing) of immunocompromised mouse is had the effect that promotes propagation, the propagation function that the prompting corn extract possibly regulated the immunocyte of body under different situations.
But the mouse spleen lymphocyte transformation experiment assessing compound that adopts in the experiment wherein is the lymphocytic propagation of stimulant reflection T with ConA to the influence of mouse lymphocyte propagation, is the propagation of stimulant reflection bone-marrow-derived lymphocyte with LPS.But tardy parasexuality reaction experiment assessing compound is to the mouse cell Immune Effects.But immunity internal organs/body weight ratio assessing compound is to mouse immune organ and whole influence.But mouse carbon clearance experiment assessing compound is to the influence of mouse monokaryon-macrophage function.
Experimental result of the present invention shows; Corn extract accrues to the splenocyte of normal mouse and immunocompromised mouse all has facilitation; It is inhibited to stimulate T lymphopoiesis down and the LPS bone-marrow-derived lymphocyte under stimulating to breed to normal mouse ConA, and T under two kinds of stimulants of immunologic hypofunction mouse are induced or bone-marrow-derived lymphocyte are bred and had the increase effect.To the not influence of immune organ index of normal mouse, and the reduction of the index and spleen index of counter-rotating immunologic hypofunction mouse.Tardy parasexuality reaction to the dinitrofluorobenzene of T cell mediated causes is inhibited.Show that corn extract has immunoregulation effect to the function of mouse lymphocyte.Corn extract shows the not influence of monokaryon-macrophage function not influence of mouse carbon clearance test.
Because body immune system is the system of defense that is made up of jointly nospecific immunity, humoral immunity and cellular immunity, humoral immunity is mainly accomplished by the B cell, and cellular immunity is mainly accomplished by the T cell.To pathogenic microorganism invasion and attack and cells in vivo variation, humoral immunity and cellular immunity produce various biological effects through forming modes such as antibody or generation cell factor to specific antigen.
Therefore corn extract of the present invention can be used to prepare the medicine that prevents and/or treats disease of immune system.
If body is by certain antigen sensibilization, when exposure phase once more with antigen the time then secondary immune response be enhanced.When the immunity of body is in high response status, then answer immune response to cross strong and cause tissue damage, be called hypersensitivity.Therefore because corn extract induces the normal mouse spleen lymphocyte proliferation of generation that significant inhibitory effect is all arranged for ConA and LPS, can be used for treating hypersensitivity.Hypersensitivity comprises the hypersensitivity of I type, the hypersensitivity of II type, the hypersensitivity of III type and the hypersensitivity of IV type.The hypersensitivity of I type comprises pollinosis, bronchial astehma, atopic dermatitis, food allergy.The hypersensitivity of II type comprises for example transfusion reaction of interindividual II type of the same race hypersensitivity, erythroblastosis fetalis, graft-rejection, and the hypersensitivity of LADA II type is autoimmune hemolytic anemia, empsyxis ephritis syndrome, LADA receptor disease for example.The hypersensitivity of III type comprises the inflammation damnification due to the local immune complex that forms, the disease due to the CIC ELISA.Hypersensitivity of IV type such as contact dermatitis, graft-rejection.
Corn extract can be treated autoimmunity disease such as system lupus erythematosus, rheumatoid arthritis, myasthenia gravis.
Corn extract has the effect that promotes propagation to the SPL (cell proliferation minimizing) of immunocompromised mouse, therefore can treat various immunologic deficiency diseases.
The dosage of extract pharmaceutical composition of the present invention according to prevent or treat the character and the order of severity of disease, the individual instances of patient or animal, method of administration and formulation etc. can have large-scale variation.In general, the appropriate dose scope of the every day of extract of the present invention is the 0.001-150mg/Kg body weight, is preferably the 0.1-100mg/Kg body weight, and more preferably the 1-60mg/Kg body weight most preferably is the 2-30mg/Kg body weight.Above-mentioned dosage can a dosage unit or is divided into several dosage unit administrations, and this depends on doctor's clinical experience and comprises the dosage regimen of using other treatment means.
Extract of the present invention or composition can be taken separately, or merge use with other treatment medicine or symptomatic drugs.When extract of the present invention and other medicine existence synergy, should adjust its dosage according to actual conditions.
Advantage of the present invention:
(1) no matter have the two-phase immunoregulation effect, be that immunologic function is crossed by force or immunologic hypofunction, all can regulate to normal level.
(2) has no toxic and side effect.
(3) preparation technology is simple, does not need complex apparatus, and cost is low.
Term and abbreviation
YM-J95: corn 95% alcoholic extract
YM-J80: corn 80% alcoholic extraction, the extract of water extraction again
YM-J50: corn 50% alcoholic extract
YM-S: corn water extract
Description of drawings
Fig. 1 corn extract YM-J50, YM-S are to the influence of male mice body weight
Fig. 2 corn extract YM-J50, YM-S are to the influence of female mice body weight
Fig. 3 corn extract YM-J50, YM-S are to the influence of male mice food ration
Fig. 4 corn extract YM-J50, YM-S are to the influence of female mice food ration
The specific embodiment
The preparation of corn extract
Take by weighing the corn that meets GB GB1353-1999, clean with flushing with clean water, adopt fashion of extrusion to pulverize, put in the refluxing extraction device and preserve moisture and soaked into 2 hours; Extract 2 times with 95% alcohol reflux, (quantity of solvent: 5 times, 4 times); Filter, merge extract, decompression recycling ethanol to rare medicinal extract shape; Its volume be equivalent to 0.4 times of raw material (extract/raw material=L/kg), or density is about 1.10, ethanol extract.
With the residue water after extracting reflux or decoct extract 2 times (quantity of solvent: 5 times, 4 times, if decoct; Then must note supplementing solvent water), filter, merge extract; Recovered under reduced pressure water causes rare medicinal extract shape, and its volume is equivalent to 0.3 times of (extract/raw material=L/kg), get water extract of raw material.
Ethanol extract and water extract are merged mixing, adopt spray pattern dry, obtain this patent corn extract.
Embodiment 2
Take by weighing the corn that meets GB GB1353-1999, clean with flushing with clean water, adopt fashion of extrusion to pulverize, put in the refluxing extraction device and preserve moisture and soaked into 1 hour; Extract 2 times with 80% alcohol reflux, (quantity of solvent: 4 times, 3 times); Filter, merge extract, decompression recycling ethanol to rare medicinal extract shape; Its volume be equivalent to 0.3 times of raw material (extract/raw material=L/kg), or density is about 1.10, ethanol extract.
With the residue water after extracting reflux or decoct extract 2 times (quantity of solvent: 5 times, 4 times, if decoct; Then must note supplementing solvent water), filter, merge extract; Recovered under reduced pressure water causes rare medicinal extract shape, and its volume is equivalent to 0.35 times of (extract/raw material=L/kg), get water extract of raw material.
Ethanol extract and water extract are merged mixing, adopt spray pattern dry, obtain this patent corn extract YM-J80.
Embodiment 3
Take by weighing the corn that meets GB GB1353-1999, clean with flushing with clean water, adopt fashion of extrusion to pulverize, put in the refluxing extraction device and preserve moisture and soaked into 1 hour; Extract 2 times with 50% alcohol reflux, (quantity of solvent: 5 times, 4 times); Filter, merge extract, decompression recycling ethanol to rare medicinal extract shape; Its volume be equivalent to 0.35 times of raw material (extract/raw material=L/kg), or density is about 1.10, ethanol extract.
With the residue water after extracting reflux or decoct extract 2 times (quantity of solvent: 5 times, 4 times, if decoct; Then must note supplementing solvent water), filter, merge extract; Recovered under reduced pressure water causes rare medicinal extract shape, and its volume is equivalent to 0.4 times of (extract/raw material=L/kg), get water extract of raw material.
Ethanol extract and water extract are merged mixing, adopt spray pattern dry, obtain this patent corn extract.
Embodiment 4
The 10kg corn is extracted 3 times with 95% alcohol reflux, and by weight, quantity of solvent is respectively 5 times of corn, 3.5 times, 3.5 times, merges extract, decompression recycling ethanol, vacuum drying.Get corn ethanol extract YM-J95.
The 10kg corn is extracted 3 times with 80% alcohol reflux, and by weight, quantity of solvent is respectively 5 times of corn, 3.5 times, 3.5 times, merges extract, decompression recycling ethanol, vacuum drying.Get corn ethanol extract YM-J80.
Embodiment 6
The 10kg corn is extracted 3 times with 50% alcohol reflux, and by weight, quantity of solvent is respectively 5 times of corn, 3.5 times, 3.5 times, merges extract, decompression recycling ethanol, vacuum drying.Get corn ethanol extract YM-J50.
Embodiment 7
With 10kg corn water refluxing extraction 3 times, by weight, quantity of solvent is respectively the quantity of solvent of corn: 5 times, 3.5 times, 3.5 times, merge extract, recovered under reduced pressure water, vacuum drying.Get corn water extract YM-S.
Embodiment 8
The 20kg corn decoct is extracted (quantity of solvent: 5 times, 5 times, 5 times) 3 times with pure water under the condition of pressurization 0.01-0.1MPa, each time is 30-60min, filters, and merges extract, reclaims solvent, to about 10L, volatilizes vacuum drying with caldron.Get corn water extract YM-S.
The 20kg corn decocted under 3 hours condition of atmospheric pressure reflux with pure water extract (quantity of solvent: 5 times, 5 times, 5 times) 3 times, each time is 30-60min, filters, and merges extract, reclaims solvent, to about 10L, volatilizes vacuum drying with caldron.Get corn water extract YM-S.
The 20kg corn decoct is extracted (quantity of solvent: 5 times, 5 times, 5 times) 3 times with pure water under the condition of pressurization 0.01-0.1MPa, each time is 30-60min; Filter, merge extract, remaining corn adds 50% ethanol atmospheric pressure reflux again and extracts each 1 hour 2 times; Filter, merge, reclaim solvent with aqueous extract; To about 10L, volatilize vacuum drying with caldron.Get corn water extract YM-S.
Embodiment 11
The evaluation of chemical composition in the corn extract:
Get corn extract 6000ml (containing extract 400g approximately through determination of solid content), 4 times (1500ml 1500ml), merges petroleum ether layer, reclaims benzinum and gets ligroin extraction 5g for 3000ml, 2000ml with petroleum ether extraction.Water layer with ethyl acetate extraction 4 times (2500ml, 1500ml, 1500ml, 1500ml) combined ethyl acetate layer reclaims solvent and gets ethyl acetate extract 3g, show in the corn extract the solvable composition of ethyl acetate seldom.Water layer is used extracting n-butyl alcohol 3 times again, and (2000ml, 1500ml 1500ml), merge n-butanol layer; With 5% sodium acid carbonate extraction 3 times (1000ml, 1000ml, 1000ml); (1500ml 1500ml), reclaims solvent and gets n-butanol extract 3.5g with distilled water washing 2 times again.Buck is partly used H
2SO
4Be acidified to subacidity, (2000ml, 1500ml), n-butanol layer is washed with water to neutrality, and (1500ml 1500ml), reclaims the extracting n-butyl alcohol acidic moiety 3g of solvent to use extracting n-butyl alcohol again 2 times.
Water layer after the extraction volatilizes and obtains about 380g
Above result shows that corn extract is a main compound with the water-soluble sugar constituents.
Get 50% corn extract, be dissolved in water,, the aqueous solution is volatilized in water-bath with extracting n-butyl alcohol 3 times, the brown solid thing.Get this solids, for dissolving, measure 1H with deuterium, 13C and DEPT NMR spectrum, must data 1HNMR (400MHz, D2O) δ 5.43 (1H, d, J=4.0Hz); 4.24 (1H, d, J=8.8Hz), 4.07 (1H, t, J=8.8Hz), 3.92 (1H, m); 3.88 (1H, m), 3.85 (4H, m), 3.78 (1H, t, J=9.6Hz); 3.58 (1H, dd, J=10.0,4.0Hz), 3.49 (1H, t, J=9.6Hz).
13CNMR(100MHz，D2O)δ：103.8(s)，92.3(d)，81.5(d)，76.6(d)，74.2(d)，72.7(d)，72.6(d)，71.2(d)，69.4(d)，62.5(t)，61.5(t)，60.3(t)。Above data are consistent with the structure of sucrose, show to contain oligosaccharides in the corn extract.
The water intaking extract is dissolved in water, with extracting n-butyl alcohol 3 times, the aqueous solution is volatilized in water-bath, the brown solid thing.Get this solids, with D
21H NMR (400MHz) spectrum is measured in the O dissolving, and the result shows that the resonance signal of 1 hydrogen of sugar is arranged in δ 4.6-5.4 scope, the resonance signal of hydrogen on sugared company's oxygen carbon is arranged in δ 3.1-4.2 scope, shows to contain the polysaccharide composition.
Get n-butanol extract 3.5g; Adding silica gel mixed sample with the methyl alcohol dissolving, is adsorbent with silica gel, chloroform-methanol in varing proportions (100: 0-100: 1-50: 1-25: 1-15: 1-10: 1-5: 1-3: 1) carry out chromatographic isolation for eluant, eluent; Every 500ml collects a, collects 80 parts altogether.Separate out white solid at the 39th part, cross and filter 50mg.It is the monoglyceride constituents of unrighted acid to detect supposition through 1HNMR.Other stream parts are not separated out solid, show the n-butanol DDGS that contains very small amount in the corn extract.
Qualitative reaction: get corn extract a little, be dissolved in the 2ml water, drip 1ml alpha-Naphthol solution, fully the vibration back drips the 2ml concentrated sulfuric acid along test tube wall, leaves standstill, and a lilac band occurs at the solution middle part, proves to contain carbohydrate content.
Above-mentioned identification reaction corn extract of the present invention mainly contains corn oligosaccharides and corn polysaccharide.
Pharmacological evaluation
Experimental example 1 corn extract YM-J50, YM-S acute toxicity test in mice
1. make a summary
The 1% sodium carboxymethylcellulose suspension 18.0g/kg of disposable orally give YM-J50 of mouse or YM-S, none death of mouse in 14 days, generally in order, the no abnormality seen reaction.Body weight and 1% sodium carboxymethylcellulose control group no significant difference relatively in the mouse 14 days.
In 14 days observation periods, except that YM-J50 18.0g/kg group male mice feed intake obviously reduced than 1% sodium carboxymethylcellulose control group after administration on the 5th day, all the other were respectively organized each day and compare no significant difference with control group.
YM-J50, YM-S 18.0g/kg group male mice liver coefficient obviously reduce (P＜0.05 than control group; P＜0.01); YM-J50 18.0g/kg group female mice thymus gland coefficient obviously increases (P＜0.01) than control group, all the other internal organs: the heart, spleen, lung, kidney, brain, testis internal organs relative weights such as (uterus) and control group be no significant difference relatively.
This dosage is 90 times of " the H22 mouse-borne tumor test of pesticide effectiveness " effective dose (200mg/kg).
2. experiment purpose
Observe and the acute toxicity of estimating corn extract YM-J50, YM-S, reference is provided for formulating the clinical application scheme.
3. receive the reagent thing
YM-J50, YM-S: provide by institute of Materia Medica,Chinese Academy of Medical Sciences, outward appearance is the brown ceramic powder shape.Test sample is preserved in 4 ℃ of refrigerators.Test sample is mixed with the suspension of 0.3g/mL with 1% sodium carboxymethylcellulose.Dosage is the 0.3mL/10g body weight, administration 2 times, twice administration midfeather 5h.
4. animal
SPF Kunming mouse (KM) mouse, 60, female, hero half and half.Provided by Beijing Vital River Experimental Animals Technology Co., Ltd., licensing is numbered SCXK (capital) 2007-0001.The animal 16h that stops eating with the animal random packet, specifies a single animal number for every animal by the back body weight of stopping eating, and ticks on the hair of animal with saturated picric acid solution (yellow), with as the numbering of discerning animal.Change the mouse cage bedding and padding every day once.
At duration of test, animal is raised in No. 1 Animal House of Chinese department of traditional Chinese medicine institute's medical experiment animal center.Raise according to State Standard of the People's Republic of China GB 14922.2-2001.Animal House is a barrier system, artificial lighting, and in 12 hours light and shade cycles, temperature is controlled at 20～22 ℃ of scopes, and relative humidity is 40～70%, 15 times/h of rate of ventilation.
Use standard mouse pellet is provided by Beijing section Austria feed corporation,Ltd that pulls together, and product license number is SCXK (capital) 2005-0007, and registration card number: (joining) word is raised No. 238 in the capital.Lot number: 07100801.
With 500ml Merlon drinking-water bottle splendid attire pure water, supply animal arbitrarily to drink, change fresh drinking-water bottle every day.
5. experimental technique
5.1 experiment is divided into groups:
Kunming mouse is weighed behind the mouse fasting 16h (can't help water) before the experiment, 60 of the healthy mices of selection 17-20g body weight, male and female half and half.The back body weight is divided into sodium carboxymethylcellulose control group (20), YM-J50 18.0g/kg group (20) to animal at random, YM-S 18.0g/kg organizes (20) totally 3 groups, male and female half and half by stopping eating.
5.2 method of administration:
Each organizes equal gastric infusion 2 times, midfeather 5h, and the 0.3ml/10g body weight, control group is irritated capacity 1% sodium carboxymethylcelluloses such as stomach.
5.3 observing time: 14 days
5.4 experimental record:
5.4.1 general state is observed: observe generally performance, mouse activity, breathing, behavior after the mouse administration, have or not situation such as convulsions, gastrointestinal reaction, feed water inlet; Record has or not dead mouse; If dead mouse is arranged, then dead mouse is carried out gross anatomy, observe the internal organs situation.Surviving animals is observed 1 time every day such as the general situation of record animal, hair color, activity, gait, expression, stool, urine etc.Observation has or not poisoning symptom to take place and time of taking place and duration, recovery situation, writes down dying animal.
5.4.2 body weight weighing: to surviving animals weighing the weight of animals 1 time every other day, the equal non-fasting of animal when weighing.16h begins fasting before zootomy, faces body weight after the when dissected weighing fasting.
5.4.3 feed consumption: viewing duration, in trough, quantitatively add feed, claim forage volume every other day 1 time, record adds and remaining forage volume respectively.The forage volume that feed consumption equals to add deducts surplus.Convert every animal consumption every day.
5.4.4 internal organs are weighed and the relative weight conversion: weighing is carried out in heart, liver, spleen, lungs, kidney, brain, thymus gland, testis, uterus, according to the body weight of every animal, the relative weight of conversion internal organs.Internal organs relative weight (g internal organs weight/100g body weight)=100 * organ weights (g)/the weight of animals (g).
6. statistical analysis: all female, the male separately ANOVA of the spss13.0 statistical software method of data such as body weight, feed consumption, internal organs relative weight is carried out statistical analysis.
7. experimental result
7.1 dead animal record and overview:
Behind the oral YM-J50 of mouse, repose, the movable minimizing, it is normal that about 15min recovers.None death of mouse was observed 14 days, mouse hair color, behavior, activity normally, do not see abnormal responses such as digestive system, nervous system, mouse feed, drinking-water, neural reflex etc. and control group no significant difference.
Behind the oral YM-S of mouse, movable normal.None death of mouse was observed 14 days, mouse hair color, behavior, activity normally, do not see abnormal responses such as digestive system, nervous system, mouse feed, drinking-water, neural reflex etc. and control group no significant difference.
7.2 mouse body weight and food ration:
Acute toxicity testing was observed during 14 days, and male mice body weight YM-J50 and YM-S 18.0g/kg group compare no significant difference with 1% sodium carboxymethylcellulose control group.The result sees table 1, Fig. 1.
Acute toxicity testing was observed during 14 days, and female mice body weight YM-J50 and YM-S 18.0g/kg group compare no significant difference with 1% sodium carboxymethylcellulose control group.The result sees table 2, Fig. 2.
Acute toxicity testing was observed during 14 days; YM-J50 18.0g/kg group male mice feed intake obviously reduced than 1% sodium carboxymethylcellulose control group after administration on the 5th day; But the feed consumption at other fates is not seen influence, so think that certain contingency is arranged.YM-J50 18.0g/kg organizes female mice and YM-S 18.0g/kg is female, the feed consumption of male mice all compares no significant difference with control group.The result sees table 3, Fig. 3.
Acute toxicity testing was observed during 14 days, and female mice feed intake YM-J50 and YM-S 18.0g/kg group compare no significant difference with 1% sodium carboxymethylcellulose control group, and the result sees table 4.Fig. 4 shows after the administration of YM-J50 and YM-S 18.0g/kg group the 3rd day, and the mouse feed intake has reduction slightly than control group, all increases to some extent than control group since the 7th day.
7.3 internal organs relative weight:
The liver relative weight of YM-J50, YM-S 18.0g/kg group buck obviously reduces (P＜0.05 than 1% sodium carboxymethylcellulose control group; P＜0.01), internal organs relative weight such as the heart, spleen, lung, kidney, brain, thymus gland, testis and control group relatively do not have significant difference.The result sees table 5.
The thymus gland relative weight of YM-J50 18.0g/kg group jenny obviously increases (P＜0.01) than 1% sodium carboxymethylcellulose control group, and internal organs relative weight such as the heart, liver, spleen, lung, kidney, brain, uterus and control group relatively do not have significant difference.The result sees table 6.
Table 1 corn extract YM-J50, YM-S are to the influence (n=10,
) of male mice body weight
Table 2 corn extract YM-J50, YM-S are to the influence (n=10,
) of female mice body weight
Table 3 corn extract YM-J50, YM-S are to the influence of male mice food ration (n=2,
g/ only/day)
Table 4 corn extract YM-J50, YM-S are to the influence of female mice food ration (n=2,
g/ only/day)
Table 5 corn extract YM-J50, YM-S are to the influence (n=10,
) of male mice internal organs relative weight
Table 6 corn extract YM-J50, YM-S are to the influence (n=10,
) of female mice internal organs relative weight
8. brief summary and conclusion
After the disposable filling stomach of mouse is tried thing YM-J50 18.0g/kg or YM-S 18.0g/kg, after administration, in 14 days observation periods, do not see animal dead, two groups of mouse all do not see toxic reaction.YM-J50 18.0g/kg group male mice feed intake reduced than control group after administration on the 5th day, but thought and be fortuitous phenomena.YM-J5018.0g/kg organizes female mice and YM-S 18.0g/kg is female, the feed consumption of male mice all compares no significant difference with control group.YM-J50 18.0g/kg, YM-S 18.0g/kg group male mice liver coefficient obviously reduce (P＜0.05 than 1% sodium carboxymethylcellulose control group; P＜0.01); YM-J50 18.0g/kg group female mice thymus gland coefficient obviously increases (P＜0.01) than 1% sodium carboxymethylcellulose control group, all the other internal organs: the heart, spleen, lung, kidney, brain, testis internal organs relative weights such as (uterus) and 1% sodium carboxymethylcellulose control group be no significant difference relatively.In addition, the body weight of YM-J50 18.0g/kg, YM-S 18.0g/kg group mouse is than 1% sodium carboxymethylcellulose control group no significant difference, and each hair color, behavior, activity, feed, drinking-water, digestive system, neural reflex etc. of organizing mouse is no abnormality seen all.The perusal heart, spleen, lung, kidney, brain, testis main organs such as (uterus) are not seen obviously unusual.
After conclusion, mouse stomach gave YM-J50 18.0g/kg or YM-S 18.0g/kg, the animal well-tolerated was not seen animal dead, did not see obvious toxic reaction.Think that tentatively YM-J50 and YM-S are safer.
The influence that experimental example 2 corn extracts transform mouse spleen lymphocyte
1, principle T cell, B cell surface have the acceptor and the mitogen receptor of identification antigen, and corresponding lymphocyte clone is bred.Concanavalin A (ConA) optionally stimulates T cell proliferation as the polyclone stimulant; Lipopolysaccharides (LPS) then stimulates B cell proliferation.Utilize mitochondria hydrolase in living cells, the particularly proliferative cell can MTT be decomposed into the principle of bluish violet crystallization, can reflect the propagation situation of cell through the OD value of measuring the bluish violet product.
2, key instrument and material
RPMI-1640 cell culture fluid (Gibco); Hyclone (PAA); ConA and LPS (Sigma); MTT (Sigma) 200 order cells sieve, 96 porocyte culture plates, carbon dioxide incubator, ELIASA
3, experimental procedure
3.1 adopt Kunming kind immunocompromised mouse (endoxan is with the modeling in 4 days of the continuous lumbar injection of 30mg/kg dosage) and normal mouse in the splenocyte suspension preparation experiment; The aseptic spleen of getting; Pulverize gently, process single cell suspension, filter through 200 eye mesh screens; With PBS washing lotion washing 2 times, each 10 minutes (1000rpm/min).At once used the erythrocyte cracked liquid splitting erythrocyte 4-5 minute, PBS washing lotion washing 2 times, each 10 minutes (1000rpm/min).Cell is suspended from 1640 culture mediums that 10ml contains 10% serum, and counting and adjustment cell concentration are 1*10
6Individual/ml.
3.2 lymphocyte proliferation assay joins splenocyte suspension in the 96 porocyte culture plates; 200 μ l/ holes; The stimulant group adds LPS (final concentration is 5 μ g/ml) or ConA (final concentration is 3 μ g/ml); The administration group adds the medicine (final concentration is 100 μ g/ml) of LPS or ConA and respective concentration, and control group adds corresponding solvent.Place 5%CO2,37 degree were cultivated 72 hours, cultivated and finished preceding 4 hours, and every hole adds 10 μ l/ hole MTT (original concentration 5mg/ml).Cultivation end back is centrifugal to be abandoned supernatant and adds cell pyrolysis liquid, and the 570nm/630nm wavelength is measured the OD value down.
4, data
Relatively adopt student ' s T check between group, P＜0.05 is judged as significant difference.
5, experimental result
The influence that the mouse lymphocyte that table 7, extract stimulate LPS transforms
(n＝3，mean±sd)
| Normal mouse (O.D.) | Immunocompromised mouse (O.D.) | |
| Negative control group | 0.205±0.018 | 0.408±0.049 |
| The LPS stimulating group | 0.352±0.039 ## | 0.282±0.038 ## |
| YM-J80(100μg/ml) | 0.23±0.005** | 0.503±0.068** |
| YM-J50(100μg/ml) | 0.295±0.008* | 0.364±0.033* |
* P＜0.05 with, * * P＜0.01 is compared with stimulating group; ##P＜0.01 is compared with negative control group.
Visible by table 7, but LPS significant stimulation normal mouse bone-marrow-derived lymphocyte propagation, and YM-J80 and YM-J50 can significantly suppress this proliferation function.Intraperitoneal injection of cyclophosphamide causes the mouse bone-marrow-derived lymphocyte multiplication capacity of immunocompromised than the remarkable decline of negative control group, and YM-J80 and YM-J50 all can significantly improve its multiplication capacity.
The influence that the mouse lymphocyte that table 8, extract stimulate ConA transforms
(n＝3，mean±sd)
| Normal mouse (O.D.) | Immunocompromised mouse (O.D.) | |
| Negative control group | 0.187±0.013 | 0.442±0.016 |
| The ConA stimulating group | 0.422±0.033 ## | 0.383±0.017 ## |
| YM-J80(100μg/ml) | 0.333±0.040** | 0.493±0.060** |
| YM-J50(100μg/ml) | 0.210±0.088** | 0.419±0.028* |
* P＜0.05 with, * * P＜0.01 is compared with stimulating group; ##P＜0.01 is compared with negative control group.
Visible by table 8, but ConA significant stimulation normal mouse T lymphopoiesis, and YM-J80 and YM-J50 can significantly suppress this proliferation function.Intraperitoneal injection of cyclophosphamide causes the mouse T lymphocyte multiplication capacity of immunocompromised than the remarkable decline of negative control group, and YM-J80 and YM-J50 all can significantly improve its multiplication capacity.
6, conclusion
Body immune system is the system of defense that is made up of jointly nospecific immunity, humoral immunity and cellular immunity, and humoral immunity is mainly accomplished by the B cell, and cellular immunity is mainly accomplished by the T cell.To pathogenic microorganism invasion and attack and cells in vivo variation, humoral immunity and cellular immunity produce various biological effects through forming modes such as antibody or generation cell factor to specific antigen.The effect of YM-J80 and YM-J50 is mainly estimated in this research through B cell and T cell proliferation.Visible by the result; YM-J80 and YM-J50 induce the normal mouse spleen lymphocyte proliferation of generation that significant inhibitory effect is arranged for ConA and LPS when 100 μ g/ml concentration; And the SPL (cell proliferation minimizing) of immunocompromised mouse is had the effect that promotes propagation, the propagation function of pointing out YM-J80 and YM-J50 possibly regulate the immunocyte of body under different situations.
Experimental example 3 corn extract YM-S are to the evaluation of effect of mouse immune system
One, purpose
Carry out the evaluation of effect of corn extract YM-S to the mouse immune system.
Two, experiment material
1, animal used as test
The C57BL/6 mouse, 6-8 week, male (the SPF level, the licensing numbering: SCXK capital 2004-0001), available from Institute of Experimental Animals, Chinese Academy of Medical Sciences
The ICR mouse, 18-20g, male, SPF level (available from Beijing dimension tonneau China animal center)
2, experiment reagent and compound
MTT, concanavaline (ConA), lipopolysaccharides (LPS) (sigma), RPMI-1640 (GiCBo), DFNB, india ink (commercially available), it is pure that other reagent is homemade analysis.Endoxan (commercially available), YM-S (providing) by Hailin seminar of the institute of Materia Medica,Chinese Academy of Medical Sciences native chemical research department Qin
3, laboratory apparatus
Low-temperature and high-speed centrifuge (Hettich EBA12R); Superclean bench (Beijing semiconductor equipment one factory); XSZ-D2 inverted microscope (optical instrument factory, Chongqing); Model 450 type ELIASAs (Bio-Rad); Constant-temperature shaking water-bath (YAMAYO BT-25); The multi-functional oscillator of GZ-I (Haidian, Beijing electronic medical instruments factory); Rco3000T-5vBA CO2gas incubator (Revco)
Three, experimental technique
1, the foundation of immunocompromised mouse model
The model group mouse was injected 4 days at administration beginning in the 3rd day intraperitoneal injection of cyclophosphamide (30mg/Kg) every day 1 time continuously, continued administration simultaneously.
2, the preparation of mouse spleen lymphocyte
Get mice spleen under the gnotobasis, place the plate that fills aseptic D-hanks liquid in right amount, tear to shreds, process single cell suspension with tweezers.Cross 200 order cells sieve, aseptic PBS washed twice, each 5ml.Cell is resuspended, add Tris-NH4CL5ml, leave standstill 5min, splitting erythrocyte, centrifuge washing twice, 1000rpm, 5min.The re-suspended cell suspension, using complete RPMI-1640 adjustment cell concentration is 5 * 10
5Individual/ml.
3, lymphproliferation response
With the every hole 100 μ l of SPL suspension, be inoculated in the 96 hole circle floor cells plates, add ConA (final concentration is 3 μ g/ml) or LPS (final concentration is 5 μ g/ml) simultaneously, 100 μ l/ holes.Other sets blank hole (not adding derivant), 37 ℃, 5%CO
2Hatch 72h, cultivate and finish preceding 4h, add MTT (5mg/ml) 20 μ l, after cultivation finishes, measure the 540nm place and measure the OD value, the calculating LSI.The stimulating group OD value medicine of LSI (SI)=stimulating group OD value/not.
4, internal organs/weight ratio pH-value determination pH
Measure thymus gland/body weight ratio and the spleen/body weight ratio of the immunocompromised mouse that corn extract induces normal mouse and endoxan.Every treated animal number average >=12, administration time is 7 days.After administration finishes, weighing mouse body weight, thymic weight, spleen weight.Thymus index=thymic weight/10g body weight, index and spleen index=spleen weight/10g body weight.
5, tardy parasexuality reaction (DTH)
Every treated animal number average >=12, administration time is 7 days.Except that the normal control group, every group of mouse web portion scraped hair, the about 3cm * 3cm of area.50ul evenly smears sensitization with DFNB solution.Evenly smear reinforcement with DNFB solution 50ul two days later.Evenly smeared with DNFB solution 10ul in the 7th day and respectively organize mouse right ear (two sides) and attack, attack back 24h cervical vertebra dislocation and put to death mouse, cut left and right sides auricular concha, take off the auricle of diameter 8mm, weigh with card punch.The degree of representing DTH with the difference of left and right sides ear weight.
6, mouse carbon is cleaned up experiment
Every treated animal number average >=12, administration time is 7 days.2h after the last administration, every mouse is by tail vein injection india ink 0.1ml/10g, then in 2min (t
1), 10min (t
10), get eye socket blood 20 μ l respectively, put and fill 2ml 0.1% sodium carbonate in vitro, in 600nm wavelength colorimetric, read optical density (OD
1, OD
10).After getting blood, draw neck to put to death, weigh respectively, liver and spleen weight, represent the ability that mouse carbon is cleaned up with phagocytic index.By formula calculate and clean up index (K value) K=(logOD1-logOD10)/(t10-t1)=[log (OD1/OD10)]/8.The K value obtains phagocytic index α value
after body weight and liver spleen heavily convert
Four, experimental result
1, corn extract influence that normal mouse and immunocompromised mouse spleen lymphocyte are accrued
As shown in table 1, YM-S accrues to normal and immunocompromised mouse lymphocyte all has obvious facilitation.
The influence that table 1 compound accrues to normal mouse and immunocompromised mouse spleen lymphocyte
n＝5，Mean±SD.
2, corn extract is to the normal mouse of conA stimulation and the influence of immunocompromised mice spleen lymphocytes proliferation
As shown in table 2, YM-S suppresses the intact animal lymphopoiesis that conA stimulates, and promotes the lymphopoiesis of immunocompromised animal.
Table 2 compound is to the mouse of ConA stimulation and the influence of immunocompromised mice spleen lymphocytes proliferation
n＝5，Mean±SD.
3, corn extract is to the normal mouse of LPS stimulation and the influence of immunocompromised mice spleen lymphocytes proliferation
As shown in table 3, YM-S can significantly suppress the propagation of the normal mouse SPL of LPS stimulation, but promotes the lymphopoiesis of immunocompromised mouse.
Table 3 compound is to the mouse of LPS stimulation and the influence of immunocompromised mice spleen lymphocytes proliferation
n＝5，Mean±SD
4, corn extract is to the influence of internal organs/body weight ratio of normal mouse and immunocompromised mouse
As shown in table 4, YM-S does not all make significant difference to the thymus index and the index and spleen index of normal mouse.Thymus index and the index and spleen index of immunocompromised mouse all significantly reduce than normal control group, and YM-S can significantly increase the index and spleen index of immunocompromised mouse, and its thymus index is not had appreciable impact.
Table 4 compound is to the influence of the organ index of normal mouse and immunocompromised mouse
N=10, Mean ± SD,
bP＜0.01vs normal group,
dP＜0.01vs model control group
5, the influence of corn extract ear's inflammation that dinitrofluorobenzene is induced
As shown in table 5, YM-S can significantly alleviate the degree of the tardy parasexuality reaction of mouse.
The influence of ear's inflammation that table 5 compound is induced dinitrofluorobenzene
n＝12，Mean±SD
6, corn extract is to the influence of mouse monokaryon-macrophage function
As shown in table 6, YM-S does not have appreciable impact to the carbon clearance ability of normal mouse.Body weight, liver weight, spleen heavily there is not appreciable impact.
Table 6 compound is to the influence of mouse monokaryon-macrophage function
N=12, Mean ± SD vs normal control group
Five conclusions
The mouse spleen lymphocyte transformation experiment that adopts in the experiment can be estimated the influence of extract to mouse lymphocyte propagation, wherein is the lymphocytic propagation of stimulant reflection T with ConA, is the propagation of stimulant reflection bone-marrow-derived lymphocyte with LPS.Tardy parasexuality reaction experiment can be estimated extract to the mouse cell Immune Effects.Immunity internal organs/body weight ratio can be estimated extract to mouse immune organ and whole influence.The experiment of mouse carbon clearance can be estimated the influence of extract to mouse monokaryon-macrophage function.
This experimental result shows; YM-S accrues to the splenocyte of normal mouse and immunocompromised mouse all has facilitation; It is inhibited to stimulate T lymphopoiesis down and the LPS bone-marrow-derived lymphocyte under stimulating to breed to normal mouse ConA, and T under two kinds of stimulants of immunologic hypofunction mouse are induced or bone-marrow-derived lymphocyte are bred and had the increase effect.To the not influence of immune organ index of normal mouse, and the reduction of the index and spleen index of counter-rotating immunologic hypofunction mouse.Tardy parasexuality reaction to the dinitrofluorobenzene of T cell mediated causes is inhibited.Show that YM-S has immunoregulation effect to the function of mouse lymphocyte.YM-S shows the not influence of monokaryon-macrophage function not influence of mouse carbon clearance test.
Claims (6)
1. corn extract is regulated the application in the immunologic function product in preparation, and the preparation method of said corn extract is:
A) take by weighing corn, clean with flushing with clean water, pulverize, put in the extraction element and preserve moisture infiltration;
B) use alcohol extract, filter, merge extract, decompression recycling ethanol to rare medicinal extract shape gets ethanol extract;
C) the residue water after the said extracted is extracted, filter, merge extract, recovered under reduced pressure water causes rare medicinal extract shape, gets water extract;
D) alcohol extract and water extract are merged mixing, drying obtains corn extract.
2. according to the application of claim 1, it is characterized in that the preparation method of said corn extract is:
A) take by weighing the corn that meets GB GB1353-1999, clean with flushing with clean water, adopt fashion of extrusion to pulverize, put in the refluxing extraction device and preserve moisture and soaked into 1-2 hour;
B) extract 2 times with the 50%-95% alcohol reflux, quantity of solvent is respectively 5 times and 4 times, filters; Merge extract, decompression recycling ethanol to rare medicinal extract shape, its volume are equivalent to 0.3-0.4 times of raw material; Said ratio is extract/raw material=L/kg, and density is 1.10, gets ethanol extract;
C) the residue water after will extracting refluxes or decocts and extracts 2 times, and quantity of solvent is respectively 5 times and 4 times, if decoction; Then must replenish the aqueous solvent of volatilization, filter, merge extract; Recovered under reduced pressure water to rare medicinal extract shape; Its volume is equivalent to 0.3-0.4 times of raw material, and said ratio is extract/raw material=L/kg, gets water extract;
D) ethanol extract and water extract are merged mixing, adopt spray pattern dry, obtain corn extract.
3. corn extract is regulated the application in the immunologic function product in preparation, and the preparation method of said corn extract is:
Corn is decocted extraction 3 times with pure water under the condition of 0.01-0.1MPa of pressurizeing, quantity of solvent is 3-6 times, and 3-6 times, 3-6 times, each time is 30-60min, filtration, and the merging extract, concentrate drying gets corn extract.
4. corn extract is regulated the application in the immunologic function product in preparation, and the preparation method of said corn extract is:
Corn is decocted extraction 3 times with pure water under the condition of 0.01-0.1MPa of pressurizeing, quantity of solvent is 3-6 times, and 3-6 times, 3-6 times, each time is 30-60min, filtration, and the merging extract gets aqueous extract;
Remaining corn is added 50% ethanol atmospheric pressure reflux again extract 2 times, each 1 hour, filter, merge with aqueous extract, concentrate drying gets corn extract.
5. according to arbitrary described application among the claim 1-4, it is characterized in that described product comprises food, health products, medicine.
6. according to arbitrary described application among the claim 1-4, it is characterized in that described product is powder, tablet, capsule, oral liquid.
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| CN111558007B (en) * | 2020-06-01 | 2022-04-26 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Corn root extract and preparation method and application thereof |
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| CN1058224A (en) * | 1990-07-12 | 1992-01-29 | 山东大学 | From maize yellow-powder, extract the food dye maize |
| CN1522596A (en) * | 2003-02-17 | 2004-08-25 | 吉林省高等院校科技开发研究中心 | Method for extracting composite corn embryo health care functional factor having corn embryo cake as raw material |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1058224A (en) * | 1990-07-12 | 1992-01-29 | 山东大学 | From maize yellow-powder, extract the food dye maize |
| CN1522596A (en) * | 2003-02-17 | 2004-08-25 | 吉林省高等院校科技开发研究中心 | Method for extracting composite corn embryo health care functional factor having corn embryo cake as raw material |
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| 李君霞.多糖功效分析及其应用研究的进展.《甘肃联合大学学报》.2007,第21卷(第5期),54-58. * |
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