CN105030562B - A kind of antiallergy complex polysaccharide composition and its preparation method and application - Google Patents
A kind of antiallergy complex polysaccharide composition and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a kind of antiallergy complex polysaccharide compositions, by including that following parts by weight component is made: Radix Astragali 10~90, cortex albiziae 10~90, Radix Glycyrrhizae 1~50.The preparation method of antiallergy complex polysaccharide composition, the steps included are as follows: (1) beat powder (2) and feed intake: the water or alcohol-water mixture (3) that 5~50 times of its weight amounts are added in medicinal powder extract: extracting under the conditions of -0.1~70MPa, 0~25 DEG C, then it is centrifuged, collects centrifugate；(4) separation concentration: centrifugate microfiltration membranes are removed into impurity, then use the ultrafiltration membrane grading purification polysaccharide of 100Da~20000Da flux.The beneficial effects of the present invention are: (1) has specific antiallergy functional component, product quality is easy to control；(2) there is too many levels to play synergistic function on sensitization target spot, has no toxic side effect, is highly-safe；(3) organic solvent-free contacts, nonhazardous residual；And preparation process is simple, easy to operate, is suitable for industrialization promotion application.
Description
Technical field
The present invention relates to a kind of complex polysaccharide compositions, are that one kind can be applied to antiallergy drug and prevent more specifically
Control the antiallergy complex polysaccharide composition and its preparation method and application in skin allergy cosmetics.
Background technique
Human allergy is a complicated process, mainly with immunity of organism globulin IgE, nuclear factor tenth of the twelve Earthly Branches second albumen,
Interleukin 8, tumor necrosis factor-alpha and 5 aspects of histamine are closely bound up, therefore in human body Typical allergic mechanism
Working in this 5 key links is more effective, direct, comprehensive prevention and treatment allergy approach.And polysaccharide is used as one kind in plant
The very wide active constituent of middle distribution, has unique advantage in antianaphylactic application field.
For the antiallergy mechanism of polysaccharide, traditional Chinese medicine thinks mainly to reach corresponding by adjusting body's immunity
Effect.And with the development of science and technology, the antiallergy mechanism of polysaccharide is gradually appeared in the newspapers in the research of molecular biosciences, cytogene
Road.As Chinese patent [open (bulletin) number CN 103467616A] discloses a kind of laver amylose obtained by extraction purification
It can increase the gene expression amount of Th1 cell factor IFN-γ, reduce the gene expression amount of Th2 cell factor IL-4.Contemporary Chinese
Reporting ophiopogonpolysaccharide energy antagonism acetylcholine and histamine mixed liquor using pharmacy [1999,16 (2)] stimulates caused normal globefish
The bronchial smooth muscle of sensitized guinea pig caused by mouse and ovalbumin shrink, inhibit the generation of sensitized guinea pig asthma and have compared with
The effect of significant anti-mouse ear xenogenesis models of passive skin irritability.Pharmacology and Clinics of Chinese Materia Medica [2014；30 (3): 86-89] report a Ji
Blazei polysaccharide can reduce the intensity of allergic inflammation reaction by inhibiting mast cell degranulation and discharging histamine.Chinese Hospitals pharmacy
Magazine [2011,31 (18): 1532-1534] reports Phellinus polysaccharide anti-allergic effects and inhibits mast cell degranulation and inflammatory
The release of meson is related.
However, human allergy is a complicated process.Mainly with immunity of organism globulin IgE, nuclear factor tenth of the twelve Earthly Branches second
Albumen, interleukin 8, tumor necrosis factor-alpha and 5 aspects of histamine are closely bound up.So occurring in human body Typical allergic
Working in 5 key links of mechanism is more effective, direct, comprehensive prevention and treatment allergy approach.But at present research be only with
Single medicinal material obtain polysaccharide carry out the one-side anti-allergic effects Mechanism Study of cellular and molecular level, not yet someone with Radix Astragali,
Cortex albiziae, Radix Glycyrrhizae are that raw material forms complex polysaccharide for the report in terms of too many levels antiallergy.
Radix Astragali, cortex albiziae, Radix Glycyrrhizae only stop most of research of their anti-allergic effects as common antiallergy Chinese medicine
It stays in the separation to its one pack system and chemical component identification, but is not attempt to the active polysaccharide of these three different types of structure
By rationally compounding to obtain the antiallergy complex polysaccharide of synergistic function, also its active ingredient screening and effect are not tested
Card.
Research shows that [Clinical Journal of Chinese Medicine, 2012 (4): 4-6.], astragalus polyose
It is that content is most in Radix Astragali, the strongest substance of immunocompetence, multi-party and playing immunological enhancement, spy can not only be enhanced
Specific immunological, moreover it is possible to enhance nospecific immunity, the immune function of normal body can not only be enhanced, moreover it is possible to enhance abnormal body
Immune function.Such as the Dendritic Cells that astragalus polyose Fiber differentiation obtains has the increment of allogeneic peripheral blood T lymphocyte
There is obvious stimulation effect, and act on enhancing with increasing for cell quantity, be in good dose-effect relationship, it was demonstrated that yellow cyclopentadienyl polysaccharide is not only
It can orient and induce cord blood monocyte-macrophage and be divided into DC, and the maturation of DC can be promoted, specific activation T cell generates thin
Born of the same parents are immune.It has been found that 8 kinds of polysaccharide components in astragalus mongolicus: astragalus polyose I, II, III, glucan AG-1, AG-2, heteroglycan AH-
1, AH-2 and a kind of water-soluble dextran.Total starches in Astragalus membranacus ward off aldehydic acid, galactolipin by glucose, arabinose, grape
The composition such as aldehydic acid.
Licorice polysaccharide has important physiological activity, and research finds it with antiviral, immunological regulation and antitumor
Activity.Reticuloendothelial system can be activated, there is anti-complement activity, directly stimulation B lymphocyte proliferation, induce IgM in human body,
The generation of IgG, moreover it is possible to induce with Stimulation effect interferon, enhance body killing cell activity.The immunological regulation of licorice polysaccharide
Effect is mainly reflected in the influence to mononuclear phagocyte system function in mouse reticuloendothelial system, can also activate static mouse
Spleen lymphocyte proliferation effect and anti-complement activity etc..Study the inhibition that licorice polysaccharide inhibits tumour, it is believed that licorice polysaccharide can
With independent or collaboration chemotherapeutic collective effect in tumour cell.The monosaccharide of composition licorice polysaccharide is generally arabinose, gala
Sugar, rhamnose and glucose, partially acidic polysaccharide also contain galacturonic acid and glucuronic acid, also contain fucose, wood
Sugar, mannose etc..There are also scholars, and a-l, the single glucan of 4- connection are isolated to from licorice.
Cortex albiziae has tranquilizing soporific, antifertility, anti-inflammatory, the inhibition multiple biological activities such as tumour and immunological regulation, decocting
The administration of agent rat oral gavage can inhibit peritoneal mast cells degranulation, can obviously inhibit antigen (horse serum) to the sensitization process of rat
And antibody generation process.There is scholar to guess that its antiallergy effective component is heat-resisting water-soluble substances.Some researches show that close simultaneously
Polysaccharide in joyous skin is the active constituent of cellular immunity, has Enhancement test mouse body hematid immunity function.
The isolation and purification method of traditional polysaccharide usually all include water mention, alcohol precipitation, grease removal, removing protein, decoloration, post separation this
A little basic steps.There is the method for protein in common Polysaccharide removing: sevag method, trifluorotrichloroethane method, trichloroacetic acid method,
The principle of these methods is that organic solvent is added to make protein denaturation precipitation and polysaccharide retains in the solution.These methods can not be kept away
Exempt from that dissolvent residual can be brought.
Summary of the invention
Object of the present invention is to places in view of the shortcomings of the prior art, provide a kind of definite ingredients, too many levels collaboration increases
The antiallergy complex polysaccharide composition with antiallergy of effect.
The second object of the present invention is to provide a kind of preparation method of antiallergy complex polysaccharide composition.
The third object of the present invention provides the application of the antiallergy complex polysaccharide composition.
In order to solve the above technical problems, the present invention adopts the following technical scheme: a kind of antiallergy complex polysaccharide composition,
By including that following parts by weight component is made: Radix Astragali 10~90, cortex albiziae 10~90, Radix Glycyrrhizae 1~50.
Preferably, the antiallergy complex polysaccharide composition, by including that following parts by weight component is made: Radix Astragali 30~
50, cortex albiziae 10~40, Radix Glycyrrhizae 10~30.
Above-mentioned antiallergy complex polysaccharide composition can be decocted using conventional alcohol water to prepare, but preparation efficiency it is not high and
Less effective.
A kind of preparation method of antiallergy complex polysaccharide composition comprising the step of it is as follows:
(1) it beats powder: Radix Astragali, cortex albiziae and Radix Glycyrrhizae is crushed；
(2) it feeds intake: the water or alcohol-water mixture of 5~50 times of its weight amounts being added in medicinal powder, stirs evenly；
(3) extract: extracted under the conditions of -0.1~70MPa, 0~25 DEG C, collect extracting solution be centrifuged, collect from
Heart liquid；
(4) separation concentration: removing solid particle, the impurity such as bacterium for centrifugate microfiltration membranes, then with 100Da~
The ultrafiltration membrane grading purification polysaccharide of 20000Da flux is finally concentrated, is dry.
In (2) step, the additional amount of water is 20~30 times of medicinal powder weight.The preferred pure water of water.
In (3) step, extraction conditions be pressure be -0.1~35MPa, Extracting temperature is 10~20 DEG C, extraction time
It is 1 time.
In (4) step, the aperture specification of microfiltration membranes is 0.2 μm~1.0 μm；Ultrafiltration membrane aperture specification be 1000Da~
10000Da。
The ethanol water that hundred number of alcohol-water mixture preferred volume is 50~95%.
In (4) step, obtained trapped fluid is concentrated to 1.03~1.08 specific gravity.
Antiallergy complex polysaccharide composition, which can be used for preparing, has effects that the cosmetics of antiallergy, additive amount for 1~
50% (hundred number of quality).The dosage form of cosmetics includes creme, lotion, aqua, gelling agent, facial mask, inuncts and lotion.
Antiallergy complex polysaccharide composition, which can be used for preparing, to be had effects that in the drug of antiallergy, additive amount for 10~
90%, dosage form includes liquid preparation (decoction, syrup, injection, mixture, tincture etc.), solid pharmaceutical preparation (powder, electuary, ball
Agent, tablet, capsule etc.), semisolid preparation (for oral administration cream, external plaster, paste etc.), gas formulation (aerosol, fumicants etc.).
After the present invention rationally compounds Radix Astragali, Radix Glycyrrhizae, cortex albiziae polysaccharide, show that the present invention mentions by efficacy validation experiment
It takes object that can inhibit IgE activity expression, prevents the release of histamine, block the activation of NF- κ B, inhibit IL-8 activity expression, inhibit
TNF-α activity expression has anti-inflammatory, antiallergy, the effect for enhancing immunity of organisms.
Compared with prior art, the beneficial effects of the present invention are:
(1) compared with existing antiallergy Chinese medicine compound prescription, the present invention has specific antiallergy functional component, and product quality is held
It is easy to control；
(2) compared with existing antiallergy Western medicine, there is extract of the present invention too many levels to play synergy on sensitization target spot
Effect, has no toxic side effect, is highly-safe；
(3) contact of process organic solvent-free, nonhazardous residual are manufactured；And preparation process is simple, easy to operate, fits
Together in industrialization promotion application.
Detailed description of the invention
Fig. 1 is preparation method process flow diagram of the invention；
Fig. 2 is histogram of the extract of the present invention to hyaluronidase inhibition assay result；
Fig. 3 is the RNA electrophoretogram of molecular gene regulating and expressing；
Fig. 4 is the amplification curve of the NFKBp65 of molecular gene regulating and expressing；
Fig. 5 is the melting curve of the NFKBp65 of molecular gene regulating and expressing；
Fig. 6 is the amplification curve of the GAPDH of molecular gene regulating and expressing；
Fig. 7 is the melting curve of the GAPDH of molecular gene regulating and expressing；
Fig. 8 is the structural schematic diagram of cryogenic high pressure difference extractor.
Specific embodiment
In order to preferably illustrate the content of present invention, it is illustrated below with several preferable specific embodiments.
Inventive principle
The present invention according to Chinese medicine compound prescription eliminating evil go because, strengthen the body resistance to consolidate the constitution, by coordinating zang-fu differentiation function to correcting yin-yang
It contains and declines partially partially, body recovery is made to carry out reasonable formula to the treatment cause of disease reason of the normal condition of yin and yang in equilibrium.Radix Astragali, cortex albiziae,
Radix Glycyrrhizae three has the active polysaccharide of different types of structure, thus in effect can difference, in specific portfolio ratio situation
Under can achieve synergistic function.This is also a major reason for instructing prescription of the present invention.
Membrane separation process is pure physical separation, and room temperature is separated, be concentrated, refined driven by pressure, is separated
Not undergoing phase transition in journey, is not had the variation of calorifics by isolated component, there will not be chemically or biological variation；
Membrane separation process process totally closed operation, not vulnerable to pollution, finished product, which is not required to addition preservative, to be stored for a long time.With low energy
The characteristics of consumption, high efficiency and normal-temperature operation.
The permission particle size range of Ultra filtration membrane particle is 0.1~0.001 μm, and the granularity of polysaccharide is 0.002~0.01 μ
M, so, it can choose suitable ultrafiltration membrane and polysaccharide isolated and purified.The principle of ultra-filtration and separation polysaccharide usually first retains
The impurity such as relative molecular weight big resin, tannin are removed, small molecular protein, amino acid etc. are then filtered out, finally obtain certain point
The active polysaccharide of son amount range.
The specific effect that the present invention has is as follows:
First, the composition that existing antiallergy Chinese medicine compound prescription includes is generally all relatively more, and composition of the invention
Three kinds of simple Chinese medicine materials are only needed, product quality is easy to control, and has specific antiallergic effect.And it is of the invention compound
It is not being simply mixed for random three kinds of raw materials, three is that there are difference in effect, is only just reached under specific portfolio ratio
To synergistic function.Also show that the anti-inflammatory too many levels that the composition has, antiallergy, enhancing body are exempted from by Efficacy experiments
Epidemic disease power effect is substantially better than single component active polysaccharide, is in particular in and is able to suppress IgE activity expression, prevents releasing for histamine
Put, the activation that blocks NF- κ B, inhibit IL-8 activity expression, inhibit TNF-α activity expression total 5 have more effectively, directly,
Comprehensive prevention and treatment allergy approach.
Second, compared with existing antiallergy Western medicine, antiallergy complex polysaccharide composition of the invention is according to Chinese medicine compound prescription "
The traditional Chinese medicine treatment cause of disease reason because of ", " strengthen the body resistance to consolidate the constitution " is gone in eliminating evil, corrected by coordinating zang-fu differentiation function it is yin or yang excess partially
It declines, makes body recovery to the normal condition of yin and yang in equilibrium.It has no toxic side effect, highly-safe while of the invention composition also has
There is too many levels to play synergistic function on sensitization target spot.
Third, traditional separation of polysaccharides purification process normally comprise water mention, alcohol precipitation, grease removal, removing protein, decoloration, column point
From these basic steps.There is the method for protein in common Polysaccharide removing: sevag method, trifluorotrichloroethane method, trichloroacetic acid
Method, principle are all that organic solvent is added to make protein denaturation precipitation and polysaccharide retains in the solution.But these above-mentioned sides
Method inevitably brings the problem of dissolvent residual.
The present invention is extracted using cryogenic high pressure difference and combines microfiltration membranes, ultra-filtration and separation technique, and manufacturing process does not contact organic molten
Agent, nonhazardous residual, preparation process is simply easy to operate, is very suitable to the popularization and application of industrialization.
Membrane separation process is pure physical separation, and room temperature is separated, be concentrated, refined driven by pressure, is separated
Not undergoing phase transition in journey, is not had the variation of calorifics by isolated component, there will not be chemically or biological variation.
Moreover, membrane separation process process is totally closed operation, not vulnerable to pollution, finished product, which is not required to addition preservative, to be deposited for a long time
It puts, has the advantages that low energy consumption, high efficiency and normal-temperature operation.
The principle of ultra-filtration and separation polysaccharide is that first retention removes the impurity such as big resin, the tannin of relative molecular weight, is then filtered out
Small molecular protein, amino acid etc. finally obtain the active polysaccharide of certain molecular weight range.The permission granularity of Ultra filtration membrane particle
Range is 0.1~0.001 μm, and the granularity of polysaccharide is 0.002~0.01 μm, as long as the ultrafiltration membrane of selection suitable particle size range,
Achieve that the polysaccharide to corresponding granularity isolates and purifies.
Embodiment 1
A kind of antianaphylactic complex polysaccharide is made of following parts by weight component: Radix Astragali 50, cortex albiziae 35, Radix Glycyrrhizae 15.
Embodiment 2
A kind of antianaphylactic complex polysaccharide, by following mass percentage composition: being made of following parts by weight component: yellow
Stilbene 40, Radix Glycyrrhizae 35, cortex albiziae 25.
Embodiment 3
A kind of antianaphylactic complex polysaccharide is made of following parts by weight component: Radix Astragali 35~45, cortex albiziae 20~30,
Radix Glycyrrhizae 15~25.
Embodiment 4
A kind of antianaphylactic complex polysaccharide is made of following parts by weight component: Radix Glycyrrhizae 40, Radix Astragali 30, cortex albiziae 30.
Embodiment 5
A kind of antianaphylactic complex polysaccharide is made of following parts by weight component: cortex albiziae 35, Radix Astragali 35, Radix Glycyrrhizae 25.
Embodiment 6
A kind of antianaphylactic complex polysaccharide is made of following parts by weight component: Radix Astragali 50, Radix Glycyrrhizae 30, cortex albiziae 20.
The preparation method of above-mentioned antiallergy complex polysaccharide, as shown in Figure 1, comprising the following steps:
(1) it beats powder: above-mentioned medicinal material is distinguished powder at 30-70 mesh, or by above-mentioned medicinal material by co-grinding after prescription weighing
It is broken into 30-70 mesh, is then uniformly mixed.
(2) feed intake: it is 60~90% that water or the volume by volume concentration that 20-40 times of medicinal powder quality is measured are added in above-mentioned medicinal powder
Ethanol solution stirs evenly；25-35 times of water better effect of medicinal powder quality is added.
(3) it extracts: being extracted under the conditions of -0.1~70MPa, 0~25 DEG C with cryogenic high pressure difference extractor, collect extracting solution
It is centrifuged；Preferably, it is -0.1~30MPa, 10~15 DEG C in the condition of cryogenic high pressure difference extractor, extracts effect in this way
Rate is higher, and active matter transfer, dissolution are more complete.Centrifuge separation using link-suspended basket centrifuge carry out, revolving speed be 2500-4500 turn/
Second.
(4) separation concentration: the centrifugate of collection microfiltration membranes are removed into the impurity such as solid particle, bacterium, then use 200Da
The ultrafiltration membrane grading purification polysaccharide of~15000Da flux, finally with nanofiltration membrane concentration, discharging.
Wherein, the microfiltration membranes can use polyvinyl chloride (PVC) microfiltration membranes and polypropylene (PP) microfiltration membranes, preferred hole
Diameter is 0.3 μm~0.8 μm.Wherein, ultrafiltration membrane is classified pure retain for the first ultrafiltration membrane for using 10000-20000Da flux first and removes
Big molecular impurity is gone, then filters out small molecular weight impurity with the ultrafiltration membrane of 100-10000Da flux.Most preferably, it uses first
The ultrafiltration membrane retention of 15000-20000Da flux removes big molecular impurity, then uses the ultrafiltration membrane filtration of 1000-10000Da flux
Except small molecular weight impurity, it is achieved in that purity of polysaccharide height, yield is high.The retention molecule of the nanofiltration membrane is 500-1500, the U.S.
The NF series nanofiltration membrane of Filmtec company or the NTR-7400 series nanofiltration membrane of Japanese day east electrician.
Wherein, the cryogenic high pressure difference extractor is as shown in figure 8, by extraction chamber 1, rewinding room 2, extracting solution tank battery 3, pressure
Contracting air station 4, feeding port 5, blender 6, solvent sprayer 7, condenser 8, freezing water station 9, solvent storage tank 10, condensate buffering tank
11, vacuum station 12, temperature set station 13, extraction slag storage tank 14, temperature detecting controller 20, pressure testing and controlling device 30 form；The extraction chamber 1
A pressure vessel with heat preservation sandwich layer is shared with the rewinding room 2 and passes through leaf filter interval, built in the extraction chamber 1
The blender 6；The extraction chamber 1 and the rewinding room 2 can be the seperated standalone for being also possible to disjunctor, the extraction
Divided between room 1 and the rewinding room 2 with filter, the filter is board-like or rolling, filled type, pillar or different shaped
The combination of formula filter；The top of the extraction chamber 1 is equipped with the feeding port 5, and installs the solvent sprayer 7, solvent spray
Day with fog 7 is connected to solvent storage tank 10 by pipeline；The rewinding room 2 is connected to extracting solution tank battery 3 by pipeline；The extraction chamber 1
Bottom is connected to by pipeline and extracts slag storage tank 14；The condensate buffering tank 11 is connected to vacuum station 12 by pipeline；The extraction
1 top of room is connected to the compressed air reservoir of air compression station 4 by pipeline；The temperature adjustment of pressure vessel interlayer and temperature set station 13
Liquid storage tank establishes circulating line connection；The extraction chamber 1 is equipped with temperature detecting controller 20 and pressure testing and controlling device 30.It is slow by condensing
The vacuum orifice rushed on tank 11 is established pipeline with the vacuum buffer tank of vacuum station 12 and is connect, and makes to make to receive as needed when operation
Vacuum degree in material room 2 is up to 90Pa；Compression by compressed air inlet and air compression station 4 at the top of extraction chamber is empty
Gas storage tank establishes pipeline connection, makes the pressure in rewinding room can be made to be up to 80MPa as needed when operation；Pass through interlayer
Temperature adjusting liquid inlet and outlet establish circulating line with the temperature adjustment liquid storage tank of temperature set station 13 and connect, temperature adjusting liquid temperature general control-
35 DEG C~50 DEG C, make to make extraction chamber and the indoor temperature control of rewinding at -25 DEG C~40 DEG C when operation as needed；Pass through
Valve on the pressure testing and controlling device 30 at the top of extraction chamber 1, temperature detecting controller 20 and pipeline establish control system information collection and
Transmission path, these information are beneficial to automatically control the procedure design of operation and application.Differential pressure extractor it is attached equipped with cleaning,
Decontamination system.
Steps are as follows when operation:
1. charging: opening feeding port 5, dry material to be extracted is put into extraction chamber by feed opening by starting stirring dress device 6
In.
2. vacuumizing: after material finishes, closing feeding port 5, starting freezing water station 9 makes chilled water enter condenser 8 and follows
Ring starts vacuum station, will be evacuated in extraction chamber 1 by condensate buffering tank 11, after the vacuum degree for reaching needs, keeps certain
Time.
3. solubilization matchmaker: quantitatively being sprayed into the solvent in solvent storage tank 10 in extraction chamber on demand by solvent sprayer 7
1, kept for certain time.
4. subtracting vacuum: starting air compression station 4, logical compressed air enter in extraction chamber 1, are gradually reduced house vacuum degree
To -0.1MPa.
5. pressurization: continue to be passed through compressed air, pressure in extraction chamber 1 made to gradually rise up to highest 70MPa, make extracting solution into
Enter in rewinding room 2.
6. operating procedure 2~5 is repeated, until extracting solution does not show selected index components identification.
It is expelled in extraction slag storage tank 14 7. opening extraction chamber's bottom valve and will extract slag.
Embodiment 7
A kind of preparation method of antianaphylactic complex polysaccharide, its step are as follows:
(1) 15 kilograms of Radix Astragali, 3 kilograms of cortex albiziae, 12 kilograms of Radix Glycyrrhizae are respectively crushed into after 80 mesh powder again by composition
Formula is uniformly mixed；
B. 150 kilograms of 50% ethanol waters of percentage by volume are added in the powder for taking step a to obtain, after mixing evenly
To liquid to be extracted；
C. the liquid pump to be extracted step b obtained enters cryogenic high pressure difference extractor, pressure is 15MPa, temperature is 10 DEG C
Under conditions of extract it is primary；
D. extracting solution step c obtained is sent into link-suspended basket centrifuge centrifuge separation, and revolving speed is 3000 revolutions per seconds；
E. the microfiltration membranes that centrifugate aperture step d obtained is 0.4 μm remove impurity；
F. it first uses the ultrafiltration membrane of 15000Da flux to retain the filtrate that step e is obtained to remove big molecular impurity, collect filter
Liquid, then small molecular weight impurity is removed with the ultrafiltration membrane retention of 3000Da flux；
G. 1.03 specific gravity first are reduced to by trapped fluid nanofiltration membrane that step f is obtained is reverse osmosis concentrated, then concentrate are sprayed dry
It is dry, obtain the extract finished product of complex polysaccharide composition.
Embodiment 8
A kind of preparation method of antianaphylactic complex polysaccharide, its step are as follows:
A. match again by composition after 11 kilograms of Radix Astragali, 6 kilograms of cortex albiziae, 13 kilograms of Radix Glycyrrhizae being respectively crushed into 40 mesh powder
Side is uniformly mixed；
B. the powder for taking step a to obtain is added 130 kilograms of pure water, obtains liquid to be extracted after mixing evenly；
C. the liquid pump to be extracted step b obtained enters cryogenic high pressure difference extractor, pressure is 35MPa, temperature is 5 DEG C
Under the conditions of extract it is primary；
D. extracting solution step c obtained removes the dregs of a decoction with flat centrifuge；
E. the microfiltration membranes that centrifugate aperture step d obtained is 0.45 μm remove impurity；
F. it first uses the ultrafiltration membrane of 20000Da flux to retain the filtrate that step e is obtained to remove big molecular impurity, collect filter
Liquid, then small molecular weight impurity is removed with the ultrafiltration membrane retention of 500Da flux；
G. the trapped fluid vacuum-concentrcted first obtained step f is dried under reduced pressure to 1.08 specific gravity, then by concentrate, is obtained
To the extract finished product of complex polysaccharide composition.
Embodiment 9
A kind of antiallergic mask is made of following weight percent raw material:
In addition it can add appropriate amount of essence and other active components.Wherein antianaphylactic complex polysaccharide preferred embodiment
4 obtained antianaphylactic complex polysaccharides.
The step of preparation method of above-mentioned antiallergic mask, is as follows:
(1) polyvinyl alcohol is added in polyvinyl acetate emulsion, stirring is mixed stand-by；
(2) sodium carboxymethylcellulose, complex polysaccharide and propylparaben are added to part (total amount 30-60%)
It is mixed in distilled water, and is heated to 80 DEG C or so, stirred 2 hours, it is cooling stand-by；
(3) tween, glycerol and remaining distilled water and essence are mixed, is stirred evenly stand-by；
(4) solution (2) is poured into solution (1), again pours into solution (3) after stirring, be stirred 3 hours up at
Product.
Embodiment 10
A kind of antiallergy granule is made of following weight percent raw material:
A kind of preparation method of antiallergy granule, the steps include: the raw and auxiliary material for weighing above-mentioned formula ratio, is sufficiently mixed
Ethanol in proper amount, low temperature drying after wet granulation are added after uniformly.
Effect experiment
It is extracted obtained by Example 7 (the antianaphylactic complex polysaccharide that remaining embodiment obtains also has similar effect) below
Object carries out efficacy validation experiment.
1) hyaluronidase Inhibition test
Experimental method: the antianaphylactic complex polysaccharide of the present invention is added in hyaluronidase after calcium ion activated, is then added
Sodium Hyaluronate is complexed under alkaline condition with acetylacetone,2,4-pentanedione after reacted, is finally developed the color with Ehrlich's reagent, in 518nm
Place measures its ultraviolet absorption value.
Experimental result: being shown in Fig. 2, illustrates that there is the antianaphylactic complex polysaccharide of the present invention good hyaluronidase to inhibit effect
Fruit.
2) cell function is tested
Experimental method: it is added after lipopolysaccharides (LPS) stimulation people's Acute Monocytic Leukemia Cell Line (THP-1) 24 hours
The antianaphylactic complex polysaccharide of the present invention of various concentration is cultivated, and IL-8 in research THP-1 cell, TNF-α, IgE are passed through
The expression that immune response, NF- κ 1 B gene regulate and control.
Experimental result: as the increase of concentration, IL-8, TNF- of LPS induction is added in the antianaphylactic complex polysaccharide of the present invention
α, lgE expression activity substantially reduce.It can inhibit IL-8, TNF-α, lgE activity expression substantially in additional amount 1.0%.Illustrate this
Invention extract has good anti-inflammatory, antiallergic effect.It see the table below 1~3.
1. test specimen IL-8 concentration calculation of table
2. test specimen TNF-α concentration calculation of table
3. test specimen IgE concentration calculation of table
3) expression analysis of molecular gene regulation
Experimental method: it after LPS stimulation is added 24 hours in people's Acute Monocytic Leukemia Cell Line THP-1, is added different dense
Antianaphylactic complex polysaccharide culture of the invention 24 hours of degree, the expression of NF- κ B in Q-PCR quantitative detection THP-1.
As the result is shown: after LPS stimulation is added in THP-1 cell, antiallergy of the present invention is added in NF- κ B expression increment 87.1%
Complex polysaccharide culture after, various concentration additional amount shows the NF- kB activity expression for inhibiting LPS stimulation, when the anti-mistake of the present invention
Quick complex polysaccharide additional amount can reduce the NF- kB activity expression of 77.4%LPS induction at 0.5%, in additional amount 1.0%
The NF- kB activity expression that LPS induction can be completely eliminated, is as a result shown in Fig. 3~7.
Polyoses content detection
[preparation of reference substance solution] takes DEXTROSE ANHYDROUS reference substance 25mg, accurately weighed, sets in 250ml measuring bottle, adds water
Appropriate dissolution, is diluted to scale, shakes up to get (containing DEXTROSE ANHYDROUS 0.1mg) in every 1ml.
[preparation of standard curve] is accurate to measure reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, respectively
It sets in tool plug test tube, respectively plus water is mended to 2.0ml, and 5% phenol solution 1ml of each accurate addition shakes up, accurate rapidly that sulfuric acid is added
5ml shakes up, and places 10 minutes, sets in 40 DEG C of water-baths and keep the temperature 15 minutes, takes out, is rapidly cooled to room temperature, is with corresponding reagent
Blank measures absorbance, using absorbance as ordinate, concentration is according to UV-VIS spectrophotometry at the wavelength of 490nm
Abscissa draws standard curve.
[sample measurement] takes the antianaphylactic complex polysaccharide powder 1.0g that the above embodiment of the present invention obtains, accurately weighed,
Add diethyl ether 100ml, is heated to reflux 1 hour, stands, lets cool, carefully discard ether solution, residue sets water-bath Back stroke and uses up ether.It is added
80% ethyl alcohol 100ml is heated to reflux 1 hour, filters while hot, and filter residue and filter are washed by several times with 80% ethyl alcohol 30ml of heat, filter residue
It is set in flask together with filter paper, adds water 150ml, be heated to reflux 2 hours.It filters while hot, with a small amount of hot water washing nozzle, merging filtrate
It with washing lotion, letting cool, moves in 250ml measuring bottle, be diluted with water to scale, appropriate dilution, precision measure dilution 1ml after shaking up,
It sets in tool plug test tube, adds water 1.0ml, the method under the preparation of sighting target directrix curve, from " 5% phenol solution of each accurate addition
1ml " rises, and measures absorbance in accordance with the law, from the weight (mg) containing glucose in test solution is read on standard curve, calculates, i.e.,
?.
The determination of polysaccharide result of the extract of the present invention of table 4
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent replacement mode, be included within the scope of the present invention.
Claims (10)
1. a kind of antiallergy complex polysaccharide composition, it is characterised in that: by including that following parts by weight component is made: Radix Astragali 10~
90, cortex albiziae 10~90, Radix Glycyrrhizae 1~50；
The steps included are as follows for preparation method:
(1) it beats powder: Radix Astragali, cortex albiziae and Radix Glycyrrhizae is crushed；
(2) it feeds intake: the water or alcohol-water mixture of 5~50 times of its weight amounts being added in medicinal powder, stirs evenly；
(3) it extracts: extracting under the conditions of -0.1~70MPa, 0~25 DEG C, be then centrifuged, collect centrifugate；
(4) separation concentration: centrifugate microfiltration membranes are removed into impurity, are then classified with the ultrafiltration membrane of 100Da~20000Da flux
Purified polysaccharide.
2. antiallergy complex polysaccharide composition according to claim 1, it is characterised in that: by including following parts by weight array
Divide and be made: Radix Astragali 30~50, cortex albiziae 10~40, Radix Glycyrrhizae 10~30.
3. a kind of preparation method of antiallergy complex polysaccharide composition, it is characterised in that: the steps included are as follows for it:
(1) it beats powder: Radix Astragali, cortex albiziae and Radix Glycyrrhizae is crushed；
(2) it feeds intake: the water or alcohol-water mixture of 5~50 times of its weight amounts being added in medicinal powder, stirs evenly；
(3) it extracts: extracting under the conditions of -0.1~70MPa, 0~25 DEG C, be then centrifuged, collect centrifugate；
(4) separation concentration: centrifugate microfiltration membranes are removed into impurity, are then classified with the ultrafiltration membrane of 100Da~20000Da flux
Purified polysaccharide.
4. the preparation method of antiallergy complex polysaccharide composition according to claim 3, it is characterised in that: (2) step
In rapid, the additional amount of water is 20~30 times of medicinal powder weight.
5. the preparation method of antiallergy complex polysaccharide composition according to claim 3, it is characterised in that: (3) step
In rapid, extraction conditions be pressure be -0.1~35MPa, Extracting temperature is 10~20 DEG C.
6. the preparation method of antiallergy complex polysaccharide composition according to claim 3, it is characterised in that: (4) step
In rapid, the aperture specification of microfiltration membranes is 0.2 μm~1.0 μm；Ultrafiltration membrane aperture specification is 1000Da~10000Da.
7. the preparation method of antiallergy complex polysaccharide composition according to claim 3, it is characterised in that: the alcohol water is mixed
Close the ethanol water that object preferred volume percentage is 50~95%.
8. the preparation method of antiallergy complex polysaccharide composition according to claim 3, it is characterised in that: (4) step
In rapid, obtained trapped fluid is concentrated to 1.03~1.08 specific gravity.
9. antiallergy complex polysaccharide composition according to claim 1, it is characterised in that: be used to prepare with antiallergy function
The cosmetics of effect, additive amount are 1~50%.
10. antiallergy complex polysaccharide composition according to claim 1, it is characterised in that: be used to prepare with antiallergy
The drug of effect, additive amount are 10~90%.
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