CN105030562A - Anti-allergic compound polysaccharide composition, preparation method and application thereof - Google Patents
Anti-allergic compound polysaccharide composition, preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to an anti-allergic compound polysaccharide composition which is prepared from following components, by weight: 10-90 parts of astragalus membranaceus, 10-90 parts of cortex albiziae and 1-50 parts of radix glycyrrhizae. A preparation method of the anti-allergic compound polysaccharide composition includes the following steps: (1) crushing the raw materials into powder; (2) material feeding: adding water or a water-alcohol mixture by 5-50 times by weight of the powder into the powder; (3) extraction: performing extraction under -0.1-70 MPa at 0-25 DEG C, and performing centrifugal separation and collecting a centrifugate; (4) separation and concentration: removing impurities from the centrifugate through a micro-filtering membrane, and performing classification purification to the polysaccharide through an ultra-filtering membrane being 100-20000 Da in flux. The compound polysaccharide composition has the following beneficial effects: (1) the composition has definite anti-allergic functional components and is easy to control in quality; (2) the composition has a synergistic effect on a sensitizing target point with multiple links, is free of toxic and side effects and is high in safety; (3) the composition is free of contact with organic solvents, is free of residue of toxicity; and the preparation method is simple in process and is easy to carry out, and is suitable for being popularized industrially.
Description
Technical field
The present invention relates to a kind of complex polysaccharide compositions, more particularly, is a kind ofly can be applicable to antiallergic complex polysaccharide composition and method of making the same in antiallergic medicine and control skin allergy cosmetics and application.
Background technology
Human allergy is a complicated process, main closely bound up with immunity of organism globulin IgE, nuclear factor second at tenth of the twelve Earthly Branches albumen, interleukin 8, tumor necrosis factor-alpha and histamine 5 aspects, therefore working in these 5 key links of human body Typical allergic mechanism is comparatively effectively, directly, comprehensively prevent and treat irritated approach.And polysaccharide is as a kind of very wide active component that distributes in plant, there is unique advantage in antianaphylactic application.
For the antiallergic mechanism of polysaccharide, Chinese medicine is thought and is mainly reached corresponding effect by conditioner body immunity function.And along with the development of science and technology, the antiallergic mechanism of polysaccharide is gradually appeared in the newspapers in the research of molecular biosciences, cytogene.As Chinese patent [open (bulletin) number CN103467616A] discloses the gene expression amount that a kind of Porphyra Polysaccharide obtained by extraction purification can increase Th1 cytokine IFN-γ, reduce the gene expression amount of Th2 cytokine IL-4.The bronchial smooth muscle that Chinese Journal of Modern Applied Pharmacy [1999,16 (2)] reports the sensitized guinea pig that Radix Ophiopogonis polysaccharide energy antagonism acetylcholine and histamine mixed liquor stimulate the normal guinea pig that causes and ovalbumin to cause shrinks, suppress the generation of sensitized guinea pig asthma and have the effect of more significant anti-mouse ear xenogenesis models of passive skin irritability.Pharmacology and Clinics of Chinese Materia Medica [2014; 30 (3): 86-89] intensity of Agaricus Blazei Murrill polysaccharide by suppressing mast cell degranulation and release histamine can alleviate allergic inflammation reaction is reported.It is relevant with suppressing the release of mast cell degranulation and inflammatory meson that Journal of Chinese Hospital Pharmacy [2011,31 (18): 1532-1534] reports phellin polysaccharides anti-allergic effects.
But human allergy is a complicated process.Main and immunity of organism globulin IgE, nuclear factor second at tenth of the twelve Earthly Branches albumen, interleukin 8, tumor necrosis factor-alpha and histamine 5 aspects are closely bound up.Comparatively effectively, directly, comprehensively prevent and treat irritated approach so work in 5 key links of human body Typical allergic mechanism.But at present research is only the polysaccharide obtained with single medical material carries out the one-side anti-allergic effects Mechanism Study of cellular and molecular level, not yet form complex polysaccharide report for too many levels antiallergic aspect with the Radix Astragali, Cortex Albiziae, Radix Glycyrrhizae for raw material by people.
The Radix Astragali, Cortex Albiziae, Radix Glycyrrhizae are as common antiallergic Chinese medicine, only rest in the separation of its one pack system and chemical composition qualification to the research of their anti-allergic effects great majority, but do not attempt the active polysaccharide of these three kinds of different types of structure to pass through the rationally composite antiallergic complex polysaccharide obtaining synergistic function, not to its active ingredient screening and efficacy validation yet.
Research shows [ClinicalJournalofChineseMedicine; 2012 (4): 4-6.]; astragalus polysaccharides is the class material that in the Radix Astragali, content at most, immunocompetence is the strongest; in many ways immunological enhancement is being played; specific immunity can not only be strengthened; can also enhancing non-specific immunity, the immunologic function of normal body can not only be strengthened, the immunologic function of abnormal body can also be strengthened.The dendritic cell that such as astragalus polysaccharides inducing culture obtains has obvious stimulation effect to the lymphocytic increment of allogeneic peripheral blood T, and increasing and act on enhancing with cell quantity, in good dose-effect relationship, confirm that astragalus polysaccharide not only orientation can induce cord blood monocyte-macrophage and be divided into DC, and the maturation of DC can be impelled, specific activation T cell, produces cellular immunization.8 kinds of polysaccharide components have been found: astragalus polysaccharides I, II, III, glucosan AG-1, AG-2, heteropolysaccharide AH-1, AH-2 in Radix Astagali, and a kind of water-soluble glucan.Total polysaccharides in Radix Astragali, is warded off aldehydic acid, galacturonic acid etc. formed by glucose, arabinose, grape.
Angelica Polysaccharide there is important physiologically active, research finds that it has antiviral, immunomodulating and antineoplastic are active.Can activate reticuloendothelial system, have anticomplementary activity, directly stimulate B lymphocyte proliferation, in induction human body, the generation of IgM, IgG, can also induce and Stimulation effect interferon, the activity of the killer cell of enhancing body.The immunoregulation effect of Angelica Polysaccharide is mainly reflected in the impact on mononuclear phagocyte system function in mice reticuloendothelial system, also can activate static mice spleen lymphocytes proliferation effect and anticomplementary activity etc.The suppression of research Angelica Polysaccharide Tumor suppression, think Angelica Polysaccharide can independent or collaborative chemotherapeutic combined effect in tumor cell.The monosaccharide of composition Angelica Polysaccharide is generally arabinose, galactose, rhamnose and glucose, and partially acidic polysaccharide, also containing galacturonic acid and glucuronic acid, also contains fucose, xylose, mannose etc.Also have scholar from licorice, be separated to the single glucosan of a-l, 4-connection.
Cortex Albiziae has the multiple biological activitys such as tranquilizing soporific, antifertility, antiinflammatory, Tumor suppression and immunomodulating, the administration of water decoction rat oral gavage can suppress peritoneal mast cells retting conditions, can obviously suppress antigen (horse serum) to the sensitization process of rat and antibody generation process.Its antiallergic effective ingredient is heat-resisting water-soluble substances to have scholar to guess.There are some researches show, the polysaccharide in Cortex Albiziae is the active component of cellular immunization simultaneously, has Enhancement test mice body hematid immunity function.
The isolation and purification method of traditional polysaccharide all comprises water extraction usually, precipitate with ethanol, grease removal, removing protein, decolouring, post are separated these basic steps.In conventional Polysaccharide removing, method of protein has: sevag method, trifluorotrichloroethane method, trichloroacetic acid method, and these side's ratio juris add that organic solvent makes protein denaturation precipitation and polysaccharide retains in the solution.These methods inevitably bring dissolvent residual.
Summary of the invention
The present invention seeks to the weak point existed for prior art, the antiallergic complex polysaccharide compositions with antiallergic effect of a kind of definite ingredients, too many levels Synergistic is provided.
The second object of the present invention is to provide a kind of preparation method of antiallergic complex polysaccharide compositions.
The third object of the present invention provides the application of described antiallergic complex polysaccharide compositions.
For solving above-mentioned technical problem, the present invention adopts following technical scheme: a kind of antiallergic complex polysaccharide compositions, makes: the Radix Astragali 10 ~ 90, Cortex Albiziae 10 ~ 90, Radix Glycyrrhizae 1 ~ 50 by comprising following parts by weight component.
Preferably, described antiallergic complex polysaccharide compositions, makes by comprising following parts by weight component: the Radix Astragali 30 ~ 50, Cortex Albiziae 10 ~ 40, Radix Glycyrrhizae 10 ~ 30.
Above-mentioned antiallergic complex polysaccharide compositions can adopt conventional alcohol soak by water to prepare, but preparation efficiency is not high and less effective.
A preparation method for antiallergic complex polysaccharide compositions, it step comprised is as follows:
(1) powder is beaten: the Radix Astragali, Cortex Albiziae and Radix Glycyrrhizae are pulverized;
(2) feed intake: in medicated powder, add its weight 5 ~ 50 times amount water or alcohol-water mixture, stir;
(3) extract: extract under-0.1 ~ 70MPa, 0 ~ 25 DEG C of condition, collect extracting solution and carry out centrifugalize, collect centrifugal liquid;
(4) separation is concentrated: centrifugal liquid micro-filtration membrane is removed the impurity such as solid particle, antibacterial, then uses the ultrafilter membrane grading purification polysaccharide of 100Da ~ 20000Da flux, finally concentrated, dry.
In described (2) step, the addition of water is 20 ~ 30 times for medicated powder weight.The preferred pure water of water.
In described (3) step, extraction conditions is pressure is-0.1 ~ 35MPa, Extracting temperature is 10 ~ 20 DEG C, extraction time is 1 time.
In described (4) step, the aperture specification of micro-filtration membrane is 0.2 μm ~ 1.0 μm; Ultrafilter membrane aperture specification is 1000Da ~ 10000Da.
Described alcohol-water mixture preferred volume hundred number is the ethanol water of 50 ~ 95%.
In described (4) step, the trapped fluid obtained is concentrated to 1.03 ~ 1.08 proportions.
Antiallergic complex polysaccharide compositions may be used for preparing the cosmetics with antiallergic effect, and addition is 1 ~ 50% (quality hundred number).The dosage form of cosmetics comprises cream, emulsion, water preparation, gel, facial film, inuncts and lotion.
Antiallergic complex polysaccharide compositions may be used for preparing in the medicine with antiallergic effect, addition is 10 ~ 90%, and dosage form comprises liquid preparation (decoction, syrup, injection, mixture, tincture etc.), solid preparation (powder, electuary, pill, tablet, capsule etc.), semi-solid preparation (cream, external plaster, paste for oral administration etc.), gas formulation (aerosol, fumicants etc.).
After the present invention is rationally composite the Radix Astragali, Radix Glycyrrhizae, Cortex Albiziae polysaccharide, shown by efficacy validation experiment, extract of the present invention can suppress IgE activity expression, stop the release of histamine, block the activation of NF-κ B, suppress IL-8 activity expression, suppress TNF-alpha active to be expressed, there is antiinflammatory, the effect of antiallergic, enhancing human body immunity power.
Compared with prior art, the invention has the beneficial effects as follows:
(1) compared with existing antiallergic Chinese medicine compound, the present invention has clear and definite antiallergic functional component, and product quality easily controls;
(2) compared with existing antiallergic Western medicine, extract of the present invention has too many levels and play synergistic function on sensitization target spot, has no side effect, safety is high;
(3) manufacture the contact of process organic solvent-free, nonhazardous remains; And preparation technology simply, easily operates, be suitable for Industry Promotion application.
Accompanying drawing explanation
Fig. 1 is preparation method process flow diagram of the present invention;
Fig. 2 is the block diagram of extract of the present invention to hyaluronidase Inhibition test 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 representation of cryogenic high pressure difference extractor.
Specific embodiment
In order to set forth content of the present invention better, be described with some preferably specific embodiments below.
Inventive principle
The present invention goes because of, strengthening the body resistance according to Chinese medicine compound is eliminating evil, yin or yang excessly partially to decline, make body recovery carry out reasonable formula to the treatment cause of disease reason of the normal condition of YIN and YANG in a relative equilibrium by coordinating zang-fu differentiation function thus correcting.The Radix Astragali, Cortex Albiziae, Radix Glycyrrhizae three have the active polysaccharide of different types of structure, so can difference to some extent in effect, can reach synergistic function in specific portfolio ratio situation.This is also the major reason instructing prescription of the present invention.
Membrane separation process is the separation of pure physical property, under pressure-driven room temperature carry out being separated, concentrate, refining, not undergoing phase transition in separation process, separated component does not have the change of calorifics, does not have chemical or biological change yet; Membrane separation process flow process totally closed operation, not vulnerable to pollution, finished product does not need to add antiseptic just can long-term storage.There is the feature of low energy consumption, high efficiency and normal-temperature operation.
The permission particle size range of Ultra filtration membrane granule is 0.1 ~ 0.001 μm, and the granularity of polysaccharide is 0.002 ~ 0.01 μm, so, suitable ultrafilter membrane can be selected to carry out separation and purification to polysaccharide.The principle of ultra-filtration and separation polysaccharide is generally first retain the large impurity such as resin, tannin of removing relative molecular weight, and then filtering small molecular protein, aminoacid etc., finally obtain the active polysaccharide of certain molecular weight scope.
The concrete effect that the present invention has is as follows:
The first, the composition that existing antiallergic Chinese medicine compound includes is general all many, and compositions of the present invention only needs three kinds of simple raw material of Chinese medicine, and product quality easily controls, and has clear and definite antiallergic effect.And compound of the present invention is not the simple mixing of random three kinds of raw materials, there is difference in three, only under specific portfolio ratio, just reaches synergistic function.Also shown by Efficacy experiments, the too many levels antiinflammatory that said composition has, antiallergic, enhancing human body immunity power successful are better than single component active polysaccharide, be in particular in can suppress IgE activity expression, stop the release of histamine, block the activation of NF-κ B, suppress IL-8 activity expression, suppress TNF-alpha active express total 5 have comparatively effectively, directly, the comprehensive irritated approach of control.
Second, compared with existing antiallergic Western medicine, antiallergic complex polysaccharide compositions of the present invention is the Chinese medicine treatment cause of disease reason according to Chinese medicine compound " eliminating evil go because of ", " strengthening the body resistance ", yin or yang excessly partially to decline by coordinating zang-fu differentiation function thus correcting, make body recovery arrive the normal condition of YIN and YANG in a relative equilibrium.Have no side effect, safety is high, and simultaneously compositions of the present invention also has too many levels play synergistic function on sensitization target spot.
3rd, traditional separation of polysaccharides purification process normally comprises water extraction, precipitate with ethanol, grease removal, removing protein, decolouring, post are separated these basic steps.In conventional Polysaccharide removing, method of protein has: sevag method, trifluorotrichloroethane method, trichloroacetic acid method, and its principle is all add that organic solvent makes protein denaturation precipitation and polysaccharide retains in the solution.But these above-mentioned methods inevitably bring the problem of dissolvent residual.
The present invention adopts cryogenic high pressure difference to extract in conjunction with micro-filtration membrane, ultra-filtration and separation technique, and manufacture process is organic solvent exposure not, and nonhazardous remains, and preparation technology simply easily operates, and is very applicable to applying of industrialization.
Membrane separation process is the separation of pure physical property, under pressure-driven room temperature carry out being separated, concentrate, refining, not undergoing phase transition in separation process, separated component does not have the change of calorifics, does not have chemical or biological change yet.And membrane separation process flow process is totally closed operation, not vulnerable to pollution, finished product does not need to add antiseptic just can long-term storage, has the advantage of low energy consumption, high efficiency and normal-temperature operation.
The principle of ultra-filtration and separation polysaccharide first retains the large impurity such as resin, tannin of removing relative molecular weight, and then filtering small molecular protein, aminoacid etc., finally obtain the active polysaccharide of certain molecular weight scope.The permission particle size range of Ultra filtration membrane granule is 0.1 ~ 0.001 μm, and the granularity of polysaccharide is 0.002 ~ 0.01 μm, as long as select the ultrafilter membrane of suitable particle size scope, just can realize carrying out separation and purification to the polysaccharide of corresponding granularity.
Embodiment 1
A kind of antianaphylactic complex polysaccharide, is made up of following parts by weight component: the Radix Astragali 50, Cortex Albiziae 35, Radix Glycyrrhizae 15.
Embodiment 2
A kind of antianaphylactic complex polysaccharide, is made up of following mass percent: be made up of following parts by weight component: the Radix Astragali 40, Radix Glycyrrhizae 35, Cortex Albiziae 25.
Embodiment 3
A kind of antianaphylactic complex polysaccharide, is made up of following parts by weight component: the Radix Astragali 35 ~ 45, Cortex Albiziae 20 ~ 30, Radix Glycyrrhizae 15 ~ 25.
Embodiment 4
A kind of antianaphylactic complex polysaccharide, is made up of following parts by weight component: Radix Glycyrrhizae 40, the Radix Astragali 30, Cortex Albiziae 30.
Embodiment 5
A kind of antianaphylactic complex polysaccharide, is made up of following parts by weight component: Cortex Albiziae 35, the Radix Astragali 35, Radix Glycyrrhizae 25.
Embodiment 6
A kind of antianaphylactic complex polysaccharide, is made up of following parts by weight component: the Radix Astragali 50, Radix Glycyrrhizae 30, Cortex Albiziae 20.
The preparation method of above-mentioned antiallergic complex polysaccharide, as shown in Figure 1, comprises the following steps:
(1) powder is beaten: after above-mentioned medical material is weighed by prescription, mixed powder is broken into 30-70 order, or above-mentioned medical material is ground into 30-70 order respectively, then mix homogeneously.
(2) feed intake: in above-mentioned medicated powder, add medicated powder quality 20-40 times amount water or volume by volume concentration are the alcoholic solution of 60 ~ 90%, stir; Add medicated powder quality 25-35 times of water effect better.
(3) extract: extract with cryogenic high pressure difference extractor under-0.1 ~ 70MPa, 0 ~ 25 DEG C of condition, collect extracting solution and carry out centrifugalize; Preferably, the condition of cryogenic high pressure difference extractor be-0.1 ~ 30MPa, 10 ~ 15 DEG C, such extraction efficiency is higher, and active matter shifts, stripping is more complete.Centrifugalize adopts link-suspended basket centrifuge to carry out, and rotating speed is 2500-4500 revolutions per second.
(4) separation is concentrated: the centrifugal liquid micro-filtration membrane of collection is removed the impurity such as solid particle, antibacterial, then uses the ultrafilter membrane grading purification polysaccharide of 200Da ~ 15000Da flux, finally, discharging concentrated by NF membrane.
Wherein, described micro-filtration membrane can adopt polrvinyl chloride (PVC) micro-filtration membrane and polypropylene (PP) micro-filtration membrane, and preferred aperture is 0.3 μm ~ 0.8 μm.Wherein, ultrafilter membrane classification is pure for first first adopting the ultrafilter membrane of 10000-20000Da flux to retain removing macromole impurity, then uses the ultrafilter membrane filtering small molecular weight impurity of 100-10000Da flux.Most preferred, first adopt the ultrafilter membrane of 15000-20000Da flux to retain removing macromole impurity, then use the ultrafilter membrane filtering small molecular weight impurity of 1000-10000Da flux, obtain purity of polysaccharide so high, yield is high.The molecule that retains of described NF membrane is 500-1500, the NF series NF membrane of Filmtec company of the U.S. or the NTR-7400 series NF membrane of Japan day east electrician.
Wherein, described cryogenic high pressure difference extractor as shown in Figure 8, is made up of extraction chamber 1, rewinding room 2, extracting solution tank battery 3, air compression station 4, dog-house 5, agitator 6, solvent aerosol apparatus 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; Described extraction chamber 1 and described rewinding room 2 share the pressure vessel of a band heat preservation sandwich layer and pass through leaf filter interval, the built-in described agitator 6 in described extraction chamber 1; Described extraction chamber 1 and described rewinding room 2 can be split also can be the standalone of disjunctor, with filter segmentation between described extraction chamber 1 and described rewinding room 2, described filter is board-like or the combination of rolling, filled type, pillar or different types filter; The top of described extraction chamber 1 is provided with described dog-house 5, and installs described solvent aerosol apparatus 7, and solvent aerosol apparatus 7 is by pipeline communication solvent storage tank 10; Described rewinding room 2 is by pipeline communication extracting solution tank battery 3; Slag storage tank 14 is extracted by pipeline communication bottom described extraction chamber 1; Described condensate buffering tank 11 passes through pipeline communication with vacuum station 12; The compressed air reservoir of described top, extraction chamber 1 and air compression station 4 passes through pipeline communication; Pressure vessel interlayer is set up circulating line with the temperature adjusting liquid storage tank of temperature set station 13 and is connected; Described extraction chamber 1 is provided with temperature detecting controller 20 and pressure testing and controlling device 30.Set up pipeline by the vacuum orifice on condensate buffering tank 11 with the vacuum buffer tank of vacuum station 12 to be connected, make the vacuum in rewinding room 2 can be made as required to be up to 90Pa during operation; Set up pipeline by the compressed air inlet at top, extraction chamber with the compressed air reservoir of air compression station 4 to be connected, make the pressure of rewinding indoor can be made as required to be up to 80MPa during operation; Set up circulating line by the temperature adjusting liquid import and export of interlayer with the temperature adjusting liquid storage tank of temperature set station 13 to be connected, temperature adjusting liquid temperature general control, at-35 DEG C ~ 50 DEG C, makes to make the temperature of extraction chamber and rewinding indoor control at-25 DEG C ~ 40 DEG C during operation as required; Set up information gathering and the transmission path of control system by the valve on the pressure testing and controlling device 30 at top, extraction chamber 1, temperature detecting controller 20 and pipeline, these information will be conducive to procedure design and the application of automatic control operation.Differential pressure extractor is attached is furnished with cleaning, decontamination system.
During operation, step is as follows:
1. reinforced: to open dog-house 5, start and stir dress device 6, dry material to be extracted is dropped in extraction chamber by charge door.
2. evacuation: after material finishes, closes dog-house 5, starts freezing water station 9 and make chilled water enter condenser 8 and circulate, and starts vacuum station, is evacuated by condensate buffering tank 11 by extraction chamber 1, after reaching the vacuum of needs, keeps certain hour.
3. add solvent: the solvent needed amount in solvent storage tank 10 quantitatively to be sprayed in extraction chamber 1 by solvent aerosol apparatus 7, keep certain hour.
4. subtract vacuum: start air compression station 4, logical compressed air enters in extraction chamber 1, makes house vacuum degree drop to-0.1MPa gradually.
5. pressurize: continue to pass into compressed air, make pressure in extraction chamber 1 rise to the highest 70MPa gradually, extracting solution is entered in rewinding room 2.
6. repeatedly carry out operating procedure 2 ~ 5, until extracting solution does not show selected index components identification.
7. open extraction chamber's bottom valve extraction slag to be expelled in extraction slag storage tank 14.
Embodiment 7
A preparation method for antianaphylactic complex polysaccharide, its step is as follows:
(1) Radix Astragali 15 kilograms, Cortex Albiziae 3 kilograms, 12 kilograms, Radix Glycyrrhizae are ground into respectively after 80 order powder again by composite formula mix homogeneously;
B. get the powder that step a obtains, add 150 kilograms of percentage by volume 50% ethanol waters, after stirring, obtain liquid to be extracted;
C. the liquid pump to be extracted obtained by step b enters cryogenic high pressure difference extractor, pressure be 15MPa, temperature carries out extracting once under being the condition of 10 DEG C;
D. the extracting solution obtained by step c sends into link-suspended basket centrifuge centrifugalize, and rotating speed is 3000 revolutions per seconds;
E. centrifugal liquid aperture steps d obtained is the micro-filtration membrane removing impurity of 0.4 μm;
F. the filtrate obtained by step e first retains removing macromole impurity with the ultrafilter membrane of 15000Da flux, collects filtrate, then retains removing small molecular weight impurity with the ultrafilter membrane of 3000Da flux;
G. the trapped fluid NF membrane first obtained by step f is reverse osmosis concentrated is reduced to 1.03 proportions, then by concentrated solution spraying dry, obtains the extract finished product of complex polysaccharide compositions.
Embodiment 8
A preparation method for antianaphylactic complex polysaccharide, its step is as follows:
A. the Radix Astragali 11 kilograms, Cortex Albiziae 6 kilograms, 13 kilograms, Radix Glycyrrhizae are ground into respectively after 40 order powder again by composite formula mix homogeneously;
B. get the powder that step a obtains, add 130 kilograms of pure water, after stirring, obtain liquid to be extracted;
C. the liquid pump to be extracted obtained by step b enters cryogenic high pressure difference extractor, pressure be 35MPa, temperature carries out extracting once under being the condition of 5 DEG C;
D. the flat centrifuge removing medicinal residues of extracting solution step c obtained;
E. centrifugal liquid aperture steps d obtained is the micro-filtration membrane removing impurity of 0.45 μm;
F. the filtrate obtained by step e first retains removing macromole impurity with the ultrafilter membrane of 20000Da flux, collects filtrate, then retains removing small molecular weight impurity with the ultrafilter membrane of 500Da flux;
G. the trapped fluid vacuum-concentrcted first obtained by step f to 1.08 proportions, then by concentrated solution drying under reduced pressure, obtains the extract finished product of complex polysaccharide compositions.
Embodiment 9
A kind of antiallergic mask, it is made up of following percentage by weight raw material:
Appropriate amount of essence and other active component can also be added in addition.The wherein antianaphylactic complex polysaccharide that obtains of antianaphylactic complex polysaccharide preferred embodiment 4.
The step of the preparation method of above-mentioned antiallergic mask is as follows:
(1) join in polyvinyl acetate emulsion by polyvinyl alcohol, stirring is mixed stand-by;
(2) sodium carboxymethyl cellulose, complex polysaccharide and propyl p-hydroxybenzoate are joined mix and blend in part (total amount 30-60%) distilled water, and be heated to about 80 DEG C, stir 2 hours, cool stand-by;
(3) by tween, glycerol and residue distilled water and essence mixing, stir stand-by;
(4) solution (2) is poured in solution (1), again solution (3) is poured into after stirring, be uniformly mixed 3 hours and get product.
Embodiment 10
A kind of antiallergic granule, it is made up of following percentage by weight raw material:
A preparation method for antiallergic granule, the steps include: the raw and auxiliary material taking above-mentioned formula ratio, adds ethanol in proper amount after abundant mix homogeneously, oven drying at low temperature after wet granulation.
Effect experimental
Example 7 (the antianaphylactic complex polysaccharide that all the other embodiments obtain also has similar effect) is extract obtained below carries out efficacy validation experiment.
1) hyaluronidase Inhibition test
Experimental technique: hyaluronidase adds the antianaphylactic complex polysaccharide of the present invention after calcium ion activated, then hyaluronate sodium is added, after reaction, use acetylacetone,2,4-pentanedione complexation in the basic conditions, finally with Ehrlich's reagent colour developing, measure its ultraviolet absorption value at 518nm place.
Experimental result: see Fig. 2, illustrates that the antianaphylactic complex polysaccharide of the present invention has good hyaluronidase inhibition.
2) cell function experiment
Experimental technique: the antianaphylactic complex polysaccharide of the present invention stimulating people's Acute Monocytic Leukemia Cell Line (THP-1) to add variable concentrations after 24 hours through lipopolysaccharide (LPS) is cultivated, by the immunoreation of IL-8, TNF-α, IgE in research THP-1 cell, the expression of NF-κ 1 B gene regulation and control.
Experimental result: along with the antianaphylactic complex polysaccharide of the present invention adds the increase of concentration, IL-8, TNF-α, lgE expression activity of LPS induction significantly reduces.Substantially IL-8, TNF-α, lgE activity expression can be suppressed when addition 1.0%.Illustrate that extract of the present invention has good antiinflammatory, antiallergic effect.See the following form 1 ~ 3.
Table 1. test specimen IL-8 concentration calculates
Table 2. test specimen TNF-α concentration calculates
Table 3. test specimen IgE concentration calculates
3) expression analysis of molecular gene regulation and control
Experimental technique: people's Acute Monocytic Leukemia Cell Line THP-1 adds after LPS stimulates 24 hours, and the antianaphylactic complex polysaccharide of the present invention adding variable concentrations cultivates 24 hours, the expression of NF-κ B in Q-PCR detection by quantitative THP-1.
Result shows: after THP-1 cell adds LPS stimulation, NF-κ B expresses increment 87.1%, after adding the antianaphylactic complex polysaccharide cultivation of the present invention, variable concentrations addition all shows the NF-kB activity suppressing LPS to stimulate and expresses, the NF-kB activity that can reduce 77.4%LPS induction when the antianaphylactic complex polysaccharide addition of the present invention 0.5% time is expressed, the NF-kB activity eliminating LPS induction when addition 1.0% is completely expressed, and the results are shown in Figure 3 ~ 7.
Polyoses content detects
[preparation of reference substance solution] gets anhydrous glucose reference substance 25mg, accurately weighed, puts in 250ml measuring bottle, and add water appropriate dissolving, is diluted to scale, shakes up, and obtains (containing anhydrous glucose 0.1mg in every 1ml).
[preparation of standard curve] precision measures reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, put in tool plug test tube respectively, add water respectively and mend to 2.0ml, each precision adds 5% phenol solution 1ml, shake up, rapid precision adds sulphuric acid 5ml, shakes up, place 10 minutes, put in 40 DEG C of water-baths and be incubated 15 minutes, take out, be cooled to room temperature rapidly, with corresponding reagent for blank, according to ultraviolet visible spectrophotometry, measuring absorbance at the wavelength place of 490nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
[sample determination] gets the antianaphylactic complex polysaccharide powder 1.0g that the above embodiment of the present invention obtains, and accurately weighed, add diethyl ether 100ml, reflux 1 hour, leaves standstill, lets cool, carefully discard ether solution, and residue puts water-bath Back stroke ether to the greatest extent.Add 80% ethanol 100ml, reflux 1 hour, filters while hot, and filtering residue and the hot 80% ethanol 30ml gradation of filter are washed, and filtering residue is put in flask together with filter paper, and add water 150ml, reflux 2 hours.Filter while hot, with a small amount of hot wash filter, merging filtrate and washing liquid, let cool, move in 250ml measuring bottle, be diluted with water to scale, shake up rear appropriate dilution, precision measures diluent 1ml, puts in tool plug test tube, add water 1.0ml, the method under the preparation of sighting target directrix curve, from " each precision adds 5% phenol solution 1ml ", measure absorbance in accordance with the law, read the weight (mg) containing glucose in need testing solution from standard curve, calculate, to obtain final product.
The determination of polysaccharide result of table 4 extract of the present invention
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be equivalent replacement mode, be included within protection scope of the present invention.
Claims (10)
1. an antiallergic complex polysaccharide compositions, is characterized in that: make by comprising following parts by weight component: the Radix Astragali 10 ~ 90, Cortex Albiziae 10 ~ 90, Radix Glycyrrhizae 1 ~ 50.
2. the preparation method of antiallergic complex polysaccharide compositions according to claim 1, is characterized in that: make by comprising following parts by weight component: the Radix Astragali 30 ~ 50, Cortex Albiziae 10 ~ 40, Radix Glycyrrhizae 10 ~ 30.
3. a preparation method for antiallergic complex polysaccharide compositions, is characterized in that: it step comprised is as follows:
(1) powder is beaten: the Radix Astragali, Cortex Albiziae and Radix Glycyrrhizae are pulverized;
(2) feed intake: in medicated powder, add its weight 5 ~ 50 times amount water or alcohol-water mixture, stir;
(3) extract: extract under-0.1 ~ 70MPa, 0 ~ 25 DEG C of condition, then carry out centrifugalize, collect centrifugal liquid;
(4) separation is concentrated: centrifugal liquid micro-filtration membrane is removed impurity, then uses the ultrafilter membrane grading purification polysaccharide of 100Da ~ 20000Da flux.
4. the preparation method of antiallergic complex polysaccharide compositions according to claim 3, is characterized in that: in described (2) step, and the addition of water is 20 ~ 30 times for medicated powder weight.
5. the preparation method of antiallergic complex polysaccharide compositions according to claim 3, is characterized in that: in described (3) step, and extraction conditions is pressure is-0.1 ~ 35MPa, Extracting temperature is 10 ~ 20 DEG C.
6. the preparation method of antiallergic complex polysaccharide compositions according to claim 3, is characterized in that: in described (4) step, and the aperture specification of micro-filtration membrane is 0.2 μm ~ 1.0 μm; Ultrafilter membrane aperture specification is 1000Da ~ 10000Da.
7. the preparation method of antiallergic complex polysaccharide compositions according to claim 3, is characterized in that: described alcohol-water mixture preferred volume hundred number is the ethanol water of 50 ~ 95%.
8. the preparation method of antiallergic complex polysaccharide compositions according to claim 3, is characterized in that: in described (4) step, the trapped fluid obtained is concentrated to 1.03 ~ 1.08 proportions.
9. antiallergic complex polysaccharide compositions according to claim 1, is characterized in that: for the preparation of the cosmetics with antiallergic effect, and addition is 1 ~ 50%.
10. antiallergic complex polysaccharide compositions according to claim 1, is characterized in that: for the preparation of having in the medicine of antiallergic effect, addition is 10 ~ 90%.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115069092A (en) * | 2022-08-08 | 2022-09-20 | 碧菲分离膜（大连）有限公司 | Membrane method for extracting traditional Chinese medicine |
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