CN110384787A

CN110384787A - A kind of Chinese medicine composition and preparation method thereof for treating allergic rhinitis

Info

Publication number: CN110384787A
Application number: CN201810356800.5A
Authority: CN
Inventors: 李友林
Original assignee: SINO-JAPANESE FRIENDSHIP HOSPITAL
Current assignee: SINO-JAPANESE FRIENDSHIP HOSPITAL; China Japan Friendship Hospital
Priority date: 2018-04-19
Filing date: 2018-04-19
Publication date: 2019-10-29

Abstract

The present invention provides a kind of Chinese medicine composition for treating allergic rhinitis, and the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 5-50 parts by weight, ramulus cinnamomi 3-27 parts by weight, flower bud of lily magnolia 2-20 parts by weight, radix saposhnikoviae 3-24 parts by weight, Rhizoma Atractylodis Macrocephalae 3-27 parts by weight, rhizoma zingiberis 2-20 parts by weight, cortex cinnamomi 1-8 parts by weight, Radix Paeoniae Alba 2-20 parts by weight, dark plum 4-30 parts by weight, Schisandra chinensis 2-20 parts by weight, rhizoma anemarrhenae 2-20 parts by weight, Radix Glycyrrhizae 2-20 parts by weight.

Description

A kind of Chinese medicine composition and preparation method thereof for treating allergic rhinitis

Technical field

The present invention relates to a kind of Chinese medicine compositions, and in particular to a kind of Chinese medicine composition for treating allergic rhinitis and its system Preparation Method belongs to field of medicaments.

Background technique

Allergic rhinitis (allergic rhinitis, AR) i.e. allergic rhinitis refer to that atopic individuals contact allergen It is mainly discharged afterwards by the medium (mainly histamine) that IgE is mediated, and there are many immunocompetent cell and cell factors etc. to participate in Schneiderian membrance non-infectious inflammatory disease.AR is one of most common anaphylactia, and disease incidence rises in obvious in recent years Trend about influences the crowd of global 10%-25%, especially higher in its national illness rate of industry prosperity.Conservative estimation Global AR patient is more than 500,000,000.11, China key city telephone questionnaire investigation display AR illness rate is about 11%.AR can lead to very More diseases and disability, have become major global public health problem.

Western medical treatment AR is based on immunotherapy and drug therapy, and there is deficiencies: the more difficult determination of immunotherapy allergen And standard is difficult consistent, works slow, the course for the treatment of is long, should not adhere to, it is difficult to cure；Drug therapy mainly has decongestant, antihistamine Medicine, glucocorticoid, anticholinergic drug, mast cell stabilizers, leukotriene receptor antagonists etc. with symptomatic treatment, alleviate disease Based on shape, although alleviation symptom is very fast, ideal radical cure method there is no at present.

Chinese traditional treatment AR, therapy concentrate on sensible, dampness removing stagnation resolvation of inducing diaphoresis etc., also slightly take into account righting.But mainly to dispel table It is not comprehensive to human righteousness's solid protection based on card；Globality and systematicness to interpretation of the cause, onset and process of an illness essence understand that deficiency, the level of understanding need to be mentioned It is high；Therapeutic effect is still weak, and is not able to satisfy clinical demand.

Summary of the invention

In order to solve the problems existing in the prior art, the present invention provide a kind of Chinese medicine composition for treating allergic rhinitis and its Preparation method.

As one aspect of the present invention, the present invention provides a kind of Chinese medicine composition for treating allergic rhinitis, the Chinese medicine Composition is made of the following raw material medicine: Radix Astragali, the flower bud of lily magnolia, radix saposhnikoviae, Rhizoma Atractylodis Macrocephalae, rhizoma zingiberis, cortex cinnamomi, Radix Paeoniae Alba, dark plum, Schisandra chinensis, is known ramulus cinnamomi Female, Radix Glycyrrhizae.

In a particular embodiment, the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 5-50 parts by weight, ramulus cinnamomi 3-27 parts by weight, flower bud of lily magnolia 2-20 parts by weight, radix saposhnikoviae 3-24 parts by weight, Rhizoma Atractylodis Macrocephalae 3-27 parts by weight, rhizoma zingiberis 2-20 parts by weight, cortex cinnamomi 1- 8 parts by weight, Radix Paeoniae Alba 2-20 parts by weight, dark plum 4-30 parts by weight, Schisandra chinensis 2-20 parts by weight, rhizoma anemarrhenae 2-20 parts by weight, Radix Glycyrrhizae 2- 20 parts by weight.

In a preferred embodiment, the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 8-40 parts by weight, ramulus cinnamomi 4-22 parts by weight, flower bud of lily magnolia 3-16 parts by weight, radix saposhnikoviae 4-18 parts by weight, Rhizoma Atractylodis Macrocephalae 4-22 parts by weight, rhizoma zingiberis 3-16 parts by weight, cortex cinnamomi 1- 6 parts by weight, Radix Paeoniae Alba 3-16 parts by weight, dark plum 6-24 parts by weight, Schisandra chinensis 3-16 parts by weight, rhizoma anemarrhenae 3-16 parts by weight, Radix Glycyrrhizae 3- 16 parts by weight.

In further preferred embodiment, the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 10-30 weight Part, ramulus cinnamomi 5-18 parts by weight, flower bud of lily magnolia 4-12 parts by weight, radix saposhnikoviae 5-14 parts by weight, Rhizoma Atractylodis Macrocephalae 5-18 parts by weight, rhizoma zingiberis 4-12 weight Part, cortex cinnamomi 2-5 parts by weight, Radix Paeoniae Alba 4-12 parts by weight, dark plum 8-20 parts by weight, Schisandra chinensis 4-12 parts by weight, rhizoma anemarrhenae 4-12 weight Part, Radix Glycyrrhizae 4-12 parts by weight.

In further preferred embodiment, the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 15-25 weight Part, ramulus cinnamomi 7-12 parts by weight, flower bud of lily magnolia 5-8 parts by weight, radix saposhnikoviae 6-9 parts by weight, Rhizoma Atractylodis Macrocephalae 7-12 parts by weight, rhizoma zingiberis 5-8 parts by weight, meat Osmanthus 2-4 parts by weight, Radix Paeoniae Alba 5-8 parts by weight, dark plum 10-15 parts by weight, Schisandra chinensis 5-8 parts by weight, rhizoma anemarrhenae 5-8 parts by weight, Radix Glycyrrhizae 5-8 parts by weight.

In most preferred embodiments, the Chinese medicine composition is made of the following raw material medicine: 21 parts by weight of Radix Astragali, ramulus cinnamomi 9 parts by weight, 6 parts by weight of the flower bud of lily magnolia, windproof 7 parts by weight, 9 parts by weight of Rhizoma Atractylodis Macrocephalae, 6 parts by weight of rhizoma zingiberis, 3 parts by weight of cortex cinnamomi, 6 weight of Radix Paeoniae Alba Part, 12 parts by weight of dark plum, 6 parts by weight of Schisandra chinensis, 6 parts by weight of rhizoma anemarrhenae, 6 parts by weight of Radix Glycyrrhizae；

Or, the Chinese medicine composition is made of the following raw material medicine: 16 parts by weight of Radix Astragali, 11 parts by weight of ramulus cinnamomi, 5 weight of the flower bud of lily magnolia Part, windproof 8 parts by weight, 8 parts by weight of Rhizoma Atractylodis Macrocephalae, 7 parts by weight of rhizoma zingiberis, 2 parts by weight of cortex cinnamomi, 7 parts by weight of Radix Paeoniae Alba, 11 parts by weight of dark plum, 7 parts by weight of Schisandra chinensis, 5 parts by weight of rhizoma anemarrhenae, 7 parts by weight of Radix Glycyrrhizae；

Or, the Chinese medicine composition is made of the following raw material medicine: 24 parts by weight of Radix Astragali, 8 parts by weight of ramulus cinnamomi, 7 weight of the flower bud of lily magnolia Part, windproof 6 parts by weight, 11 parts by weight of Rhizoma Atractylodis Macrocephalae, 5 parts by weight of rhizoma zingiberis, 4 parts by weight of cortex cinnamomi, 5 parts by weight of Radix Paeoniae Alba, 14 parts by weight of dark plum, 5 parts by weight of Schisandra chinensis, 7 parts by weight of rhizoma anemarrhenae, 5 parts by weight of Radix Glycyrrhizae.

As preferred embodiment, the Radix Astragali is made a living Radix Astragali；The Rhizoma Atractylodis Macrocephalae is rhizoma atractylodis macrocephalae；The Radix Glycyrrhizae is made a living sweet Grass.

Chinese medicine composition of the present invention can be the composition mixed after each bulk pharmaceutical chemicals crush, can also mix or The active component that the extract or the further polishing purification of extract obtained after individually extracting obtains, can also be addition Regular dosage form made of pharmaceutically acceptable auxiliary material.

Wherein, the extracting method includes decocting extraction, refluxing extraction, Soakage extraction, ultrasonic extraction, seepage pressure effects, micro- Wave extraction etc.；The purification process includes water extract-alcohol precipitation, alkali soluble acid sinks and various column chromatography purification process, such as macroreticular resin Column, silicagel column, gel column, reversed-phase column etc.；The regular dosage form include but is not limited to injection, capsule, tablet, granule, Gelling agent, sustained release agent, oral solution, pill or nanometer formulation；The pharmaceutically acceptable auxiliary material include: filler, disintegrating agent, Lubricant, suspending agent, adhesive, sweetener, corrigent, preservative, matrix etc..Filler include: starch, pregelatinized starch, Lactose, mannitol, chitin, microcrystalline cellulose, sucrose etc.；Disintegrating agent includes: starch, pregelatinized starch, microcrystalline cellulose, carboxylic Methyl starch sodium, crosslinked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, croscarmellose sodium etc.；Lubricant packet It includes: magnesium stearate, lauryl sodium sulfate, talcum powder, silica etc.；Suspending agent includes: polyvinylpyrrolidone, crystallite fibre Tie up element, sucrose, agar, hydroxypropyl methyl cellulose etc.；Adhesive includes starch slurry, polyvinylpyrrolidone, hydroxypropyl methyl Cellulose etc..

As the second aspect of the invention, the present invention provides the Chinese medicine composition in preparation treatment allergic rhinitis Application in drug.

As the third aspect of the invention, the present invention provides the medicine that the Chinese medicine composition has anti-inflammatory effect in preparation Application in object.

As the fourth aspect of the invention, the present invention, which provides the Chinese medicine composition, has anti-allergic effects in preparation Application in drug.

As the fifth aspect of the invention, the present invention provides the preparation method of the Chinese medicinal composition granules, described Method includes:

Composition extract powder is taken, with excipient in 3: the ratio of (1~2) mixes, and it is dry using wet granulation, obtain particle Agent；

Wherein, the composition extract powder is the extract obtained after each bulk pharmaceutical chemicals extract according to a conventional method, or is mentioned The active component for taking the further polishing purification of object to obtain.The extracting method include decoct extraction, refluxing extraction, Soakage extraction, Ultrasonic extraction, seepage pressure effects, Microwave Extraction etc.；The purification process includes water extract-alcohol precipitation, alkali soluble acid sinks and various column chromatographys Purification process, such as macroporous resin column, silicagel column, gel column, reversed-phase column.

In a particular embodiment, the excipient in dextrin, lactose, soluble starch any one or it is several Kind, preferably dextrin；The ethanol solution that the wetting agent is 90%~95%, preferably 94% ethanol solution.The combination Object extract powder and excipient ratios are preferably 3: 2.

In a particular embodiment, in the pelletization envionmental humidity below 74%.

In a preferred embodiment, the composition extract powder is prepared as follows:

Step a takes ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, four taste of the flower bud of lily magnolia to obtain volatile oil using extraction by steam distillation volatile oil A, the aqueous B and dregs of a decoction C after distillation；

Step b will volatilize oily A and be included with betadex；Dregs of a decoction C extracting in water, obtains extracting solution D；

Step c, takes Radix Astragali, radix saposhnikoviae, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, dark plum, Schisandra chinensis, rhizoma anemarrhenae, eight taste of Radix Glycyrrhizae, and extracting in water must extract Liquid E；

Step d merges the aqueous B after extracting solution D, E and step a distillation, and adding ethyl alcohol to alcohol content is 50%~70%, Filtrate recycling ethanol is taken, dry, pulverize, is mixed with betadex inclusion compound to get composition extract powder.

Wherein, the ratio of step b volatile oil and betadex is 1: (5~10)；It is preferred that 1: (6~9)；Most preferably 1: 8； Step d adds ethyl alcohol to alcohol content to be 60%.

As the sixth aspect of the invention, the present invention provides the detection method of the Chinese medicinal composition granules, described Method includes:

Assay A:

Determination condition: using octadecylsilane key and silica gel as filler；With acetonitrile-water (25~35): (65~75) are Mobile phase；

The preparation of reference substance solution: taking Astragaloside IV reference substance, and organic solvent dissolution obtains reference substance solution；

The preparation of test solution: taking Chinese medicinal composition granules, and organic solvent extracts, and obtains test solution；

Measuring method: drawing reference substance solution and test solution respectively, injects liquid chromatograph, measurement.

Assay B:

Determination condition: using octadecylsilane key and silica gel as filler；With methanol-water (10~20): (80~90) are Mobile phase；

The preparation of reference substance solution: taking Paeoniflorin reference substance, and organic solvent dissolution obtains reference substance solution；

In a particular embodiment, the organic solvent be selected from methanol, acetone, n-butanol, ethyl acetate, petroleum ether, Any one in chloroform；The extracting method is selected from ultrasonic extraction, refluxing extraction, cold soaking extraction, seepage pressure effects, Microwave Extraction The methods of.

In a preferred embodiment, it in the preparation of assay A test solution, is extracted using methanol eddy；More into one Step, test solution the preparation method is as follows: take Chinese medicinal composition granules, add methanol cold soaking to stay overnight, refluxing extraction, filtrate is returned It receives solvent and is concentrated to dryness, residue is dissolved in water, and is saturated with water n-butanol shaking and extracts, n-butanol extracting liquid liquid is washed with ammonia solution Wash, ammonia solution discards, and the n-butanol liquid after washing is evaporated, residue add methanol make dissolution to get.

In a preferred embodiment, mobile phase is acetonitrile-water (31: 69) in assay A；It is flowed in assay B Mutually it is methanol-water (15: 85).

In a particular embodiment, the detection method further includes one or more of following identification:

Identify A:

Test solution preparation: taking Chinese medicinal composition granules, and organic solvent extracts, and obtains test solution；

Control medicinal material solution preparation: taking Radix Astragali control medicinal material, and organic solvent extracts, and obtains control medicinal material solution；

Detection: drawing test solution and control medicinal material solution respectively, and point is on same silica gel plate, with chloroform-first Alcohol-water (10~15): (5~9): (1~3) is solvent, and expansion is heated to the colour developing of 10% ethanol solution of sulfuric acid at 105 DEG C Spot development is clear；

Identify B:

Reference substance solution preparation: taking Paeoniflorin reference substance, and organic solvent dissolution obtains reference substance solution；

Detection: drawing test solution and control solution respectively, and point is on same silica gel plate, with chloroform-acetic acid Ethyl ester-methyl alcohol-formic acid (5~10): (1~3): (2~7): (0.1~0.3) is solvent, and expansion is molten with 5% vanillin-sulfuric acid Liquid colour developing, it is clear to be heated to spot development at 105 DEG C；

Identify C:

Control medicinal material solution preparation: taking windproof control medicinal material, and organic solvent extracts, and obtains control medicinal material solution；

Reference substance solution preparation: taking 5-O- methyl visamminol glycosides reference substance, and it is molten to obtain reference substance for organic solvent dissolution Liquid；

Detection: drawing test solution, control medicinal material solution and control medicinal material solution respectively, point on same silica gel plate, With chloroform-methanol (3~8): (0.5~2) is solvent, and expansion is set and inspected under ultraviolet lamp (254nm)；

Identify D:

Control medicinal material solution preparation: taking Schisandra chinensis control medicinal material, and organic solvent extracts, and obtains control medicinal material solution；

Reference substance solution preparation: taking schizandrin reference substance, and organic solvent dissolution obtains reference substance solution；

Detection: drawing test solution, control medicinal material solution and control medicinal material solution respectively, point on same silica gel plate, With petroleum ether-Ethyl formate-formic acid (12~18): (4~8): (0.5~2) is solvent, and expansion is set under ultraviolet lamp (254nm) It inspects；

Identify E:

Control medicinal material solution preparation: extracting liquorice control medicinal material, organic solvent extract, and obtain control medicinal material solution；

Detection: drawing test solution and control medicinal material solution respectively, and point is on same silica gel plate, with ethyl acetate-first Acid-glacial acetic acid-water (12~18): (0.5~2): (0.5~2): (1~3) is solvent, and expansion is molten with 10% sulfuric acid ethyl alcohol It is clear to be heated to spot development at 105 DEG C for liquid；

In a preferred embodiment, identify A sample solution preparation method are as follows: take Chinese medicinal composition granules, add first Alcohol ultrasonic extraction, filtrate recycling design are simultaneously concentrated to dryness, and residue is dissolved in water, and are saturated with water n-butanol shaking and are extracted, n-butanol Extracting solution liquid is washed with ammonia solution, and ammonia solution discards, and the n-butanol liquid after washing is evaporated, residue add methanol make dissolution to get.

Each bulk pharmaceutical chemicals of the present invention are recorded in " Chinese Pharmacopoeia " 2015 editions, and concocting method is " pharmacopeia " record method.

Radix Astragali in prescription of the present invention, sweet, warm, return lung, the spleen channel, invigorating qi for strengthening superficies, strengthening spleen and tonifying lung；Ramulus cinnamomi, pungent, sweet, warm is returned The heart, lung, bladder meridian, warming meridian, helps yang transforming qi；Two medicines are total to the function of figure warming Yang, enlivening spleen benefit lung, with for monarch drug in a prescription.Ministerial drug one is (pungent It is smooth), it is pungent, warm, it attaches to the lung and stomach meridians, function is apt to expelling cold, eliminating, clearing the nasal passage, nasal obstruction, the key medicine of nasosinusitis；Ministerial drug two (radix saposhnikoviae), it is pungent, sweet, micro- Temperature returns bladder, liver, the spleen channel, expels pathogenic wind from the body surface, and tool rises the property of clear eliminating dampness, can be used for the insufficiency of the spleen with overabundance of dampness, lucid yang failing to raise；Ministerial drug three is (white Art), bitter, sweet, warm, returns spleen, stomach meridian, strengthening the spleen and replenishing qi；Ministerial drug four (rhizoma zingiberis), pungent, hot, returns spleen, stomach, kidney, the heart, lung channel, middle benefit gas dissipate It is cold, recover Yang and smooth venation, eliminating dampness dissolving phlegm；Ministerial drug five (cortex cinnamomi), pungent, sweet, big heat.Return kidney, spleen, the heart, Liver Channel, mends fire supporing yang, cold dispelling stops Bitterly, warming meridians and promoting circulation of qi, let the fire back to its origin；Five medicines or walk to express key or warming meridians and promoting circulation of qi or strengthening the spleen and replenishing qi to help lung be ministerial drug altogether.Adjutant one (Radix Paeoniae Alba), it is bitter, sour, slightly cold, return liver, the spleen channel, yin, the analgesic of soft liver, suppressing liver-YANG are held back in blood-nourishing, and astringing lung-QI yin consolidates lung body, smooth mechanism of qi benefit Qi and blood；Adjutant two (dark plum), it is sour, puckery, it puts down, returns liver, spleen, lung, large intestine channel, astringe the lung to stop cough, relieving diarrhea with astringents promotes the production of body fluid to quench thirst, adjutant Three (Schisandra chinensis), acid, sweet, warm, return lung, the heart, kidney channel, astringency inducing, supplementing qi and promoting the production of body fluid, tonifying kidney and calming nerves are kind to control long-last cough and virtual asthma, saliva wound It is thirsty, feeble pulse of losing heart, Heat Diabetes, palpitation and insomnia；Adjutant four (rhizoma anemarrhenae), it is bitter, sweet, it trembles with fear, return lung, stomach, kidney channel, clearing heat-fire, Fluid dryness, the anti-hot and suffocating interior life of deficiency of both lung and spleen；Above-mentioned four medicine, convergence gas yin, facilitates strengthening spleen and tonifying lung, reaches the same goal by different routes and mutually shares as assistant Medicine.Radix Glycyrrhizae, sweet and neutral, return heart, lung, spleen, stomach meridian are invigorated the spleen and benefited qi, clearing heat and detoxicating, expelling phlegm and arresting coughing, coordinating the drug actions of a prescription, with to make medicine.Entirely " gentle pungent Jin Peiben " principle is closed by side, and acridness and sweetness activating yang, sour-sweet herbs nourishing yin, cold temperature, which dissipate to receive, to be used in combination, and is searched for the primary cause of disease in treatment, play altogether strengthening spleen and tonifying lung, The function of warm tonneau key.

Clinical research show 100 AR patients after the traditional chinese medicine composition of the invention is treated, effective 76, effective 15, nothing Effect 9, effective percentage 91%.Symptom and sign integral, VAS scale integral and the former total IgE of serum anaphylaxis are aobvious before relatively treating after treatment Writing reduces (P < 0.01).Show that the traditional chinese medicine composition of the invention can improve the clinical symptoms and sign of AR patient, reduces patient's blood The clear total IgE of anaphylactogen is horizontal, significant in efficacy.

Detailed description of the invention

Fig. 1 is Astragaloside IV reference substance solution chromatogram

Fig. 2 is Chinese medicine composition test solution chromatogram

Fig. 3 is Radix Astragali feminine gender test solution chromatogram

Fig. 4 is Paeoniflorin reference substance solution chromatogram

Fig. 5 is Chinese medicine composition test solution chromatogram

Fig. 6 is Radix Paeoniae Alba feminine gender test solution chromatogram

Specific embodiment

1 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 21g, ramulus cinnamomi 9g, flower bud of lily magnolia 6g, radix saposhnikoviae 7g, Rhizoma Atractylodis Macrocephalae 9g, rhizoma zingiberis 6g, cortex cinnamomi 3g, Radix Paeoniae Alba 6g, dark plum 12g, Schisandra chinensis 6g, rhizoma anemarrhenae 6g, Radix Glycyrrhizae 6g；

Preparation method: taking each bulk pharmaceutical chemicals in proportion, and water boiling and extraction obtains Chinese medicine composition decoction.

2 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 16g, ramulus cinnamomi 11g, flower bud of lily magnolia 5g, radix saposhnikoviae 8g, Rhizoma Atractylodis Macrocephalae 8g, rhizoma zingiberis 7g, cortex cinnamomi 2g, Radix Paeoniae Alba 7g, dark plum 11g, Schisandra chinensis 7g, rhizoma anemarrhenae 5g, Radix Glycyrrhizae 7g；

3 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 24g, ramulus cinnamomi 8g, flower bud of lily magnolia 7g, radix saposhnikoviae 6g, Rhizoma Atractylodis Macrocephalae 11g, rhizoma zingiberis 5g, cortex cinnamomi 4g, Radix Paeoniae Alba 5g, dark plum 14g, Schisandra chinensis 5g, rhizoma anemarrhenae 7g, Radix Glycyrrhizae 5g；

4 Chinese medicinal composition granules of embodiment

Preparation method:

Ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, the flower bud of lily magnolia extract volatile oil 10 hours, and volatile oil is included with betadex, the water after distillation Liquid is spare, and the dregs of a decoction add 8 times of amount water to extract 1 hour, and filtration, the another device of filtrate stores for future use；

Eight taste such as remaining Radix Astragali, Radix Paeoniae Alba adds water to cook three times, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned Two liquid merge, and are concentrated into the clear cream that relative density is 1.15~1.20 (60 DEG C), add ethyl alcohol to make alcohol content up to 60%, stirring is quiet It sets overnight, filtrate recycling ethanol, filtrate is concentrated into right amount, is dried under reduced pressure, and fine powder is ground into, with above-mentioned betadex inclusion compound It mixes, obtains composition extract powder；

Composition extract powder is uniformly mixed with dextrin by 3: the 2 auxiliary ratio of medicine, wet granulation, is wetting with 94% ethyl alcohol Agent, 74% hereinafter, preparing particle, dry, packaging gets product particle for envionmental humidity control.

The research of the invention finds that wanting strictly controlled environment relative humidity 74% hereinafter, if relatively wet in pelletization Degree is higher than 74%, and it will cause the material moisture absorption, granulation is difficult.

5 Chinese medicinal composition granules of embodiment

Preparation method is the same as embodiment 4.

6 Chinese medicinal composition granules of embodiment

Preparation method is the same as embodiment 4.

7 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 18g, ramulus cinnamomi 10g, flower bud of lily magnolia 5g, radix saposhnikoviae 9g, Rhizoma Atractylodis Macrocephalae 7g, rhizoma zingiberis 8g, cortex cinnamomi 3g, Radix Paeoniae Alba 5g, dark plum 12g, Schisandra chinensis 5g, rhizoma anemarrhenae 8g, Radix Glycyrrhizae 5g；

Preparation method:

Ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, the flower bud of lily magnolia extract volatile oil 12 hours, and volatile oil is included with betadex, the water after distillation Liquid is spare, and the dregs of a decoction add 10 times of amount water to extract 1 hour, and filtration, the another device of filtrate stores for future use；

Eight taste such as remaining Radix Astragali, Radix Paeoniae Alba adds water to cook three times, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned Two liquid merge, and are concentrated into the clear cream that relative density is 1.15~1.20 (60 DEG C), add ethyl alcohol to make alcohol content up to 70%, stirring is quiet It sets overnight, filtrate recycling ethanol, filtrate is concentrated into right amount, is dried under reduced pressure, and fine powder is ground into, with above-mentioned betadex inclusion compound It mixes, obtains composition extract powder；

Composition extract powder is uniformly mixed with dextrin by 3: the 1 auxiliary ratio of medicine, wet granulation, is wetting with 94% ethyl alcohol Agent, 74% hereinafter, preparing particle, dry, packaging gets product particle for envionmental humidity control.

8 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 23g, ramulus cinnamomi 7g, flower bud of lily magnolia 8g, radix saposhnikoviae 6g, Rhizoma Atractylodis Macrocephalae 10g, rhizoma zingiberis 5g, cortex cinnamomi 3g, Radix Paeoniae Alba 8g, dark plum 10g, Schisandra chinensis 8g, rhizoma anemarrhenae 5g, Radix Glycyrrhizae 8g；

Preparation method is the same as embodiment 7.

9 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 12g, ramulus cinnamomi 15g, flower bud of lily magnolia 4g, radix saposhnikoviae 12g, Rhizoma Atractylodis Macrocephalae 6g, rhizoma zingiberis 10g, cortex cinnamomi 2g, Radix Paeoniae Alba 10g, crow Plum 9g, Schisandra chinensis 10g, rhizoma anemarrhenae 4g, Radix Glycyrrhizae 10g；

Preparation method is the same as embodiment 7.

10 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 27g, ramulus cinnamomi 6g, flower bud of lily magnolia 10g, radix saposhnikoviae 5g, Rhizoma Atractylodis Macrocephalae 15g, rhizoma zingiberis 4g, cortex cinnamomi 5g, Radix Paeoniae Alba 4g, dark plum 18g, Schisandra chinensis 4g, rhizoma anemarrhenae 10g, Radix Glycyrrhizae 4g；

Preparation method:

Ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, the flower bud of lily magnolia extract volatile oil 8 hours, and volatile oil is included with betadex, the water after distillation Liquid is spare, and the dregs of a decoction add 15 times of amount water to extract 1 hour, and filtration, the another device of filtrate stores for future use；

Eight taste such as remaining Radix Astragali, Radix Paeoniae Alba adds water to cook three times, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned Two liquid merge, and are concentrated into the clear cream that relative density is 1.15~1.20 (60 DEG C), add ethyl alcohol to make alcohol content up to 50%, stirring is quiet It sets overnight, filtrate recycling ethanol, filtrate is concentrated into right amount, is dried under reduced pressure, and fine powder is ground into, with above-mentioned betadex inclusion compound It mixes, obtains composition extract powder；

Composition extract powder is uniformly mixed with dextrin by 3: the 2 auxiliary ratio of medicine, wet granulation, is wetting with 95% ethyl alcohol Agent, 74% hereinafter, preparing particle, dry, packaging gets product particle for envionmental humidity control.

11 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 9g, ramulus cinnamomi 20g, flower bud of lily magnolia 3g, radix saposhnikoviae 16g, Rhizoma Atractylodis Macrocephalae 4g, rhizoma zingiberis 14g, cortex cinnamomi 1g, Radix Paeoniae Alba 14g, dark plum 7g, Schisandra chinensis 14g, rhizoma anemarrhenae 3g, Radix Glycyrrhizae 14g；

The preparation method is the same as that of Example 10.

12 Chinese medicinal composition granules of embodiment

Prescription: Radix Astragali 35g, ramulus cinnamomi 4g, flower bud of lily magnolia 14g, radix saposhnikoviae 4g, Rhizoma Atractylodis Macrocephalae 20g, rhizoma zingiberis 3g, cortex cinnamomi 6g, Radix Paeoniae Alba 3g, dark plum 22g, Schisandra chinensis 3g, rhizoma anemarrhenae 14g, Radix Glycyrrhizae 3g；

The preparation method is the same as that of Example 10.

13 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 7g, ramulus cinnamomi 25g, flower bud of lily magnolia 2g, radix saposhnikoviae 22g, Rhizoma Atractylodis Macrocephalae 3g, rhizoma zingiberis 18g, cortex cinnamomi 1g, Radix Paeoniae Alba 18g, dark plum 5g, Schisandra chinensis 18g, rhizoma anemarrhenae 2g, Radix Glycyrrhizae 18g；

14 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 45g, ramulus cinnamomi 3g, flower bud of lily magnolia 18g, radix saposhnikoviae 3g, Rhizoma Atractylodis Macrocephalae 25g, rhizoma zingiberis 2g, cortex cinnamomi 7g, Radix Paeoniae Alba 3g, dark plum 26g, Schisandra chinensis 2g, rhizoma anemarrhenae 18g, Radix Glycyrrhizae 2g；

15 Chinese medicine composition decoction of embodiment

Prescription: Radix Astragali 15g, ramulus cinnamomi 12g, flower bud of lily magnolia 5g, radix saposhnikoviae 9g, Rhizoma Atractylodis Macrocephalae 7g, rhizoma zingiberis 8g, cortex cinnamomi 2g, Radix Paeoniae Alba 8g, dark plum 10g, Schisandra chinensis 8g, rhizoma anemarrhenae 5g, Radix Glycyrrhizae 8g；

16 Chinese medicine composition tablet of embodiment

Prescription: Radix Astragali 25g, ramulus cinnamomi 7g, flower bud of lily magnolia 8g, radix saposhnikoviae 6g, Rhizoma Atractylodis Macrocephalae 12g, rhizoma zingiberis 5g, cortex cinnamomi 4g, Radix Paeoniae Alba 5g, dark plum 15g, Schisandra chinensis 5g, rhizoma anemarrhenae 8g, Radix Glycyrrhizae 5g；

Preparation method:

Tablet is made in composition extract powder addition conventional tablet auxiliary material.

17 Chinese medicine composition tablet of embodiment

Prescription: Radix Astragali 30g, ramulus cinnamomi 5g, flower bud of lily magnolia 12g, radix saposhnikoviae 5g, Rhizoma Atractylodis Macrocephalae 18g, rhizoma zingiberis 4g, cortex cinnamomi 5g, Radix Paeoniae Alba 4g, dark plum 20g, Schisandra chinensis 4g, rhizoma anemarrhenae 12g, Radix Glycyrrhizae 4g；

The preparation method is the same as that of Example 16.

18 Chinese medicine composition tablet of embodiment

Prescription: Radix Astragali 10g, ramulus cinnamomi 18g, flower bud of lily magnolia 4g, radix saposhnikoviae 14g, Rhizoma Atractylodis Macrocephalae 5g, rhizoma zingiberis 12g, cortex cinnamomi 2g, Radix Paeoniae Alba 12g, crow Plum 8g, Schisandra chinensis 12g, rhizoma anemarrhenae 4g, Radix Glycyrrhizae 12g；

The preparation method is the same as that of Example 16.

Embodiment 19

Prescription: Astragalus Root P.E 21g, Ramulus Cinnamomi extract 9g, Flos Magnoliae extract 6g, Radix Saposhnikoviae extract 7g, Rhizoma Atractylodis Macrocephalae extract 9g, Rhizoma Zingiberis extract 6g, cinnamomum cassia extract 3g, radix paeoniae alba extraction 6g, smoked plum extractive 12g, Schisandra chinens P.E 6g, rhizoma anemarrhenae mention Take object 6g, licorice 6g.

Embodiment 20

Prescription: Astragalus Root P.E 16g, Ramulus Cinnamomi extract 11g, Flos Magnoliae extract 5g, Radix Saposhnikoviae extract 8g, Rhizoma Atractylodis Macrocephalae extract 8g, Rhizoma Zingiberis extract 7g, cinnamomum cassia extract 2g, radix paeoniae alba extraction 7g, smoked plum extractive 11g, Schisandra chinens P.E 7g, rhizoma anemarrhenae mention Take object 5g, licorice 7g.

Embodiment 21

Prescription: Astragalus Root P.E 24g, Ramulus Cinnamomi extract 8g, Flos Magnoliae extract 7g, Radix Saposhnikoviae extract 6g, Rhizoma Atractylodis Macrocephalae extract 11g, Rhizoma Zingiberis extract 5g, cinnamomum cassia extract 4g, radix paeoniae alba extraction 5g, smoked plum extractive 14g, Schisandra chinens P.E 5g, rhizoma anemarrhenae Extract 7g, licorice 5g.

Embodiment 22

Prescription: Astragalus Root P.E 18g, Ramulus Cinnamomi extract 10g, Flos Magnoliae extract 5g, Radix Saposhnikoviae extract 9g, Rhizoma Atractylodis Macrocephalae extract 7g, Rhizoma Zingiberis extract 8g, cinnamomum cassia extract 3g, radix paeoniae alba extraction 5g, smoked plum extractive 12g, Schisandra chinens P.E 5g, rhizoma anemarrhenae mention Take object 8g, licorice 5g.

Embodiment 23

Prescription: Astragalus Root P.E 23g, Ramulus Cinnamomi extract 7g, Flos Magnoliae extract 8g, Radix Saposhnikoviae extract 6g, Rhizoma Atractylodis Macrocephalae extract 10g, Rhizoma Zingiberis extract 5g, cinnamomum cassia extract 3g, radix paeoniae alba extraction 8g, smoked plum extractive 10g, Schisandra chinens P.E 8g, rhizoma anemarrhenae Extract 5g, licorice 8g.

Ramulus Cinnamomi extract described in above embodiments 19-23, Rhizoma Zingiberis extract, cinnamomum cassia extract, Flos Magnoliae extract are respectively Ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, the flower bud of lily magnolia extractive of volatile oil；The Astragalus Root P.E, Radix Saposhnikoviae extract, Rhizoma Atractylodis Macrocephalae extract, Radix Paeoniae Alba mention Take object, smoked plum extractive, Schisandra chinens P.E, anemarrhena asphodefoides extract, licorice be respectively Radix Astragali, radix saposhnikoviae, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, The extract that dark plum, Schisandra chinensis, rhizoma anemarrhenae, Radix Glycyrrhizae prepare through aqueous extraction-alcohol precipitation technology, the alcohol precipitation concentration are 60%.

The experiment of 24 clinical effectiveness of embodiment

1 data and method

1.1 general information

Control group: it goes to a doctor in October, 2009 in July, 2010 in China-Japan Friendship Hospital, meet syndrome of qi deficiency (deficiency of both lung and spleen) AR patient cases data 20, the age (42.1 ± 12.3) year, wherein male 12, female 8.

Experimental group: it goes to a doctor in October, 2010 in October, 2014 in China-Japan Friendship Hospital, meet syndrome of qi deficiency (deficiency of both lung and spleen) AR patient cases data 100, the age (44.2 ± 14.1) year, wherein male 57, female 43.

1.2 diagnostic criteria

The diagnostic criteria of AR is referring to Chinese Journal of Otorhinolaryngology Head And Neck Surgery editorial board, Chinese Medical Association's otolaryngology-head and neck " diagnosis of allergic rhinitis and the treatment guidelines (2009) " that surgery branch nasology group is formulated are drafted.Chinese medical discrimination diagnostic criteria It combines and faces referring to National Standard of the People's Republic of China GB/T16751.2-1997 " tcm clinical practice diagnosis and treatment term syndrome part " Bed is studied and accumulates Consensus of experts triplicity formulation, specific manifestation are as follows: nasal obstruction caused by deficiency of both lung and spleen, the failure of lung key, sneeze frequency Make, runny nose is like water；Or with fatigue and weakness, or with cold, anemophobia, or with skin wheal or Na Gu be not fragrant or abdominal distention after meal, or Companion defecates, and uncomfortable, half congealed, discharge is powerless, or sleep not firm or dreaminess or dry or dry hardship, or white glutinous with cough, or with coughing up Phlegm；Tongue is fat to have an indentation greatly, and tongue fur is white or white micro- greasy or Bai Weihuang；Arteries and veins is thin and delicate or heavy thin.

It is included in standard

1. being diagnosed as AR and meeting the card type person of Chinese medicine syndrome of qi deficiency (deficiency of both lung and spleen)；2. the age between 18-70 years old, gender It is unlimited；3. the course of disease was at 1 year or more.

Exclusion criteria

1. gestation or breast feeding women；2. with other serious disease patients；3. merging mental disease or serious neurological official It can disease patient；4. subjective malaise symptoms person cannot be expressed；5. receiving to suck corticosteroid treatment simultaneously, or there is nose other are different The often person that may interfere with curative effect evaluation.

1.3 curative effect index

1.3.1 curative effect index

Doctor records patient symptom integral joint observation patient sign integral (the two integral is added, to treat total mark).

1.3.1.1 symptom integral standard: being shown in Table 1.

1 symptom integral standard of table

Classification is scored	Rhiocnesmus	Sneeze *	Runny nose * *	Nasal obstruction
					0 point	Nothing	Nothing	Nothing	Nothing
1 point	Interruption	3-5	< 4	Feel when conscious air-breathing
					2 points	Ant row sense, but can endure	6-10	5-9	Intermittent or interactivity
3 points	Ant row sense, unbearably	> 11	> 10	Almost whole day mouth breathing

Note: * 1 continuous sneeze number；* blows nose number daily.

1.3.1.2 somatic feature score standard: being shown in Table 2.

2 somatic feature score standard of table

1.3.2 secondary efficacy index

1.3.2.1 patient voluntarily assesses the visual analogue scale scale (visual analog scale, VAS) of its symptom Score: advising patient according to nearest 1 week Symptoms, mark the resulting score of the symptom on scale, detailed programs be rhiocnesmus, Nasal obstruction, sneeze, runny nose, eye symptom include that eye itches, is envious and swells, shed tears, ophthalmodynia, feel suffocated, wheeze, coughing, pressure in chest.

Slide calliper rule shape is made in VAS, and respectively there is the lateral scale of 10cm long on two sides, and slide calliper rule one side has smile and pain at the both ends of horizontal line Bitter two kinds of different cartoon expressions, slide calliper rule another side horizontal line one end mark 0 indicate asymptomatic；The other end is 10, indicates that symptom is extremely tight Weight；Sandwich digit indicates different degrees of.When questionnaire survey, slide calliper rule card expression one is facing patient by researcher, allow patient self Rhinitis and related symptoms severity, the shifting mark on mobile slide calliper rule are assessed, researcher records the corresponding numerical value in the slide calliper rule other side.Meter Calculate the sum of each symptom VAS (Multi-VAS) scoring.(Uni-VAS, total symptom VAS are that patient exists to the another record total symptom VAS of patient Its overall sense of discomfort of self-assessment is marked on the slide calliper rule of 0-10, patient's " your disease of your common sensation is generally inquired by researcher Sick distress level is how " and allow patient that the number on slide calliper rule is specified to assess).

1.3.2.2 the former total IgE measurement of serum anaphylaxis:

The former total IgE of pretherapy and post-treatment serum anaphylaxis is detected by China-Japan Friendship Hospital's division of respiratory disease control laboratory, < 60kU/L It is judged as negative；>=60kU/L is judged as positive.

1.4 curative effect determinate standard

Referring to the curative effect determinate standard of 2009 " diagnosis of allergic rhinitis and treatment guidelines (2009) ".According to pretherapy and post-treatment The summation scored, curative effect are evaluated by following equation: the preceding total mark of (total mark after total mark-treatment before treating)/treatment × 100%.>=51% is effective, 50%-21% be it is effective ,≤20% is invalid.

1.5 method

1.5.1 drug

Control drug: raw Radix Astragali 21g, ramulus cinnamomi 9g, windproof 6g, flower bud of lily magnolia 6g, rhizoma atractylodis macrocephalae 9g, rhizoma zingiberis 6g, Radix Paeoniae Alba 6g, licorice 6g；

Experimental drug: Radix Astragali 21g, ramulus cinnamomi 9g, flower bud of lily magnolia 6g, radix saposhnikoviae 7g, Rhizoma Atractylodis Macrocephalae 9g, rhizoma zingiberis 6g, cortex cinnamomi 3g, Radix Paeoniae Alba 6g, crow Plum 12g, Schisandra chinensis 6g, rhizoma anemarrhenae 6g, Radix Glycyrrhizae 6g.

1.5.2 instructions of taking

Daily 1 dose, 2 clothes of decocting point, 7d is 1 course for the treatment of, even served 2 courses for the treatment of, total 14d.

1.5.3 statistical method

It counted, analyzed using 17.0 pairs of SPSS every data, parameters are indicated with x- ± s, using pairing Wilcoxon signed rank sum test is that difference is statistically significant with P < 0.05.

2 experimental results

2.1 efficient results

It the results are shown in Table 1.

3 clinical efficacy result of table

	Sample size	Obvious effective rate	It is efficient
				Control group	20	4 (20%)	15 (75%)
Experimental group	100	76 (76%)	15 (15%)

The result shows that obvious effective rate 20% in the experiment of 20 people of control group, effective percentage 75%；Experimental group expands sample size, Prescription is optimized simultaneously, more obvious improvement and alleviation of the improvement prescription to patient clinical symptom, wherein 100 patients Middle obvious effective rate is 76%, total effective rate 91%.

The pretherapy and post-treatment symptom integral result of 2.2 experimental groups

It the results are shown in Table 4.

The pretherapy and post-treatment shape somatic feature score of 4 experimental group of table, 100 patients compares (x- ± s divides)

Project	Before treatment	After treatment
			Rhiocnesmus	2.58±0.50	1.20±0.55**
Sneeze	1.97±0.86	0.08±0.27**
			Runny nose	2.72±0.53	0.06±0.23**
Nasal obstruction	2.44±0.77	0.76±0.71**
			Sign score	2.04±0.27	0.89±0.72**

Note: compared with before treatment, * * P < 0.01.Following table is same.

The pretherapy and post-treatment VAS total mark of 2.3 experimental groups, total symptom VAS compare

It the results are shown in Table 5.Before VAS total mark and total symptom VAS are substantially less than treatment after treatment, difference is statistically significant (P < 0.01).

51 00 patients of table pretherapy and post-treatment VAS and the total IgE of anaphylactogen compare (x- ± s divides)

Project	Before treatment	After treatment
			VAS total mark	16.43±2.63	5.21±2.57**
Total symptom VAS	8.24±1.90	3.17±3.20**
			IgE	77.56±11.16	59.63±9.63*

The total IgE value of the pretherapy and post-treatment anaphylactogen of 2.4 experimental groups compares.

It is significantly reduced after the total IgE treatment of anaphylactogen, or even normal level (< 60kU/L) can be down to, difference has statistics meaning Adopted (P < 0.01).

Actual situation cold-heat jumble is presented in AR interpretation of the cause, onset and process of an illness essence, and Characteristics of Syndromes is related to the function of multiple internal organs based on deficiency of both lung and spleen It is deficient, so, only progress " using lung spleen as the internal organs integral syndrome differentiation of core ", with the treatment principle of " gentle pungent Jin Peiben ", AR could thoroughly be cured.This research is by illustrating Chinese medicine of the present invention to 100 AR patient itself cross-reference clinical research observations Composition treatment AR clinical efficacy is preferable, and before primary and secondary wants curative effect index to be superior to treatment after treatment, total effective rate reaches 91%； Organic conception and the syndrome differentiation of zang-fu viscera for sufficiently inheriting Chinese medicine are theoretical, reflect AR patient and lose by the viscera function of core of lung spleen It adjusts, the essential problem of physical function state entire lowering；It compensates for current Chinese patent drug mostly to set about from real example, is to dispel exterior syndrome It is main, righting is slightly taken into account, it is overall to the incomplete defect of human righteousness's solid protection.

25 pharmacodynamic study of embodiment

This research be based on the traditional chinese medicine composition of the invention effect and AR pathogenesis, establish cavy AR model caused by TDI, The passive skin mistake of AR model caused by egg protein, xylene-induced ear swelling in mice model, swollen hyperplasia of rat granuloma model, allotransplantation in rats Quick reaction model and mouse ear xenogenesis passive cutaneous anaphylaxis model, observation the traditional chinese medicine composition of the invention make the treatment of AR With to provide experimental basis for its further clinical application.

The influence of cavy allergic rhinitis caused by 1 couple of TDI

AR guinea pig model is established using 10%TDI olive oil solution nasal drip, after model success, after successive administration 14d, is seen It examines each group behaviouristics and scores, enzyme linked immunosorbent assay (ELISA) detects SERUM IgE, IFN-γ, IL-12, IL-4, IL- 6, nasal mucosa IL-6, TNF-α, IFN-γ detect in TNF-α and the content of schneiderian membrance HIS, immunohistochemistry staining method Content.Optical microphotograph microscopic observation nasal mucosa Pathologic changes and pathological score, and carry out eosinophil (EOS) meter Number.

1.1 materials and methods

1.1.1 material

1.1.1.1 animal and grouping

SPF grades of cavys, 70 (Beijing Vital River Experimental Animals Technology Co., Ltd.), animal credit number is SCXK (capital) 2012-0001,220~240g of weight, be randomly divided into normal group, model group, rhinitis group (0.41g/kg), chlorine thunder he Determine group (0.91mg/kg), Chinese medicine composition high dose group (3.04g/kg), middle dose group (1.52g/kg), low dose group (0.76g/kg), every group 10, male and female each 5.

1.1.1.2 drug and reagent

Loratadine tablet (Xi'an Yang Sen, batch number: 050218967)；

Rhinitis piece (Chinese medicines group, batch number: 14272)；

The traditional chinese medicine composition of the invention granule (is prepared, 1g is equivalent to 5.7577g crude drug amount, distilled water by 4 method of embodiment It is spare to be configured to 0.304g/mL)；

2,4- toluene di-isocyanate(TDI)s (TDI, sigma company, bw-d0007)；

Olive oil (Chinese medicines group, batch number: 20140725)；

Cavy IgE, IL-4, IL-6, IL-12, TNF-α, IFN-γ, HIS kit (Lan Ji Biotechnology Co., Ltd, Batch number: 20150724)；

Ethyl alcohol (Beijing chemical plant, batch number: 20140607)；

Formalin (Tianjin good fortune morning chemical reagent factory, batch number: 20141020)；

Dimethylbenzene (Tong Guang fine chemistry industry company, Beijing, batch number: 20140719)；

Breathe out Rui Shi haematoxylin dye liquor (Beijing Shiji Heli Biological Technology Co., Ltd., batch number: 20141203)；

Breathe out Rui Shi eosin stain (Beijing Shiji Heli Biological Technology Co., Ltd., product batch: 20141203)；

DAB (Dako, batch number: K5007)；

Anti-TNF alpha antibody (abcam, batch number: ab6671)；

IL-6Polyclonal Antibody (proteintech, batch number: 21865-1-AP)；

Interferon gamma Polyclonal Antibody (proteintech, batch number: 15365-1- AP)。

1.1.1.3 laboratory apparatus

Microscope (Japanese OLYMPUS company), image analysis software (MIAS)；

Adopt figure software Nikon NIS-Elements D3.2 (Japanese Nikon company)；

ASP300 takes off full-automatic multi-functional microplate reader: MULTISKAN MK3 (Thermo company)；

GL-21M high speed freezing centrifuge (Shanghai Lu Xiangyi centrifuge Instrument Ltd.)；

BX51 water dispenser (German LEICA company)；

EG1150H embedding machine (German LEICA company)；

RM2235 slicer (German LEICA company)；

H I1 210 spreads out piece machine (German LEICA company).

1.2 method

1.2.1 model foundation

Using 10%TDI olive oil solution to cavy bilateral nostril collunarium (every 5 μ L of side), one time a day, it is changed to after continuous 7d The next day 1 time.After the completion of daily collunarium, score according to following standards of grading: 1. rhiocnesmus: slight: dab nose several times, 1 point；Severe: It is more than to scratch nasal surface, 2 points.2. sneeze: 1~3 1 point, 4~10 2 points, 11 or more 3 points.3. runny nose: it flow to prenaris, 1 Point；Beyond prenaris, 2 points；Runny nose is had one's face covered with 3 points.The allergic conditions of each group cavy in 30min are observed, each symptom integral is superimposed, Total score is greater than 5 points of persons for modeling success.Normal group substitutes 10%TDI olive oil solution with olive oil, and method and steps is the same as cause Quick group.

1.2.2 grouping and administration

Modeling the 8th day, according to neurological deficit score as a result, being grouped gastric infusion again, successive administration 14d, one time a day, TDI The next day of being changed to collunarium with maintain allergic symptom until experiment terminate, continue to observe the symptoms after collunarium and score.Model group and normal Group gives the distilled water stomach-filling of equivalent.

1.2.3 Testing index and method

1.2.3.1 animal behavioral study

The allergic conditions of each group cavy, and the record that scores are observed after administration 14d.

1.2.3.2 biochemical and amynologic index detection

After last excitation for 24 hours, 10% chloraldurate (300mg/kg) intraperitoneal injection of anesthesia, abdominal aortic blood, 3500r/ Min is centrifuged 10min.Serum is separated, is placed under -80 DEG C of refrigerators and saves.Serum is detected according to ELISA kit specification step IgE, IFN-γ, IL-4, IL-6, IL-12, TNF-α are horizontal.Take cavy schneiderian membrance physiological saline be homogenized, ELISA method detection its HIS content.

1.2.3.3 nasal mucosa pathological observation

5 cavys of every group of selection, open rapidly bridge of the nose, peel off upper jaw osseous part skin, the vestibular wall on the outside of the nasal septum of right side It cuts, opens exposed nasal septum and bilateral nasal cavity upwards, take bilateral mucous membrane of nasal septum immediately later, be fixed on 10% neutral formalin It is fixed in solution, paraffin embedding, slice, HE dyeing, observation schneiderian membrance pathological change, and carry out EOS counting.

HE staining procedure: the fixed sample of 10% neutral formalin, conventional alcohol dehydration, dimethylbenzene processing, stone Wax embedding, 3 μm of serial section are dried spare.Dimethylbenzene I, II dewaxing 15min.Dehydrated alcohol I, dehydrated alcohol II, 95% second Each 2min of aquation, flowing water rinse step by step for alcohol, 80% ethyl alcohol, and haematoxylin dyeing 10min washes 3min, color separation liquid 10s, washing 3 Secondary, 0.25% returns blue liquid 40s, washes 3 times, 1% eosin stains 1min, washes 3 times.95% ethyl alcohol I, 95% ethyl alcohol II take off step by step Water 1min, dehydrated alcohol I, II are dehydrated 2min, dimethylbenzene I2min, the transparent 5min of dimethylbenzene II, III, neutral gum envelope step by step Gu.

Pathological score standard: 0 grade, cilium is completely neat, and columnar epithelium is normal, a small amount of goblet cell, remembers 0 point；I grades, cup Shape cytosis, the inflammatory cell that minority is dispersed in, the glandular hyperplasia of film lamina propria accounts for the 1/3 of mucosa lamina propria hereinafter, 1 point of note； II grades, goblet cell gathers, and cilium lodging is irregular, accidental to fall off, and more acidophic cell is swum out of, and the body of gland of film lamina propria increases It is raw to account for the 1/3~2/3 of mucosa lamina propria, remember 2 points；III level, goblet cell gather, and cilium largely falls off, epithelium damage layer or group Texture disorder is knitted, a large amount of acidophic cells, which are swum out of, accounts for 2/3 or more of whole epithelial inflammation cells, and the glandular hyperplasia of film lamina propria accounts for 2/3 or more of mucosa lamina propria remembers 3 points.

1.2.3.4 immunohistochemical staining

Immunohistochemical staining (IHC) method detects nasal mucosa IFN-γ, IL-6, TNF-α expression.

Operating procedure: 2 μm of schneiderian membrance paraffin section are used to water with distillation washing 2 times through conventional dewaxing (dyeing with HE) PBS (phosphate buffer) is rinsed 1 time, each 5min, pressure cooker antigen retrieval, and PBS is rinsed 2~3 times, each 5min；3% H2O2 is stored at room temperature 10min；It is rinsed 2~3 times with PBS, each 5min.37 DEG C of primary antibody incubation 2h, PBS rinsing 3 times is added dropwise, every time 5min；Secondary antibody is added dropwise and is incubated at room temperature 50min, DAB (diaminobenzidine) 3~5min of colour developing grasps dyeing journey under the microscope Degree, tap water rinse, haematoxylin redyeing 2min, and hydrochloride alcohol differentiation, dehydration, dimethylbenzene is transparent, neutral gum mounting.

Every picture randomly selects 5 visuals field under high power lens, and brown particle is positive mark in endochylema, using Image- Pro Plus Version 6.0 analyzes software, analyzes image area according to formula (IOD/area) and it is close to acquire positive expression light Angle value.

1.2.4 statistical method

All data are with mean ± standard deviationIt indicates, handled with 9.3 statistical software of SAS, data obey normal state Distribution uses variance analysis, compares examined using t two-by-two, and data are disobeyed normal distribution or heterogeneity of variance and examined using nonparametric It tests, wherein guinea-pig nasal non-cancer lesion degree is analyzed using Ridit method, is that difference has statistics meaning with P < 0.05 or P < 0.01 Justice.

1.3 experimental result

1.3.1 on the ethological influence of AR cavy

Occur rhiocnesmus, sneeze symptom after sensitized guinea pig 1d collunarium, in 30min, performance degree weight is different, but without bright Aobvious runny nose symptom, since 4d, with the increase of collunarium number, there is clear nasal discharge in each group, and behaviouristics symptom is also more and more brighter Aobvious, then without evident act symptom, hints model replicates successfully Normal group.It is administered since the 8th day, after 14d is administered, removes Outside low dose group, each administration group symptom compared with model group is substantially reduced (P < 0.01 or P < 0.05).It the results are shown in Table 6.

6 each group cavy nose behaviouristics symptom score of table

Note: compared with normal group, ▲ P < 0.05, ▲ ▲ P < 0.01；Compared with model group, * P < 0.05, * * P < 0.01。

1.3.2 to the influence of AR guinea pig serum biochemical immunity index

1.3.2.1 to the influence of AR guinea pig serum IgE

Compared with normal group, model group serum IgE level is significantly raised (P < 0.05).Compared with model group, each administration group Serum IgE level is substantially reduced (P < 0.01).It the results are shown in Table 7.

The variation of 7 ELISA method detection SERUM IgE of table

1.3.2.2 influence of the traditional chinese medicine composition of the invention to AR guinea pig serum IFN-γ

Compared with normal group, model group guinea pig serum IFN-γ level is remarkably decreased (P < 0.05).Compared with model group, High, middle dose group serum I FN- γ level has significant raising (P < 0.05 or 0.01), the results are shown in Table 8.

8 ELISA method detection serum I FN- γ of table variation

Group	Dosage (g/kg)	IFN-γ(pg/mL)
			Normal group	-	119.03±41.32
Model group	-	82.99±21.87^▲
			Loratadine group	0.00091	116.02±41.35^*
Rhinitis group	0.405	123.79±22.87^**
			Chinese medicine composition high dose group	3.04	107.80±13.36^**
Chinese medicine composition middle dose group	1.52	107.93±22.03^*
			Chinese medicine composition low dose group	0.76	89.04±30.11

Note: compared with normal group, ▲ P < 0.05, ▲ ▲ P < 0.01；Compared with model group, * P < 0.05, * * P < 0.01.

1.3.2.3 influence of the traditional chinese medicine composition of the invention to AR guinea pig serum IL-12

Compared with normal group, model group guinea pig serum IL-12 level is remarkably decreased (P < 0.05).Compared with model group, this The horizontal significant raising (P < 0.01) of invention Chinese medicine composition high dose group guinea pig serum IL-12, the results are shown in Table 9.

The variation of 9 ELISA method detection serum IL -12 of table

Group	Dosage (g/kg)	IL-12(pg/mL)
			Normal group	-	120.85±45.36
Model group	-	77.32±23.15^▲
			Loratadine group	0.00091	123.37±47.77^*
Rhinitis group	0.405	123.97±25.43^**
			Chinese medicine composition high dose group	3.04	111.25±19.09^**
Chinese medicine composition middle dose group	1.52	99.83±32.28
			Chinese medicine composition low dose group	0.76	86.00±34.71

1.3.2.4 to the influence of AR guinea pig serum IL-4

Compared with Normal group, model group guinea pig serum IL-4 level has raising trend, but without significant difference (P > 0.05).Compared with model group, each dosage group levels of IL-4 is substantially reduced (P < 0.01 or P < 0.05).It the results are shown in Table 10.

The variation of 10 ELISA method detection serum IL -4 of table

Group	Dosage (g/kg)	n	IL-4(pg/mL)
				Normal group	_	9	673.71±74.31
Model group	_	10	683.91±77.51
				Loratadine group	0.00091	10	631.83±85.98
Rhinitis group	0.405	10	603.26±82.55^*
				Chinese medicine composition high dose group	3.04	10	502.14±1116.91^**
Chinese medicine composition middle dose group	1.52	10	555.37±121.31^*
				Chinese medicine composition low dose group	0.76	10	507.89±90.96^**

1.3.2.5 to the influence of AR guinea pig serum IL-6

Compared with normal group, the horizontal significant raising (P < 0.01) of model group guinea pig serum IL-6.Compared with model group, respectively Administration group Serum IL-6 levels are substantially reduced (P < 0.01 or P < 0.05).It the results are shown in Table 11.

The variation of 11 ELISA method detection blood serum IL-6 of table

Group	Dosage (g/kg)	n	IL-6(pg/mL)
				Normal group	-	9	326.10±96.20
Model group	-	10	449.09±57.58^▲▲
				Loratadine group	0.00091	10	342.84±52.10^**
Rhinitis group	0.405	10	336.00±103.38^**
				Chinese medicine composition high dose group	3.04	10	350.48±70.70^**
Chinese medicine composition middle dose group	1.52	10	364.32±94.72^*
				Chinese medicine composition low dose group	0.76	10	292.24±54.60^**

1.3.2.6 to the influence of AR guinea pig serum TNF-α

Compared with normal group, model group serum TNF-cc level is significantly raised (P < 0.01).Compared with model group, the present invention Each administration group serum TNF-cc level of Chinese medicine composition is substantially reduced (P < 0.01 or P < 0.05).It the results are shown in Table 12.

The variation of 12 ELISA method detection Serum TNF-α of table

Group	Dosage (g/kg)	n	TNF-α(pg/mL)
				Normal group	_	9	281.20±71.23
Model group	_	10	364.32±70.09^▲
				Loratadine group	0.00091	10	319.42±83.85
Rhinitis group	0.405	10	302.96±67.95
				Chinese medicine composition high dose group	3.04	10	302.99±61.55^*
Chinese medicine composition middle dose group	1.52	10	277.03±83.57^**
				Chinese medicine composition low dose group	0.76	10	266.06±42.30^**

1.3.2.7 to the influence of AR cavy schneiderian membrance HIS

Compared with normal group, model group schneiderian membrance HIS level has raising trend, but difference is not significant (P > 0.05)；With Model group compares, and the traditional chinese medicine composition of the invention group schneiderian membrance HIS level has certain decreasing trend, but unobvious (the P > of difference 0.05).It the results are shown in Table 13.

13 ELISA method detection schneiderian membrance HIS of table variation

Group	Dosage (g/kg)	Schneiderian membrance HIS (ng/mL)
			Normal group	_	5.23±1.42
Model group	_	7.03±1.98
			Loratadine group	0.00091	6.59±0.99
Rhinitis group	0.405	6.02±0.77
			Chinese medicine composition high dose group	3.04	5.74±1.06
Chinese medicine composition middle dose group	1.52	4.97±0.72
			Chinese medicine composition low dose group	0.76	5.97±2.03

1.3.3 to the influence of AR guinea-pig nasal Mucosal inflammatory index

1.3.3.1 to the influence of AR cavy schneiderian membrance IFN-γ expression

Light microscopic observation guinea-pig nasal mucosa-immune histochemical staining slice, the region for being dyed to brown granular is IFN-γ Positive expression region.The results show that there are a large amount of brown granulars between normal rats nasal epithelial cells, IFN-γ is prompted Largely it is present between normal nasal epithelial cells, and cavy schneiderian membrance IFN-γ positive expression amount is apparently higher than model group (P < 0.05)；Compared with model group, each administration group cavy schneiderian membrance IFN-γ expression quantity significantly increases (P < 0.05)；It the results are shown in Table 14。

The influence that 14 the traditional chinese medicine composition of the invention of table expresses AR cavy schneiderian membrance IFN-γ

Group	Dosage (g/kg)	OD value
			Normal group	-	0.156±0.080
Model group	-	0.046±0.041▲
			Loratadine group	0.00091	0.151±0.016**
Rhinitis group	0.405	0.198±0.080**
			Chinese medicine composition high dose group	3.040	0.113±0.043*
Chinese medicine composition middle dose group	1.520	0.159±0.048*
			Chinese medicine composition low dose group	0.760	0.115±0.029*

1.3.3.2 to the influence of AR cavy schneiderian membrance IL-6 expression

Light microscopic observation guinea-pig nasal mucosa-immune histochemical staining slice, the region for being dyed to brown granular is IL-6 sun Property expression region.Compared with normal group, model group cavy schneiderian membrance IL-6 OD value significantly increases (P < 0.05)；With model Group compares, and (P < 0.05, the results are shown in Table 15 for each administration group schneiderian membrance IL-6 positive expression OD value significant decrease.

The influence that 15 the traditional chinese medicine composition of the invention of table expresses AR cavy schneiderian membrance IL-6

Group	Dosage (g/kg)	OD value
			Normal group	-	0.048±0.034
Model group	-	0.149±0.064^▲
			Loratadine group	0.00091	0.067±0.027^*
Rhinitis group	0.405	0.020±0.008^**
			Chinese medicine composition high dose group	3.040	0.061±0.018^*
Chinese medicine composition middle dose group	1.520	0.063±0.029^*
			Chinese medicine composition low dose group	0.760	0.050±0.014^*

Note: compared with normal group, ▲ P < 0.05, ▲ ▲ P < 0.01；Compared with model group, * P < 0.05, * * P < 0.01

1.3.3.3 to the influence of AR cavy schneiderian membrance TNF-α expression

Light microscopic observation guinea-pig nasal mucosa-immune histochemical staining slice, the region for being dyed to brown granular is TNF-α Positive expression region.Compared with normal group, the visible apparent positive expression product of model group cavy schneiderian membrance TNF-α, optical density Value is significant to increase (P < 0.05)；Compared with model group, each administration group schneiderian membrance TNF-α positive expression OD value is significantly reduced (P < 0.05) shows that the traditional chinese medicine composition of the invention can reduce positive expression of the TNF-α in schneiderian membrance, the results are shown in Table 16.

The influence that 16 the traditional chinese medicine composition of the invention of table expresses AR cavy schneiderian membrance TNF-α

Group	Dosage (g/kg)	OD value
			Normal group	-	0.042±0.038
Model group	-	0.312±0.210^▲
			Loratadine group	0.00091	0.032±0.029^*
Rhinitis group	0.405	0.033±0.022^*
			Chinese medicine composition high dose group	3.040	0.027±0.019^*
Chinese medicine composition middle dose group	1.520	0.210±0.099^*
			Chinese medicine composition low dose group	0.760	0.041±0.038^*

1.3.4 on the pathological influence of AR cavy nasal mucosa

1.3.4.1 schneiderian membrance EOS is counted

Every slice randomly selects 5 high power fields (× 400), carries out EOS counting.Compared with normal group, model group EOS Infiltrate obvious (P < 0.01)；Compared with model group, in addition to the traditional chinese medicine composition of the invention low dose group, other each groups EOS infiltrates feelings Condition mitigates (P < 0.01 or P < 0.05).Show that the traditional chinese medicine composition of the invention can reduce AR cavy schneiderian membrance EOS infiltration, subtracts Light schneiderian membrance inflammatory reaction, the results are shown in Table 17.

17 schneiderian membrance EOS weighted mean value of table

Group	Dosage (g/kg)	EOS counts (a)
			Normal group	-	2.28±1.28
Model group	-	17.04±7.20^▲▲
			Loratadine group	0.00091	2.32±0.84^**
Rhinitis group	0.405	3.64±2.17^**
			Chinese medicine composition high dose group	3.04	5.48±2.30^**
Chinese medicine composition middle dose group	1.52	7.12±4.39^*
			Chinese medicine composition low dose group	0.76	9.80±5.24

1.3.4.2 schneiderian membrance pathological observation and scoring

Normal group nasal mucosa is thin by pseudostratified ciliated columnar epithelium cell, goblet cell, sertoli cell and substrate The composition such as born of the same parents, the dense fibrous connective tissue of submucosa is very thin, mucosal epithelium structural integrity, marshalling, cilium thickness one It causes, has no obvious inflammatory cell infiltration；Model group nasal mucosa epithelium is imperfect, there is obscission, and glandular hyperplasia is obvious, sticks Film lamina propria is shown in a large amount of EOS；Loratadine group mucosal epithelium is substantially intact, a small amount of inflammatory cell infiltration, on rhinitis group mucous membrane Skin portion falls off, and part glandular hyperplasia has no inflammatory cell infiltration；The traditional chinese medicine composition of the invention high dose group mucosal epithelium is completeer It is good, it is seen that part inflammatory cell infiltration；Middle dose group mucous membrane is substantially intact, there is a small amount of inflammatory cell infiltration；Low dose group mucous membrane Partial exfoliation, glandular hyperplasia is obvious, and inflammatory cell infiltration is more apparent.

Compared with normal group, model group pathological score is significantly raised (P < 0.01).Compared with model group, Loratadine group, The scoring of rhinitis group is substantially reduced (P < 0.05), and after the traditional chinese medicine composition of the invention treats 14d, high dose group schneiderian membrance pathology is commented Divide and is substantially reduced (P < 0.05).It the results are shown in Table 18.

The scoring of 18 cavy schneiderian membrance pathological grading of table

The influence of allergic rhinitis rat caused by 2 pairs of egg proteins

AR rat model is prepared using egg protein injection and schneiderian membrance stimulus method, after model is successfully established, observes each group row To learn and scoring, ELISA method detection SERUM IgE, IFN-γ, IL-4, IL-6, TNF-α, the content of IL-17, immuning tissue Chemical dyeing method detects nasal mucosa IL-6, TNF-α, IFN-γ expression, carries out nasal secretion EOS meter under optical microscopy Number observes nasal mucosa Pathologic changes and scores.

2.1 materials and methods

2.1.1 material

2.1.1.1 animal and grouping

SPF grades SD rat 70, half male and half female, 200~300g of weight is purchased from Beijing dimension tonneau China experimental animal technology Co., Ltd, animal credit number: SCXK (capital) 2012-0001.Animal is divided by normal group, model group, chlorine using randomized Lei Tading group (1.17mg/kg), rhinitis group (0.6g/kg), the traditional chinese medicine composition of the invention high dose group (4.6g/kg), this hair Bright Chinese medicine composition middle dose group (2.3g/kg), the traditional chinese medicine composition of the invention low dose group (1.15g/kg), every group 10.

2.1.1.2 drug and reagent

Egg protein (Ovalbumin, OVA, lot number: SLBK6455V, sigma company, the U.S.)；

The traditional chinese medicine composition of the invention granule (is prepared into, 1g is equivalent to 5.7577g crude drug amount, distillation by 4 method of embodiment It is spare that water is configured to 0.304g/mL)；

Loratadine tablet (Xi'an Yang Sen, batch number: 050218967)；

Rhinitis piece (Foshan Dezhong Pharmaceutical Co., Ltd., batch number: 14272)；

Gel aluminum hydroxide (Thermo company, batch number: pc 197962)；

Sodium chloride injection (Shijiazhuang Siyao Co., Ltd, batch number: 1511023204)；

Rat IgE ELISA kit (Xin Bosheng, batch number: R160322-117b)；

Rat IL-4ELISA kit (Xin Bosheng, batch number: R160415-002a)；

Rat IL-17A ELISA kit (Xin Bosheng, batch number: R160415-17a)；

Rat TNF-α ELISA kit (Xin Bosheng, batch number: R160415-102a)；

Rat IFN-γ ElISA kit (Xin Bosheng, batch number: R160415-101a)；

Rat IL-6 kit (CSB, batch number: D10033395)；

Anti-TNF alpha antibody (abcam, batch number: ab6671)；

IL-6Polyclonal Antibody (proteintech, batch number: 21865-1-AP)；

Interferon gamma Polyclonal Antibody (proteintech, batch number: 15365-1- AP):

Ethyl alcohol (Beijing chemical plant, batch number: 20140607)；

Breathe out Rui Shi eosin stain (Beijing Shiji Heli Biological Technology Co., Ltd., batch number: 20141203)；

DAB (Dako, batch number: K5007).

2.1.1.3 laboratory apparatus

Microscope (Japanese OLYMPUS company), image analysis software (MIAS)；

Adopt figure software Nikon NIS-Elements D 3.2 (Japanese Nikon company)；

BX51 water dispenser (German LEICA company)；

EG1150H embedding machine (German LEICA company)；

RM2235 slicer (German LEICA company)；

H I1 210 spreads out piece machine (German LEICA company).

2.2 method

2.2.1 model copy

2.2.1.1 basic sensitization

Experimental group: being dissolved in 1mL physiological saline for 1mg OVA and 30mg Al (OH) 3 Gel Adjuvant and be made into suspension, in 0.2mL OVA adjuvant suspension is injected in rat two sides groin, toes behind two sides, abdominal cavity by 1d, 5d, 10d respectively. Normal group substitutes sensitization adjuvant, the same experimental group of method and steps with same amount of normal saline.

2.2.1.2 local sensitization

Except for the normal group, in 15d, with 10%OVA normal saline solution bilateral collunarium, every side 50uL, collunarium 7d.Just Normal group replaces 10%OVA suspension, the same experimental group of method and steps with same amount of normal saline.

2.2.2 administration

After model success, 1 collunarium maintains the stimulation to schneiderian membrance the next day model group, and administration group 1h before daily collunarium is filled Stomach administration.14d is administered in continuous gavage, and every 7d observation is primary, records symptom and scores.

2.2.3 observation index

2.2.3.1 animal behavioral study

Sniffle is observed after local excitation in 30min, calculating behaviouristics total score by standards of grading, (being greater than 5 points is animal Model success).Standards of grading: 1. 0 point: no sneeze scratches nose and nasal discharge；2. 1 point: sneeze < 4, slightly scratching nose, prenasal There is nasal discharge in hole；3. 2 points: sneeze 4~10, frequently scratching nose, nasal discharge is more than prenaris；4. 3 points: sneeze > 11, Nose, friction are persistently scratched, face is covered with nasal discharge.

2.2.3.2 biochemical and immune index detection

After last excitation for 24 hours, 10% chloraldurate (350mg/kg) intraperitoneal injection of anesthesia, abdominal aortic blood, 3500r/ Min is centrifuged 10min.Serum is separated, is placed under -80 DEG C of refrigerators and saves.According to ELISA kit specification step detect IgE, IFN-γ, IL-4, IL-6, TNF-α, IL-17 are horizontal.

2.2.3.3 nasal secretion EOS is counted

Bridge of the nose is opened rapidly, peels off upper jaw osseous part skin, and vestibular wall is cut on the outside of the nasal septum of right side, opens exposure upwards Nasal septum and bilateral nasal cavity, first dip secretion with cotton swab, are applied on glass slide in the same direction, and row HE dyes (every group of selection 5 Only), (400) randomly select 5 visuals field (size is 677.3mm × 508mm) under optical microscopy, carry out EOS counting.

2.2.3.4 observing nasal mucosa Pathologic changes

Rats with bilateral mucous membrane of nasal septum is taken, is fixed in 10% neutral formalin solution, paraffin embedding, HE dyeing observation nose is glutinous Film pathological change simultaneously scores (the same first part of operating procedure).

2.2.3.5 immunohistochemical staining

Immunohistochemical staining (IHC) method detects nasal mucosa IFN-γ, IL-6, TNF-α expression.Every figure under high power lens Piece randomly selects 5 visuals field, analyzes software using Image-Pro Plus Version 6.0, image area is analyzed, according to public affairs Formula (IOD/area), acquires positive expression OD value.The same first part of operating procedure.

2.2.4 statistical analysis

Experimental data is with mean ± standard deviationIt indicates, is handled using 9.3 statistical software of SAS.If each group is obeyed Normal distribution and variance is neat, using one-way analysis of variance, two groups are compared and are examined using t；If each group disobey normal distribution or Heterogeneity of variance, using non-parametric test.

2.3 experimental result

2.3.1 influence of the traditional chinese medicine composition of the invention to AR rat behavior

Before administration, experimental group rat is compared with normal group, and sneeze, scratching nose, runny nose etc., behaviouristics symptom score obviously increases (P < 0.01), hints model replicates successfully.After drug treatment 14d, compared with normal group, neurological deficit score obviously increases model group Add (P < 0.01)；For each dosage group of the traditional chinese medicine composition of the invention compared with model group, neurological deficit score is substantially reduced (P < 0.01), Prompt the traditional chinese medicine composition of the invention can obviously improve AR rat sniffle.It the results are shown in Table 19.

Neurological deficit score record sheet after table 19 is administered

2.3.2 influence of the traditional chinese medicine composition of the invention to AR rat biochemistry and amynologic index

2.3.2.1 influence of the traditional chinese medicine composition of the invention to AR rat blood serum IgE

After drug treatment 14d, model group is compared with normal group, the horizontal obvious rising (p < 0.01) of rat blood serum IgE；This Compared with model group, rat blood serum IgE level is substantially reduced (p < 0.01 or 0.05) invention Chinese medicine composition high dose group, knot Fruit is shown in Table 20.

The variation of 20 ELISA method detection SERUM IgE of table

2.3.2.2 influence of the traditional chinese medicine composition of the invention to AR rat blood serum IFN-γ

After drug treatment 14d, for model group compared with normal group, rat blood serum IFN-γ level is decreased obviously (P < 0.01)； The traditional chinese medicine composition of the invention high and low dose group compared with model group, rat blood serum IFN-γ it is horizontal it is significantly raised (P < 0.01 or 0.05) 21, be the results are shown in Table.

21 ELISA method detection serum I FN- γ of table variation

Group	Dosage (g/kg)	n	IFN-γ(pg/mL)
				Normal group	-	10	12.99±5.79
Model group	-	10	7.92±1.53^▲▲
				Loratadine group	0.00091	9	9.35±0.76^**
Rhinitis group	0.405	10	11.06±3.63^*
				Chinese medicine composition high dose group	3.04	9	10.404±2.05^**
Chinese medicine composition middle dose group	1.52	10	8.65±1.50
				Chinese medicine composition low dose group	0.76	10	10.884±2.24^**

2.3.2.3 influence of the traditional chinese medicine composition of the invention to AR rat blood serum IL-4

After drug treatment 14d, model group is compared with normal group, the horizontal obvious rising (P < 0.01) of rat blood serum IL-4；This For invention Chinese medicine composition middle dose group compared with model group, rat blood serum IL-4 level is decreased obviously (P < 0.05), the results are shown in Table 22。

The variation of 22 ELISA method detection serum IL -4 of table

Group	Dosage (g/kg)	n	IL-4(pg/mL)
				Normal group	_	10	5.33±1.97
Model group	_	10	10.33±5.78^▲▲
				Loratadine group	0.00091	10	5.86±2.10^**
Rhinitis group	0.405	10	8.00±2.30
				Chinese medicine composition high dose group	3.04	10	6.74±1.49
Chinese medicine composition middle dose group	1.52	10	5.91±1.97^*
				Chinese medicine composition low dose group	0.76	10	9.60±4.42

2.3.2.4 influence of the traditional chinese medicine composition of the invention to AR rat blood serum IL-6

After drug treatment 14d, model group is compared with normal group, and rat blood serum IL-6 level is in rising trend, but without significant Sex differernce (P > 0.05)；Each dosage group of the traditional chinese medicine composition of the invention is compared with model group, and rat blood serum IL-6 level is without conspicuousness Change (P > 0.05), the results are shown in Table 23.

The variation of 23 ELISA method detection blood serum IL-6 of table

Group	Dosage (g/kg)	n	IL-6(pg/mL)
				Normal group	_	10	17.78±3.94
Model group	_	10	21.15±6.21
				Loratadine group	0.00091	10	18.91±2.88
Rhinitis group	0.405	10	20.58±7.87
				Chinese medicine composition high dose group	3.04	10	20.59±4.47
Chinese medicine composition middle dose group	1.52	10	22.63±6.58
				Chinese medicine composition low dose group	0.76	10	25.22±8.18

2.3.2.5 influence of the traditional chinese medicine composition of the invention to AR rat blood serum TNF-α

After drug treatment 14d, model group is compared with normal group, and ascendant trend is presented in rat blood serum TNF-α level, but without aobvious It writes difference (P > 0.05)；Compared with model group, rat blood serum TNF-α is horizontal obvious for the traditional chinese medicine composition of the invention height, middle dose group Decline (P < 0.05 or 0.01), the results are shown in Table 24.

The variation of 24 ELISA method detection Serum TNF-α of table

Group	Dosage (g/kg)	n	TNF-α(pg/mL)
				Normal group	_	10	74.80±14.13
Model group	_	10	86.68±9.64
				Loratadine group	0.00091	9	76.49±9.28^*
Rhinitis group	0.405	10	65.49±16.54^**
				Chinese medicine composition high dose group	3.04	9	74.29±12.92^*
Chinese medicine composition middle dose group	1.52	10	72.25±9.26^**
				Chinese medicine composition low dose group	0.76	10	77.18±17.32

2.3.2.6 influence of the traditional chinese medicine composition of the invention to AR rat blood serum IL-17

After drug treatment 14d, model group is compared with normal group, the horizontal obvious rising (P < 0.01) of rat blood serum IL-17； The traditional chinese medicine composition of the invention low dose group is compared with model group, and rat blood serum IL-17 level is decreased obviously (P < 0.05), as a result It is shown in Table 25.

The variation of 25 ELISA method detection serum IL -17 of table

Group	Dosage (g/kg)	n	IL-17(pg/mL)
				Normal group	_	10	8.45±3.17
Model group	_	10	14.91±7.58^▲
				Loratadine group	0.00091	9	9.03±3.55^*
Rhinitis group	0.405	10	11.76±2.55
				Chinese medicine composition high dose group	3.04	9	14.33±4.89
Chinese medicine composition middle dose group	1.52	10	11.29±3.16
				Chinese medicine composition low dose group	0.76	10	9.204±3.88^*

2.3.3 the influence that the traditional chinese medicine composition of the invention expresses AR rat schneiderian membrance IFN-γ, IL-6, TNF-α

2.3.3.1 the influence that the traditional chinese medicine composition of the invention expresses AR rat schneiderian membrance IFN-γ

Light microscopic observation rat schneiderian membrance immunohistochemical staining slice, the results show that compared with normal group, model group rats Schneiderian membrance IFN-γ positive expression OD value is substantially reduced (P < 0.05)；Compared with model group, Chinese medicine high and low dose group is big Mouse schneiderian membrance OD value significantly increases (P < 0.05 or 0.01)；It the results are shown in Table 26.

The influence that 26 the traditional chinese medicine composition of the invention of table expresses AR rat schneiderian membrance IFN-γ

Group	Dosage (g/kg)	OD value
			Normal group	-	0.126±0.058
Model group	-	0.044±031^▲
			Loratadine group	0.0017	0.056±0.02^*
Rhinitis group	0.60	0.168±0.014^**
			Chinese medicine composition high dose group	4.60	0.161±0.026^**
Chinese medicine composition middle dose group	2.30	0.128±0.062
			Chinese medicine composition low dose group	1.15	0.160±0.028^**

2.3.3.2 the influence that the traditional chinese medicine composition of the invention expresses AR rat schneiderian membrance IL-6

Light microscopic observation rat schneiderian membrance immunohistochemical staining slice, the results show that compared with normal group, model group rats Schneiderian membrance IL-6 positive expression OD value significantly increases (P < 0.01)；Compared with model group, Chinese medicine is high, middle dose group nose is glutinous Film IL-6 OD value significantly reduces (P < 0.01), the results are shown in Table 27.

The influence that 27 the traditional chinese medicine composition of the invention of table expresses AR rat schneiderian membrance IL-6

Group	Dosage (g/kg)	Average optical density value
			Normal group	-	0.052±0.031
Model group	-	0.142±0.029^▲▲
			Loratadine group	0.0017	0.057±0.013^**
Rhinitis group	0.60	0.039±0.033^**
			Chinese medicine composition high dose group	4.60	0.050±0.014^**
Chinese medicine composition middle dose group	2.30	0.042±0.022^**
			Chinese medicine composition low dose group	1.15	0.086±0.042

2.3.3.3 the influence that the traditional chinese medicine composition of the invention expresses AR rat schneiderian membrance TNF-α

Light microscopic observation rat schneiderian membrance immunohistochemical staining slice, the results showed that, compared with normal group, model group rats Schneiderian membrance TNF-α positive expression OD value significantly increases (P < 0.01)；Compared with model group, Chinese medicine high dose group schneiderian membrance TNF-α positive expression OD value significantly reduces (P < 0.05 or 0.01), shows that the traditional chinese medicine composition of the invention can reduce TNF- Positive expression of the α in schneiderian membrance, the results are shown in Table 28.

The influence that 28 the traditional chinese medicine composition of the invention of table expresses AR rat schneiderian membrance TNF-α

Group	Dosage (g/kg)	OD value
			Normal group	-	0.013±0.009
Model group	-	0.084±0.029^▲▲
			Loratadine group	0.0017	0.010±0.014^**
Rhinitis group	0.60	0.027±0.021^**
			Chinese medicine composition high dose group	4.60	0.037±0.016^**
Chinese medicine composition middle dose group	2.30	0.051±0.023
			Chinese medicine composition low dose group	1.15	0.049±0.025

2.3.4 the traditional chinese medicine composition of the invention is on the pathological influence of AR rat nasal mucosa

2.3.4.1 influence of the traditional chinese medicine composition of the invention to AR rat nasal secretion EOS

After 14d is administered, for model group compared with normal group, nasal secretion EOS quantity is significantly raised (P < 0.01).This hair For bright each dosage group of Chinese medicine composition compared with model group, EOS quantity is substantially reduced (P < 0.05 or 0.01).Loratadine group, nose Scorching Kang Zuyu model group compares, and EOS infiltration is decreased obviously (P < 0.05 or 0.01), the results are shown in Table 29.

29 rat nasal secretion EOS of table is counted

Group	Dosage (g/kg)	EOS is counted
			Normal group	-	3.92±0.99
Model group	-	8.80±2.24▲▲
			Loratadine group	0.0017	4.88±1.56*
Rhinitis group	0.60	3.92±1.03**
			Chinese medicine composition high dose group	4.60	4.04±1.09**
Chinese medicine composition middle dose group	2.30	3.53±1.25**
			Chinese medicine composition low dose group	1.15	5.12±1.15*

2.3.4.2 the traditional chinese medicine composition of the invention is on the pathological influence of AR rat tissue

After HE dyeing it is in the form of a column arrangement, has no the congestion of blood vessel and tissue as it can be seen that normally group schneiderian membrance epithelium is intact under light microscopic Oedema, inflammatory cell infiltration, glandular hyperplasia；Mucosal epithelium damage in model group part falls off, and goblet cell is more, lamina propria blood vessel Dilatation and congestion, with more eosinophil and other inflammatory cell infiltrations, glandular hyperplasia under mucous membrane, compared with normal group, disease Reason scoring has significant difference (P < 0.01)；Loratadine group, rhinitis group mucosal epithelium are substantially intact, and a small amount of inflammation is thin Born of the same parents infiltrate, and body of gland has slight hyperplasia under mucous membrane, and compared with model group, pathological score is decreased obviously (P < 0.05 or 0.01)；This hair Bright each dosage group mucosal epithelium of Chinese medicine composition is substantially intact, partially visible glandular hyperplasia, blood vessel mild hyperaemia, a small amount of acidophilia Granulocyte infiltration, compared with model group, pathological score is decreased obviously (P < 0.05 or 0.01), the results are shown in Table 30.

The scoring of 30 rat schneiderian membrance pathological grading of table

Note: compared with normal group,^▲P < 0.05,^▲▲P < 0.01；Compared with model group,^*P < 0.05,^**P < 0.01

3 the traditional chinese medicine composition of the invention are anti-inflammatory, anti-allergic effects are studied

Method is formed using xylene-induced ear swelling in mice method and swollen hyperplasia of rat granuloma, studies the traditional chinese medicine composition of the invention Anti-inflammatory effect；This hair is observed using allotransplantation in rats passive cutaneous anaphylaxis method and mouse ear xenogenesis passive cutaneous anaphylaxis method The anti-allergic effects of bright Chinese medicine composition.

3.1 materials and methods

3.1.1 experimental material

3.1.1.1 animal

Selection SPF grades kunming mice 100, half male and half female, 18~22g of weight.SPF grades SD rat 110, male and female are each Half, 180~200g of weight.The above animal is purchased from Beijing Vital River Experimental Animals Technology Co., Ltd., animal credit number: SCXK (capital) 2012-0001.

3.1.1.2 drug and reagent

The traditional chinese medicine composition of the invention granule (is prepared, 1g is equivalent to 4.8804g crude drug amount) by 4 method of embodiment；

Loratadine (Xian-Janssen Pharmaceutical Ltd., lot number: 150923784)；

Dexamethasone acetate tablets (Tianjin Lisheng Pharmaceutical Co., Ltd., lot number: 1501006)；

Dimethylbenzene (Beijing Chemical Plant, lot number: 20150506)；

Dual anti-(CORNING, the suspension of mycillin, lot number: 30-002-CI)；

Egg protein (sigma company, the U.S., lot number: SLBK6455V)；

It adsorbs DPT vaccine (Changchun Changsheng Biological Technology Co., Ltd, lot number: 20150918-02)；

Evans blue (Sinopharm Chemical Reagent Co., Ltd., lot number: wc20070607)；

Acetone (Beijing Chemical Plant, lot number: 20150316)；

Potassium hydroxide powder (Beijing Chemical Plant, lot number: 20141118)；

Phosphoric acid (Sinopharm Chemical Reagent Co., Ltd., lot number: 20160127).

3.1.1.3 instrument

Labofuge refrigerated centrifuge (German Heraeus company)；

Punch (diameter 6.5mm)；

Electronic analytical balance (Ohaus company, the U.S.)；

WFJ2000 type ultraviolet-uisible spectrophotometer (Shanghai You Nike Instrument Ltd.)；

QL-901 vortex oscillator (its woods Bell's instrument manufacturing Co., Ltd, Haimen City)；

Electric heating constant-temperature blowing drying box (Hengfeng Medical Instruments Co., Ltd., Huangsih City)；

Vertical pressure steam sterilization pan (Medical Equipment Plant, Shanghai Boxun Industrial Co., Ltd.).

3.2 experimental method

3.2.1 xylene-induced ear swelling in mice is tested

Animal is divided into control group, Dexamethasone group (5mg/kg), the traditional chinese medicine composition of the invention high dose group at random (7.86g/kg), middle dose group (3.93g/kg), low dose group (1.96g/kg), every group 10, half male and half female.By above-mentioned grouping Mode, successive administration 5d, 1 times/day.After the 5th day administration 45min, 50 μ L caused by dimethylbenzene xylene are uniformly smeared on mouse right ear two sides Inflammation, after smearing 20min, cervical dislocation is lethal, and two ears of left and right, the left and right ear of same area is beaten with punch under auricle edge scissor Piece is weighed immediately after.Using the difference of two auricle weight as swelling, the inhibition percentage of drug is calculated.Inhibiting rate (%)= (control group swelling-medication group swelling)/control group swelling × 100%.Control group gives the distilled water of equivalent, method and The same experimental group of step.

3.2.2 swollen hyperplasia of rat granuloma is tested

Be randomly divided into control group, the traditional chinese medicine composition of the invention agent high dose group (4.6g/kg), middle dose group (2.3g/kg), Low dose group (1.15g/kg), Dexamethasone group (0.79mg/kg), every group 10, half male and half female.Rat chloraldurate An opening is made in (350mg/kg) anesthesia, the disinfection of 75% ethyl alcohol abdomen among abdomen, removes skin to groin with haemostatic clamp Place, at preparatory high pressure sterilization and 50mg cotton balls (dual anti-100 μ L is added) implantation rat groin, skin suture is clear to animal Start to be administered after waking up.In addition to distilled water is given in control group stomach-filling, relative medicine is given in other each administration group stomach-fillings, one time a day, 1mL/100g, continuous 7d.1h after the last administration, cervical dislocation put to death rat, open skin of abdomen, carefully remove cotton balls, be placed in electricity It weighs on sub- balance, as cotton balls weight in wet base.Cotton balls is placed in 60 DEG C of baking ovens, dry 12h takes out and weighs as dry weight.

3.2.3 models of passive skin irritability of rats reaction experiment

3.2.3.1 the preparation of rat antisera

SD rat male and female each 5, every 5% egg protein normal saline solution 0.2mL of mouse hind leg intramuscular injection are chosen in experiment, simultaneously DPT vaccine 0.2mL is injected intraperitoneally.After normal raising 13d, anesthesia, abdominal aortic blood, 3500r/min is centrifuged 15min, quiet After setting 2h, rat blood serum is separated, -20 DEG C of refrigerators is placed in and saves.Prepare anti-egg protein rat blood serum.

3.2.3.2 passive cutaneous anaphylaxis

SD rat is divided into control group, the traditional chinese medicine composition of the invention high dose group (4.6g/kg), middle dose group (2.3g/kg), Low dose group (1.15g/kg), Loratadine group (1.16mg/kg), every group 10, male and female each 5.By above-mentioned group technology, chlorine 7d is administered in Lei Tading group and Chinese medicine each group continuous gavage, one time a day, antigen attack is given after not secondary administration 45min.Control group fills Stomach distilled water.It is administered the 6th day, dorsal line two sides is cut off under the anesthesia of chloraldurate away from hair (every side at 1.5~2cm of backbone 3cm × 3cm), and by above-mentioned antiserum normal saline dilution at 1: 2 and 1: 6, intracutaneous injection is in rat spine two sides shaving portion Point, each dilution injects two o'clock, every 0.1mL.It is administered the 7th day, tail vein injection ovalbumin (2%) after last dose 1h With Evans blue (0.5%) physiological saline mixed solution 1mL.

3.2.3.3 collection of specimens and Indexs measure

Sacrificed by decapitation after 30min, rat dorsum skin is overturn, and is cut back locus coeruleus skin, is shredded, with 5mL acetone-life It manages salt water (7: 3) to impregnate, is placed in 37 DEG C of water-baths and heats 48h, be centrifuged, supernatant taken, with ultraviolet-uisible spectrophotometer in wave Long 620nm measures OD value.

3.2.4 mouse ear xenogenesis passive cutaneous anaphylaxis

Mouse is randomly divided into control group, the traditional chinese medicine composition of the invention agent high dose group (7.86g/kg), middle dose group (3.93g/kg), low dose group (1.96g/kg), Loratadine group (2mg/kg), every group 10, half male and half female.Continuous gavage is given Medicine 7d, administered volume 0.2mL/10g, give antigen attack after last stomach-filling 45min by 1 times/day.It is administered the 6th day, mouse two Auricle respectively injects 25 μ L of rat antisera.Antigen attack after for 24 hours, 2% ovalbumin of tail vein injection and 0.5% Evans blue are molten Liquid 0.25mL.After 30min, mouse cervical dislocation is put to death, cuts Mice Auricle (indigo plant dye position).Cut auricle is placed in test tube In, the potassium hydroxide solution (1mol/L) that 0.75mL is added impregnates, and 37 DEG C are digested overnight.Next day adds 3.75mL phosphoric acid (0.6mol/L) and acetone mixture (5: 13, v/v), shakes through vortex oscillator, then is centrifuged 15min with 2500r/min, will be upper Clear liquid takes out, and ultraviolet-uisible spectrophotometer measures OD value at wavelength 640nm, calculates the inhibiting rate of drug.Inhibiting rate (%) =(control group OD value-medication group OD value)/control group OD value × 100%.

3.2.5 statistical method

Experimental data usesIt indicates, is handled using 9.3 statistical software of SAS.If each group Normal Distribution and variance Qi Ze uses variance test, compares examined using LSD two-by-two, use non-parametric test if each group disobeys normal distribution.With P < 0.05 is that difference is statistically significant.

3.3 experimental result

3.3.1 the traditional chinese medicine composition of the invention paraxylene causes the influence of mouse ear swelling

Compared with the control group, Dexamethasone group and the high, medium and low dosage group of the traditional chinese medicine composition of the invention can significantly reduce diformazan Benzene causes mice auricle swelling (P < 0.05 or 0.01), shows that the traditional chinese medicine composition of the invention can inhibit dimethylbenzene induced mice auricle Inflammatory reaction.It the results are shown in Table 31.

The influence of 31 the traditional chinese medicine composition of the invention paraxylene of table cause mouse ear swelling

Note: compared with the control group, * P < 0.05, * * P < 0.01.

3.3.2 the influence that the traditional chinese medicine composition of the invention agent forms swollen hyperplasia of rat granuloma

Compared with the control group, Dexamethasone group can significantly lower granuloma induced by implantation of cotton pellets weight (P < 0.01), Chinese medicine group of the present invention Each dosage group rat cotton balls dry weight of object, weight in wet base are closed without significant difference (P > 0.05), prompts the traditional chinese medicine composition of the invention to cotton balls institute It causes rat granulation tissue hyperplasia without apparent inhibiting effect, the results are shown in Table 32.

Influence of 32 the traditional chinese medicine composition of the invention of table to swollen hyperplasia of rat granuloma

Group	Dosage	Cotton balls weight in wet base/mg	Cotton balls dry weight/mg
				Control group	-	1416.0±907.3	215.9±25.4
Dexamethasone group	0.79mg/kg	1128.0±575.2^**	173.1±12.7^**
				Chinese medicine composition high dose group	4.60g/kg	1363.8±177.1	212.8±29.7
Chinese medicine composition middle dose group	2.30g/kg	1395.6±794.4	216.5±20.3
				Chinese medicine composition low dose group	1.15g/kg	1398.7±103.3	212.9±16.2

Note: compared with the control group, * * p < 0.01.

3.3.3 influence of the traditional chinese medicine composition of the invention to allotransplantation in rats passive cutaneous anaphylaxis

Compared with the control group, Loratadine group, the traditional chinese medicine composition of the invention each group can obviously reduce Locus Coeruleus In The Rat OD value (P < 0.05 or 0.01) shows that the traditional chinese medicine composition of the invention has allotransplantation in rats passive cutaneous anaphylaxis caused by egg protein Certain inhibiting effect.It the results are shown in Table 33.

The influence that 33 the traditional chinese medicine composition of the invention of table reacts models of passive skin irritability of rats

Group	Dosage	OD	Inhibiting rate
				Control group	-	0.134±0.081	-
Loratadine group	1.16mg/kg	0.059±0.039^*	55.97%
				Chinese medicine composition high dose group	4.6g/kg	0.036±0.018^*	73.13%
Chinese medicine composition middle dose group	2.3g/kg	0.048±0.045^*	64.18%
				Chinese medicine composition low dose group	1.15g/kg	0.024±0.020^**	82.09%

Note: compared with the control group, * P < 0.05, * * P < 0.01.

3.3.4 influence of the traditional chinese medicine composition of the invention to mouse ear xenogenesis passive cutaneous anaphylaxis

Compared with the control group, Loratadine group can substantially reduced Mice Auricle locus coeruleus OD value (P < 0.01), Chinese medicine of the present invention Composition high dose group can significantly reduce Mice Auricle indigo plant dye degree (P < 0.05), prompt the traditional chinese medicine composition of the invention can be obvious Inhibit the xenogenesis passive cutaneous anaphylaxis of egg protein induced mice ear, the results are shown in Table 34.

Influence of 34 the traditional chinese medicine composition of the invention of table to mouse ear xenogenesis passive cutaneous anaphylaxis

Note: compared with the control group, * P < 0.05, * P < 0.01.

26 process for producing granula of embodiment is investigated

1 dry granulation

The big advantage of the one of dry granulation is to reduce the dosage of auxiliary material, and then reduce patient's dose, therefore take appropriate combination Object extract powder adds dextrin, lactose respectively, is uniformly mixed with 3: 1 ratio, carries out dry-pressing granulation with GZL20 type dry granulating machine, Dry granulation the results are shown in Table 35.

Composition extract powder is prepared as follows:

Eight taste such as remaining Radix Astragali, Radix Paeoniae Alba adds water to cook three times, 1.5 hours every time, collecting decoction, filtration, filtrate with it is above-mentioned Two liquid merge, and are concentrated into the clear cream that relative density is 1.15~1.20 (60 DEG C), add ethyl alcohol to make alcohol content up to 60%, stirring is quiet It sets overnight, filtrate recycling ethanol, filtrate is concentrated into right amount, is dried under reduced pressure, and fine powder is ground into, with above-mentioned betadex inclusion compound It mixes, obtains composition extract powder.

35 dry granulation experimental result of table

Excipient	Roll wheel speed (HZ)	Roll wheel pressure (MPa)	One-pass molding rate (%)
				Extract powder: dextrin	15	+4	44.85
Extract powder: lactose	15	+4	36.00

The experimental results showed that roll wheel pressure be 4MPa when, granule fines are more, and one-pass molding rate is lower, respectively less than 50, and Extract powder viscosity is stronger, occurs gluing in pelletization and rolls wheel phenomenon, pelletizes more difficult, be unable to meet production requirement, therefore use instead wet Method granulation.

2 wet granulations

The selection of 2.1 wetting agents

The common wetting agent of wet granulation is ethyl alcohol, therefore selects ethyl alcohol for wetting agent, and investigate different ethanol concentration to system The influence of grain situation.

2.2 supplementary product kinds are investigated

Extract powder and dextrin, lactose, soluble starch are pressed into 3: 1 mixings respectively, 95% alcohol granulation is sprayed into, with softwood Shape, the ratio of briquetting of particle, granulation complexity and particle color, fine powder, bonding situation etc. be evaluation index, the results are shown in Table 36。

36 supplementary product kind of table is investigated

Excipient	Softwood character	Granulation complexity	Ratio of briquetting (%)
				Extract powder: dextrin	Loosely, no agglomerate	Sieve can be passed through completely	60.08
Extract powder: lactose	Loosely, no agglomerate	Sieve can be passed through completely	47.20
				Extract powder: soluble starch	Loosely, no agglomerate	Sieve can be passed through completely	53.74

As can be seen from the results, three kinds of auxiliary materials can make grain forming, and granulation is easy, but when dextrin is added particle ratio of briquetting most Height, and dextrin is cheap compared to lactose and soluble starch, stability is good, good fluidity, therefore select dextrin for auxiliary material.

The dosage of 2.3 auxiliary materials and the investigation of wetting agent concentration of alcohol

Appropriate extract powder and dextrin are uniformly mixed in 3: 1,3: 2,1: 1 ratio respectively, spraying appropriate concentration respectively is 90%, 94%, 95% alcohol granulation, granulation situation the results are shown in Table 37.

37 granule moulding process optimization test result of table

As can be seen from the above results, extract powder is to wet more sensitive, and when to select 90% ethyl alcohol be wetting agent, agglomerate is more, Granulation is difficult；Selecting 95% ethyl alcohol is wetting agent, particle is made loosely, fine powder is more, and ratio of briquetting is lower；Select 94% ethyl alcohol When for wetting agent, it is easy granulation, grain graininess is uniform, and ratio of briquetting is higher.When the auxiliary ratio of medicine is 3: 1,3: 2, particle can be made, Therefore selecting hydroscopicity is index, further determines that the auxiliary ratio of best medicine.

The determination of the auxiliary ratio of 2.4 medicines

Using the hygroscopicity of particle as evaluation index, the auxiliary ratio of optimal medicine of particle is screened, extract powder and appropriate dextrin are weighed, It mixes according to 3: 1 and 3: 2, pelletizes using 94% ethyl alcohol as wetting agent respectively, measure its Moisture percentage, it is bent to draw moisture absorption Line.

The hygroscopicity that can be seen that particle after pelletizing from the hygroscopicity result of particle is improved compared to extract powder, Compare medicine it is auxiliary than be 3: 1 and 3: 2 particle, medicine it is auxiliary than 3: 1 made from particle hydroscopicity it is higher, and 3: 1 granulation more difficult, agglomerates More, when the auxiliary ratio of medicine is 3: 2, granulation is easy, and compared with medicine is auxiliary than 3: 1, dose difference is little, therefore the auxiliary ratio of final choice medicine is 3: 2,94% ethyl alcohol is wetting agent, and granulation is easy, and grain forming rate is high.

The thin layer of 27 Chinese medicinal composition granules of embodiment identifies

Identify A:

The preparation of test solution: Chinese medicinal composition granules 4g prepared by Example 4, it is finely ground, add methanol 60mL, surpasses Sonication 30 minutes, filtration, filtrate was set and is evaporated on water-bath, and residue adds water 50mL to dissolve, and is saturated with water n-butanol shaking and extracts 4 Secondary, each 50mL merges n-butanol liquid, is washed twice with ammonia solution, each 50mL, ammonia solution discards, and n-butanol liquid is evaporated, residual Slag adds methanol 5mL to make to dissolve, as test solution.

The preparation of control medicinal material solution: taking Radix Astragali control medicinal material 4g, and water 200mL is added to decoct 1 hour, filtration, and filtrate is appropriate It is concentrated (about 20mL), is saturated with water n-butanol shaking and extracts 4 times, each 20mL, merge n-butanol liquid, wash two with ammonia solution Secondary, each 20mL, ammonia solution discards, and n-butanol liquid is evaporated, and residue adds methanol 5mL to dissolve, as control medicinal material solution.

The preparation of negative control solution: in prescription ratio, autogamy is free of group's medicine of Radix Astragali, and negative sky is made by preparation process Negative control solution is made in white preparation, the preparation method with test solution.

Indentification by TLC: drawing above-mentioned each 10 μ L of three kinds of solution, is put respectively on same silica gel g thin-layer plate, with trichlorine Lower layer's solution of methane-methanol-water (13: 7: 2) is solvent, is unfolded, and takes out, dries, and is sprayed with 10% ethanol solution of sulfuric acid, In 105 DEG C to be heated to spot development clear, sets and inspects under ultraviolet lamp (365nm).

Test result: in sample chromatogram, on position corresponding with reference medicine chromatography, show identical brown under daylight Color spot point negative sample does not interfere with.

Identify B:

The preparation of test sample solution: Chinese medicinal composition granules 1g prepared by Example 4, it is finely ground, add methanol 20mL, ultrasound Processing 20 minutes, filtration, filtrate are set and are evaporated on water-bath, and residue adds methanol 1mL to make to dissolve, as test solution.

The preparation of reference substance solution: taking Paeoniflorin reference substance, adds methanol that solution of every 1mL containing 0.6mg is made, as control Product solution.

The preparation of negative control solution: in prescription ratio, autogamy is free of group's medicine of Radix Paeoniae Alba, and negative sky is made by preparation process Negative control solution is made in white preparation, the preparation method with test solution.

Indentification by TLC: draw 4 μ L of reference substance solution, each 10 μ L of test solution, negative control solution, put respectively in On same silica gel g thin-layer plate, with chloroform-acetate-methanol-formic acid (8: 2: 5: 0.2) for solvent, it is unfolded, takes out, It dries, sprays with 5% vanillin-sulfuric acid solution, it is clear to be heated to spot development at 105 DEG C.

Test result: in sample chromatogram, at the position corresponding to the chromatogram of the reference substance, the spot of same color is shown；Yin Property sample does not interfere with.

Identify C

The preparation of test solution: Chinese medicinal composition granules 4g prepared by Example 4, it is finely ground, add acetone 20mL, surpasses Sonication 20 minutes, filtration；Filtrate is set and is evaporated on water-bath, and residue adds methanol 1mL to make to dissolve, as test solution.

The preparation of reference substance solution: taking 5-O- methyl visamminol glycosides reference substance, adds methanol that every 1mL is made containing 0.5mg Solution, as reference substance solution.

The preparation of control medicinal material solution: taking windproof control medicinal material 2g, adds acetone 20mL, is ultrasonically treated 20 minutes, filters, filter Liquid is set and is evaporated on water-bath, and residue adds methanol 1mL to make to dissolve, as control medicinal material solution.

The preparation of negative control solution: in prescription ratio, negative sky is made by preparation process in group's medicine of the autogamy without radix saposhnikoviae Negative control solution is made in white preparation, the preparation method with test solution.

Indentification by TLC: 4 μ L of reference substance solution is drawn, control medicinal material solution, test solution, negative control solution are each 10 μ L put respectively on same silica GF254 lamellae, with chloroform-methanol (5:1) for solvent, are unfolded, take out, dry in the air It is dry, it sets and is inspected under ultraviolet lamp (254nm).

Test result: in sample chromatogram, on position corresponding with reference substance and reference medicine chromatography, same color is shown Spot；Negative sample does not interfere with.

Identify D

The preparation of test solution: Chinese medicinal composition granules 2g prepared by Example 4, it is finely ground, add chloroform 25mL is ultrasonically treated 30 minutes, and filtration, filtrate sets and be evaporated on water-bath, and the residue 1mL that adds methylene chloride makes to dissolve, as examination Product solution.

The preparation of reference substance solution: taking schizandrin reference substance, adds methanol that solution of every 1mL containing 0.5mg is made, as Reference substance solution.

The preparation of control medicinal material solution: taking Schisandra chinensis control medicinal material 1g, adds chloroform 25mL, is ultrasonically treated 30 minutes, Filtration, filtrate sets and is evaporated on water-bath, and the residue 1mL that adds methylene chloride makes to dissolve, as control medicinal material solution.

The preparation of negative control solution: in prescription ratio, autogamy is free of group's medicine of Schisandra chinensis, feminine gender is made by preparation process Negative control solution is made in blank formulation, the preparation method with test solution.

Indentification by TLC: drawing above-mentioned each 10 μ L of three kinds of solution, puts respectively on same silica GF254 lamellae, with Petroleum ether (30~60 DEG C)-Ethyl formate-formic acid (15: 6: 1) upper solution is solvent, is unfolded, and takes out, hangs ultraviolet lamp It is inspected under (254nm).

Identify E

The preparation of test solution: Chinese medicinal composition granules 3g prepared by Example 4, it is finely ground, add methanol 30mL, surpasses Sonication 30 minutes, filtration, filtrate was set and is evaporated on water-bath, and residue adds water 20mL to dissolve, and was extracted 2 times with ethyl acetate shaking, Each 20mL, combined ethyl acetate liquid, is evaporated, and residue adds methanol 10mL to make to dissolve, as test solution.

The preparation of control medicinal material solution: extracting liquorice control medicinal material 2g adds water 200mL to decoct 1 hour, filtration, and filtrate is appropriate It is concentrated (about 20mL), is extracted 2 times, each 20mL with ethyl acetate shaking, combined ethyl acetate liquid is evaporated, and residue adds methanol 10mL makes to dissolve, as control medicinal material solution.

The preparation of negative control solution: in prescription ratio, taking the extract of scarce Radix Glycyrrhizae to mix, negative blank formulation be made, Negative control solution is made in preparation method with test solution.

Indentification by TLC: 5 μ L of control medicinal material solution, test solution and each 10 μ L of negative control solution are drawn, respectively Point with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for solvent, is unfolded, takes out on same silica gel g thin-layer plate, It dries, sprays with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C.

Test result: in sample chromatogram, on position corresponding with reference medicine chromatography, the fluorescent spot of same color is shown Point；Negative sample does not interfere with.

The present invention is attempted using cinnaldehydrum as Index Establishment detection method, but ramulus cinnamomi and the principle active component of cortex cinnamomi are equal For cinnaldehydrum, specificity is not strong, therefore does not set up the detection method；Refer in addition, the present invention has also investigated camphor in the flower bud of lily magnolia and has been used as Ingredient is marked, but discovery has negative interference, specificity is not strong, due to the more difficult determination of index components or it is difficult to obtain reference substance, The present invention does not set up corresponding detection method；Ramulus cinnamomi, rhizoma zingiberis, cortex cinnamomi, four taste of the flower bud of lily magnolia, the spot of aobvious same color in test sample Point, the principle active component atractylone of Rhizoma Atractylodis Macrocephalae do not show spot, and corresponding other ingredients have negative interference, therefore above-mentioned a few taste medicines are not made For the index for detecting the traditional chinese medicine composition of the invention.

The assay of 28 Chinese medicinal composition granules of embodiment

The foundation of 1 Astragaloside IV chromatographic condition

The foundation of 1.1 chromatographic conditions

Chromatographic condition: Agilent Zorbax SB-C18 chromatographic column (4.6mm × 250mm, 5 μm), acetonitrile-water (31: 69) For mobile phase, flow velocity 1.0mLmin^-1, 30 DEG C of column temperature, ELSD parameter: 100 DEG C of drift tube temperature, carrier gas (compressed air) flow velocity 2.7L·min^-1, 10 μ L of sample volume.

The preparation and measurement of 1.2 solution

(1) preparation of reference substance solution

Take Astragaloside IV reference substance appropriate, it is accurately weighed, add methanol that the molten of every 1mL 0.30225mg containing Astragaloside IV is made Liquid to get.

(2) preparation of test solution

(2.1) investigation of extracting method

It was found that, being easy when index components Astragaloside IV is directly dissolved with water in test sample with when extracting n-butyl alcohol There is emulsion, and common breaking method cannot be such that emulsion layer eliminates well, present invention trial is first extracted with methanol To remove a part of impurity, then when with extracting n-butyl alcohol, i.e., without emulsion.

The influence that circumfluence method and ultrasonic method extract Astragaloside IV is investigated, concrete operations are as follows:

Circumfluence method: taking Chinese medicinal composition granules 4g, accurately weighed, sets in stuffed conical flask, adds methanol 60mL, cold soaking mistake Night refluxing extraction 1 hour, lets cool, and filtering, filtrate is concentrated to dryness, and residue adds water 40mL to make to dissolve, and is saturated with water n-butanol shaking It extracts 4 times, each 50mL, merges n-butanol liquid, washed 2 times, each 50mL with ammonia solution, n-butanol liquid is evaporated, and residue adds first Alcohol makes to dissolve, and is settled to 5mL volumetric flask, shake up to get.

Ultrasonic method: taking Chinese medicinal composition granules 4g, accurately weighed, sets in stuffed conical flask, adds methanol 60mL, ultrasound mentions 30 minutes, 45 minutes are taken, are let cool, are filtered, remaining operates same circumfluence method.

Each 10 μ L of above two test solution is taken respectively, is measured in accordance with the law, is calculated the content of Astragaloside IV, the results are shown in Table 38。

38 extracting method of table is investigated

Method	Astragaloside IV mg/g
		Reflux 1 hour	0.44
Ultrasound 30 minutes	0.39
		Ultrasound 45 minutes	0.34

As a result it is found that circumfluence method extract Astragaloside IV content be higher than ultrasonic method, and ultrasound 30 minutes when be higher than 45 minutes, The content of the prompt too long Astragaloside IV of ultrasonic time declines instead, therefore determines that the extracting method of Astragaloside IV is circumfluence method.

(2.2) investigation of extraction time

To extract the Astragaloside IV in Chinese medicinal composition granules to greatest extent, return time is further investigated to Radix Astragali Reflux 1 hour, 2 hours, 3 hours have been investigated in the influence of glucoside content, experiment, remaining operates same circumfluence method, it is molten to prepare test sample Liquid, 10 μ L of sample introduction, measures in accordance with the law, calculates the content of Astragaloside IV, the results are shown in Table 39.

39 extraction time of table is investigated

Extraction time (hour)	Astragaloside IV mg/g
		Reflux 1 hour	0.44
Reflux 1 hour	0.45
		Reflux 3 hours	0.47

As a result: refluxing extraction 3 hours, the content highest of Astragaloside IV determined the preparation side that refluxing extraction 3 hours are sample Method.

The content of Astragaloside IV, measurement result are aobvious in precipitating after also having investigated reflux, ultrasound in above-mentioned experimentation Showing may be due to the too low chromatographic peak for failing to detect Astragaloside IV of content in precipitating.

To sum up, determine test solution solution the preparation method is as follows: takes this product, finely ground, takes about 4g, accurately weighed, sets In stuffed conical flask, add methanol 60mL, cold soaking is overnight, refluxing extraction 3 hours, lets cool, and filters, with a small amount of methanol (about 30mL) point Secondary washing container and residue, merge washing lotion and filtrate, recycling design are simultaneously concentrated to dryness, and residue adds water 40mL to make to dissolve, full with water It is extracted 4 times, each 50mL with n-butanol shaking, merges n-butanol liquid, sufficiently washed 2 times with ammonia solution, each 50mL, ammonia solution It discards, merges n-butanol liquid, be evaporated, residue adds methanol to make to dissolve and be transferred in 5mL measuring bottle, with methanol constant volume to scale, shakes It is even, filtration, take subsequent filtrate to get.

The preparation of negative test solution: self-control is free of the blank sample of Radix Astragali, is prepared into negative test solution.

Measuring method: accurate respectively to draw 10 μ L of reference substance solution, 20 μ L, 10 μ L of test solution injects liquid chromatograph, It measures in accordance with the law.

1.3 methodological study

(1) specificity is tested

Accurate absorption 10 μ L of reference substance solution, 20 μ L respectively, test solution 10 μ L, negative 10 μ L of test solution, HPLC measurement.The result is shown in Figure 1-3.

The result shows that negative test solution has no chromatographic peak at retention time identical with Astragaloside IV reference substance, Feminine gender does not interfere with.

(2) linear relationship is investigated

The preparation of reference substance solution: taking Astragaloside IV reference substance appropriate, accurately weighed, adds methanol that every 1mL is made containing Radix Astragali The solution of first glycosides 1.209mg is to get reference substance stock solution.Precision absorption Astragaloside IV stock solution is appropriate, sets 10mL volumetric flask, adds Methanol dilution at concentration be respectively 0.0755625mg/mL, 0.151125mg/mL, 0.30225mg/mL, 0.43524mg/mL, The solution of 0.6045mg/mL, 0.7254mg/mL.It is accurate respectively to draw 20 μ L, high performance liquid chromatograph is injected, peak area is recorded, With Astragaloside IV peak area logarithm (Y) for ordinate, Astragaloside IV sample volume logarithm (X) is that abscissa does figure, obtains Astragaloside IV Regression equation be y=1.6908x+11.362 (R²=0.9993).The result shows that Astragaloside IV 0.0755625mg/mL~ Linear relationship is good in 0.7254mg/mL.

(3) precision test

Precision draws 20 μ L of Astragaloside IV reference substance solution, and continuous sample introduction measures 6 times, measures the peak area result of reference substance It is shown in Table 40.

40 Precision test result of table (n=6)

The result shows that: relative standard deviation RSD < 3% shows that precision is good.

(4) stability test

Chinese medicinal composition granules (finely ground) is taken, about 4g is taken, it is accurately weighed, Chinese medicinal composition granules are prepared in accordance with the law for examination Product solution, 0,2,4,6,8,12,24 after preparation hour measures in accordance with the law respectively, and Astragaloside content is averaged in 24 hours Value is 0.4518mg/g, RSD=2.76%, it was demonstrated that Chinese medicinal composition granules sample is good in 24 hours internal stabilities, as a result It is shown in Table 41.

41 stability test result of table

(5) reappearance test

It takes with a collection of Chinese medicinal composition granules sample, according to Chinese medicinal composition granules sample solution preparation method, Test solution is prepared, 6 parts of operation repetitive, is analyzed into HPLC, corresponding peak area is recorded, calculates Astragaloside content, as a result see Table 42.

42 reproducible test results of table (n=6)

The result shows that: the content average value for measuring Astragaloside IV in test sample is 0.4865mg/g, and RSD < 3% shows Reproducibility is good.

(6) sample recovery rate is tested

The Chinese medicinal composition granules sample of 6 parts of known contents is taken, finely ground, every part of about 2g is accurately weighed, is separately added into dense Degree is the Astragaloside IV reference substance solution 1.0mL of 0.976mg/mL, prepares test solution, measures content, calculates sample-adding recycling Rate the results are shown in Table 43.

43 sample recovery rate test result (n=6) of table

The result shows that: the average recovery rate of Astragaloside IV is 98.05%, RSD=2.93%, n=6, sample recovery rate symbol It closes and requires.

Conclusion: to sum up, the precision of Astragaloside IV method, linear, reproducibility, stability, accuracy meet the requirements, It is feasible to illustrate that the content assaying method is stablized.

The foundation of 2 Paeoniflorin chromatographic conditions

The foundation of 2.1 chromatographic conditions

Chromatographic condition: Waters symmetryshield^TMRP18 chromatographic column (4.6mm × 150mm, 5 μm)；Methanol-water It (15: 85) is mobile phase, flow velocity 1.0mLmin^-1, 30 DEG C of column temperature, sample volume is 10 μ L, Detection wavelength: 230nm.

The preparation and measurement of 2.2 solution

The preparation of reference substance solution: taking Paeoniflorin reference substance appropriate, accurately weighed, adds methanol that every 1mL first containing Radix Astragali is made The solution of glycosides 0.04112mg to obtain the final product.

The preparation of test solution solution: taking this product, finely ground, takes about 1g, accurately weighed, sets in stuffed conical flask, accurate Methanol 50mL, close plug is added, weighed weight is ultrasonically treated (power 100W, frequency 40KHZ) 30 minutes, lets cool, then weighed heavy Amount, the weight of less loss is supplied with methanol, is shaken up, filter, take subsequent filtrate to get.

The preparation of negative test solution: in prescription taste of traditional Chinese medicine ratio, autogamy is free of group's medicine of Radix Paeoniae Alba, by preparation process system It is standby to prepare at blank formulation, then by sample solution preparation method to get negative test solution.

Measuring method: it is accurate respectively to draw reference substance solution, test solution and each 10 μ L of negative test solution, inject liquid Chromatography, measurement.

Paeoniflorin content measuring method is found in establishing, and uses the lower content assaying method of 2015 editions " Chinese Pharmacopoeia " Radix Paeoniae Albas When measuring Paeoniflorin, the hangover of Paeoniflorin peak；Different mobile phase condition (1. Wondasil TM C18 chromatographies have been attempted afterwards and respectively Column (250mm × 4.6mm, 5 μm), -0.1% phosphoric acid solution of acetonitrile are mobile phase, gradient elution, Detection wavelength is respectively 230, 286nm, flow velocity 1.0mlmin^-1；2. Eclipse XDB-C18 chromatographic column (250mm × 4.6mm, 5 μm), -0.1% phosphorus of acetonitrile Aqueous acid (14: 86) is mobile phase, flow velocity: 1.0mlmin^-1, Detection wavelength: 230nm；3. Agilent ZORBAXSBC18 (4.6mm × 250mm, 5 μm) chromatographic column, -0.1% phosphoric acid water of acetonitrile are mobile phase, gradient elution, flow velocity 1.0mlmin^-1, 25 DEG C of column temperature, Detection wavelength: 230nm), different test samples prepare impurity-removing method, as a result, it has been found that the hangover of Paeoniflorin peak also without To improvement；Finally discovery uses Waters symmetryshield^TMWhen RP18 chromatographic column (4.6mm × 150mm, 5 μm), flowing Mutually acid adding can also not improve the tailing factor at Paeoniflorin peak, therefore final choice Waters symmetryshield^TMRP18 is measured Paeoniflorin.

2.3 methodological study

(1) specificity is investigated

It is accurate respectively to draw reference substance solution, negative test solution and 10 μ L of test solution, liquid chromatograph is injected, Measurement.As a result see Fig. 4-6.

The result shows that negative sample is in retention time identical with Paeoniflorin reference substance, no chromatographic peak, therefore think negative It does not interfere with.

(2) the relational investigation of line

The preparation of reference substance solution: taking Paeoniflorin reference substance appropriate, accurately weighed, adds methanol that every 1mL is made containing Paeoniflorin The solution of 0.1028mg is to get reference substance stock solution.Precision absorption Paeoniflorin stock solution is appropriate, sets 10mL volumetric flask, adds methanol Be diluted to concentration be respectively 0.01028mg/mL, 0.02056mg/mL, 0.04112mg/mL, 0.049344mg/mL, The standard solution of 008224mg/mL, 0.1028mg/mL.It is accurate respectively to draw 10 μ L, high performance liquid chromatograph is injected, is measured, Paeoniflorin standard curve is drawn, the regression equation for obtaining Paeoniflorin is y=10⁷x-19813(R²=0.9999).The result shows that Chinese herbaceous peony Glycosides linear relationship in 0.01028mg/mL~0.1028mg/mL is good.

(3) precision test

Precision draws 10 μ L of Paeoniflorin reference substance solution, continuous sample introduction 6 times, measures the peak area of reference substance, the results are shown in Table 44。

44 Precision test result of table (n=6)

(4) stability test

Prepare Chinese medicinal composition granules test solution, respectively after preparation 2,4,6,8,10,12,14,16,18, Sample introduction is analyzed within 24 hours, and the average value of paeoniflorin content is 1.86mg/g, RSD=2.55% in 24 hours, shows that sample exists Internal stability is good within 24 hours, the results are shown in Table 45.

45 stability test result of table

(5) reappearance test

It takes and prepares 6 parts of test solution according to preparation method of test article with a collection of Chinese medicinal composition granules sample, measure Peak area calculates paeoniflorin content, the results are shown in Table 46.

46 reproducible test results of table (n=6)

The result shows that: the content average value for measuring Paeoniflorin in test sample is 1.7184mg/g, RSD < 3%, shows weight Existing property is good.

(6) sample recovery rate is tested

The Chinese medicinal composition granules sample of 6 parts of known contents is taken, finely ground, every part of about 0.5g is accurately weighed, is separately added into Paeoniflorin reference substance 0.879mg is prepared into test solution, measures content, calculates sample recovery rate, the results are shown in Table 47.

47 sample recovery rate test result (n=6) of table

The result shows that: the mean sample recovery rate of Paeoniflorin is 102.27, RSD=2.28%, n=6, shows that the method is loaded The rate of recovery meets the requirements.

Conclusion: to sum up, the methodology of Paeoniflorin meets the requirements, and it is feasible to illustrate that the content assaying method is stablized.

Three batches of sample assays

Three batches of traditional Chinese medicine particle samples are prepared, content is measured by the content assaying method of above-mentioned foundation, calculates Radix Astragali The content of first glycosides and Paeoniflorin.It the results are shown in Table 48.

48 3 batches of sample assay results of table

Batch	Astragaloside IV mg/g	Paeoniflorin mg/g
			20150528	0.54	1.68
20150628	0.49	1.88
			20150728	0.50	1.81
Average value	0.51	1.79

As a result: in present composition particle, the average content of Astragaloside IV is 0.51mg/g, the average content of Paeoniflorin For 1.79mg/g.

It draws up and determines content limit are as follows: the every 1g of this product is containing Radix Astragali with Astragaloside IV (C₃₁H₆₈O₁₄) meter, 0.41mg must not be less than.

The every 1g of this product is containing Radix Paeoniae Alba with Paeoniflorin (C₂₃H₂₈O₁₁) meter, 1.43mg must not be less than.

Claims

1. a kind of Chinese medicine composition for treating allergic rhinitis, which is characterized in that the Chinese medicine composition is by the following raw material medicine system At: Radix Astragali, ramulus cinnamomi, the flower bud of lily magnolia, radix saposhnikoviae, Rhizoma Atractylodis Macrocephalae, rhizoma zingiberis, cortex cinnamomi, Radix Paeoniae Alba, dark plum, Schisandra chinensis, rhizoma anemarrhenae, Radix Glycyrrhizae.

2. Chinese medicine composition as described in claim 1, which is characterized in that the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 5-50 parts by weight, flower bud of lily magnolia 2-20 parts by weight, radix saposhnikoviae 3-24 parts by weight, Rhizoma Atractylodis Macrocephalae 3-27 parts by weight, are done ramulus cinnamomi 3-27 parts by weight Ginger 2-20 parts by weight, Radix Paeoniae Alba 2-20 parts by weight, dark plum 4-30 parts by weight, Schisandra chinensis 2-20 parts by weight, are known cortex cinnamomi 1-8 parts by weight Female 2-20 parts by weight, Radix Glycyrrhizae 2-20 parts by weight.

3. Chinese medicine composition as claimed in claim 2, which is characterized in that the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 8-40 parts by weight, flower bud of lily magnolia 3-16 parts by weight, radix saposhnikoviae 4-18 parts by weight, Rhizoma Atractylodis Macrocephalae 4-22 parts by weight, are done ramulus cinnamomi 4-22 parts by weight Ginger 3-16 parts by weight, Radix Paeoniae Alba 3-16 parts by weight, dark plum 6-24 parts by weight, Schisandra chinensis 3-16 parts by weight, are known cortex cinnamomi 1-6 parts by weight Female 3-16 parts by weight, Radix Glycyrrhizae 3-16 parts by weight.

4. Chinese medicine composition as claimed in claim 3, which is characterized in that the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 10-30 parts by weight, ramulus cinnamomi 5-18 parts by weight, flower bud of lily magnolia 4-12 parts by weight, radix saposhnikoviae 5-14 parts by weight, Rhizoma Atractylodis Macrocephalae 5-18 parts by weight, Rhizoma zingiberis 4-12 parts by weight, cortex cinnamomi 2-5 parts by weight, Radix Paeoniae Alba 4-12 parts by weight, dark plum 8-20 parts by weight, Schisandra chinensis 4-12 parts by weight, Rhizoma anemarrhenae 4-12 parts by weight, Radix Glycyrrhizae 4-12 parts by weight.

5. Chinese medicine composition as claimed in claim 4, which is characterized in that the Chinese medicine composition is made of the following raw material medicine: Radix Astragali 15-25 parts by weight, flower bud of lily magnolia 5-8 parts by weight, radix saposhnikoviae 6-9 parts by weight, Rhizoma Atractylodis Macrocephalae 7-12 parts by weight, are done ramulus cinnamomi 7-12 parts by weight Ginger 5-8 parts by weight, cortex cinnamomi 2-4 parts by weight, Radix Paeoniae Alba 5-8 parts by weight, dark plum 10-15 parts by weight, Schisandra chinensis 5-8 parts by weight, rhizoma anemarrhenae 5-8 parts by weight, Radix Glycyrrhizae 5-8 parts by weight.

6. Chinese medicine composition as claimed in claim 5, which is characterized in that the Chinese medicine composition is made of the following raw material medicine: 21 parts by weight of Radix Astragali, 9 parts by weight of ramulus cinnamomi, 6 parts by weight of the flower bud of lily magnolia, windproof 7 parts by weight, 9 parts by weight of Rhizoma Atractylodis Macrocephalae, 6 parts by weight of rhizoma zingiberis, cortex cinnamomi 3 parts by weight, 6 parts by weight of Radix Paeoniae Alba, 12 parts by weight of dark plum, 6 parts by weight of Schisandra chinensis, 6 parts by weight of rhizoma anemarrhenae, 6 parts by weight of Radix Glycyrrhizae；

Or, the Chinese medicine composition is made of the following raw material medicine: 16 parts by weight of Radix Astragali, 11 parts by weight of ramulus cinnamomi, 5 parts by weight of the flower bud of lily magnolia, Windproof 8 parts by weight, 8 parts by weight of Rhizoma Atractylodis Macrocephalae, 7 parts by weight of rhizoma zingiberis, 2 parts by weight of cortex cinnamomi, 7 parts by weight of Radix Paeoniae Alba, 11 parts by weight of dark plum, the five tastes Sub 7 parts by weight, 5 parts by weight of rhizoma anemarrhenae, 7 parts by weight of Radix Glycyrrhizae；

Or, the Chinese medicine composition is made of the following raw material medicine: 24 parts by weight of Radix Astragali, 7 parts by weight of the flower bud of lily magnolia, are prevented at 8 parts by weight of ramulus cinnamomi 6 parts by weight of wind, 11 parts by weight of Rhizoma Atractylodis Macrocephalae, 5 parts by weight of rhizoma zingiberis, 4 parts by weight of cortex cinnamomi, 5 parts by weight of Radix Paeoniae Alba, 14 parts by weight of dark plum, the five tastes Sub 5 parts by weight, 7 parts by weight of rhizoma anemarrhenae, 5 parts by weight of Radix Glycyrrhizae.

7. Chinese medicine composition as claimed in any one of claims 1 to 6, which is characterized in that the Chinese medicine composition is each bulk pharmaceutical chemicals The composition mixed after crushing or the mixing of each bulk pharmaceutical chemicals or after individually extracting obtained extract or extract into The active component that one one-step refining purifies, or regular dosage form made of pharmaceutically acceptable auxiliary material is added.

8. application of the Chinese medicine composition as claimed in any one of claims 1 to 6 in the drug of preparation treatment allergic rhinitis.

9. application of the Chinese medicine composition as claimed in any one of claims 1 to 6 in the drug that preparation has anti-inflammatory effect.

10. application of the Chinese medicine composition as claimed in any one of claims 1 to 6 in the drug that preparation has anti-allergic effects.