CN102085248A

CN102085248A - Traditional Chinese medicinal composition for treating cervix diseases, method for preparing same and method for detecting same

Info

Publication number: CN102085248A
Application number: CN 200910252151
Authority: CN
Inventors: 朱飞锦; 左之文; 罗杰英; 胡随瑜
Original assignee: Zhuzhou Qianjin Pharmaceutical Co Ltd
Current assignee: Zhuzhou Qianjin Pharmaceutical Co Ltd
Priority date: 2009-12-07
Filing date: 2009-12-07
Publication date: 2011-06-08
Anticipated expiration: 2029-12-07
Also published as: CN102085248B

Abstract

The invention relates to a traditional Chinese medicinal composition for treating cervix diseases. The traditional Chinese medicinal composition comprises the following components in part by weight: 35 to 45 parts of bark of tree of heaven, 25 to 35 parts of lightyellow sophora root, 25 to 35 parts of tree peony bark, 25 to 35 parts of olibanum and 0.8 to 1.2 parts of borneol, wherein the lightyellow sophora root has the same weight as the tree peony bark and the olibanum. The invention also provides a method for detecting the traditional Chinese medicinal composition and a method for preparing a gelling agent of the traditional Chinese medicinal composition. In the traditional Chinese medicinal composition, on a basis of ensuring that the lightyellow sophora root has the same content as the tree peony bark and the olibanum, the content of bark of tree of heaven and the content of borneol are adjusted, so that the traditional Chinese medicinal composition fully utilizes the effects of various raw materials to achieve the effects of clearing away heat, eliminating dampness, removing blood stasis and promoting regeneration of the tissue and has more exact efficacy. In the method for preparing the gelling agent of the traditional Chinese medicinal composition, all components are extracted respectively to prepare the gelling agent, so that the method is simple and suitable for industrial production, has high extraction rate of drug effect ingredients and low cost. In addition, the invention also provides the method for detecting the traditional Chinese medicinal composition, which can effectively identify components and content of the traditional Chinese medicinal composition and has little disturbance and high result repeatability.

Description

A kind of Chinese medicine composition for the treatment of cervical disease and preparation method thereof and detection method

Technical field

The present invention relates to the field of Chinese medicines, relate to a kind of Chinese medicine composition for the treatment of cervical disease and preparation method thereof and detection method specifically.

Background technology

Cervicitis is clinical one group of common clinical disease.Cervical erosion is the main local patholoic change in the chronic cervicitis, is commonly encountered diseases, the frequently-occurring disease of gynecological.Its sickness rate accounts for more than 50% of gynaecopathia.Show after deliberation, cervical erosion and cervical cancer important relationship arranged.The cervical cancer variability of patients of cervical ruin is 2.5%, and more non-patients of cervical ruin is high 6 times.

Doctor trained in Western medicine thinks that cervical erosion is the modal clinical manifestation of chronic cervicitis.Its cause of disease mostly is long-term chronic mechanical irritation and damage, as childbirth, miscarriage or surgical injury etc. is the main root inducement, causes the pathogen intrusion and causes that acute infection period is active treatment not, gradually form chronic cervicitis, make cervix uteri mainly produce rotten to the corn pathological change.Think it mainly is that the squamous epithelial cancer on cervix uteri surface comes off on the cervical erosion pathology, by columnar epithelium is replaced, the result that last veins beneath the skin appears.Research in recent years show again cervical erosion and whole body immunologic mechanism, great relation is arranged.The result shows: being related of the infection of HPV 16 (HPV-16), human herpesvirus 2's type (HSV-2) cytomegalovirus (CMV) and chlamydia trachomatis (CT) and cervical erosion is close.Behind the viral infection body cell, can directly exert an influence to immunity of organism, this influence comprises two aspects: the one, immunosuppressant, the 2nd, immunological enhancement, and based on immunosuppressant.

Western medical treatment cervical erosion, secondly first-selected physiotherapy is external used medicine.The prolonged application physiotherapy can produce a lot of side effect, and treats easily recurrence of back, and contraindication is arranged again simultaneously; The Western medicine medicine for external use mostly is corrosive medicine, and the focus shallow to comparison sheet, that area is smaller has certain effect, but treatment is how not thorough, and long-term prescription and naturopathy can cause many side effect, and removes and stop easily recurrence of back.

Motherland's medical science thinks that it is to invade in the noxious dampness and the damp-heat gluing each other multiple abscess part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels that cervical erosion forms reason, and it is not solid to cause conception vessel, and dai channel is failed to keep an appointment, and vessels of the uterus is qi-blood disharmony, the local QI and blood retardance of cervix uteri, and the evil stasis of blood causes corruption.Studies show that through modern Chinese medicine, cervical erosion cause of disease complexity, but it mainly be damp is main, sick position is at uterus, and is dirty in close relations with spleen kidney two, and pathogenesis is mainly the damp damage and appoints, is with two arteries and veins, vessels of the uterus is qi-blood disharmony, and with the passing of time the then damp and hot stasis of blood in cervical orifice of uterus part is long-pending, the pyretic toxicity heap soil or fertilizer over and around the roots stagnates, blood stasis causes corruption.Modern simultaneously also have the expert to think that cervical erosion and liver also have certain relation.Dampness-heat of the liver is made a bet, but coup injury is appointed the band passages through which vital energy circulates, and causes primary disease.As fully visible, damp has important effect in the pathogenic process of cervical erosion, so the treatment of cervical erosion is engaged in damp eliminating.In recent years, traditional Chinese medical science worker has carried out a large amount of observations to the effectiveness of Chinese medicine cervical erosion, the result shows, Chinese medicine has therapeutical effect preferably for cervical erosion, and can reduce the side effect and the palindromia rate of physiotherapy, Chinese traditional treatment cervical erosion becomes one of method that generally adopts clinically thus.

Because its sick bit comparison is special, the endo-medicine effect is slower, is not easy to through sick position, so the traditional Chinese medical science in recent years mostly is external treatment to the treatment of cervical erosion.Conclude bibliographical information in recent years, the Chinese medicine of external treatment cervical erosion commonly used roughly is divided into following a few class: 1. removing dampness, subside a swelling, hold back skin ulcer, putrefaction removing, granulation promoting medicine; 2. heat clearing away, detoxifcation, dampness, blood-cooling drugs; 3. invigorate blood circulation, blood stasis dispelling, swollen medicine looses.Dosage form mostly is powder, suppository, pill, tablet, unguentum, paste etc.This shows that treatment cervical erosion medication will be based on dampness or damp eliminating medicine, and is aided with promoting blood circulation and cooling blood, the promoting tissue regeneration and ulcer healing medicine is main.

The invention provides a kind of Chinese medicine composition that can treat cervical disease and preparation method thereof and detection method.

Summary of the invention

The purpose of this invention is to provide a kind of Chinese medicine composition for the treatment of cervical disease;

Another object of the present invention provides the preparation method of Chinese medicine composition gel;

The present invention also provides the detection method of this Chinese medicine composition.

Chinese medicine composition of the present invention contains the component of following weight portion: Cortex Ailanthi: 35-45; Radix Sophorae Flavescentis: 25-35; Cortex Moutan: 25-35; Olibanum: 25-35; Borneolum Syntheticum: 0.8-1.2, wherein Radix Sophorae Flavescentis and Cortex Moutan, Olibanum equivalent.

The component that preferably contains following weight portion: Cortex Ailanthi: 40; Radix Sophorae Flavescentis: 30; Cortex Moutan: 30; Olibanum: 30; Borneolum Syntheticum: 1, wherein Radix Sophorae Flavescentis and Cortex Moutan, Olibanum equivalent.

Cortex Ailanthi: " pharmacopeia ": property hardship, puckery, cold, return large intestine, stomach, Liver Channel.The function heat clearing and damp drying, astringent therapy leukorrhagia stopping, antidiarrheal, hemostasis.Be used for leucorrhea with red and white discharge, damp-heat dysentery, the chronic diarrhea chronic dysentery is had blood in stool, metrorrhagia etc." Treatise on Dietetic Therapy ": " woman's metrorrhagia and puerperal blood more than, menses is too much, also ends under the leucorrhea with bloody discharge." Compendium of Material Medica: " and the Chinese toon root white skin, cool in nature and can puckery blood.Believing damp and hot is disease, the turbid band of dysentery, and all cards of smart sliding nocturnal emission are used it invariably." " Thunder God is concocted the property of medicine and is separated ": " it is puckery to drown." " herbal readability ": " and kill all as the infantile malnutrition worm, ascarid, tuberculosis passes corpse, the disease that red and white leukorrhea is turbid.”

Radix Sophorae Flavescentis: " pharmacopeia ": the property hardship, cold.GUIXIN, liver, stomach, large intestine, urinary bladder channel.Function heat clearing away bath is wet, and parasite killing must be urinated.Be used for hematodiarrhoea, have blood in stool, jaundice urine retention, leucorrhea with red and white discharge, swelling of the vulva pudendal pruritus, eczema, eczema, skin pruritus, scabies leprosy; External treatment trichomonal vaginitis." herbal classic ": " main stagnation of QI in chest and abdomen, note of the ancient Chinese abdominal mass is gathered, jaundice, drowning has the heeltap, relieves oedema or abdominal distension through diuresis or purgation, and removes carbuncle, invigorating middle warmer, arresting lacrimation makes eye bright." " not Lu ": " and peace will benefit is smart for nourishing the liver courage, settling five organs, nine orifices, remove the latent heat intestinal and ward off, quench the thirst, sober up, yellowish or reddish urine, it is evil to treat the malignant boil bottom, flat gastric qi is made us having a liking for food and is made light of one's life by commiting suicide." " the southern regions of the Yunnan Province book on Chinese herbal medicine ": " and removing heat from blood is separated pyretic toxicity, mange, and the impetigo poison is the best.Treat skin pruritus, the tinea corporis scabies is respectively arranged, naughtiness is quiet in vain, and discharging fresh blood stool is had blood in stool.Pathogenic wind expelling, detumescence poison, clearing away phlegm poison." modern pharmacological research shows, this product has antiinflammatory, promotes effects such as immunity, resisting pathogenic microbes (" China is herbal ").

Cortex Moutan: " pharmacopeia ": property hardship, suffering are slightly cold.GUIXIN, liver, kidney channel.The function clearing away heat and cooling blood, blood circulation promoting and blood stasis dispelling.Be used for maculae caused by violent heat pathogen, hematemesis and epistaxis, night fever abating at dawn, lossless hectic fever due to YIN-deficiency, the amenorrhea dysmenorrhea, carbuncle sore tumefacting virus falls and pounces on the pain of injury." herbal classic ": " main cold brimful of tears, apoplexy clonic convulsion, infantile convulsion pathogen are removed the hard blood stasis of note of the ancient Chinese and are stayed stomach, and settling five organs is treated the carbuncle skin ulcer." " property of medicine opinion ": " control cold air, all pains of loosing.It is obstructed to control woman's passages through which vital energy circulates, blood drop pain in the lumbar region." " solar corona ": " and remove pathogen, please color, the natural fibre line of meat blood vessels of being open to the custom, evacuation of pus, logical menstruation disappears and pounces on the damage blood stasis, and reuniting the fractured tendons and bones removes migratory arthralgia, the tire lower eyelid that falls, the cold and hot vim and vigour of all woman in puerperal." Zhang Jiegu: " can rush down the fire among the cloudy born of the same parents." " detailed outline ": ", control the blood fire second of the three ten-day periods of the hot season, attached dysphoria with smothery sensation with blood, hemopoietic, removing heat from blood." modern pharmacological research shows, that this product has is antibiotic, promote effects such as immunity, hemorrhage.(" China's book on Chinese herbal medicine ").

Olibanum: property suffering, bitter, tepor.GUIXIN, liver, spleen channel.The function blood-activating and qi-promoting, inducing menstruation to relieve menalgia, detumescence and promoting granulation.Be used for trusted subordinate's pain, rheumatic arthralgia, amenorrhea dysmenorrhea, traumatic injury stasis of blood pain, swollen ulcer drug, acute appendicitis, the sore being festered and unhealed etc." not Lu ": " it be swollen to treat the edema due to wind pathogen poison, removes bad odor.Treat the gas urticaria poison of itching." " solar corona ": " therapeutic method to keep the adverse QI flowing downwards is beneficial smart, mends the waist knee joint, controls kidney qi, ends cholera, dashes pathogen in the evil, cardiac and abdominal pain chronic infectious disease gas.Soft extracts, pain relieving is put on flesh." detailed outline ": " promoting blood circulation ends large intestine dysentery pain and the stasis of married woman's QI rising in reverse order, and the trusted subordinate has a pain." supplement to the Herbal: " and treat deaf, the apoplexy locked mouth, married woman's vim and vigour can be sent out wine, manage air-cooledly, and only large intestine is let out Bleach, treats all skin ulcers and makes internal diabetes." modern pharmacological research shows, this product has antiinflammatory, promotes effects such as immunity, cholesterol reducing and analgesia (" China's book on Chinese herbal medicine ").

Borneolum Syntheticum: property suffering, hardship, cold.Function is opened surreptitiously refreshment, the heat radiation pain relieving, and removing nebula makes eye bright.Cure mainly the apoplexy locked mouth, sick coma brimful of tears, infantile convulsion expectorant fan, deafness with qi stagnation, conjunctivitis, sore throat, aphtha, carbuncle, hemorrhoid, enterobiasis." Haiyao Bencao, Oversea Materia Medica ": " external oculopathy eye in main, three worms are controlled five hemorrhoid, make eye bright relieving palpitation, secret essence." " Newly Revised Canon of Materia Medica ": " and main trusted subordinate's pathogen, rheumatism is gathered, and deafness makes eye bright, and goes conjunctival congestion skin nebula." " medical center is usurped and is wanted the property of medicine ": " granulation promoting pain relieving." " the first life bud of book on Chinese herbal medicine ": " the heat of the diaphragm plug of being open to the custom must close and stop up wind saliva, and the interior external oculopathy of point is looked thing and do not seen, and moves back conjunctival congestion with ophthalmalgia, and the skin nebula is invaded eyeball.Relieving convulsion the making eye bright of energy, the kind secret essence of calming the nerves.Control trusted subordinate's pathogen, the curing rheumatism deafness." modern pharmacological research shows, this product has good antiinflammatory, antibacterial action (" China's book on Chinese herbal medicine ").

The present invention is a monarch drug with the Cortex Ailanthi clearing heat and expelling damp; With the Radix Sophorae Flavescentis of the heat clearing and damp drying of equivalent and the Cortex Moutan of clearing away heat and cooling blood is ministerial drug, and auxiliary monarch drug is to strengthen the removing damp-heat effect; Olibanum, Borneolum Syntheticum are adjuvant drug, blood stasis dispelling, and pain relieving, heat clearing away, granulation promoting improves therapeutic effect.

Chinese medicine composition of the present invention can adopt this area method commonly used to make various dosage forms, for using conveniently preferred gel.

The present invention also provides the preparation method of the gel of above-mentioned Chinese medicine composition:

1) Cortex Ailanthi adds 2-9 times of water gaging, decocts with water 1-3 time, decocts 1-2.5 hour at every turn, and collecting decoction filters, and filtrate decompression is concentrated into the Cortex Ailanthi clear paste that relative density is 1.05～1.10 (60 ℃);

2) Radix Sophorae Flavescentis adds 6-12 times of water gaging, decocts with water 1-3 time, soaks 0.5-1.5 hour before decocting for the first time, the each decoction 0.5-3 hour, collecting decoction filters, filtrate decompression is concentrated into the Radix Sophorae Flavescentis clear paste of relative density 1.10～1.15 (60 ℃), put coldly, add ethanol and make and contain alcohol amount and reach 60-70%, left standstill 8-24 hour, filter, filtrate recycling ethanol merges with the Cortex Ailanthi clear paste to there not being the alcohol flavor, is concentrated into the clear paste of relative density 1.05～1.10 (60 ℃);

3) Olibanum adds 6-10 times of water gaging, distills 5-9 hour, collects volatile oil;

4) Cortex Moutan distills with steam distillation, and collecting quality is the distillate of 4-8 times of medical material amount, places crystallize, filters, and the paeonol crystallization is put in addition; The aqueous solution redistillation, collecting quality is the distillate of 2-4 times of medical material amount, places crystallize, filters, and merges the paeonol crystallization;

5) get it filled and join in the mixing clear paste of Cortex Ailanthi and Radix Sophorae Flavescentis, stir evenly, place, make abundant swelling with sodium carboxymethyl cellulose, medicinal glycerin;

6) get the volatile oil that Borneolum Syntheticum adds Olibanum, the paeonol crystallization with dissolve with ethanol after, add ethyl hydroxybenzoate and Tween-80, after stirring makes dissolving, join in the swollen solution that step 5) obtains, add the water mixing, place and made abundant swelling in 12-36 hour, stir, packing promptly gets gel.

Wherein sodium carboxymethyl cellulose, medicinal glycerin, ethyl hydroxybenzoate and Tween-80 are gel pharmaceutic adjuvants commonly used, and its consumption can be adjusted according to those skilled in the art's requirement; Preferably, the consumption of sodium carboxymethyl cellulose, medicinal glycerin is respectively the 0.6-1.5 weight portion of used Borneolum Syntheticum weight, and the consumption of ethyl hydroxybenzoate and Tween-80 is respectively the 0.01-0.1 weight portion of used Borneolum Syntheticum weight.

The concentration of effective ingredient can be adjusted according to amount of water in the preparation process and adjuvant amount in the described gel, and the gained gel is the 0.1-0.2 weight portion of used raw material of Chinese medicine medicine gross weight, preferred 0.15 weight portion.

Chinese medicine composition heat clearing and damp drying of the present invention, expelling the stasis for the new is used for profuse leukorrhea due to the dampness and heat stasis, yellow skin or white, or be pus; The waist abdomen weighs down and expands, or the puckery pain of dribbling urination; Or pudendal pruritus, bitter taste in the mouth and dry throat, red tongue, yellow fur or greasy, stringy pulse or cunning, and cervicitis has above-mentioned proposita.

In addition, the present invention also provides the detection method of above-mentioned Chinese medicine composition, comprises discrimination method and content assaying method:

The described discrimination method of Chinese medicine composition of the present invention comprises following one or more methods:

(1) get the 1g sample, add water 30ml, fully stir, room temperature was placed 30 minutes, supersound process 5 minutes, and the dislocation separatory funnel adds 10-30ml ether extraction 1-3 time, merges ether solution, and room temperature volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the paeonol reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (10: 0.5) is developing solvent, launch, take out, dry, spray is with the acid 5% ferric chloride alcoholic solution of hydrochloric acid, and 105 ℃ to be heated to the speckle colour developing clear;

(2) according to discrimination method (1) preparation sample solution, get 0.5ml, add methanol to 5ml as need testing solution; Other gets the Borneolum Syntheticum reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-chloroform-ethyl acetate (10: 3: 1) is developing solvent, launch, take out, dry, spray develops the color with 5% phosphomolybdic acid ethanol test solution, puts 105 ℃ and is heated to clear spot;

(3) get the 1g sample, add water 30ml, stir evenly, placed 30 minutes, supersound process 5 minutes adds 1mol/L sodium hydroxide solution 0.3ml, stirs evenly, move in the separatory funnel, with 10-30ml chloroform extraction 1-3 time, merge chloroform liquid, put evaporate to dryness in the water-bath, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Radix Sophorae Flavescentis control medicinal material 1g, adds chloroform 25ml, strong ammonia solution 0.5ml, close plug, and jolting was constantly placed 24 hours, filtered, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with benzene-acetone-ethyl acetate-strong ammonia solution (2: 3: 4: 0.2) be developing solvent, launch, take out, dry, spray is with the bismuth potassium iodide test solution of new system, and it is clear to be heated to the speckle colour developing;

In the described authentication method (3), the reference substance of Radix Sophorae Flavescentis also can be the matrine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Or select for use Radix Sophorae Flavescentis control medicinal material and matrine to make control medicinal material solution and reference substance solution simultaneously, carry out thin layer chromatography and detect.

Utilize above-mentioned thin layer liquid chromatography to differentiate, the good separating effect of the effective ingredient of Chinese medicine composition of the present invention, negative interference is little, the test sample chromatograph with control medicinal material and the corresponding position of reference substance chromatograph on, can show the same color speckle, so said method can be used to differentiate Chinese medicine composition of the present invention.

(4) acidity is differentiated: get 1.0 gram samples, add water 50ml, after stirring made dispersion, room temperature was placed 30 minutes, and supersound process 5 minutes is measured (an appendix VI of Chinese Pharmacopoeia version in 2005 H) in accordance with the law, and pH value is 4.5～6.0.

The described content assaying method of Chinese medicine composition of the present invention is to adopt high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D) to measure the content of active component, comprising:

1) preparation of reference substance solution: precision takes by weighing through 10-14 hour matrine reference substance of phosphorus pentoxide drying under reduced pressure and oxymatrine reference substance, adds mobile phase and makes the solution that every 1ml contains matrine, each 40 μ g of oxymatrine respectively;

2) preparation of need testing solution: sample thief, fully grind well, the accurate title, decided 1.5-2.5g, puts in the 100ml beaker, adds water 50ml, stir evenly, room temperature was placed 30 minutes, supersound process (power 300W, frequency 25KHz) 5 minutes, add 1mol/L sodium hydroxide solution 0.3ml, stir evenly, move in the separatory funnel, use 20-40ml chloroform extraction 1-3 time, merge chloroform liquid, evaporate to dryness, residue add methanol 10ml makes dissolving, moves in the 50ml measuring bottle, and add mobile phase and be diluted to scale, as need testing solution A (mensuration oxymatrine); Precision is measured A solution 5ml, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, as need testing solution B (mensuration matrine);

3) measure: accurate respectively reference substance solution and each 10 μ l of need testing solution A, B of drawing, inject chromatograph of liquid, measure;

Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; Acetonitrile-0.1% trifluoroacetic acid solution (3.5: 96.5) is a mobile phase; The detection wavelength is 220nm.Number of theoretical plate should be not less than 2000 by matrine.

In the Chinese medicine composition of the present invention, every 1g contains Radix Sophorae Flavescentis with matrine (C ₁₅H ₂₄N ₂O) and oxymatrine (C ₁₅H ₂₄N ₂O ₂) the total amount meter, must not be less than 5.0mg.

By adjustment to content, Chinese medicine composition of the present invention is at the content that guarantees to have adjusted on the Radix Sophorae Flavescentis basis identical with Cortex Moutan content Cortex Ailanthi and Borneolum Syntheticum, curative effect is more definite, all medicines and closing, play heat clearing and damp drying altogether, the merit of expelling the stasis for the new, compare with existing Chinese toon breast, can be in all directions at adapting to disease and syndrome characteristics, meet theory of Chinese medical science and clinical practice, Pharmacodynamic test of active extract result shows, Chinese medicine composition of the present invention has the effect of rising for the Cavia porcellus itch-threshold due to the histamine phosphate, illustrate that it has itching-relieving action preferably, and for Candida albicans, beta hemolytic streptococcus, gold Portugal bacterium, form staph, escherichia coli, Bacillus proteus has the good in-vitro inhibitory action; The present invention also at the characteristics of each component, extracts respectively each component, the production procedure of attached gel agent, proposed the preparation method of the gel of Chinese medicine composition of the present invention, method is simple, is fit to suitability for industrialized production, active ingredient extraction ratio height is saved cost; The effective ingredient content foot that makes, good effect; The present invention also provides the detection method of this Chinese medicine composition in addition, and its good separating effect negative disturbs for a short time, and good reproducibility as a result can effectively be differentiated the composition and the content of this Chinese medicine composition.

Description of drawings

Fig. 1 causes the influence of pain effect to the mice hot plate method for Chinese medicine composition gel of the present invention.

Fig. 2 is the HPLC figure of determination of matrine.

The HPLC figure that Fig. 3 measures for oxymatrine concentration.

The specific embodiment

Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.Specialize as non-, the accepted standard product are from Nat'l Pharmaceutical ﹠ Biological Products Control Institute among the embodiment.

Embodiment 1

[prescription]

Cortex Ailanthi 2000g Radix Sophorae Flavescentis 1500g Cortex Moutan 1500g

Olibanum 1500g Borneolum Syntheticum 50g

[method for making]

1) Cortex Ailanthi decocts with water 2 times, adds 6 times of water gagings for the first time, decocts 2 hours, adds 4 times of water gagings for the second time, decocts 1 hour, and collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.05 (60 ℃)

2) Radix Sophorae Flavescentis decocts with water secondary, adds 9 times of water gagings for the first time and soaks 1 hour, decocts 2 hours, for the second time add 7 times of water gagings, decocted collecting decoction 1 hour, filter, filtrate decompression is concentrated into the clear paste of relative density 1.10 (60 ℃), puts cold, adding ethanol makes and contains alcohol amount and reach 65%, standing over night filters, and filtrate recycling ethanol is to there not being the alcohol flavor, merge with above-mentioned Cortex Ailanthi clear paste, be concentrated into the clear paste of relative density 1.05 (60 ℃);

3) Olibanum adds the water of 8 times of amounts, distills 7 hours, collects volatile oil, and is standby;

4) Cortex Moutan distills with steam distillation, collects the about 9000ml of distillate, places crystallize, filters, and the paeonol crystallization is put in addition; The aqueous solution redistillation is collected the about 4500ml of distillate, places crystallize, filters, and merges the paeonol crystallization, and is standby;

5) get it filled in addition and join in the mixing clear paste of above-mentioned Cortex Ailanthi and Radix Sophorae Flavescentis, stir evenly, place, make abundant swelling with sodium carboxymethyl cellulose 40g, medicinal glycerin 50g;

6) get Borneolum Syntheticum and add Olibanum volatile oil, the paeonol crystallization adds ethanol 50ml, add ethyl hydroxybenzoate 2g and Tween-80 2g, after stirring makes dissolving, join in the above-mentioned swollen solution, adding distil water is to 1000g, stirred 30 minutes, place and made abundant swelling in 24 hours, stir, packing promptly gets gel.

Embodiment 2

[prescription]

Cortex Ailanthi 2250g Radix Sophorae Flavescentis 1250g Cortex Moutan 1250g

Olibanum 1250g Borneolum Syntheticum 40g

[method for making]

1) extract: Cortex Ailanthi decocts with water 3 times, adds 9 times of water gagings for the first time, decocts 2.5 hours, adds 6 times of water gagings for the second time, decocted 1 hour, and added 2 times of water gagings for the third time, decocted 1 hour, collecting decoction filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.10 (60 ℃);

2) Radix Sophorae Flavescentis decocts with water 3 times, adds 12 times of water gagings for the first time and soaks 1.5 hours, decocts 3 hours, for the second time add 9 times of water gagings, decocted 1 hour, add 6 times of water gagings for the third time, decocted 0.5 hour, collecting decoction filters, filtrate decompression is concentrated into the clear paste of relative density 1.15 (60 ℃), puts coldly, adds ethanol and makes and contain alcohol amount and reach 60%, left standstill 24 hours, and filtered, filtrate recycling ethanol is to there not being the alcohol flavor, merge with above-mentioned clear paste, be concentrated into the clear paste of relative density 1.10 (60 ℃);

3) Olibanum adds the water of 6 times of amounts, distills 9 hours, collects volatile oil, and is standby;

4) Cortex Moutan distills with steam distillation, collects the about 12000ml of distillate, places crystallize, filters, and the paeonol crystallization is put in addition; The aqueous solution redistillation is collected the about 6000ml of distillate, places crystallize, filters, and merges the paeonol crystallization, and is standby;

5) get it filled and join in the above-mentioned clear paste, stir evenly, place, make abundant swelling with sodium carboxymethyl cellulose 30g, medicinal glycerin 75g;

6) get Borneolum Syntheticum and add above-mentioned volatile oil, the paeonol crystallization adds ethanol 60ml, add ethyl hydroxybenzoate 0.5g, Tween-80 0.5g, after stirring makes dissolving, join in the above-mentioned swollen solution, adding distil water is to 1000g, stirred 30 minutes, and placed and made abundant swelling in 24 hours, stir, packing promptly gets gel.

Embodiment 3

[prescription]

Cortex Ailanthi 1750g Radix Sophorae Flavescentis 1750g Cortex Moutan 1750g

Olibanum 1750g Borneolum Syntheticum 60g

[method for making]

1) extract: Cortex Ailanthi decocts with water 1 time, adds 9 times of water gagings, decocts 2.5 hours, filters, and filtrate decompression is concentrated into the clear paste that relative density is 1.05 (60 ℃);

2) Radix Sophorae Flavescentis decocts with water 1 time, add 12 times of water gagings and soaked 1.5 hours, decocted 3 hours, filter, filtrate decompression is concentrated into the clear paste of relative density 1.10 (60 ℃), put coldly, add ethanol and make and contain alcohol amount and reach 60%, left standstill 8 hours, filter, filtrate recycling ethanol merges with above-mentioned clear paste to there not being the alcohol flavor, is concentrated into the clear paste of relative density 1.05 (60 ℃);

3) Olibanum adds the water of 10 times of amounts, distills 5 hours, collects volatile oil, and is standby;

4) Cortex Moutan distills with steam distillation, collects the about 6000ml of distillate, places crystallize, filters, and the paeonol crystallization is put in addition; The aqueous solution redistillation is collected the about 3000ml of distillate, places crystallize, filters, and merges the paeonol crystallization, and is standby;

5) get it filled and join in the above-mentioned clear paste, stir evenly, place, make abundant swelling with sodium carboxymethyl cellulose 75g, medicinal glycerin 30g;

6) get Borneolum Syntheticum and add above-mentioned volatile oil, the paeonol crystallization adds ethanol 60ml, add ethyl hydroxybenzoate 5g, Tween-80 5g, after stirring makes dissolving, join in the above-mentioned swollen solution, adding distil water is to 1000g, stirred 30 minutes, and placed and made abundant swelling in 24 hours, stir, packing promptly gets gel.

Embodiment 4

(1) gets embodiment 1 and 2 each 1g of gel that prepare, be mixed with need testing solution in accordance with the following methods: add water 30ml, fully stir, room temperature was placed 30 minutes, supersound process 5 minutes, dislocation separatory funnel, 3 times (30ml, 20ml, 10ml) extracted in the jolting that adds diethyl ether, merge ether solution, room temperature volatilizes, and residue adds methanol 1ml makes dissolving;

Other gets the paeonol reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (10: 0.5) is developing solvent, launch, take out, dry, spray is with the acid 5% ferric chloride alcoholic solution of hydrochloric acid, and 105 ℃ to be heated to the speckle colour developing clear.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show same color and speckle.

(2) get each 0.5ml of discrimination method (1) preparation need testing solution, add methanol to 5ml as need testing solution.Other gets the Borneolum Syntheticum reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-chloroform-ethyl acetate (10: 3: 1) is developing solvent, launch, take out, dry, spray develops the color with 5% phosphomolybdic acid ethanol test solution, puts 105 ℃ and is heated to clear spot.In the test sample chromatograph, with reference substance chromatograph relevant position on, show the speckle of two same colors.

(3) get embodiment 1 and 2 each 1g of gel that prepare, be mixed with need testing solution in accordance with the following methods: add water 30ml, stir evenly, placed 30 minutes, supersound process 5 minutes adds 1mol/L sodium hydroxide solution 0.3ml, stirs evenly, move in the separatory funnel, extract 3 times (30ml, 20ml, 20ml) with the chloroform jolting, merge chloroform liquid, put evaporate to dryness in the water-bath, residue adds methanol 1ml makes dissolving, as need testing solution.

Other gets Radix Sophorae Flavescentis control medicinal material 1g, adds chloroform 25ml, strong ammonia solution 0.5ml, close plug, and jolting was constantly placed 24 hours, filtered, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.It is an amount of to get the matrine reference substance again, adds dehydrated alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution.

Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-acetone-ethyl acetate-strong ammonia solution (2: 3: 4: 0.2) be developing solvent, launch, take out, dry, spray is with the bismuth potassium iodide test solution of new system, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the same color speckle.

(4) acidity detects: get each 1.0 gram of gel of embodiment 1 and 2 preparations, add water 50ml respectively, after stirring makes dispersion, room temperature was placed 30 minutes, supersound process 5 minutes is measured (an appendix VI of Chinese Pharmacopoeia version in 2005 H) in accordance with the law, and pH value is respectively 5.1 and 5.3.

Embodiment 5

Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D).

The preparation of reference substance solution: precision takes by weighing through 12 hours matrine reference substance of phosphorus pentoxide drying under reduced pressure, oxymatrine reference substance an amount of, adds mobile phase and makes the solution that every 1ml contains matrine, each 40 μ g of oxymatrine respectively, promptly.

The preparation of need testing solution: get the content of the gel of embodiment 1 preparation, put in the mortar, fully grind well, the accurate title, decided 2.120g, puts in the 100ml beaker, adds water 50ml, stir evenly, room temperature was placed 30 minutes, supersound process (power 300W, frequency 25KHz) 5 minutes, add 1mol/L sodium hydroxide solution 0.3ml, stir evenly, move in the separatory funnel, with 3 (40ml of chloroform extraction, 30ml, 30ml), merge chloroform liquid, evaporate to dryness, residue adds methanol 10ml makes dissolving, move in the 50ml measuring bottle, and add mobile phase and be diluted to scale, as need testing solution A (mensuration oxymatrine); Precision is measured 5ml need testing solution A, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, as need testing solution B (mensuration matrine).

Accurate respectively reference substance solution and each the 10 μ l of need testing solution A, B of drawing of algoscopy inject chromatograph of liquid, measure;

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; Acetonitrile-0.1% trifluoroacetic acid solution (3.5: 96.5) is a mobile phase; The detection wavelength is 220nm.Number of theoretical plate is by matrine 2400.The results are shown in Figure 2 and Fig. 3.

Experimental example

Test material:

1. experimental animal:

50 of Cavia porcelluss, body weight 250-300g, female, animal housing of Jiangxi Medical College provides.The quality certification number: 021-9602.

Mice Kunming kind 18-22g, ♀ ♂ dual-purpose, the Jiangxi College of Traditional Chinese Medicine Experimental Animal Center provides, the quality certification number: 021-9601.

Rat SD is female, and body weight is that animal housing of Jiangxi Medical College provides about 200-280 gram, the quality certification number: 021-9602.

Rabbit: the male and female dual-purpose, body weight (2.5 ± 0.2) kg, animal housing of Jiangxi Medical College provides, the quality certification number: 021-9603.

2. medicine and reagent:

(1) gel of Chinese medicine composition of the present invention (being called for short the Chinese toon breast), specification: 21.83g crude drug/ml, lot number: 020424; Provide by Xiangye No. 2 Hospital of Central South University clinical pharmacy research department.

Excipient: 0.4% sodium carboxymethyl cellulose, face with preceding and be made into 0.4% sodium carboxymethyl cellulose with distilled water, provide by Xiangye No. 2 Hospital of Central South University clinical pharmacy research department.

The dosage of Cavia porcellus is:

The 10ml Chinese toon breast stock solution of Chinese toon breast high dose: embodiment 1 and 2 preparations adds substrate (0.4% sodium carboxymethyl cellulose) and gets final product to 20ml, and concentration is 10.92g crude drug/ml; Chinese toon Ruzhong dosage: 5ml Chinese toon breast stock solution adds substrate and gets final product to 20ml, and concentration is 5.46g crude drug/ml; Chinese toon breast low dosage: 2.5ml Chinese toon breast stock solution adds substrate and gets final product to 20ml, and concentration is 2.73g crude drug/ml;

Mice with dosage is:

Chinese toon breast high dose: 10ml Chinese toon breast stock solution adds substrate and gets final product to 20ml, and concentration is 10.92g crude drug/ml; Chinese toon Ruzhong dosage: 5ml Chinese toon breast stock solution adds substrate and gets final product to 20ml, and concentration is 5.46g crude drug/ml; Chinese toon breast low dosage: 2.5ml Chinese toon breast stock solution adds substrate and gets final product to 20ml, and concentration is 2.73g crude drug/ml;

Rat with dosage is:

Chinese toon breast high dose: 20ml Chinese toon breast stock solution adds substrate and gets final product to 40ml, and concentration is 10.92g crude drug/ml; Chinese toon Ruzhong dosage: 10ml Chinese toon breast stock solution adds substrate and gets final product to 40ml, and concentration is 5.46g crude drug/ml; Chinese toon breast low dosage: 5ml Chinese toon breast stock solution adds substrate and gets final product to 40ml, and concentration is 2.73g crude drug/ml;

The rabbit dosage is:

Chinese toon breast high dose: 50ml Chinese toon breast stock solution adds substrate and gets final product to 100ml, and concentration is 10.92g crude drug/ml; Chinese toon Ruzhong dosage: 25ml Chinese toon breast stock solution adds substrate and gets final product to 100ml, and concentration is 5.46g crude drug/ml; The low high dose of Chinese toon breast: 12.5ml Chinese toon breast stock solution adds substrate and gets final product to 100ml, and concentration is 2.73g crude drug/ml.

(2) glacial acetic acid: 500ml/ bottle, AR, Zhuzhou nine divisions of China in remote antiquity chemical reagent factory, lot number: 990114

0.6% acetic acid: get the 0.6ml glacial acetic acid and add normal saline to 100ml,

(3) dolantin: specification: 50mg * 1ml, Qinghai Pharmaceutic Plant, lot number: 010408, get 1 of dolantin, add substrate to 50ml, be 1mg/ml, 20mg/Kg;

(4) FUFANG HUANGSONG XIYE: Yuan An Tang Pharmaceutical Co., Ltd., Guangxi, the 250ml/ bottle, washout usefulness, lot number: 200204162, use its stock solution during use;

(5) histamine phosphate: 5 gram/bottles, Chinese Academy of Sciences Shanghai east wind Biochem Technology, INC. of biochemical institute, lot number: 9303044; The time spent precision takes by weighing histamine phosphate 0.05g, and adding distil water is made into 0.1% histamine phosphate 50ml to 50ml;

Get 0.1% histamine phosphate 1,2,3...8,9ml, adding distil water are the 0.01%-0.09% histamine phosphate to 10ml.

(6) dimethylbenzene, CA, Hubei University chemical plant, lot number: 960818

(7) prednisone: 5mg * 100 slice, Xianju, Zhejiang pharmaceutical Co. Ltd produces, lot number 020502;

(8) golden Portugal bacterium liquid: derive from Jiangxi Province healthcare hospital for women ﹠ children, patient's vaginal secretions is cultivated recently, faces with before being made into 10 ⁸Individual/L;

(9) Candida albicans: derive from the women and children of Jiangxi Province and protect establishment separation recently from vaginal secretions, face with before provoking a little lawn and inoculate;

(10) staphylococcus epidermidis, colon bacillus, Bacillus proteus, bacillus pyocyaneus, group B streptococcus, gonococcus: all derive from the women and children of Jiangxi Province and protect establishment separation recently from vaginal secretions;

(11) the phenol red meat soup prescription of glucose: meat soup 100ml (PH7.6); Glucose 1g; 1% phenol red solution 0.5ml is used for golden Portugal bacterium, form staph, colon bacillus, Bacillus proteus and bacillus pyocyaneus and cultivates;

(12) serum broth prescription: meat soup 100ml (PH7.6); Glucose 1g; 1% phenol red solution 0.5ml, the aseptic serum 1ml of rabbit is used for group B streptococcus and gonococcus and cultivates;

(13) the husky Ruo Shi liquid culture based formulas of protecting: peptone 1g, glucose 4g, distilled water 100ml is used for the cultivation of Candida albicans.

3. administration capacity: the rabbit vagina administration is 0.5ml/Kg; Cavia porcellus instep coating capacity is every of 0.2ml/; Mouse vagina administration capacity is every of 0.2ml/, and rat vagina administration capacity is every of 0.2ml/.

4. instrument and equipment

722 spectrophotometers: Shanghai the 3rd analytical tool factory;

The OLYMPUS microscope: Taiwan produces;

The digital display thermostat water bath, Jintan City, Jiangsu Medical Instruments factory;

PYX-DSH-40 * 50 type water isolation type electro-heating standing-temperature cultivators, the Shanghai medical apparatus and instruments factory of making a leapleap forward;

The portable pressure steam sterilizer of YXQ.SG41.020 type electric heating, Shanghai Medical Nuclear Instrument Factory;

SWCG-1C type clean bench, SuZhou Antai Air Tech Co., Ltd. of Su Jing group.

5. statistical disposition

Experimental data is carried out statistical disposition with the SPSS system, checks (normal state) according to the distribution situation of data with t, or t ' (abnormal) check, to draw corresponding conclusion.

Test method and result:

One, Chinese medicine composition gel of the present invention (Chinese toon breast) is to the influence of Cavia porcellus itch-threshold due to the histamine phosphate

Test method and result:

1. animal grouping: be divided into 5 groups at random, every group 10, be Chinese toon curdling colloid low dose group (2.73g crude drug/ml), middle dosage group (5.46g crude drug/ml), high dose group (10.92g crude drug/ml), FUFANG HUANGSONG XIYE group (stock solution), and matrix group (0.4% sodium carboxymethyl cellulose).

2. matched group Cavia porcellus, FUFANG HUANGSONG XIYE group Cavia porcellus, the right back instep of the Chinese toon basic, normal, high dosage group Cavia porcellus of breast are shaved hair and are coated with relative medicine 1 time, shave gross area and be about 1cm2, the coating capacity is about 0.2ml, be coated in the skin of unhairing surface equably with the mouse stomach syringe needle, with gauze parcel immobilization with adhesive tape contact 4 hours; Matched group, FUFANG HUANGSONG XIYE group, the right back instep of the Chinese toon basic, normal, high dosage group Cavia porcellus of breast all abraded with coarse sandpaper in second day, the degree of being with the oozing of blood (is wiped the wounded and is not participated in grouping, do not know the group of Cavia porcellus), and be coated with relative medicine 1 time, the coating capacity is about 0.2ml, is coated in the skin of unhairing surface equably with the mouse stomach syringe needle; After 30 minutes, clean wound surface, dry the affected part gently with gauze, with 0.01%, 0.02%, the 0.03%... progressive concentration, every three minutes once, be 0.05ml/ only, when Cavia porcellus occurring and lick metapedes at every turn, the histamine phosphate total amount that record gives is an itch-threshold, and each is organized the Cavia porcellus itch-threshold and sees the following form:

Table 1: a thousand pieces of gold Chinese toon curdling colloid is to the influence of histamine phosphate induced mice itch-threshold

Adopt the t check, compare with matrix group, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

As can be seen from the above table, except that Chinese toon curdling colloid low dose group, the Chinese toon curdling colloid of middle and high dosage group and FUFANG HUANGSONG XIYE group all can improve Cavia porcellus itch-threshold due to the histamine phosphate, illustrate that this medicine has tangible itching-relieving action.

Two, to the influence (writhing method) of mice analgesic activity:

1. animal grouping: animal is divided into 6 groups at random, and 10 every group, i.e. the basic, normal, high dosage group of Chinese toon curdling colloid, dolantin group, model group and matched group.

2. observation index: model group and matched group give 0.4% sodium carboxymethyl cellulose, and other are respectively organized equal vagina and give relative medicine 0.2ml, seal vaginal orifice with vaseline; After the administration 60 minutes, remove the matched group intraperitoneal injection of saline, the equal lumbar injection 0.6% acetic acid 0.2ml of each Mus observes that writhing response mice number of elements (abdominal part indent, stretching, extension hind leg, buttocks are raised) and number of times appear in each group in 10 minutes, the results are shown in following table.

Table 2 a thousand pieces of gold Chinese toon curdling colloid is to the influence of mice analgesic activity

The result adopts the t check, compares with matrix group, ^△P＜0.001; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01, ^{▲ ▲ ▲}P＜0.001.

As can be seen from the above table, lumbar injection 0.6% glacial acetic acid can cause mice and pain reaction occurs.Model group mouse writhing reaction times showed increased, illustrate that modeling is successful, remove the dolantin group and can obviously reduce mouse writhing reaction times and number of elements, the basic, normal, high dosage group of Chinese toon breast all can not obviously reduce mouse writhing reaction number of elements and number of times, illustrates that its analgesic effect is not obvious.

Three, to the influence (hot plate method) of mice analgesic activity:

1. animal grouping: animal is divided into 5 groups at random, and 14 every group, i.e. the basic, normal, high dosage group of Chinese toon curdling colloid, dolantin group, matched group (matrix group).

2. observation index: 90 of female mices, water bath with thermostatic control is adjusted to 55 ± 0.5 ℃, the contact water surface at the bottom of the metal dish, the heating back is as thermostimulation, certainly drop into hot plate to the pain threshold of the time of metapedes occurring licking with stopwatch record mice as this Mus, is qualified with pain threshold in second at 5-30, sieve totally 70 of mices, carry out random packet by its pain threshold, it is the same to divide into groups, and matched group gives 0.4% sodium carboxymethyl cellulose, and other are respectively organized equal vagina and give relative medicine 0.2ml, seal vaginal orifice with vaseline, be administered once every day, for three days on end, and after the last administration 30,60,120, surveyed the mice pain threshold in 240 minutes respectively, calculate its threshold of pain and improve percentage rate, the formula calculating down of threshold of pain raising rate.The results are shown in Table 3, table 4 and Fig. 1.

Table 3: a thousand pieces of gold Chinese toon curdling colloid causes the influence of pain effect to the mice hot plate method

Compare with the normal saline group, ^△P＜0.05, ^{△ △}P＜0.01.

Table 4: a thousand pieces of gold Chinese toon curdling colloid is proposed the influence of rate to the mice hot plate method threshold of pain

Compare with matrix group, ^△P＜0.05, ^{△ △}P＜0.01.

From table 3,4 and Fig. 1 can find out, the dolantin group just can obviously improve the mice pain threshold in 30 minutes after administration, in the time of 60,90,120 minutes, act on obviously, and Chinese toon breast various dose group does not have tangible raising effect to the mice pain threshold, illustrates that the analgesic activity of Chinese toon breast is not obvious yet.

Four, to the influence (auricle edema method) of mouse corrosion disease effect

1. animal grouping: animal is divided into 6 groups at random, and 10 every group, i.e. the basic, normal, high dosage group of Chinese toon curdling colloid, prednisone group, FUFANG HUANGSONG XIYE group and matched group.

2. observation index: matched group gives 0.4% sodium carboxymethyl cellulose, and other are respectively organized equal vagina and give relative medicine 0.2ml, seal vaginal orifice with vaseline; Left auricle only is coated with 100% dimethylbenzene 0.02ml/ after 30 minutes, after 4 hours, mice is pulled cervical vertebra put to death, and cuts two ears along the auricle baseline, lays round auricle at same position respectively with the straight card punch of 8mm, and torsion balance is weighed; Every Mus left side auricle deducts the auris dextra sheet and heavily is the swelling degree.The results are shown in following table.

Table 5 a thousand pieces of gold Chinese toon curdling colloid is to the influence of mice auricle swelling

The result adopts the t check, compares with the normal saline group, ^△P＜0.001; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01, ^{▲ ▲ ▲}P＜0.001.

As can be seen from the above table, except that having obviously, prednisone group mice auricle swelling degree alleviates, other differentiations of respectively organizing mice auricle swelling degree and matrix group are not obvious, illustrate that basic, normal, high three dosage groups of Chinese toon breast and FUFANG HUANGSONG XIYE group xylol induced mice auricle inflammation do not have the obvious suppression effect.

Five, to the influence of rat bacterial vaginitis due to the golden Portugal bacterium

Test objective: add the golden Portugal of inoculation bacterium by the injury rats vaginal wall and cause the scorching model of rat vagina, give medicine, observe medicine to integral status, vaginal congest edema, vaginal smear examination leukocyte and the antibacterial of animal, and the change of pathological examination results, weigh the therapeutical effect of medicine bacterial vaginitis.

1. animal grouping: animal is divided into 6 groups at random, and 10 every group, i.e. the basic, normal, high dosage group of Chinese toon curdling colloid, FUFANG HUANGSONG XIYE group, model group and normal control group.

2. observation index: each organize rat all with No. 7 blunt syringe needles on vaginal wall from inside to outside every Mus draw 30 times, by one man operation, to guarantee the dynamics uniformity, except that matched group, other each groups are all injected the 108/L gold bacterium liquid 0.2ml of Portugal, seal vaginal orifice with vaseline; Saw all under all the other each arrangements of mirrors that obvious WBC increased in second day except that matched group, a large amount of golden Portugals bacterium is arranged in the smear, vaginal secretions increases, vaginal mucosa hyperemia, swelling, surface coverage adularescent thing; Rose in the 3rd day, matched group gives 0.4% sodium carboxymethyl cellulose, and other are respectively organized equal vagina and give relative medicine 0.2ml, seal vaginal orifice with vaseline; Integrality, the vagina of each treated animal of perusal every day have or not congested swelling, and inflammatory cell and antibacterial in its vaginal secretions of plate coating checking, and the vaginal secretions smear is made in administration in the 7th day after 5 hours, see WBC and golden Portugal bacterium; Take off cervical vertebra and put to death rat, take out its vagina, observe its vaginal mucosa and have or not hyperemia, edema situation, again vagina is put into the BolinShi fixative, send and do check pathological section;

3, the therapeutic evaluation of formulation vaginitis is criterion with the pathological changes gross examination of skeletal muscle of vagina tissue, sees the following form.

Table 6 gold Portugal bacterium property vaginitis rat standards of grading

4, result:

(1) overview: occur the death except that 1 routine rat after the experimental rat moulding, all the other states still can, diet and drain normally, hair color is smooth.

(2) vagina local patholoic change perusal: significantly congested, swelling appear in 1-3 days vaginas after the rat moulding, and secretions rolls up, a large amount of white viscous secretions of surface coverage.At 1-3 days vaginal secretions smears, as seen a large amount of inflammatory cells and golden Portugal bacterium are arranged, comprehensive scores does not have notable difference between each group of modelling.In modeling beginning in the 3rd day administration, dosage group big or middle is in administration beginning in the 3rd day, vagina contrafluxion, edema, secretions, mucosal erosion alleviate, slight congested swelling was arranged in 4-5 days, all the other all recover normally the 7th day vagina pathological changes except that individually mild hyperaemia swelling being arranged, and comprehensive scores and model group, vehicle group relatively have utmost point significant difference.Small dose group and positive drug group pathological changes in the time of administration 4-5 days also obviously alleviates, at the 7th day, and most basic the recoveries normally of vagina local patholoic change, comprehensive scores and model group, normal control group more also have the inspection significant difference, specifically see Table 7.

(3) vaginal smear examination: the vaginal secretions smear all is positive behind each assembly molding, as seen a large amount of golden Portugal bacterium is arranged and come off epithelial cell and inflammatory cell.After the administration the 7th day, as seen model group 10 examples, normal control group 0 routine smear had golden Portugal bacterium and a large amount of inflammatory cell and the necrosis epithelial cell that comes off.As seen the administration group has the golden Portugal bacterium except that Chinese toon curdling glue small dose group 3 example and positive drug, and the big-and-middle dosage group of Chinese toon curdling glue only has 1 and 2 examples that golden Portugal bacterium is arranged, rarely seen epithelial cell that a small amount of inflammatory cell is arranged and come off, and concrete condition sees Table 8.

(4) histological examination: each layer tissue structure of matrix group is normal, and epithelium does not have breakage, erosion, and mucosa is not seen hyperemia and edema, tela submucosa and flesh layer no abnormality seen; Obvious congestion and edema appears in model group vaginal mucosa lower floor and flesh layer, epithelium visible part incompleteness; Low dose group mucosa and tela submucosa congestion and edema alleviate, and 1 routine epithelium incompleteness is arranged; The middle visible congested swelling of dosage group part rat vagina mucosa, the portion of tissue structure recovery is normal; High dose group rat part vaginal mucosa has the mild hyperaemia edema, and it is normal that the portion of tissue structure is recovered fully; Positive drug group rat part vaginal mucosa has the mild hyperaemia edema, and it is normal that the portion of tissue structure is recovered fully.

Table 7 Chinese toon curdling colloid is to the influence of golden Portugal bacterium property vaginitis pathological changes comprehensive scores (X ± SD)

The result adopts the t check, compares with the normal control group, ^△P＜0.05, ^{△ △}P＜0.01, ^{△ △ △}P＜0.001; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01, ^{▲ ▲ ▲}P＜0.001.

The influence of table 8 pair vaginal secretions smear (WBC and golden Portugal bacterium)

The result adopts X 2 test, compares with the normal saline group, ^△P＜0.05, ^{△ △}P＜0.01, ^{△ △ △}P＜0.001; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01, ^{▲ ▲ ▲}P＜0.001.

Six, to the influence [5] of rabbit colpitis mycotica due to the Candida albicans

Test objective: add the inoculation Candida albicans by damage rabbit vagina wall and cause rabbit colpitis mycotica model, give medicine, observe medicine to integral status, vaginal congest edema, vaginal smear examination leukocyte and the antibacterial of animal, and the change of pathological examination results, weigh the therapeutical effect of medicine colpitis mycotica.

Test method and result:

1. animal grouping: animal is divided into 7 groups at random, and 8 every group, i.e. the basic, normal, high dosage group of Chinese toon curdling colloid, FUFANG HUANGSONG XIYE group, model group and normal control group and blank group (cut is not inoculated mycete).

2. observation index: each organizes rabbit all with Inoculating needle cut gently on the rabbit vaginal wall, damage being arranged but not hemorrhage be degree, and strokes 30 times, provoke a little Candida albicans with inoculating loop and be inoculated in injury region, wherein a blank cut is not inoculated Candida albicans; Rose in second day except that the blank group, see all under all the other each arrangements of mirrors that obvious WBC increases, a large amount of Candida albicans are arranged in the smear, vaginal secretions increases, vaginal mucosa hyperemia, edema, surface coverage adularescent thing; Wherein the blank group also has the vagina mild hyperaemia, in the smear leukocyte is arranged also but adularescent candidiasis not; Rose in the 3rd day, blank group and model group give 0.9% normal saline, and other are respectively organized equal vagina and give relative medicine 0.2ml, seal vaginal orifice with vaseline; Integrality, the vagina of observing each treated animal every day have or not congested swelling, and inflammatory cell and mycete in its vaginal secretions of plate coating checking, and the vaginal secretions smear is made in administration in the 6th day after 5 hours, see WBC and Candida albicans; Air embolism is put to death rat, takes out its vagina, observes its vaginal mucosa and has or not hyperemia, edema situation, vagina is put into the BolinShi fixative again, send and does check pathological section;

Table 9 mouldiness vaginitis rabbit standards of grading

4, result:

(1) overview: except that blank is shone the group, all the other respectively organize relatively poor after the experimental rabbit moulding, and diet and drainage are normal, can recover normal after the administration.

(2) vagina local patholoic change perusal: significantly congested, swelling appear in 1-3 days vaginas after the experimental rabbit moulding, and secretions rolls up, and a large amount of white viscous secretions of surface coverage are the bean curd slag specimen.At 1-3 days vaginal secretions smears, as seen a large amount of inflammatory cells and Candida albicans are arranged, comprehensive scores does not have notable difference between each group of modelling.And blank group (cut is not inoculated the mycete group) vaginal congest swelling in the time of the 3rd day disappears substantially, illustrates that not inoculating the slight damage of mycete person can oneself recover, and a series of performances that the modeling group occurs are because due to the fungal infection.In modeling beginning in the 3rd day administration, high, medium and low dosage group and positive drug group are in administration beginning in second day, vagina contrafluxion, edema alleviate, secretions reduces, slight congested swelling was arranged in 4-5 days, all the other all recover normally the 6th day vagina pathological changes except that individually mild hyperaemia swelling being arranged, and comprehensive scores and model group, matrix group relatively have utmost point significant difference, and concrete condition sees Table 10.

(3) vaginal smear examination: the vaginal secretions smear all is positive behind each assembly molding, as seen a large amount of Candida albicans is arranged and come off epithelial cell and inflammatory cell.After the administration the 6th day, model group 8 examples, matrix group 8 examples, the visible adularescent candidiasis of normal control group 0 routine smear and more inflammatory cell.The administration group is except that Chinese toon curdling glue low dose group 2 example and positive drug 2 examples as seen the adularescent candidiasises, and Chinese toon curdling glue senior middle school dosage group only has 1 routine adularescent candidiasis, and rarely seen have a small amount of inflammatory cell, and concrete condition sees Table 11.

(4) histological examination: model group and vehicle group vaginal mucosa surface have come off, irregular, cell infiltration under each layer congestion and edema, the mucosa, intracavity is full of inflammatory cell and Candida albicans.Remove low dosage and organize tela submucosa mild hyperaemia, edema, outside a small amount of inflammatory cell, other vaginal mucosa epithelial cells and normal vagina tissue indistinction (pathological replacement and photo are attached).

Table 10 Chinese toon curdling colloid is to the influence of candida albicans vaginitis pathological changes comprehensive scores (X ± SD)

Table 11 pair vagina Candida albicans and leukocytic influence

Seven, extracorporeal bacteria inhibitor test [7]

Test objective: adopt the certain culture base to isolating vaginal bacteria or mycete are cultivated the external bacteriostasis of observation medicine recently.

(1) preparation of the selection of the selection of pathogenic bacterium culture medium and bacterium liquid and bacterium liquid

1, beta hemolytic streptococcus is with 1% serum (fresh defiber rabbit anteserum) broth bouillon, and the secondary experimental bacteria that goes down to posterity inclined-plane (blood agar) is evenly scraped and got the lawn subcultivation in the 1ml serum broth with connecing collarium, puts calorstat and cultivates 18 hours for 37 ℃.The reuse serum broth was by 1: 10 diluted for use.

2, the Candida albicans fresh inclined-plane of secondary that goes down to posterity is inoculated in the husky Ruo Shi of guarantor family name culture medium, placed 37 ℃ of constant temperature culture 6 hours, and is standby.

3, other bacterium is all used the phenol red nutrient broth medium of glucose, the experimental bacteria inclined-plane evenly scraped with inoculating loop gets the lawn subcultivation and cultivated 6 hours for mid-37 ℃ in the phenol red nutrient broth medium of glucose, more than except that Hemolytic streptococcus the reuse nutrient broth press 1: 1000 diluted for use.

(two `) is subjected to reagent to adopt test tube doubling dilution [8] [9] to measure medicine minimum inhibitory concentration (MIC)

(1: 4-1: 1024) each strain bacterium is all cooked parallel test respectively to be subjected to the reagent thing with corresponding culture medium by the multiple dilution, every pipe 1ml inoculation test bacterium liquid 0.1ml, fully the rearmounted 37 ℃ of incubator 18h of mixing take out, observe the bacterial growth situation, do sterility test again, every effective inoculating loop get culture fluid be inoculated in put on Nutrient agar (blood, the Sha Shi) flat board 37 ℃ cultivate 18h after this pipe antibacterial of no bacterial growth decidable all killed by medicine, this manages drug level as the minimum bactericidal concentration (MBC) of this medicine to this bacterium.

The result: Chinese toon curdling glue is 1.36g crude drug/ml to golden Portugal bacterium, form staph, escherichia coli, Candida albicans MIC; To Bacillus proteus MIC is 5.46g crude drug/ml; Bacillus pyocyaneus there is not effect; To beta hemolytic streptococcus MIC is 2.73g crude drug/ml; Substrate (0.4% sodium carboxymethyl cellulose) does not have effect to bacterial growth; FUFANG HUANGSONG XIYE is 1: 32 stock solution to the MIC of golden Portugal bacterium; His-and-hers watches Portugal, the white MIC that reads are 1: 16 stock solution; To large intestine, be deformed into 1: 16 stock solution, be 1: 4 to the MIC of beta hemolytic streptococcus.

Table 12 Chinese toon curdling glue is to the result of 7 kinds of external bacteriostasis of bacterial strain

Annotate: ^*The aseptic medicine contrast of medicine is arranged, ^*There is bacterium not have the antibacterial contrast (as follows) of medicine

-expression asepsis growth ,+expression has bacteria growing

Table 13 FUFANG HUANGSONG XIYE is to the result of 7 kinds of external bacteriostasis of bacterial strain

-expression asepsis growth ,+expression has bacteria growing

Conclusion

1, (5.46g crude drug/ml, 10.92g crude drug/ml) have the effect of rising for the Cavia porcellus itch-threshold due to the histamine phosphate to a thousand pieces of gold Chinese toon curdling colloid, illustrate that it has itching-relieving action preferably;

2, each dosage group of a thousand pieces of gold Chinese toon curdling colloid is for the mice threshold of pain due to thermostimulation (hot plate) and the chemical stimulation (glacial acetic acid) with turn round that the body number of times not have to reduce or the effect of reduction, and analgesic activity was not obvious when its vagina administration was described;

3, the basic, normal, high dosage group of a thousand pieces of gold Chinese toon curdling colloid does not have the obvious suppression effect for the mice auricle swelling rate due to the dimethylbenzene, illustrates that its antiinflammatory action is not obvious;

4, a thousand pieces of gold Chinese toon curdling colloid low (2.73g crude drug/ml), in ((10.92g crude drug/ml) has the obvious suppression effect for the rat vagina inflammation due to the golden Portugal bacterium for 5.46g crude drug/ml), high dose medicament, can obviously alleviate rat vagina hyperemia, swelling due to the golden Portugal bacterium, can reduce or eliminate vagina partial golden Portugal bacterium and inflammatory cell, and can improve the vaginitis rat vagina pathological state due to the golden Portugal bacterium;

5, a thousand pieces of gold Chinese toon curdling colloid low (2.73g crude drug/ml), in ((10.92g crude drug/ml) has the obvious suppression effect for the rabbit vagina inflammation due to the Candida albicans for 5.46g crude drug/ml), high dose medicament, can obviously alleviate rabbit vagina hyperemia, swelling due to the Candida albicans, can reduce or eliminate vagina partial golden Portugal bacterium and inflammatory cell, can improve the vaginitis rabbit vagina pathological state due to the Candida albicans;

6, a thousand pieces of gold Chinese toon curdling colloid has the good in-vitro inhibitory action in Candida albicans, beta hemolytic streptococcus, golden Portugal bacterium, form staph, escherichia coli, Bacillus proteus.

Claims

1. Chinese medicine composition for the treatment of cervical disease contains the component of following weight portion: Cortex Ailanthi: 35-45; Radix Sophorae Flavescentis: 25-35; Cortex Moutan: 25-35; Olibanum: 25-35; Borneolum Syntheticum: 0.8-1.2, wherein Radix Sophorae Flavescentis and Cortex Moutan, Olibanum equivalent.

2. Chinese medicine composition as claimed in claim 1 is characterized in that, contains the component of following weight portion: Cortex Ailanthi: 40; Radix Sophorae Flavescentis: 30; Cortex Moutan: 30; Olibanum: 30; Borneolum Syntheticum: 1.

3. Chinese medicine composition as claimed in claim 1 or 2 is characterized in that, it is a gel.

4. the preparation method of the gel of a Chinese medicine composition as claimed in claim 3 comprises:

1) Cortex Ailanthi adds 2-9 times of water gaging, decocts with water 1-3 time, decocts 1-2.5 hour at every turn, and collecting decoction filters, and it is 1.05～1.10 Cortex Ailanthi clear paste that filtrate decompression is concentrated into relative density;

2) Radix Sophorae Flavescentis adds 6-12 times of water gaging, decocts with water 1-3 time, soaks 0.5-1.5 hour before decocting for the first time, the each decoction 0.5-3 hour, collecting decoction filters, filtrate decompression is concentrated into the Radix Sophorae Flavescentis clear paste of relative density 1.10～1.15, put coldly, add ethanol and make and contain alcohol amount and reach 60-70%, left standstill 8-24 hour, filter, filtrate recycling ethanol merges with the Cortex Ailanthi clear paste to there not being the alcohol flavor, is concentrated into the clear paste of relative density 1.05～1.10;

5. preparation method as claimed in claim 4, it is characterized in that, the consumption of sodium carboxymethyl cellulose, medicinal glycerin is respectively the 0.6-1.5 weight portion of used Borneolum Syntheticum weight, and the consumption of ethyl hydroxybenzoate and Tween-80 is respectively the 0.01-0.1 weight portion of used Borneolum Syntheticum weight; The gained gel is the 0.1-0.2 weight portion of used raw material of Chinese medicine medicine gross weight, preferred 0.15 weight portion.

6. the detection method as the arbitrary described Chinese medicine composition of claim 1-3 comprises discrimination method, and one or more methods were differentiated below described discrimination method comprised employing:

(1) get the 1g sample, add water 30ml, fully stir, room temperature was placed 30 minutes, supersound process 5 minutes, and the dislocation separatory funnel adds 10-30ml ether extraction 1-3 time, merges ether solution, and room temperature volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the paeonol reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that 10: 0.5 petroleum ether-ethyl acetate is developing solvent, launch, take out, dry, spray is with the acid 5% ferric chloride alcoholic solution of hydrochloric acid, and 105 ℃ to be heated to the speckle colour developing clear;

(2) according to discrimination method (1) preparation sample solution, get 0.5ml, add methanol to 5ml as need testing solution; Other gets the Borneolum Syntheticum reference substance, adds methanol and makes the solution that every 1ml contains 5mg, in contrast product solution; Test according to thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that petroleum ether-chloroform-ethyl acetate of 10: 3: 1 is developing solvent, launch, take out, dry, spray develops the color with 5% phosphomolybdic acid ethanol test solution, puts 105 ℃ and is heated to clear spot;

(3) get the 1g sample, add water 30ml, stir evenly, placed 30 minutes, supersound process 5 minutes adds 1mol/L sodium hydroxide solution 0.3ml, stirs evenly, move in the separatory funnel, with 10-30ml chloroform extraction 1-3 time, merge chloroform liquid, put evaporate to dryness in the water-bath, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Radix Sophorae Flavescentis control medicinal material 1g, adds chloroform 25ml, strong ammonia solution 0.5ml, close plug, and jolting was constantly placed 24 hours, filtered, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with volume ratio is 2: 3: 4: benzene-acetone of 0.2-ethyl acetate-strong ammonia solution is developing solvent, launch, take out, dry, spray is with the bismuth potassium iodide test solution of new system, and it is clear to be heated to the speckle colour developing;

(4) acidity is differentiated: get 1.0 gram samples, add water 50ml, after stirring made dispersion, room temperature was placed 30 minutes, supersound process 5 minutes, and measuring pH value is 4.5～6.0.

7. detection method as claimed in claim 6 is characterized in that, in the described authentication method (3), the reference substance of Radix Sophorae Flavescentis also can be the matrine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution; Or select for use Radix Sophorae Flavescentis control medicinal material and matrine to make control medicinal material solution and reference substance solution simultaneously, carry out thin layer chromatography and detect.

8. detection method as claimed in claim 6 is characterized in that, also comprises content assaying method, and described content assaying method is the content that adopts high effective liquid chromatography for measuring Radix Sophorae Flavescentis active component.

9. detection method as claimed in claim 8 is characterized in that, comprising:

2) preparation of need testing solution: sample thief, fully grind well, the accurate title, decided 1.5-2.5g, puts in the 100ml beaker, add water 50ml, stir evenly, room temperature was placed 30 minutes, supersound process (power 300W, frequency 25KHz) 5 minutes adds 1mol/L sodium hydroxide solution 0.3ml, stir evenly, move in the separatory funnel, use 20-40ml chloroform extraction 1-3 time, merge chloroform liquid, evaporate to dryness, residue add methanol 10ml makes dissolving, move in the 50ml measuring bottle, and add mobile phase and be diluted to scale, as need testing solution A; Precision is measured A solution 5ml, puts in the 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, as need testing solution B;

Chromatographic condition is: be filler with the octadecylsilane chemically bonded silica; With volume ratio is that 3.5: 96.5 acetonitrile-0.1% trifluoroacetic acid solution is a mobile phase; The detection wavelength is 220nm, and number of theoretical plate is not less than 2000 by matrine.

10. as the arbitrary described Chinese medicine composition of claim 1-3, it is characterized in that through measuring as the arbitrary described detection method of claim 7-9, described Chinese medicine composition contains Radix Sophorae Flavescentis with C ₁₅H ₂₄N ₂O and C ₁₅H ₂₄N ₂O ₂The total amount meter, be no less than 5.0mg/g.