CN1903290B - Traditional Chinese medicine composition for treating cervicitis and its preparation method - Google Patents

Abstract

A Chinese medicine in the form of capsule, tablet, oral liquid, particle, aerosol, ointment, etc for treating morbid leukorrhea, cervicitis, cervical erosion, etc is prepared from motherwort and Guangdong beautyberry leaf. Its preparing process and quality control method are also disclosed.

Description

Chinese medicine composition of a kind for the treatment of cervicitis and preparation method thereof

Technical field

The present invention relates to a kind of Chinese medicine composition and preparation method thereof and quality determining method, particularly a kind of Chinese medicine composition for the treatment of cervicitis and preparation method thereof and quality determining method.

Background technology

This prescription be tcm clinical practice through proved recipe, always with decoction in clinical practice, for more widely-used, facilitate patients, be easy to carry, be convenient to the transportation, further reduce taking dose, carry out the development of patent medicine.Cervicitis is in clinical genus chronic disease, and easily outbreak repeatedly, thus need to select oral solid formulation, comparatively suitable.The capsule steady quality, and production technology is simple, the mechanization degree height, production cost is low, is convenient to store, transport, carry and take, and production cost is low, and the patient is acceptant, so select to make capsule.

Herba Leonuri has promoting blood flow to regulate menstruation, and the effect of inducing diuresis to remove edema is used for menoxenia, dysmenorrhea, amenorrhea, lochiorrhea, edema oliguria.Mainly contain alkaloid, flavonoid, iridoids, Diterpenes, tannin, fatty acid, Saponin, cardiotonic glycoside and volatilization wet goods composition.According to modern pharmacology research report Herba Leonuri there is excitation in the uterus, the Herba Leonuri total alkali has excitation to the guinea pig in vitro uterus.Platelet aggregation, microthrombusis, fibrinous thrombus are formed Herba Leonuri and erythrocytic aggregation all has inhibitory action, thrombus formation time is prolonged, thrombosis contraction in length, leonurine have the effect of significant blood viscosity lowering, are a kind of good drug for invigorating blood circulation and eliminating stasis.Preliminary experiment studies show that the Herba Leonuri total alkali can obviously increase the stripped mobility that reaches at the body animal uterus, and low dosage 8mg shows as the amplitude and the frequency of uterine activity and accelerates tension variation not obvious (not causing tonic spasm); Heavy dose then shows as inhibitory action.The Herba Leonuri total alkali can increase the Cervical blood flow of whole animal simultaneously.

Its extraction conditions is mainly water extract-alcohol precipitation or certain density ethanol extraction according to the literature.The total alkaloids water solublity is big in this experimental basis Herba Leonuri, easily with sour salifiable physicochemical property, determines to carry with decocting in water, be concentrated into dried, diluted acid dissolving, the further purified process route of strong acid ion exchange resin.

The chemical constituent of Callicarpa kwangtungensis Chun is not appeared in the newspapers, and the trial test result shows flavonoid, the chemical reaction of condensed tannin, phenoloid, saccharide.The pharmacological action of Callicarpa kwangtungensis Chun does not also appear in the newspapers, and generic Folium Callicarpae Formosanae, callicarpa cathayana Chang, callicarpa pedunculata, Callicarpa nudiflora (Callicarpa nudiflora Hook.et Arn.) are reported more.Folium Callicarpae Formosanae has tangible anastalsis and increased platelets counts effect, and its ethanol extract has significant analgesia role to mice.Callicarpa nudiflora can be accelerated wound healing, reduce cicatrization and have function in delaying senility, the Callicarpa nudiflora sheet has in various degree bacteriostasis to staphylococcus aureus, salmonella typhi, streptococcus pneumoniae, is pharmacological action antibiosis anti-inflammatory drug more widely.Callicarpa cathayana Chang and callicarpa pedunculata have the bacteriostasis of height to staphylococcus aureus and Salmonella, to Candida albicans, coldly hinder bacillus and shigella flexneri has stronger bacteriostasis.Jiang Hui antimony etc. has been measured Folium Callicarpae Formosanae in this platymiscium, callicarpa cathayana Chang, Pi the antioxidation in vitro effect of leaf Folium Callicarpae Formosanae (C.kochiana), Japanese Folium Callicarpae Formosanae, Folium Callicarpae Macrophyllae, Radix Callicarpae Giraldii (C.giraldii).Bibliographical information: the toxicity of callicarpa cathayana Chang, callicarpa pedunculata and Callicarpa nudiflora is less, and clinical oral administration drug safety scope is bigger.Callicarpa kwangtungensis Chun hardship, puckery, cool is returned liver, lung, stomach warp, the effect of Bearberry Extract hemostasis, heat-clearing and toxic substances removing.Clinical various hemorrhage and laryngopharynx swelling and pain, pathopyretic ulcer such as be used for spitting blood, have blood in stool.Be usually used in treating gynaecopathias such as cervical erosion is hemorrhage, vaginitis, cervicitis.Character in, the ethanol water-soluble according to flavone compound determines that precipitate with ethanol is removed impurity with decocting in water; Macroporous resin is made with extra care flavone compound, reduces dose in order to improve curative effect, so select the flavone compound in the further refining Callicarpa kwangtungensis Chun of macroporous resin.

Summary of the invention

The object of the invention is to provide the preparation method of a kind of Chinese medicine composition and preparation thereof, and another purpose of the present invention is to provide the quality determining method and the purposes of this Chinese medicinal composition preparation.

The present invention seeks to be achieved through the following technical solutions:

The crude drug of Chinese medicine composition of the present invention is composed as follows:

Herba Leonuri 7-60 weight portion Callicarpa kwangtungensis Chun 7-60 weight portion

The preferred weight proportioning of above-mentioned raw materials medicine is as follows:

Herba Leonuri 7 weight portion Callicarpa kwangtungensis Chuns 55 weight portions

The preferred weight proportioning of above-mentioned raw materials medicine is as follows:

Herba Leonuri 60 weight portion Callicarpa kwangtungensis Chuns 15 weight portions

The preferred weight proportioning of above-mentioned raw materials medicine is as follows:

Herba Leonuri 40 weight portion Callicarpa kwangtungensis Chuns 40 weight portions

Traditional Chinese medicinal composition raw materials of the present invention, add conventional adjuvant, according to common process, make clinical acceptable forms, as: capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray.

The concrete preparation technology of the present composition is as follows:

Get the Herba Leonuri medical material, add 8-12 times of water gaging, extract 2-4 time, each 1-2h, merge extractive liquid,, filter, be concentrated into the thick paste of 50 ℃ of-70 ℃ of relative density 1.00-1.15, add the 0.1mol/L-1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1-3, be diluted to the 1mL medicinal liquid with 0.1mol/L-1mol/L hydrochloric acid then and be equivalent to the 1-2g medical material, i.e. 50 ℃ of-70 ℃ of relative densities 1.00～1.20, through centrifugal or filter after, get strong acid ion exchange resin post on the supernatant, absorption 0.5-2h, it is colourless to be eluted to effluent with distilled water, with ammonia alkalization resin PH8-9, with 30-95% ammonia ethanol or 2mol ammonia eluting, make effluent pH8-12, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 8-12 times of water gaging, extract each 1-2h, merge extractive liquid, 2-4 time, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 50 ℃ of-70 ℃ of relative densities are 1.00～1.02, add 95% ethanol and make the alcohol amount of containing be 50-70%, 24h is placed in cold preservation, filters, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 50 ℃ of-70 ℃ of relative densities are 1.00～1.02, and medicinal liquid is crossed macroporous resin, absorption 0.5-2h, it is closely colourless to be eluted to effluent with distilled water, uses the 10%-80% ethanol elution, collects the 5BV eluent, reclaim ethanol, be concentrated into dried, vacuum drying, dry extract;

Dry extract is added conventional adjuvant, make capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray according to a conventional method.

The preferred for preparation technology of the present composition is as follows:

Get the Herba Leonuri medical material, add 10 times of water gagings, extract 3 times, each 1.5h, merge extractive liquid, filters, be concentrated into the thick paste of 60 ℃ of relative densities 1.05, add the 0.1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1, make the 1mL medicinal liquid be equivalent to the 1g medical material with 0.1mol/L hydrochloric acid then, promptly 60 ℃ of relative densities are 1.00～1.02, after centrifugal, get 001 * 7 type ion exchange resin column on the supernatant, absorption 1h, it is colourless to be eluted to effluent with distilled water, with ammonia alkalization resin PH8-9,75% or 95% ammonia ethanol or 2mol ammonia eluting, make effluent pH9-10, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 10 times of water gagings, extract each 1.5h, merge extractive liquid, 3 times, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.00～1.02 to add 95% ethanol and make that to contain the alcohol amount be 60% or 70%, 24h is placed in cold preservation, filter, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.00～1.02, medicinal liquid is crossed HPD 100, HPD 400 or HPD 300 macroporous resins, absorption 1h, it is closely colourless to be eluted to effluent with distilled water, with 50%, 60% or 70% ethanol elution, collect 5 times of column volume eluents, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract;

Dry extract is added conventional adjuvant, make capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray according to a conventional method.

Quality determining method of the present invention comprises following discriminating and/or assay

Discrimination method comprises one or more in the following discriminating:

1/3 (being equivalent to crude drug amount 10g) of A. getting this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, add ethanol 5ml, and supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get 1/3 (being equivalent to crude drug amount 30g) of this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, with 4: 1: 0.5 alcohol-water-glacial acetic acid was developing solvent, launch, take out, dry, spray is with the 1%AlCl3 alcoholic solution, and hot blast blows to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay in the quality determining method is as follows:

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column is with mobile phase 0.05mol/LNa ₂HPO ₄-0.05mol/L NaH ₂PO ₄, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500; The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution; The preparation of need testing solution: with 1/3 (being equivalent to crude drug amount 10g) of this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, put in the tool plug triangular flask, add ethanol 25ml, claim to decide weight, supersound extraction 30ml is put coldly, claims to decide weight again, supply the weight of being lost with ethanol, shake up, filter; Get subsequent filtrate 10mL, water bath method with 0.05mol/LNa2HPO4 10mL dissolving, adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/LNa2HPO4 solution to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; The hydrochloric stachydrine C of pharmaceutical composition per unit preparation (pressing every of capsule, conventional formulation unit's meters such as every in tablet, every bag of granule, per 10 milliliters of liquid oral) ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

Drug combination preparation Coming-of-Age Day taking dose of the present invention is equivalent to crude drug amount 30g.

The composition capsule pharmacodynamic experiment shows: composition capsule can obviously improve the cervix uteri pathology damage due to phenol and the escherichia coli, and the local inflammation reaction is eased, and alleviates the rat vagina occult blood that the phenol stimulation causes.Composition capsule can obviously reduce above-mentioned cervicitis animal pattern IL-1 β, IL-2, TNF α, the blue albumen of copper, PGE ₂, etc. with the inflammatory mediator and the cytokine content of inflammation-related. composition capsule is external to have for multiple cervicitis pathogenic bacterium and suppresses or killing action. the composition capsule xylol causes mice ear, carrageenin causes rat paw edema and the bullate formation of rat granuloma all has remarkable inhibitory action, showing that it has remarkable antiinflammatory action. composition capsule can obviously suppress acetic acid and cause the mouse writhing reaction, having analgesic activity. composition capsule can reduce the blood viscosity of the cervicitis rat model due to the escherichia coli, shows to have certain function of promoting blood circulation to disperse blood clots.

Following experiment and embodiment are used to further specify but are not limited to the present invention.

The experiment of experimental example 1 Herba Leonuri extraction process

1, water is carried the preferred of condition

Adopt the method for orthogonal test, select four factors, three horizontal quadrature test cards, with amount of water, decocting time, decoction number of times is three factors, three levels of every factor design, the percentage composition that extracts with paste-forming rate, total alkaloids is an index, adopt L9 (34) orthogonal array experiment arrangement, factor level sees Table 1

Table 1 Herba Leonuri water is carried the factor level table

Extracting method:

According to the experimental design in the orthogonal table, every part takes by weighing medical material 50g and decocts, and extracting solution filters, and concentrates, and vacuum drying is weighed, and measures content.

Table 2 Herba Leonuri total alkali orthogonal experiment preferred result

The analysis of variance table of table 3 paste-forming rate

Analysis of variance table is 3 results show, factor A, B do not have the significance influence to paste-forming rate, and factor C has significance influence (P＜0.05), and each factor is followed successively by from big to small to the influence degree of paste-forming rate: C＞B＞A;

A3＞A2 in the A factor＞A1; B3＞B2 in the B factor＞B1; C2＞C3 in the C factor＞C1.

The analysis of variance table of table 4 total alkaloids

Analysis of variance table is 4 results show: factor A, B do not have the significance influence to paste-forming rate, and factor C has significance influence (P＜0.05), and each factor is followed successively by from big to small to the influence degree of paste-forming rate: C＞A＞B; A in the A factor ₂＞A ₃＞A ₁B in the B factor ₂＞B ₃＞B ₁C in the C factor ₃＞C ₂＞C ₁.Comprehensive above two The results of analysis of variance, in conjunction with the concrete operations condition, the process route that preferred Herba Leonuri water is carried is A ₂B ₂C ₃, promptly amount of water is 10 times, decocts 1.5 hours, decocts 3 times.

2, the technical study of the refining total alkaloids of strongly acidic cation-exchange

The selection of different model cation exchange resin:

Get various resin 20ml respectively, 3 parts on every type, wet method are packed in the identical post of specification, add sample liquid 20ml (the 1ml medicinal liquid is equivalent to the 1.0g medical material), place 30min, earlier closely colourless to effluent with distilled water flushing, with the ammonia 40ml alkalization resin of 2mol, making the effluent pH value is 9-10, with ammonia ethanol (PH9-10) eluting, making elution rate is 1-2ml/min, collects coloured eluent 100ml.

Table 5 different model ion exchange resin is to the adsorbance (n=3) of total alkaloids

Experimental result shows: under the same conditions, 007 type strong acid ion exchange resin is to the adsorbance maximum of total alkaloids, so select 007 type strong acid ion exchange resin.

The selection of eluting solvent

Get strong-acid type cation exchange tree type 7 type 20ml upper props, add sample liquid 10ml (the 1ml medicinal liquid is equivalent to the 1g medical material), after placing 0.5h, it is closely colourless to be washed till effluent with distilled water, water liquid discards, with ammonia alkalization resin PH8-9, use 2mol ammonia successively, ammonia ethanol 30%, 50%, 70%, 95% eluting, making discharge rate is 1-2ml/min, collect the each several part eluent, behind the evaporate to dryness,, respectively add the 0.5g active carbon with 0.molM hydrochloric acid 10ml dissolving, behind the heating in water bath 1min, filter, respectively with the improvement bismuth potassium iodide, the silico-tungstic acid test solution, freshly prepared Reinecke salt test solution is made identification experiment.Experimental result sees Table 6.

The investigation of table 6 eluting solvent

Annotate :-represent negative ,+represent the positive

According to experimental result, selecting 95% ammonia ethanol is eluting solvent

The investigation of ammonia ethanol elution volume

Get strong-acid type cation exchange tree type 7 type 20ml upper props, add sample liquid 10ml (the 1ml medicinal liquid is equivalent to 1g medical material sample liquid), after placing 0.5h, it is closely colourless to be washed till effluent with distilled water, water liquid discards, with ammonia alkalization resin PH8-9, ammonia alcohol 95 % eluting, making discharge rate is 1-2ml/min, it is a collecting the every 25ml of eluent, collects 6 parts continuously, behind the evaporate to dryness, dissolve with 0.1M hydrochloric acid 10ml, respectively add the 0.5g active carbon, behind the heating in water bath 1min, filter, with improvement bismuth potassium iodide, silico-tungstic acid test solution, freshly prepared Reinecke salt test solution is checked respectively.Experimental result sees Table 7.

The investigation of table 7 ethanol elution volume

Experimental result shows that ammonia ethanol elution volume is 100ml, 10ml/g

The investigation of best swap time

Get 4 parts in 007 type ion exchange resin, each 20ml, during wet method is packed the identical post of specification into respectively, all add sample liquid 10ml (the 1ml medicinal liquid is equivalent to the 1g medical material), place 0.5,1,2 respectively, behind the 6h, earlier closely colourless to effluent with distilled water flushing, the ammonia alkalization resin of reuse 2mol, making the effluent pH value is 9-10, uses alkaline ethanol (pH9-10) eluting then, make elution rate 1-2ml/min, collect coloured eluent 100ml.The preparation of sample liquid and total alkaloid contents are measured the same.Experimental result sees Table 8.

The investigation of table 8 best swap time

Experimental studies results shows, time is short or oversize, all make measurement result on the low side, trace it to its cause, swap time is short, the part biological basic ion has little time with resin-bonded or combines insecure, easily be distilled water elution and go down, swap time is oversize, and part biological basic ion and resin-bonded are very firm, be difficult for being eluted, thereby the post recovery rate was reduced.Therefore, make spent ion exchange resin, it is very crucial to grasp swap time, and according to experimental studies results, selection swap time is 1h.

The mensuration of maximal absorptive capacity

Get 6 parts of 7 type strongly acidic cation-exchanges, each 20ml, wet method is packed in the identical post of specification, adds sample liquid 6ml, 8ml, 10ml, 12ml, 14ml, 16ml (the 1ml medicinal liquid is equivalent to the 1g medical material) respectively, behind the 1h, earlier closely colourless to effluent with distilled water flushing, water liquid discards, with the ammonia alkalization resin of 2mol, making the effluent pH value is 9-10, with ammonia ethanol (PH9-10) eluting, make elution rate 1-2ml/min, collect coloured eluent 100ml.

The preparation of sample liquid and total alkaloid contents are measured the same.Experimental result sees Table 9.

The investigation of table 9 maximal absorptive capacity

Through measuring, volume is that the exchange capacity of the 7 type strong-acid cation-exchange resin maximums of 20ml is 104.55mg, just the alkaloid in the commutative 0.7g medical material of 1ml resin.Be 1.4/g promptly than applied sample amount.

Brief summary

Through strongly acidic cation-exchange being exchanged the influence factor's of Herba Leonuri total alkaloids investigation, experimental result shows: the performance of 7 type strongly acidic cation-exchanges exchange Herba Leonuri total alkaloids is best, its eluting solvent is 95% ammonia ethanol, be 1h best swap time, every milliliter of maximal absorptive capacity is the 0.7g medical material, be 1.4/g than applied sample amount promptly, with this understanding, the upper prop retention rate is 98.05%.

The experiment of experimental example 2 Callicarpa kwangtungensis Chun Study on extraction process

1, the extraction process condition is preferred

For preferred Callicarpa kwangtungensis Chun extraction process condition, be three factors with amount of water, decocting time, decoction number of times, three levels of every factor design, the percentage composition that extracts with paste-forming rate, total flavones is an index, adopts L ₉(3 ⁴) the orthogonal array experiment arrangement, the results are shown in Table 10.

Table 10 total flavones water is put forward the orthogonal experiment gauge outfit

Extracting method

According to the experimental design in the orthogonal table, every part takes by weighing medical material 50g and carries out water and carry, and extracting solution filters,

Concentrate, vacuum drying is weighed.

Orthogonal optimum seeking experimental result table 11:

The Orthogonal experiment results of table 11 Callicarpa kwangtungensis Chun total flavone

The analysis of variance table of table 12 paste-forming rate

Analysis of variance table is 12 results show, factor A, B and C do not have the significance influence to paste-forming rate, and each factor is followed successively by from big to small to the influence degree of paste-forming rate: C＞B＞A; A3＞A1 in the A factor＞A2; B2＞B1 in the B factor＞B3; C3＞C2 in the C factor＞C1.

Table 13 is the analysis of variance table of index with total flavones percentage composition in medical material

Analysis of variance table is 13 results show, factor B does not have the significance influence to the total alkaloids extraction rate, and factor A, C have significance influence (P＜0.05), and each factor is followed successively by from big to small to the influence degree of paste-forming rate: C＞A＞B; A2＞A1 in the A factor＞A3; B2＞B3 in the B factor＞B1; C3＞C2 in the C factor＞C1.The The results of analysis of variance of consolidated statement 25,26 in conjunction with the concrete operations condition, determines that it is A2B2C3 that the best has extraction process, and promptly amount of water is 10 times, decocts 1.5 hours, decocts 3 times.

2, the investigation of alcohol precipitation process

The investigation of different alcohol precipitation concentrations:

The material 200g that gets it filled extracts by selected extraction process, is concentrated into 200ml, (the 1ml medicinal liquid is equivalent to the 1g medical material) is divided into four parts, every part of 50ml, and the ethanol with 95% makes and contains alcohol amount and be respectively 50%, 60%, 70%, 80%, place after 24 hours, filter, reclaim ethanol, concentrate, vacuum drying, the accurate title, decide, and preserves standby in the sealed plastic bag of packing into behind the pulverize.Get the about 1g of dry extract (parallel two parts) of each alcohol precipitation concentration, put in the apparatus,Soxhlet's, be extracted into colourlessly with 70% ethanol 80ml, and standardize solution is in the 100ml volumetric flask.Precision is drawn 5ml in the 50ml volumetric flask, the distilled water standardize solution.Content assaying method is pressed under the 3.2.1.2 item under each precipitate with ethanol condition of mensuration content of total flavone in the gained dry extract, and experimental result sees Table 14

The investigation of the different alcohol precipitation concentrations of table 14

Annotate: the percentage composition of total flavones contains the percentage composition of total flavones in every 50g medical material

The result of the percentage composition of total flavones in medical material shows that 60% precipitate with ethanol condition is better, selects 70% precipitate with ethanol condition, the total flavones of loss 0.09%, but paste-forming rate reduces by 1.54%, determines that the precipitate with ethanol condition is more reasonable for containing alcohol amount 70%.

The investigation of different concentrating degree:

Take by weighing medical material 150g, extract by selected condition, decocting liquid is merged filter, be concentrated into 150ml, be divided into 3 parts, wherein two parts are concentrated into the 1g medicinal liquid more respectively and are equivalent to 1.5,2.0 medical materials.Add 95% ethanol respectively and make that to contain alcohol amount be 70%, put and place in the refrigerator after 24 hours, filter, reclaim ethanol, concentrate, vacuum drying accurately claims to decide weight.Get the about 1g of dry extract (parallel two parts) of each alcohol precipitation concentration, the accurate title, decide.Put in the apparatus,Soxhlet's, be extracted into colourlessly with 70% ethanol 80ml, standardize solution is in the 100ml volumetric flask.Precision is drawn 5ml in the 50ml volumetric flask, the distilled water standardize solution.Content assaying method is pressed time-and-motion study under the 3.2.1.2 item.Experimental result sees Table 15.

The different concentrating degree of table 15 are to the investigation result of general flavone content influence

Annotate: the percentage composition of total flavones contains the percentage composition of total flavones in every 50g medical material

The result shows, is evaluation index with the paste-forming rate, concentrated ratio be 1: 2 o'clock minimum, but the loss total flavones more; With the content of total flavone is evaluation index, and concentrated ratio is that 1: 1 content is the highest.The ratio that select to concentrate is 1: 1.5, and its paste-forming rate is more close than 1: 1 o'clock low 1.2% and 1: 2 o'clock, and general flavone content is close with 1: 1 o'clock, so the concentrated ratio of selection is 1: 1.5.

3, the research of macroporous resin column adsorption cleaning technology

Producing of medicinal liquid:

Extract by the preferred extraction conditions in front, both amount of water was 10 times, each 1.5 hours, decoct collecting decoction 3 times, be concentrated into the 1ml medicinal liquid and be equivalent to the 1.5g medical material, add 95% ethanol and make that to contain alcohol amount be 70%, put and place in the refrigerator after 24 hours, filter, reclaim ethanol, be concentrated into and make the 1ml medicinal liquid be equivalent to the 1.5g medical material.

The macroporous resin of different model is to the investigation of Callicarpa kwangtungensis Chun total flavone adsorbance:

Measure and handle standby various macroporous resin well, every type 20ml wet method is packed in the post of same size, every post reaches excessive adsorption when adding sample liquid 15ml, closely colourless with distilled water flushing to effluent, water liquid discards, ethanol elution with 70%, regulate discharge rate 1-2ml/min, collect eluent. concentrate the eluent of collecting an amount of, dissolve with ethanol with 70%, and standardize solution is got 5ml in the 50ml volumetric flask in the 100ml volumetric flask, the distilled water standardize solution. get 5ml in the 25ml volumetric flask, press time-and-motion study general flavone content under the 3.2.1.2 item.

Table 16 different model macroporous resin is to the adsorbance of Callicarpa kwangtungensis Chun

According to experimental studies results, select HPD-300 type macroporous resin.

The investigation of macroporous resin ethanol elution concentration

4, qualitative investigation

Experimental selection water, 10% ethanol, 30% ethanol, 50% ethanol, 70% ethanol, 95% ethanol successively eluting be adsorbed on total flavones on the resin, collect each eluent, be concentrated into dried, 95% dissolve with ethanol, hydrochloric acid magnesium powder reaction, aluminum chloride ethanol liquid detect.Experimental result sees Table 17.

Table 17 ethanol elution concentration is to the qualitative examination of total flavones eluting power

Annotate :+expression positive reaction ,-expression negative reaction,

Experimental result shows: 70% ethanol can all elute total flavones.

5, quantitative expedition

Measure the macroporous resin 20ml that handles standby HPD-300 well, wet method dress post, behind distilled water flushing, last sample 10ml (1ml is equivalent to the 1.5g medical material), place 30min,, closely colourless with distilled water flushing to effluent, use 10%, 30%, 50%, 70%, 95% ethanol elution successively, make elution rate 1-2ml/min collect eluent 100ml.The eluent of collecting is concentrated in right amount, and the ethanol standardize solution with 70% is got 5ml in the 50ml volumetric flask in the 100ml volumetric flask, and distilled water is settled to scale.Get 5ml in the 25ml volumetric flask, measure general flavone content.Experimental result sees Table 18.

Table 18 ethanol elution concentration is to the quantitative check of total flavones eluting power

Determine that according to experimental studies results 70% ethanol is eluting solvent.

The investigation of macroporous resin ethanol elution volume:

Measure and handle standby macroporous resin HPD-300 type 20ml well, wet method dress post, last sample 10ml (1ml is equivalent to the 1.5g medical material), closely colourless to effluent behind the placement 30min with distilled water flushing, discard water liquid, with 70% ethanol elution 200ml eluting, make elution rate 1-2ml/min, every collection 50ml is a, collects 4 parts, altogether 200ml.The eluent collected is dissolved in the 100ml volumetric flask surely with 70% ethanol, gets 5ml in the 50ml volumetric flask, distilled water is settled to scale.Content of total flavone is measured the same, and experimental result the results are shown in Table 19.

The investigation of table 19 eluting solvent consumption

Result of study shows that 70% ethanol 100ml can elute the Callicarpa kwangtungensis Chun total flavone of 20ml macroporous resin adsorption fully, is equivalent to the ethanol elution of 5BV70%.

The investigation of adsorption time:

Measure and handle standby macroporous resin HPD-300 type 20ml well, get 4 parts, wet method dress post, last sample liquid 10ml (1ml is equivalent to the 1.5g medical material) makes and placed 0.5,1.0,2.0,6.0 hour, makes elution rate 1-2ml/min closely colourless to effluent with distilled water flushing, water elution liquid discards, and with 70% ethanol elution eluting, collects eluent 100ml respectively, water-bath concentrates an amount of, 70% ethanol standardize solution is in the 100ml volumetric flask, and precision is measured 10ml, concentrates, behind the vacuum drying, weight decided in accurate title.Get 5ml respectively in addition in the 50ml volumetric flask, distilled water is fixed molten.Content of total flavone is measured the same.Experimental result sees Table 20.

The investigation of table 20 adsorption time

Annotate: the percentage composition of total flavones contains the percentage composition of total flavones in every 15g medical material

Experimental studies results is analyzed: adsorption time is long more, and paste-forming rate is low more, and the loss of corresponding total flavones is also big more.Adsorption time is 1h, and its paste-forming rate is lower by 0.1% than 0.5, and general flavone content is very approaching, and adsorption time is long more, though its paste-forming rate reduces, the total flavones loss is bigger, so the selection adsorption time is 1h.

The investigation of maximum applied sample amount:

Measure 6 parts on the standby macroporous resin HPD-300 type of handling well, every part of 20ml, wet method dress post, go up sample 4 respectively, 6,8,10,12,14ml sample liquid (1ml is equivalent to the 1.5g medical material), all adsorb 1.0 hours after, closely colourless with distilled water flushing to effluent, water liquid discards, with 70% ethanol 100ml eluting, make elution rate 1-2ml/min, collect eluent respectively, water-bath concentrates an amount of, 70% ethanol standardize solution is in the 100ml volumetric flask, and precision is measured 10ml, concentrates, behind the vacuum drying, weight decided in accurate title. get 5ml respectively in addition in the 50ml volumetric flask, the distilled water standardize solution. and content of total flavone is measured the same, and experimental result sees Table 21.

The investigation of table 21 macroporous resin HPD-300 type maximal absorptive capacity

Annotate: the total flavones percentage composition is equivalent to contain the percentage composition of total flavones in the medical material

Experimental studies results is analyzed: along with the increase of applied sample amount, the proportional increase of macroporous resin adsorption amount, to applied sample amount be 12, during 14ml, the increase with applied sample amount does not increase, and the content of total flavones in the medical material amplitude that descended is bigger, shows that total flavones has greater loss.Applied sample amount is 10ml, and the result shows the basic free of losses of total flavones, is 10ml (the 1m medicinal liquid is equivalent to the 1.5g medical material) so select maximum applied sample amount, is equivalent to every ml macroporous resin adsorption total flavones amount and is equivalent to total flavones contained in the 0.75g medical material.Be 1.33ml/g promptly than applied sample amount.

Experimental example 3 Pyrogentisinic Acids cause the experiment of cervicitis rat model protective effect

1. the Pyrogentisinic Acid causes the influence that cervicitis rat model vagina is occulted blood

The model group animal has tangible vaginal mucosa hemorrhage, and the positive reaction degree of occulting blood significantly strengthens, and compares P＜0.001 with matched group.Be subjected to reagent object height, middle dosage group can obviously suppress the animal vagina and occult blood, the positive reaction degree is alleviated, compare P＜0.05 with model group.Dexamethasone also reduces the animal vagina positive reaction degree of occulting blood, with model group P＜0.05 relatively.It is not obvious that KANGGONGYAN PIAN suppresses the vagina effect of occulting blood.The results are shown in Table 22.

Table 22 medicament composition capsule Pyrogentisinic Acid causes the influence that cervicitis rat model vagina occults blood (x ± s)

2. the Pyrogentisinic Acid causes the influence of cervicitis rat model cervix uteri form

Observed result shows under the mirror, the normal control group: cervical mucosa and each layer of neck tube mucosa clear in structure, cervical mucosa reach the interior a small amount of lymphocyte of neck tube mucosa down and are dispersed in infiltration; Model group: a large amount of edema of the squamous cell of cervical mucosa, cervical mucosa reach volume lymphocyte and a small amount of intiltration of acidophilic leukocyte in the neck tube mucosa down, also see histiocyte and vascular reactivity hypertrophy; Dexamethasone and KANGGONGYAN PIAN group: the squamous epithelial cancer edema of cervical mucosa alleviates, and cervical mucosa reaches neck tube mucosa endolymph cell down obviously to be reduced, and accompanies a little intiltration of acidophilic leukocyte; 12g/kg dosage group: the squamous epithelial cancer edema of cervical mucosa is obvious, and cervical mucosa has than multi-lymphocytes and a small amount of intiltration of acidophilic leukocyte down and in the neck tube mucosa, also sees histiocyte and vascular reactivity hypertrophy; 6,3g/kg dosage group: the squamous epithelial cancer edema of cervical mucosa obviously alleviates, and the subregion disappears, and cervical mucosa reaches a little lymphocyte and intiltration of acidophilic leukocyte in the neck tube mucosa down, and histiocyte and vascular reaction are not obvious; 1.5g/kg organize: the squamous epithelial cancer edema zone of cervical mucosa is obvious, and it is interior than multi-lymphocytes and intiltration of acidophilic leukocyte that cervical mucosa reaches the neck tube mucosa down, and histiocyte and vascular reaction sexually revise more obvious. the results are shown in Table 23.

Table 23 medicament composition capsule Pyrogentisinic Acid causes the influence of cervicitis rat cervix uteri inflammation degree

Kruskal-Wallis H assay is as follows:

Kruskal-Wallis?Test

3. the influence of Pyrogentisinic Acid's rat model ceruloplasmin vigor

The blue protein vigor of model group copper is with respect to blank group significantly raise (P＜0.01); Dexamethasone group and KANGGONGYAN PIAN group all can significantly suppress the rising (P＜0.05) of the blue protein vigor of copper; Pharmaceutical composition 1.5,12.0g/Kg gastric infusion all can significantly reduce the blue protein vigor (P＜0.05) of copper, wherein with 12.0g/Kg effect the strongest (P＜0.001), 1.5g/Kg take second place, and 3.0,6.0g/Kg group all do not have remarkable influence to the blue protein vigor of copper.The results are shown in Table 24.

Table 24 pharmaceutical composition Pyrogentisinic Acid causes the influence (x ± s) of the blue protein vigor of cervicitis rat plasma copper (CP)

Annotate: compare with model control group ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001

4. the Pyrogentisinic Acid causes cervicitis rat model blood plasma PGE ₂The influence of content

Compare PGE in the model group animal blood slurry with matched group ₂Obviously raise, medicament composition capsule is than PGE in the model group blood plasma ₂Obviously reduce, each dosage group all has significant difference (p＜0.001); Dexamethasone group and KANGGONGYAN PIAN group also all have significant difference (p＜0.001).The results are shown in Table 25.

Table 25 medicament composition capsule Pyrogentisinic Acid causes PGE in the cervicitis rat model blood plasma ₂The influence of content (X ± SD)

Compare with model group: * p＜0.05; * p＜0.01; * * p＜0.001.

5. the Pyrogentisinic Acid causes the influence of IL-1 β content in the cervicitis rat model serum

Compare with matched group, model group serum il-1 β obviously raises, and significant difference (p＜0.05) is arranged; KANGGONGYAN PIAN optimization prescription group height, middle dosage group are than model group serum il-1 β zero difference; Middle low dose group and low dose group reduce than model group serum il-1 β, and be variant, and wherein middle low dose group has than significant difference (p＜0.01); Dexamethasone group also tool than significant difference (p＜0.01); The KANGGONGYAN PIAN group has significant difference (p＜0.05) than model group.The results are shown in Table 26.

Table 26 medicament composition capsule Pyrogentisinic Acid causes the influence (X ± SD) of IL-1 β content in the cervicitis rat model serum

Compare with model group: * p＜0.05; * p＜0.01; * * p＜0.001.

6. the Pyrogentisinic Acid causes the influence of TNF content in the cervicitis rat model serum

Model group and matched group compare, and the model group serum TNF significantly raises, and notable statistics difference (p＜0.01) is arranged; Optimize high dose group and the minimizing of middle low dose group serum TNF content in the prescription group, than model group significant difference (p＜0.05) is arranged, wherein high dose group has significant difference (p＜0.01); Dexamethasone group also has significant difference (p＜0.05).See Table 27.

Table 27 medicament composition capsule Pyrogentisinic Acid causes the influence (X ± SD) of TNF content in the cervicitis rat model serum

Compare with model group: * p＜0.05; * p＜0.01.

Discuss and conclusion

Local mucous membrane damage and cause pathogeny imcrobe infection are the two big main causes that cause cervicitis, this result of the test shows, chemical stimulation substance, phenol or Escherichia coli transvaginal are given repeatedly and similar human cervicitis sample pathological change can be brought out, and with the abnormal change of inflammatory mediator and relevant cell factor.

This result of the test shows that the cervical tissue pathological change that medicament composition capsule Pyrogentisinic Acid and Escherichia coli cause all has obvious inhibitory action, and can reduce the generation of inflammatory mediator and relevant cell factor.For its clinical treatment cervicitis provides foundation.

Experimental example 4 antiinflammatory action experimentatioies

1, the medicament composition capsule xylol causes the influence of mice auricle swelling

Compare with model group, be subjected to reagent object height dosage group, middle dosage group that the effect (p＜0.01) of significant inhibition mice ear is arranged; In low dose group effect take second place (p＜0.05); Low dose group, KANGGONGYAN PIAN group are not observed the drug action that suppresses mice ear, and being subjected to has dose-effect relationship between the reagent thing group.Illustrate and be subjected to the reagent thing to have the drug action that suppresses mice ear.The results are shown in Table 28.

Table 28 KANGGONGYAN PIAN optimization prescription xylol causes the influence (X ± SD) of mice auricle swelling

Annotate: compare * * p＜0.01, * p＜0.05 with model control group

2, the medicament composition capsule on Carrageenan causes the influence of rat paw edema

Behind the rat paw intradermal injection carrageenin, early stage acute inflammatory reactions such as that the Mus pawl is is red, swollen, hot, pain. positive control drug KANGGONGYAN PIAN and dexamethasone all can obviously reduce carrageenin cause scorching rat paw edema degree and swelling rate (P＜0.05-0.001). pharmaceutical composition 3.0,6.0 and 12.0g/Kg gastric infusion all can significantly alleviate rat carrageenan and cause the scorching acute inflammatory reaction of 1h, 2h, 4h, 6h and 8h afterwards, make swelling degree of the paw and swelling rate significantly reduce (P＜0.05-0.001), except the 3.0g/Kg rat carrageenan cause the swelling degree of the paw of scorching back 4h; 1.5g/Kg rat paw edema degree and swelling rate are not made significant difference (P＞0.05). the results are shown in Table 29.

3, pharmaceutical composition is to the swollen influence that forms of rat granuloma

At the subcutaneous implantation sterilization of rat both sides groin cotton balls, producing the granulation hyperplasia phenomenon after 14 days. positive control drug dexamethasone and KANGGONGYAN PIAN all can significantly reduce dry weight, the weight in wet base (P＜0.001) of cotton balls, antiinflammatory action is remarkable. and pharmaceutical composition 1.5,3.0,6.0 and 12.0g/Kg gastric infusion all can significantly suppress the swollen formation of rat granuloma, alleviate chronic inflammatory reaction, the dry weight and the weight in wet base of cotton balls significantly reduced (P＜0.05), wherein with 6.0 and 12.0g/Kg the most remarkable. the results are shown in Table 30.

Table 30 pharmaceutical composition is to the swollen influence that forms of rat granuloma (x ± s)

Annotate: compare with model control group ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001

Discuss and conclusion

Result of the test shows that medicament composition capsule all has inhibitory action to acute and chronic experimental inflammation, and improving symptom for the clinical treatment cervicitis provides foundation.

Experimental example 4 analgesic activity experimentatioies

Be subjected to the reagent thing consistent with positive control drug effect trend.It is proper to illustrate that this trial drug dosage is selected, but does not have dose-effect relationship, but KANGGONGYAN PIAN is not observed positive findings.Be subjected in the reagent thing low dose group drug action the strongest, compare, utmost point significance statistical discrepancy (p＜0.01) is arranged with model control group; Other each group also all has significance statistical discrepancy (p＜0.05).The result shows: had by the reagent thing and alleviate the drug action that the acetic acid induced mice is turned round the body number of times, the effect of certain analgesic is promptly arranged.The results are shown in Table 31.

The analgesic activity of table 31 medicament composition capsule (X ± SD)

Annotate: each administration group and model control group be * * p＜0.01 relatively; * p＜0.05

Result of the test shows that pharmaceutical composition can reduce the number of times that acetic acid causes the mouse writhing reaction, shows to have certain analgesic activity, and improving pain symptom for the clinical treatment cervicitis provides foundation.

Experimental example 5: to the experimentation of hemorheology influence

The model group whole blood viscosity is apparently higher than normal control group (P＜0.01).Pharmaceutical composition 1.5,3.0,12.0g/Kg gastric infusion all can significantly reduce the whole blood viscosity (P＜0.05～0.01) of rat.The positive control drug KANGGONGYAN PIAN can significantly reduce the whole blood viscosity (P＜0.05) of rat.The results are shown in Table 32.

32 pharmaceutical compositions are to the influence of rat whole blood viscosity (x ± s)

Annotate: compare with model control group ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001

Discuss and conclusion

The result shows that medicament composition capsule can reduce escherichia coli and cause cervicitis rat model blood viscosity, to improving the partial supply of blood flow of inflammation, promoting the absorption of inflammation to have the certain significance.

All by embodiment 1 preparation capsule, following embodiment all can realize the effect of above-mentioned experimental example to the pharmaceutical composition experimental drug of above-mentioned experimental example.

The specific embodiment

Embodiment 1: the preparation of capsule

Herba Leonuri 7kg Callicarpa kwangtungensis Chun 55kg

Get the Herba Leonuri medical material, add 10 times of water gagings, extract 3 times, each 1.5h, merge extractive liquid, filters, be concentrated into the thick paste of 60 ℃ of relative densities 1.05, add the 0.1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1, make the 1mL medicinal liquid be equivalent to the 1g medical material with 0.1mol/L hydrochloric acid then, promptly 60 ℃ of relative densities are 1.02, after centrifugal, get 001 * 7 type ion exchange resin column on the supernatant, absorption 1h, it is colourless to be eluted to effluent with distilled water, with the ammonia resin that alkalizes, with the ammonia ethanol elution, make effluent pH9-10, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 10 times of water gagings, extract each 1.5h, merge extractive liquid, 3 times, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.02 to add ethanol and make that to contain the alcohol amount be 70%, 24h is placed in cold preservation, filter, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.02, medicinal liquid is crossed HPD 300 macroporous resins, absorption 1h, it is closely colourless to be eluted to effluent with distilled water, use 70% ethanol elution, collect 5 times of column volume eluents, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract;

To add starch 100kg in the dry extract, pulverize, cross sieve No. 5, get dry extract, add the ethanol moistening, make capsule.

Embodiment 2: the preparation of granule

Herba Leonuri 60kg Callicarpa kwangtungensis Chun 15kg

Get the Herba Leonuri medical material, add 8 times of water gagings, extract 4 times, each 2h, merge extractive liquid, filters, be concentrated into the thick paste of 50 ℃ of relative densities 1.00, add the 0.1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1, make the 1mL medicinal liquid be equivalent to the 1g medical material with 0.1mol/L hydrochloric acid then, promptly 70 ℃ of relative densities are 1.02, after centrifugal, get 001 * 7 type ion exchange resin column on the supernatant, absorption 2h, it is colourless to be eluted to effluent with distilled water, with the ammonia resin that alkalizes, with the ammonia ethanol elution, make effluent pH10, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 12 times of water gagings, extract each 2h, merge extractive liquid, 4 times, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 70 ℃ of relative densities are 1.00 to add ethanol and make that to contain the alcohol amount be 70%, 24h is placed in cold preservation, filter, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 70 ℃ of relative densities are 1.00, medicinal liquid is crossed HPD 300 macroporous resins, absorption 1h, it is closely colourless to be eluted to effluent with distilled water, use 80% ethanol elution, collect the 5BV eluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract;

To add starch 100kg in the dry extract, pulverize, cross sieve No. 5, get dry extract, add the ethanol moistening, add conventional adjuvant, make granule according to a conventional method.

Embodiment 3: the preparation of pill

Herba Leonuri 40kg Callicarpa kwangtungensis Chun 40kg

Get the Herba Leonuri medical material, add 12 times of water gagings, extract 2 times, each 2h, merge extractive liquid, filters, be concentrated into the thick paste of 50 ℃ of relative densities 1.15, add the 0.1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1, make the 1mL medicinal liquid be equivalent to the 1g medical material with 0.1mol/L hydrochloric acid then, promptly 70 ℃ of relative densities are 1.02, after centrifugal, get 001 * 7 type ion exchange resin column on the supernatant, absorption 2h, it is colourless to be eluted to effluent with distilled water, with the ammonia resin that alkalizes, with the ammonia ethanol elution, make effluent pH9, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 12 times of water gagings, extract each 2h, merge extractive liquid, 4 times, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 70 ℃ of relative densities are 1.02 to add ethanol and make that to contain the alcohol amount be 70%, 24h is placed in cold preservation, filter, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 70 ℃ of relative densities are 1.02, medicinal liquid is crossed HPD 300 macroporous resins, absorption 2h, it is closely colourless to be eluted to effluent with distilled water, use 60% ethanol elution, collect 5 times of column volume eluents, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract;

To add starch 100kg in the dry extract, pulverize, cross sieve No. 5, get dry extract, add the ethanol moistening, add conventional adjuvant, make pill according to a conventional method.

Embodiment 4: the preparation of tablet

Herba Leonuri 8kg Callicarpa kwangtungensis Chun 60kg

Add conventional adjuvant, make tablet according to a conventional method.

Embodiment 5: the preparation of oral liquid

Herba Leonuri 10kg Callicarpa kwangtungensis Chun 50kg

Add conventional adjuvant, make oral liquid according to a conventional method.

Embodiment 6: the discrimination method in the quality testing

Get the composite preparation of embodiment 1 and do discriminating:

A. get 2 composition capsule contents, take by weighing 0.5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get 2 composition capsule contents, take by weighing 0.5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, with 4: 1: 0.5 alcohol-water-glacial acetic acid was developing solvent, launch, take out, dry, spray is with the 1%AlCl3 alcoholic solution, and hot blast blows to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Embodiment 7: the content assaying method in the quality testing

Get the composite preparation of embodiment 1 and do assay:

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column, with mobile phase 0.05mol/LNa2HPO4-0.05mol/L NaH2PO4, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500; The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution; The preparation of need testing solution: the powder 0.5g with under the medicament composition capsule agent content uniformity item, put in the tool plug triangular flask, add ethanol 25ml, claim decide weight, supersound extraction 30ml is put coldly, claims to decide weight again, supplies the weight of being lost with ethanol, shakes up filtration; Get subsequent filtrate 10mL, water bath method with the 0.05mol/LNa2HPO410mL dissolving, adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/LNa2HPO4 solution to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; Every hydrochloric stachydrine C of medicament composition capsule ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

Embodiment 8: quality monitoring method

Get the composite preparation of embodiment 1 and do quality testing:

A. get 2 composition capsule contents, take by weighing 0.5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get 2 composition capsule contents, take by weighing 0.5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, with 4: 1: 0.5 alcohol-water-glacial acetic acid was developing solvent, launch, take out, dry, spray is with the 1%AlCl3 alcoholic solution, and hot blast blows to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column is with mobile phase 0.05mol/LNa ₂HPO ₄-0.05mol/L NaH ₂PO ₄, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500; The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution; The preparation of need testing solution: the powder 0.5g with under the medicament composition capsule agent content uniformity item, put in the tool plug triangular flask, add ethanol 25ml, claim decide weight, supersound extraction 30ml is put coldly, claims to decide weight again, supplies the weight of being lost with ethanol, shakes up filtration; Get subsequent filtrate 10mL, water bath method with the 0.05mol/LNa2HPO410mL dissolving, adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/LNa2HPO4 solution to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; Every hydrochloric stachydrine C of medicament composition capsule ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

Embodiment 9: quality monitoring method

Get the composite preparation of embodiment 2 and do quality testing:

A. get composition granule, take by weighing 5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get composition granule, take by weighing 5g, take by weighing 0.5g, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, with 4: 1: 0.5 alcohol-water-glacial acetic acid was developing solvent, launch, take out, dry, spray is with the 1%AlCl3 alcoholic solution, and hot blast blows to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column is with mobile phase 0.05mol/LNa ₂HPO ₄-0.05mol/L NaH ₂PO ₄, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500; The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution; The preparation of need testing solution: get composition granule, take by weighing 5g, put in the tool plug triangular flask, add ethanol 25ml, claim decide weight, supersound extraction 30ml is put coldly, claims to decide weight again, supplies the weight of being lost with ethanol, shakes up filtration; Get subsequent filtrate 10mL, water bath method with 0.05mol/LNa2HPO4 10mL dissolving, adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/LNa2HPO4 solution to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; Every hydrochloric stachydrine C of medicament composition capsule ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

Embodiment 10: quality monitoring method

Get the composite preparation of embodiment 4 and do quality testing:

A. get 2 in this pharmaceutical composition tablet, 0.7g adds ethanol 5ml, and supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get 2 in this pharmaceutical composition tablet, 0.7g takes by weighing 0.5g, adds ethanol 5ml, and supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, with 4: 1: 0.5 alcohol-water-glacial acetic acid was developing solvent, launch, take out, dry, spray is with the 1%AlCl3 alcoholic solution, and hot blast blows to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column is with mobile phase 0.05mol/LNa ₂HPO ₄-0.05mol/L NaH ₂PO ₄, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500; The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution; The preparation of need testing solution: get 2 in this pharmaceutical composition tablet, 0.7g puts in the tool plug triangular flask, adds ethanol 25ml, claims decide weight, and supersound extraction 30ml is put coldly, claims to decide weight again, supplies the weight of being lost with ethanol, shakes up filtration; Get subsequent filtrate 10mL, water bath method with 0.05mol/LNa2HPO4 10mL dissolving, adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/LNa2HPO4 solution to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; Every hydrochloric stachydrine C of medicament composition capsule ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

Embodiment 11:

Get Herba Leonuri 10kg, Callicarpa kwangtungensis Chun 55kg adds adjuvant, and technology is made tablet routinely, every 0.35g.

Embodiment 12:

Get Herba Leonuri 55kg, Callicarpa kwangtungensis Chun 10kg adds adjuvant, and technology is made granule routinely, every bag of 6g.

Embodiment 13:

Get Herba Leonuri 20kg, Callicarpa kwangtungensis Chun 45kg adds adjuvant, and technology is made capsule routinely, every 0.35g.

Embodiment 14:

Get Herba Leonuri 30kg, Callicarpa kwangtungensis Chun 35kg adds adjuvant, and technology is made vagina effervescence routinely, every 0.5g.

Embodiment 15:

Get Herba Leonuri 40kg, Callicarpa kwangtungensis Chun 25kg adds adjuvant, and technology is made oral liquid routinely, every 10ml.

Embodiment 16:

Get Herba Leonuri 50kg, Callicarpa kwangtungensis Chun 15kg adds adjuvant, and technology is made suppository routinely, every 5g.

Claims (11)

1. Chinese medicine composition for the treatment of cervicitis is characterized in that the crude drug of this Chinese medicine composition consists of:

Herba Leonuri 7-60 weight portion Callicarpa kwangtungensis Chun 7-60 weight portion.

2. Chinese medicine composition as claimed in claim 1 is characterized in that this traditional Chinese medicinal composition raw materials weight proportion is as follows:

Herba Leonuri 7 weight portion Callicarpa kwangtungensis Chuns 55 weight portions.

3. Chinese medicine composition as claimed in claim 1 is characterized in that this traditional Chinese medicinal composition raw materials weight proportion is as follows:

Herba Leonuri 60 weight portion Callicarpa kwangtungensis Chuns 15 weight portions.

4. Chinese medicine composition as claimed in claim 1 is characterized in that this traditional Chinese medicinal composition raw materials weight proportion is as follows:

Herba Leonuri 40 weight portion Callicarpa kwangtungensis Chuns 40 weight portions.

5. as the arbitrary described Chinese medicine composition of claim 1-4, it is characterized in that this Chinese medicine composition makes capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray.

6. as the preparation method of claim 1,2,3 or 4 described Chinese medicine compositions, it is characterized in that this method is:

Get the Herba Leonuri medical material, add 8-12 times of water gaging, extract 2-4 time, each 1-2h, merge extractive liquid,, filter, be concentrated into the thick paste of 50 ℃ of-70 ℃ of relative density 1.00-1.15, add the 0.1mol/L-1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1-3, be diluted to the 1mL medicinal liquid with 0.1mol/L-1mol/L hydrochloric acid then and be equivalent to the 1-2g medical material, i.e. 50 ℃ of-70 ℃ of relative density 1.00-1.20, through centrifugal or filter after, get strong acid ion exchange resin post on the supernatant, absorption 0.5-2h, it is colourless to be eluted to effluent with distilled water, with ammonia alkalization resin PH8-9, with 30-95% ammonia ethanol or 2mol ammonia eluting, make effluent pH8-12, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 8-12 times of water gaging, extract each 1-2h, merge extractive liquid, 2-4 time, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 50 ℃ of-70 ℃ of relative densities are 1.00-1.02, add 95% ethanol and make the alcohol amount of containing be 50-70%, 24h is placed in cold preservation, filters, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 50 ℃ of-70 ℃ of relative densities are 1.00-1.02, and medicinal liquid is crossed macroporous resin, absorption 0.5-2h, it is closely colourless to be eluted to effluent with distilled water, uses the 10%-80% ethanol elution, collects the 5BV eluent, reclaim ethanol, be concentrated into dried, vacuum drying, dry extract;

Dry extract is added conventional adjuvant, make capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray according to a conventional method.

7. the preparation method of Chinese medicine composition as claimed in claim 6 is characterized in that this method is:

Get the Herba Leonuri medical material, add 10 times of water gagings, extract 3 times, each 1.5h, merge extractive liquid, filters, be concentrated into the thick paste of 60 ℃ of relative densities 1.05, add the 0.1mol/L dissolving with hydrochloric acid, transferring pH with concentrated hydrochloric acid is 1, make the 1mL medicinal liquid be equivalent to the 1g medical material with 0.1mol/L hydrochloric acid then, promptly 60 ℃ of relative densities are 1.00-1.02, after centrifugal, get 001 * 7 type ion exchange resin column on the supernatant, absorption 1h, it is colourless to be eluted to effluent with distilled water, with ammonia alkalization resin PH8-9,75% or 95% ammonia ethanol or 2mol ammonia eluting, make effluent pH9-10, collect 10 times of column volume effluent, reclaim ethanol, be concentrated into dried, vacuum drying gets dry extract, and is standby;

Get the Callicarpa kwangtungensis Chun medical material, the section of being ground into adds 10 times of water gagings, extract each 1.5h, merge extractive liquid, 3 times, filter, be concentrated into the 1mL medicinal liquid and be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.00-1.02, add 95% ethanol and make that to contain alcohol amount be 60% or 70%, 24h is placed in cold preservation, filters, reclaim ethanol, make the 1mL medicinal liquid be equivalent to the 1.5g medical material, promptly 60 ℃ of relative densities are 1.00-1.02, and medicinal liquid is crossed HPD 100, HPD 400 or HPD 300 macroporous resins, absorption 1h, it is closely colourless to be eluted to effluent with distilled water, with 50%, 60% or 70% ethanol elution is collected 5 times of column volume eluents, reclaim ethanol, be concentrated into dried, vacuum drying, dry extract;

Dry extract is added conventional adjuvant, makes capsule, pill, tablet, granule, liquid oral, oral pastes, suppository, externally used paste, effervescent tablet, external-use lotion, aerosol or spray according to a conventional method.

8. the quality determining method of Chinese medicine composition as claimed in claim 5 is characterized in that this method comprises one or more in the following discriminating:

A. get 1/3 of this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds ethanol and makes the solution that every 1ml contains 5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution, each 5 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 4: 1: 0.5 n-butyl alcohol-hydrochloric acid-water, launch, take out, airing, spray is with rare bismuth potassium iodide test solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get 1/3 of this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, add ethanol 5ml, supersound extraction 30min filters, and filtrate evaporate to dryness, residue add ethanol 0.5ml makes dissolving, as need testing solution; Other got the Callicarpa kwangtungensis Chun powder 1g of 60 mesh sieves, with 70% ethanol 50mL reflux, extract, 0.5h, extracting solution filters, water bath method, residue add the 5ml distilled water makes dissolving, divides 2 extractions with the water saturated n-butyl alcohol of 10ml, extract merges, behind the reclaim under reduced pressure n-butyl alcohol, residue adds 510mL methanol makes dissolving, in contrast medical material solution; According to the thin layer chromatography test, draw need testing solution, each 2 μ l of reference substance solution, put respectively on same polyamide lamellae, be developing solvent with 4: 1: 0.5 alcohol-water-glacial acetic acid, launch, take out, to dry, spray is with 1%AlCl ₃Alcoholic solution, hot blast blow to speckle displaing yellow fluorescence under the 365nm wavelength; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

9. the quality determining method of Chinese medicine composition as claimed in claim 5 is characterized in that this method comprises following assay:

Chromatographic condition and system suitability test: 3.5 μ m, 250 * 4.6mm LC-SCX chromatographic column is with mobile phase 0.05mol/LNa ₂HPO ₄-0.05mol/L NaH ₂PO ₄, pH5.5, flow velocity: 1mL/min, the detection wavelength is 192nm, column temperature is 35 ℃; Number of theoretical plate calculates by the stachydrine hydrochloride peak should be not less than 1500;

The preparation of reference substance solution: it is an amount of to take by weighing 3 hours stachydrine hydrochloride reference substance of 105 ℃ of dryings, adds 0.05mol/LNa ₂HPO ₄Make the solution that every 1ml contains 0.5mg, in contrast product solution;

The preparation of need testing solution: with 1/3 of this pharmaceutical composition solid preparation Coming-of-Age Day taking dose, put in the tool plug triangular flask, add ethanol 25ml, claim decide weight, supersound extraction 30ml is put coldly, claims to decide weight again, supplies the weight of being lost with ethanol, shakes up filtration; Get subsequent filtrate 10mL, water bath method is used 0.05mol/L Na ₂HPO ₄The 10mL dissolving adds active carbon 0.5g, heating in water bath half a minute, stir, and filter, filtrate places the 25mL measuring bottle, adds 0.05mol/L Na ₂HPO ₄Solution is to scale; Algoscopy: accurate respectively reference substance solution 5 μ l and need testing solution 5～10 μ l of drawing, inject high performance liquid chromatograph, measure, calculate, promptly; The hydrochloric stachydrine C of pharmaceutical composition unit formulation ₇H ₁₃NO ₂Meter must not be less than 0.03mg.

10. as the application of the arbitrary described Chinese medicine composition of claim 1-4 in preparation treatment cervicitis medicine.

11. the application of Chinese medicine composition as claimed in claim 5 in preparation treatment cervicitis medicine.