CN1943680A

CN1943680A - A Chinese traditional medicinal composition for treatment of endometriosis and its preparation method

Info

Publication number: CN1943680A
Application number: CN 200510108013
Authority: CN
Inventors: 任明非; 杨世林; 万方; 王卫华
Original assignee: Chengdu Huanshen Group Co Ltd
Current assignee: Chengdu Huashen Technology Group Co., Ltd
Priority date: 2005-10-09
Filing date: 2005-10-09
Publication date: 2007-04-11
Anticipated expiration: 2025-10-09
Also published as: CN100500188C

Abstract

This invention provides a medicinal composition for treatment of endometriosis and its preparation method and its controlling method. This medicinal composition is prepared by using the medical ingredients of common peony root, Chinese angelica root, cattail pollen, chuanxiong ligusticum rhizome, cassiabarktree twig, yanhusuo rhizome, Chinese thorowax root, membranous milkvetch root, malaytea scurfpea fruit and myrrh. Curative effectiveness of this medical composition in treating endometriosism is definitude with fast action but without toxin and side effects.

Description

Endometriotic Chinese medicine composition of a kind of treatment and preparation method thereof

Technical field

The present invention relates to the preparation method and the method for quality control of a kind of Chinese medicine composition and said composition, particularly a kind ofly be used for the treatment of endometriotic Chinese medicine composition, relate to the preparation method and the method for quality control of said composition simultaneously.

Background technology

Endometriosis is one of common gynecological disease, frequently-occurring disease, and sickness rate obviously increases in recent years, and sickness rate is 10-20% among the women of child-bearing age, and about 40% endometriosis merges infertile.Endometriosis is having a strong impact on women's physical and mental health.Though endometriosis is found the history in existing more than 100 year, its etiology and pathogenesis is also not fully aware of, and treatment does not have ideal method yet.In recent decades, doctor trained in Western medicine is being explored application hormone medicine treatment endometriosis always.The method that pseudo pregnancy therapy, pseudo menopause therapy, the moving agent therapy of gonadotropin releasing hormone is arranged at present and oppositely add treatment on this basis.These Therapeutic Method have certain curative effect to recent relief of symptoms.But the relapse rate height, side effect is big, and difficult popularization that cost an arm and a leg.Conservative operation in the operative treatment, postoperative easily recurs, and radical operation is improper to young woman, and menopause, the syndrome incidence rate was higher.For all the foregoing reasons, more thorny in the treatment.The Chinese medicine endometriosis is better than Western medicine, and determined curative effect is not found obvious toxic-side effects, and Chinese medicine do not suppress the ovulation, favourable to gestation.And safety, taking convenience, low price, steady quality.

Summary of the invention

One object of the present invention is to disclose the endometriotic Chinese medicine composition of a kind of treatment; Another object of the present invention is to disclose a kind of preparation method for the treatment of endometriotic Chinese medicine composition; The 3rd purpose of the present invention is to disclose the method for quality control that this treats endometriotic Chinese medicine composition.

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

The crude drug composition and the proportioning of pharmaceutical composition of the present invention are as follows:

Radix Paeoniae Rubra 7-8 weight portion Radix Angelicae Sinensis 5-7 weight portion Pollen Typhae 4-6 weight portion

Rhizoma Chuanxiong 4-6 weight portion Ramulus Cinnamomi 2-4 weight portion Rhizoma Corydalis 7-8 weight portion

Radix Bupleuri 4-6 weight portion Radix Astragali 9-11 weight portion Fructus Psoraleae 4-6 weight portion

Herba Asari 1-2 weight portion Myrrha (processed) 2-4 weight portion

The optimum ratio of above-mentioned raw materials medicine is:

Radix Paeoniae Rubra 7.5 weight portion Radix Angelicae Sinensis 6 weight portion Pollen Tyjphae 5 weight portions

Rhizoma Chuanxiong 5 weight portion Ramulus Cinnamomi 3 weight portion Rhizoma Corydalis 7.5 weight portions

Radix Bupleuri 5 weight portion Radix Astragali 10 weight portion Fructus Psoraleaes 5 weight portions

Herba Asari 1.5 weight portion Myrrha (processed)s 3 weight portions

The optimum ratio of above-mentioned raw materials medicine can also for:

Radix Paeoniae Rubra 7 weight portion Radix Angelicae Sinensis 7 weight portion Pollen Tyjphae 4 weight portions

Rhizoma Chuanxiong 6 weight portion Ramulus Cinnamomi 2 weight portion Rhizoma Corydalis 8 weight portions

Radix Bupleuri 4 weight portion Radix Astragali 11 weight portion Fructus Psoraleaes 4 weight portions

Herba Asari 2 weight portion Myrrha (processed)s 2 weight portions

The optimum ratio of above-mentioned raw materials medicine can also for:

Radix Paeoniae Rubra 8 weight portion Radix Angelicae Sinensis 5 weight portion Pollen Tyjphae 6 weight portions

Rhizoma Chuanxiong 4 weight portion Ramulus Cinnamomi 4 weight portion Rhizoma Corydalis 7 weight portions

Radix Bupleuri 6 weight portion Radix Astragali 9 weight portion Fructus Psoraleaes 6 weight portions

Herba Asari 1 weight portion Myrrha (processed) 4 weight portions

Press practice of pharmacy, the above-mentioned raw materials medicine can be prepared into various clinical acceptable forms, include but not limited to a kind of in the middle of the following dosage form: as solid preparations such as tablet, pill, capsule, granule, drop pill, soft capsule and suppositorys, liquid preparations such as suspensoid, oral liquid, enema, preferred capsule formulation, tablet or granule dosage form.

This preparation of drug combination method:

Get it filled compositions crude drug ten simply by proportioning dosage, and Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis are doubly measured the alcohol heating reflux of 50-70% with 5-8, extract 2-4 time, and each 1.5-3 hour, merge ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 3-7 times of water gaging, soaked 0.5-3 hour, and water vapour distillation 2-8 hour, collect volatile oil, aqueous solution and medicinal residues are standby; Get the water that betacyclodextrin that volatile oil doubly measures with 4-8 and 40-80 doubly measure and carry out inclusion, 30-60 ℃ was stirred 0.5-1.5 hour, and cold preservation 10-18 hour, filter, inclusion complex 30-55 ℃ cold drying is ground into fine powder, and is standby; Radix Bupleuri, Radix Paeoniae Rubra, the Pollen Typhae and the Radix Astragali add 7-10 times of water gaging, extract 2-4 time, each 0.5-1.5 hour, medicinal residues such as last extracting solution and above-mentioned Ramulus Cinnamomi are closed fry in shallow oil, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, when being concentrated into 50 ℃, surveys amalgamation liquid relative density 1.10-1.15, add ethanol and make and contain alcohol amount and reach 60-80%, cold preservation is spent the night, and filters, when reclaiming ethanol to 50 ℃ respectively, surveys medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis relative density 1.15-1.20, drying, with above-mentioned volatile oil beta cyclodextrin inclusion complex fine powder and dextrin by weight 1: the 0.5-1.5 mixing, make granule, drying is made capsule, tablet or granule.

The method of quality control that this compositions is made medicament comprises discriminating and/or assay.

The discrimination method of method of quality control comprises a kind of and/or several in the following method:

A. get this drug combination preparation of 1/6 times of phase taking dose on the same day, porphyrize adds methanol 15-25ml, and supersound process 10-30 minute, filter, filtrate is as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 0.3-0.6mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 5-15 μ l, reference substance solution 5-15 μ l, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, chloroform-ethyl acetate-methanol-formic acid with 35-45: 4-6: 8-12: 0.15-0.25 is developing solvent, launch, take out, dry, spray is with the 45-55% phosphoric acid ethanol solution of 3-6% vanillin, and it is clear to be heated to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get psoralen and isopsoralen reference substance, add methanol respectively and make the solution that every 1ml contains 0.2-0.4mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 5-15 μ l, reference substance solution 4-10 μ l under the above-mentioned discriminating A item, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 7-9: 1.5-2.5 normal hexane-ethyl acetate is developing solvent, launch, take out, dry, spray is put under the 365nm ultra-violet lamp and is inspected with the 5-15% sodium hydrate methanol solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

C. get this drug combination preparation of 5/24 times of phase taking dose on the same day, porphyrize adds ethanol 10-20ml, and supersound process 0.5-1.5 hour, filter, filtrate is as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1.5-2.5g of Rhizoma Chuanxiong control medicinal material, and each adds ethanol 8-12ml, and supersound process is 0.5-1.5 hour respectively, filters, and filtrate is medical material solution in contrast; Test according to thin layer chromatography, draw each 5-15 μ 1 of above-mentioned need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 8-10: 0.8-1.2 cyclohexane extraction-ethyl acetate is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

D. get this drug combination preparation of 1/2 times of phase taking dose on the same day, porphyrize adds ammonia solution 1.5-2.5ml and chloroform 25-35ml, supersound process 30-50 minute, filter, filtrate is reclaimed chloroform to doing, residue adds ethanol 4-6ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methanol and makes the solution that every 1ml contains 0.15-0.25mg, in contrast product solution; Test according to thin layer chromatography, draw each 10-15 μ l of above-mentioned two kinds of solution, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 4-6: 3-5: 0.3-0.5 normal hexane-ethyl acetate-ammonia is developing solvent, launch, take out, dry, smoked clear with iodine vapor to the speckle colour developing, inspect under the daylight; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

E. get this drug combination preparation of phase taking dose on the same day, porphyrize is put in the round-bottomed flask of 200-300ml, add entry 40-60ml, connect volatile oil extractor, add water from the determinator upper end and make and be full of scale, 60～90 ℃ add petroleum ether 1.0-2.0ml, connect reflux condensing tube then; Flask contents is heated to boils, and kept 1.5-2.5 hour, stop heating, get petroleum ether layer as need testing solution; Other gets Herba Asari control medicinal material 8-12g and extracts volatile oil, adds chloroform 25-35ml and makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 10-20 μ l, reference substance solution 2-5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 92-98: 4-6 benzene-ethyl acetate is developing solvent, launch, take out, dry, spray is with 0.8-0.12% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

F. get this drug combination preparation of 1/4 times of phase taking dose on the same day, porphyrize, add kieselguhr 1.5-2.5g, mixing adds the methanol solution 15-25ml of 2% sodium hydroxide, heating and refluxing extraction 0.5-1.5 hour, filter, the filtrate evaporate to dryness adds water 25-35ml and makes dissolving, each with the water saturated n-butanol extraction of 35-45ml, extract 2-4 time, merge n-butyl alcohol liquid, with water washing 1-3 time, discard water liquid, with 0.8-1.2% potassium dihydrogen phosphate washing 1-2 time, discard water liquid, n-butyl alcohol liquid evaporate to dryness again, residue adds methanol 1.5-2.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.2-0.4mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 10-15 μ l, reference substance solution 5-7 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 18-22: 6-8: lower floor's solution of 1.5-2.5 chloroform-methanol-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be baked to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

This pharmaceutical composition content assaying method comprises following method:

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 65-75: 425-435 acetonitrile-water is mobile phase; The detection wavelength is 220-240nm; Theoretical cam curve is calculated by peoniflorin should be not less than 3000;

It is an amount of that the preparation of reference substance solution takes by weighing the peoniflorin reference substance, adds methanol and make the solution that every 1ml contains 0.2-1.0mg, promptly;

Content under this drug combination preparation content uniformity item is got in the preparation of need testing solution, and porphyrize is got 1.0-1.5g, put in the tool plug conical flask, it is fixed to claim, adds methanol 15-25ml, claim to decide weight, supersound process 20-30 minute, take out, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter with the 0.40-0.50um microporous filter membrane, get subsequent filtrate, promptly; Algoscopy is drawn above-mentioned reference substance solution 5-15 μ l, need testing solution 14-20 μ l respectively, injects chromatograph of liquid, measures, promptly;

This pharmaceutical composition per diem taking dose contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 69.0mg.

Wherein the day taking dose at the drug combination preparation of the present invention described in the method for quality control is equivalent to day take the about 61.5g of crude drug amount.

In this preparation of drug combination technical study experiment, by to decoction pieces soak time, amount of water to the Herba Asari volatile oil optimization experiment, guarantee to extract the volatile oil content height; As investigating index, from the orthogonal experiments table, select the optimization experiment of alcohol extraction condition with the content of psoralen, isopsoralen; With the peoniflorin is to investigate index, and the optimised process of selecting decocting to extract is; Under the different determining alcohol conditions of ethanol, measure the paeoniflorin content in each extract powder, select the best experiment of separation, purification and precipitate with ethanol; Ratio, different concentration ethanol solution by different dry extracts and dextrin are granulated as wetting agent, select preferred proportion and concentration.

Supersound extraction is saved time and the energy than reflux in this drug combination preparation assay, and is simple to operate.The blank assay of assay shows that blank solution not showing chromatographic peak with the corresponding retention time of peoniflorin reference substance place, separates noiseless.The RSD that peoniflorin is calculated in the precision test is 0.9948%, illustrates that the precision of instrument is good; It is stable in 24 hours that study on the stability gets peoniflorin.It is 1.5880% that replica test calculates RSD, illustrates that content assaying method is stable, and repeatability is better; It is 99.05% that recovery test calculates average recovery rate, and RSD is 0.9356%.Illustrate that content assaying method is feasible.The peak area and the sample size of the investigation explanation peoniflorin of linear relationship are good linear relationship.

By to the influence of rabbit animal model of endometriosis test, find that follicule-stimulating hormone (FSH) (FSH) content significantly reduces, lutropin (LH) has reduction, prolactin antagonist (PRL), estradiol (E ₂) content significantly reduces, the Ectopic Endometrium volume obviously dwindles and Ectopic Endometrium weight obviously alleviates, the endometriosis lesion degree is lighter than model group, improve the rheol trend of animal blood; By influence test, find the obviously reduction of FSH content, LH, PRL, E equally to rat endometrium dystopy disease animal model ₂Content all has downward trend, Ectopic Endometrium volume all obviously to dwindle and Ectopic Endometrium weight obviously alleviates, metrecotpia inner membrance lesion degree is lighter than model group; By rat is tested in the uterotonic influence of body, find that the agent of this medicament composition granule causes that to oxytocin the strong contraction in uterus has the effect of inhibitory action, reduction uterine activity amplitude and inhibition uterine motility; By the microcirculation test, find that the agent of this medicament composition granule improves the microcirculatory effect of mice mesentery, local disturbance of blood circulation is had protective effect; By antiinflammatory action test, find that the agent of this medicament composition granule has obvious inhibitory action, obviously suppresses the pedal swelling effect rat granuloma inflammatory model; By the analgesic activity test, find that the agent of this medicament composition granule can improve pain threshold, has significant analgesia role; By the immunologic hypofunction mice being gulped down the influence test of having a liking for function, find that the agent of this medicament composition granule gulps down immunologic hypofunction mouse monokaryon cell and have a liking for function tangible potentiation is arranged; By influence test, find that the agent of this medicament composition granule has potentiation to the cell immune response of immunologic hypofunction mice to the tardy paraphilia reflection of immunologic hypofunction mice.

Following experimental example is used to further specify but is not limited to the present invention.

Definite experiment of experimental example 1 degree of grinding and extraction time

Take by weighing each three parts of Ramulus Cinnamomi 60g, Herba Asari 30g, Myrrha (processed) 60g according to pharmaceutical composition proportioning dosage, three parts specification is respectively decoction pieces, 10 orders, 20 orders.Add 6 times of water gagings respectively, carry out steam distillation.The results are shown in Table 1:

The investigation of table 1 Herba Asari volatile oil extraction time

Extraction time (hour)	1	2	3	4	5	6	7	8	9
Extraction time (hour)										Different size volatilization (ml)
Decoction pieces										Different size volatilization (ml)	1.20	2.50	2.80	2.80	3.10	3.20	3.25	3.30	3.30
Decoction pieces	10 orders	0.90	1.50	2.00	2.50	2.70	2.90	2.90	3.00	3.00	1.20	2.50	2.80	2.80	3.10	3.20	3.25	3.30	3.30
20 orders	10 orders	0.90	1.50	2.00	2.50	2.70	2.90	2.90	3.00	3.00	1.20	2.30	3.00	3.20	3.50	3.65	3.82	3.85	3.85

By above result as can be seen, it is minimum that 10 orders are put forward oil mass, and 20 order medical material powder cause easily aborning and are burned, and the oil extracting rate of decoction pieces is compared with 20 orders, has reached 85.7%, and few one procedure, saves the energy, so select directly to carry oil with decoction pieces.Extract and carried substantially in 8 hours to the greatest extent, it is fuel-displaced the fastest to add thermal distillation 1 hour, can carry substantially to the greatest extent in 30%, 8 hour that reaches total amount, so the optimum extraction time of volatile oil is elected 8 hours as.

The experiment of experimental example 2 Herba Asari volatile oil inclusion optimal processes

Ratio (A), amount of water (B), enclose temperature (C), mixing time (D) with volatile oil and beta-schardinger dextrin-are the investigation factor, select varying level, according to L9 (3 ⁴) orthogonal table carries out inclusion, extracts with micro-volatile oil extractor respectively again, calculates the volatile oil utilization rate, carries out variance analysis, the orthogonal polynomial regression method is adopted in variance analysis.

Table 2 volatile oil inclusion factor level table

Factor level	A volatile oil: β-CD (ml/g)	B amount of water (ml)	C enclose temperature (℃)	D mixing time (h)
Factor level	A volatile oil: β-CD (ml/g)	B amount of water (ml)	C enclose temperature (℃)	D mixing time (h)	1	1∶4	60	40	0.5
2	1∶6	80	60	1.0	1	1∶4	60	40	0.5
2	1∶6	80	60	1.0	3	1∶8	100	80	1.5

Table 3 volatile oil inclusion technology Orthogonal experiment results

	A volatile oil: β-CD	The B amount of water	C enclose temperature	The D mixing time	Volatile oil utilization rate (%)
	A volatile oil: β-CD	The B amount of water	C enclose temperature	The D mixing time	Volatile oil utilization rate (%)	1	1	1	1	1	90
2	1	2	2	2	80	1	1	1	1	1	90
2	1	2	2	2	80	3	1	3	3	3	79
4	2	1	2	3	92	3	1	3	3	3	79
4	2	1	2	3	92	5	2	2	3	1	82
6	2	3	1	2	90	5	2	2	3	1	82
6	2	3	1	2	90	7	3	1	3	2	85
8	3	2	1	3	90	7	3	1	3	2	85
8	3	2	1	3	90	9	3	3	2	1	82
K1	249	267	270	254		9	3	3	2	1	82
K1	249	267	270	254		K2	264	252	254	255	∑＝770
K3	257	251	246	261	CT＝65877.78	K2	264	252	254	255	∑＝770
K3	257	251	246	261	CT＝65877.78	Q/3	65915.33	65931.33	65977.33	65887.33	Q＝K12+K22+K32
R	15	16	24	7	S＝Q/3-CT	Q/3	65915.33	65931.33	65977.33	65887.33	Q＝K12+K22+K32
R	15	16	24	7	S＝Q/3-CT	S	37.55	53.55	99.55	9.55

Table 4 volatile oil inclusion technology The results of analysis of variance

F _1-0.05(1，3)＝10.13 F _1-0.01(1，3)＝34.12

Annotate: " △ " merging is as the test error quadratic sum." * * " expression has significant differences, and " * " expression has significant difference.By in the table 4 as can be known, A factor, B factor, C factor have the highly significant influence, select A from the orthogonal experiments table ₂B ₁C ₁Be optimum condition, the D factor is significant factors not, and selecting mixing time is 1 hour.By above-mentioned test as can be known: the optimum condition of volatile oil inclusion is: be about to 6g β-CD and add fully dissolving in the 60ml water, under 40 ℃ of conditions, add 1ml volatile oil and carry out inclusion, mixing time is 1 hour.

The discrimination test of experimental example 3 drug combination preparations

The thin layer feature of Radix Paeoniae Rubra differentiates that peoniflorin is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 0736-9811.Once adopted Radix Paeoniae Rubra thin-layer developing system chloroform-ethyl acetate-methanol-formic acid (40: 5: 10: 0.2) and the upper solution of ethyl acetate-methanol-water (8: 3: 2.5) launch, it is undesirable to find that the latter launches effect, and the peoniflorin clear spot that the former obtains is concentrated, and is noiseless with the negative control product of method preparation.Developer is original to adopt 5% vanillin sulfuric acid solution, but this developer long slightly speckle heat time heating time is blackening, the bad grasp of developing time, after change 50% phosphoric acid ethanol solution of 5% vanillin into, developer sensitivity, specificity are strong, the long-time heating speckle does not turn black.Through sample and negative control test, confirm that method is feasible in the technical scheme in this way.

The thin layer of Fructus Psoraleae is differentiated: psoralen is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 0739-9706.Isopsoralen is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute and is provided, and is numbered 738-8802.In the process of test, because laboratory does not have potassium hydroxide, replace with sodium hydroxide, find that color developing effect is also good, so developer uses 10% sodium hydrate methanol solution.Negative control product with the method preparation are noiseless.Through sample and negative control test, confirm that method is feasible in the technical scheme in this way.

The thin layer of Radix Angelicae Sinensis, Rhizoma Chuanxiong is differentiated: the Radix Angelicae Sinensis control medicinal material is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 927-9303, and the Rhizoma Chuanxiong control medicinal material is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute and provided, and is numbered 918-9202.When finding with ethanol extraction Radix Angelicae Sinensis, Rhizoma Chuanxiong control medicinal material and sample in experiment, speckle clearly, and is better than ether and ethyl acetate.Radix Angelicae Sinensis feminine gender, Rhizoma Chuanxiong feminine gender and Radix Angelicae Sinensis Rhizoma Chuanxiong negative control product with the method preparation are noiseless.Through sample and negative control test, confirm that method is feasible in the technical scheme.

The thin layer of Rhizoma Corydalis is differentiated: tetrahydropalmatine is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 0726-9605.Once adopted normal hexane-chloroform-methanol (10: 6: 1), n-butyl alcohol-concentrated hydrochloric acid-water development systems such as (4: 1: 0.5) to launch, it is undesirable to find to launch effect, using normal hexane-ethyl acetate-ammonia (5: 4: 0.4) again is development system, finds that clear spot is concentrated.Negative control product with the method preparation are noiseless.Through sample and negative control test, confirm that method is feasible in the technical scheme.

The thin layer of Herba Asari volatile oil is differentiated: the Herba Asari control medicinal material is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 121204-0101.Be that solvent carries out supersound process to finished product once, and got filtrate, but volatile oil also there is relatively large loss when flinging to chloroform, so use instead with the negative control product of method preparation noiseless as need testing solution with the 25ml chloroform.Through sample and negative control test, confirm that method is feasible in the technical scheme.

The thin layer of astragaloside is differentiated: astragaloside is purchased in Nat'l Pharmaceutical ﹠ Biological Products Control Institute, is numbered 781-200009.Once adopted the thin layer discrimination method of the contained Radix Astragali of pharmacopeia to carry out thin layer chromatography, but sample chromatogram becomes band on the lamellae, has covered the speckle of astragaloside, and the Rf value of astragaloside reference substance is less.So handle sample with alkaline extraction, the astragaloside clear spot that obtains is orange-yellow speckle, and is noiseless with the negative control product of method preparation, confirms that method is feasible in the technical scheme.

The selection experiment of experimental example 4 assay mobile phases

Methanol-0.05mol/L potassium dihydrogen phosphate (pharmacopeia method), methanol-water-glacial acetic acid (30: 70: 0.5) and second eyeball-water (14: 86) etc. have been compared, peoniflorin peak hangover under methanol-water-glacial acetic acid (30: 70: 0.5) system, pharmacopeia method and second eyeball-water system peoniflorin peak separates fair, but the former compares with the latter, certain acidity is arranged, chromatographic column is easily caused damage, and second eyeball-water (14: 86) peak shape symmetry, baseline is steady, retention time is also suitable, so optimal flow was selected second eyeball-water for use 14: 86 mutually.

The selection experiment of 5 assay supersound process times of experimental example:

Take by weighing 4 parts in sample, it is fixed to claim, adds methanol 20ml, claims to decide weight, and supersound process is 15 minutes, 20 minutes, 25 minutes, 30 minutes respectively.

The selection of table 5 methanol ultrasonic time

Ultrasonic time	The peoniflorin peak area			Content of paeoniflorin (mg/g)
	The peoniflorin peak area				First pin	Second pin	Meansigma methods
	15 minutes	12042257	11924571		First pin	Second pin	Meansigma methods	11983414	5.2888
20 minutes	15 minutes	12042257	11924571	12185582	12193314	12189448	5.3402	11983414	5.2888
20 minutes	25 minutes	12200381	11959685	12185582	12193314	12189448	5.3402	12080033	5.3403
30 minutes	25 minutes	12200381	11959685	11963666	12021401	11992533.5	5.3012	12080033	5.3403

Above result shows, supersound process 25 minutes, and content is higher relatively, so select methanol supersound process 25 minutes.

Sample size is measured:

According to the method for technical scheme assay, measure three batches of finished products, take by weighing 3 duplicate samples, every bag heavy 8g the results are shown in Table 6.

Table 6 sample determination result

The sample lot number	Peoniflorin average content (mg/ bag)
The sample lot number	Peoniflorin average content (mg/ bag)	030401	53.21
030402	53.07	030401	53.21
030402	53.07	030403	54.12

6 medicament composition granule agent of experimental example are to the influence of rabbit uterus endometrium ectopia animal model endocrine level

Get healthy new zealand white rabbit 3% Nembutal vein anesthetic, back of the body position is fixing, and the abdominal part unhairing is with 2% iodine tincture and 75% ethanol routine disinfection skin of abdomen.Open the abdominal cavity, find out the uterus, downcut one section in uterus, left side, row end end-to-end anastomosis art.Uterus kind of calliper with downcutting is cut into 0.5cm * 0.5cm, plants respectively near left and right ovary and left and right cornua uteri, and stitching thread is No. 3 nylon monofilament lines, and sham operated rats is opened the abdominal cavity and do not planted endometrium, sews up stomach wall then.Take out 12 sacrifice of animal after 1 month at random, open the abdominal cavity, find out the uterus, peel off the plantation inner membrance, fix with 10% formalin, paraffin embedding, HE dyeing is observed under optical microscope, and histopathologic examination confirms.

(1) to the influence of follicule-stimulating hormone (FSH) (FSH) content

Set up model after 1 month through the endometrium transplanting, each is organized FSH and obviously raises, and with sham operated rats significant difference (P＜0.001) is arranged relatively; Perform the operation after 4 months, it is more remarkable that model group FSH raises, with before the art, postoperative relatively had significant difference in 1 month.The effect that three dosage groups of this medicament composition granule agent and danazol all have obvious inhibition FSH to raise, administration is after 3 months, and the FSH content of three dosage groups and model group comparing difference be (P＜0.05～0.001 significantly

(2) to the influence of lutropin (LH) content

The operation modeling after 1 month LH content obviously raise, model group with the operation before significant difference (P＜0.001) is relatively arranged.LH still keeps higher level after 4 months; After the administration 3 months, three dosage groups of this medicament composition granule agent have the effect that the LH value raises that suppresses, and the effect that heavy dose group inhibition LH raises is comparatively obvious, with the preceding relatively no significant difference of operation.With more also no significant difference of model group

(3) to the influence of prolactin antagonist (PRL) content

Back 1 month PRL content of performing the operation obviously raises, and before each group and the modeling significant difference (P＜0.01) is arranged more all.Performing the operation, model group PRL content significantly raises after 4 months, with comparison (P＜0.001) before the modeling.After the administration 3 months, in the agent of this medicament composition granule, heavy dose of group and danazol group PRL content obviously reduces, and with model group significant difference all (P＜0.001) arranged relatively.Modeling 1 month self relatively, in the agent of this medicament composition granule, heavy dose of group and danazol group PRL content also obviously reduces all (P＜0.001).

(4) to estradiol (E ₂) influence of content

E after the operation modeling ₂Value raises, and before each group and the modeling significant difference (P＜0.001) is arranged more all.Modeling is model group E after 4 months ₂Content significantly raises, and significant difference (P＜0.001) is arranged relatively before the modeling.After the administration 3 months, with model group relatively, in the agent of this medicament composition granule, heavy dose of group and danazol group E ₂Content obviously reduces (P＜0.01～＜0.001).With modeling after 1 month relatively, in the agent of this medicament composition granule, heavy dose of group and danazol group E ₂Content is also seen obvious reduction (P＜0.05～0.0001).

Table 7: the agent of this medicament composition granule is to the influence of rabbit uterus endometrium ectopia model endocrine level (X ± SD)

	Group	Dosage (g/kg)	FSH (mIu/ml)	LH (mIu/ml)	PRL (uIu/ml)	E ₂ (Pg/ml)
	Group	Dosage (g/kg)	FSH (mIu/ml)	LH (mIu/ml)	PRL (uIu/ml)	E ₂ (Pg/ml)	N=12 before the modeling	The heavy dose of group of dosage group in the sham-operation group model group danazol group small dose group	0.04 1.5 3 6	9.12±2.29 8.80±1.77 8.73±134 9.48±2.09 8.17±2.08 8.77±1.66	10.16±3.61 8.54±1.79 8.84±3.86 7.75±2.09 10.16±3.61 9.40±2.57	118.96±17.32 135.70±23.20 125.54±11.87 122.86±14.54 124.55±12.08 129.14±25.17	15.92±2.79 16.33±3.44 19.93±5.41 23.98±17.73 21.66±11.89 16.87±3.27
1 month n=12 of modeling	The heavy dose of group of dosage group in the sham operated rats model group danazol group small dose group	2ml 2ml 0.04 1.5 3 6	10.43±4.46 ^* 14.21±3.18### 14.16±3.55### 15.88±3.42### 14.30±3.13### 14.19±3.61###	12.33±2.62 13.76±4.23### 18.21±11.9# 16.34±5.22### 16.23±5.10## 15.34±4.72##	136.91±24.56 ^** 248.82±112.29### 220.24±52.75### 295.72±169.36## 218.24±71.59### 238.64±57.48###	17.06±2.86 ^*** 82.48±36.31### 94.26±37.35### 80.58±33.67### 78.72±25.68### 85.76±28.38###	N=12 before the modeling		0.04 1.5 3 6	9.12±2.29 8.80±1.77 8.73±134 9.48±2.09 8.17±2.08 8.77±1.66
1 month n=12 of modeling		2ml 2ml 0.04 1.5 3 6					3 months n=0 of administration	The heavy dose of group of dosage group in the sham operated rats model group danazol group small dose group	2ml 2ml 0.04 1.5 3 6	9.21±2.67 ^* 25.58±6.68△△△### 9.19±2.32 ^△△△ 18.26±8.97 ^## 16.61±4.56 ^### 13.05±3.11 ^##	10.15±3.39 ^* 14.69±4.56### 11.82±3.76 14.57±4.68### 13.77±2.66# 11.89±5.62	122.26±13.92 ^* 260.54±82.9### 123.97±14.05 ^△△△ 253.72±157.06# 129.83±17.98 ^△△△ 123.80±21.29 ^*△△△	15.09±5.56 ^* 83.33±25.88### .33.34±16.91 ^△△△# 72.59±31.89### 47.30±27.92 ^△# 37.15±27.32 ^*△△△#

Annotate: compare with model group: _*P＜0.05; _*P＜0.01; _{* *}P＜0.001.With before the modeling relatively: _#P＜0.05; _##P＜0.01; _###P＜0.001.

With 1 month relatively after the modeling: _△P＜0.05; _{△ △}P＜0.01; _{△ △ △}P＜0.001.

7 medicament composition granule agent of experimental example are to the hemorheological influence experiment of rabbit uterus endometrium ectopia animal model

Whole blood viscosity under the high shear rate of the heavy dose of group of this medicament composition granule agent is starkly lower than model group (P＜0.05).Cut the whole blood viscosity of cutting in 1 under 2 low shear rates in three dosage groups of this medicament composition granule agent reduction trend is arranged.

The agent of this medicament composition granule of table 8. is to the rheol influence of rabbit blood (n=10 X ± SD)

Group	Dosage (g/kg)	Height is cut	In cut 1	In cut 2	Low cutting	Plasma viscosity
Group	Dosage (g/kg)	Height is cut	In cut 1	In cut 2	Low cutting	Plasma viscosity			3.64±0.38	4.18±0.49	5.60±0.82	8.32±1.53	1.70±0.22
Model group		4.00±0.80	4.75±1.36	6.84±3.22	11.09±7.62	1.61±0.21			3.64±0.38	4.18±0.49	5.60±0.82	8.32±1.53	1.70±0.22
Model group		4.00±0.80	4.75±1.36	6.84±3.22	11.09±7.62	1.61±0.21	The danazol group	0.04	3.32±0.24 ^*	3.70±0.29 ^*	4.68±0.47	6.51±0.85	1.52±0.23
Small dose group	1.5	3.86±0.55	4.59±1.03	6.63±2.60	10.79±6.26	1.61±012	The danazol group	0.04	3.32±0.24 ^*	3.70±0.29 ^*	4.68±0.47	6.51±0.85	1.52±0.23
Small dose group	1.5	3.86±0.55	4.59±1.03	6.63±2.60	10.79±6.26	1.61±012	Middle dosage group	3	3.47±0.31	4.01±0.64	5.13±0.85	7.44±1.95	1.63±0.21
Heavy dose of group	6	3.35±0.17 ^*	3.79±0.21	4.93±0.41	7.06±0.87	1.45±0.16	Middle dosage group	3	3.47±0.31	4.01±0.64	5.13±0.85	7.44±1.95	1.63±0.21

Annotate: compare with model group: ^*P＜0.05.

8 medicament composition granule agent of experimental example are to the influence of rabbit uterus inner membrance dystopy volume and weight

In the agent of this medicament composition granule, heavy dose of group and danazol group Ectopic Endometrium volume obviously dwindle, weight obviously alleviates, and relatively distinguishes (P＜0.05, P＜0.01, P＜0.001) with model group.

The agent of this medicament composition granule of table 9. is to the influence of rabbit uterus Ectopic Endometrium volume and weight (X ± SD)

Group	Dosage (g/kg)	Animal (only)	Ectopic Endometrium area (cm ³)	Ectopic Endometrium weight in wet base (mg)
Group	Dosage (g/kg)	Animal (only)	Ectopic Endometrium area (cm ³)	Ectopic Endometrium weight in wet base (mg)	Sham operated rats		10	0.04±0.01 ^***	59.90±36.28 ^***
Model group		10	0.45±0.12	656.90±274.65	Sham operated rats		10	0.04±0.01 ^***	59.90±36.28 ^***
Model group		10	0.45±0.12	656.90±274.65	The danazol group	0.04	10	0.18±0.03 ^***	407.50±101.96 ^*
This medicament composition granule agent group	1.5	10	0.35±0.16	524.50±315.24	The danazol group	0.04	10	0.18±0.03 ^***	407.50±101.96 ^*
This medicament composition granule agent group	1.5	10	0.35±0.16	524.50±315.24	This medicament composition granule agent group	3	10	0.26±0.11 ^**	418.50±195.88 ^*
This medicament composition granule agent group	6	10	0.18±0.05 ^***	409.30±132.62 ^*	This medicament composition granule agent group	3	10	0.26±0.11 ^**	418.50±195.88 ^*

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

9 medicament composition granule agent of experimental example are tested rabbit endometriosis histopathologic examination

Sham operated rats: do not see ectopic inner membrance around uterus, ovary and the fallopian tube.Model group: visible part fiber, smooth muscle and interior film component in the fibrofatty tissue.The visible form of intimal epithelium differs, have flat, cube, low column or high column.The theca interna body of gland differs in size, and part is intensive, and part is sparse, and some theca interna is because hypertrophy is excessively charged into intracavity, and in the form of sheets or the island shape, indivedual bodies of gland expansions are pencil.

The danazol group: most of dystopy body of gland as seen, local dense, ectopic intimal epithelium is flat, broken, come off indivedual dystopy body of gland pyknosis.

This medicament composition granule agent small dose group: in the Ectopic Endometrium zone, theca interna is thinner, the visible zigzag of subregion epithelium, and epithelium has fragmentation, comes off, and theca interna body of gland major part is not obvious, indivedual visible pyknosis, fragmentation.

Dosage group in the agent of this medicament composition granule: Ectopic Endometrium as seen, but theca interna is thinner, glandular epithelium part growing way still can, be huge dentalation, the part exuviation, the theca interna body of gland is not obvious.The heavy dose of group of this medicament composition granule agent: mirror down with in, small dose group, the theca interna attenuation, epithelial cell cube or flat, part has and comes off visible single body of gland in the fibrous tissue.

10 medicament composition granule agent of experimental example are tested in the uterotonic influence of body rat

Get 70 of healthy rats, random packet was got healthy unpregnancy female rats subcutaneous injection diethylstilbestrol (2mg/kg body weight) in 24-48 hour before the experiment, manually impelled animal to be in proestrus or rutting period, to improve the sensitivity of uterus to medicine.With pentobarbital sodium intraperitoneal anesthesia (50mg/kg), back of the body position is fixing, opens the abdominal cavity and finds out the uterus, with line one Aconitum carmichaeli Debx. palace is mentioned with pressure transducer and is connected partial insulation during experiment.Write down (10 minutes) behind one section normalized curve, inject oxytocin (3u/ only) from femoral vein, simultaneously be subjected to the reagent thing by duodenum, model group (both the oxytocin group is given normal saline 10ml/kg) is observed the uterotonic frequency in administration front and back, amplitude and uterine motility (frequency * amplitude).

Table 10: the agent of this medicament composition granule is to the influence of normal uterine motility (n=10, X ± SD)

Group	Dosage g/kg	Before the administration (gram)	Energy after the administration (gram)
			Energy after the administration (gram)				30min	40min	50min	60min
			Matched group		0.42 ± difference	0.46±0.14 0.03±0.17**	30min	40min	50min	60min	0.50±0.15 0.07±0.16**	0.48±0.12 0.05±0.14***	0.47±0.17 0.04±0.15***
Model group		0.45 ± difference	Matched group		0.42 ± difference	0.46±0.14 0.03±0.17**	1.35±0.74 ^** 0.90±0.79 ^△△	1.96±1.42 ^** 1.51±1.49 ^△△	1.91±0.77 ^** 1.46±0.79 ^△△△	2.42±1.53 ^*** 1.97±1.52 ^△/	0.50±0.15 0.07±0.16**	0.48±0.12 0.05±0.14***	0.47±0.17 0.04±0.15***
Model group		0.45 ± difference	The indometacin group	0.014	0.40 ± difference	0.32±0.11 ^* -0.08±0.11**	1.35±0.74 ^** 0.90±0.79 ^△△	1.96±1.42 ^** 1.51±1.49 ^△△	1.91±0.77 ^** 1.46±0.79 ^△△△	2.42±1.53 ^*** 1.97±1.52 ^△/	0.29±0.11 ^ -0.11±0.10	0.20±0.06 ^* -0.20±0.08*	0.20±0.09* -0.20±0.09*
The danazol group	0.05	0.39 ± difference	The indometacin group	0.014	0.40 ± difference	0.32±0.11 ^* -0.08±0.11**	0.49±0.22 0.10±0.26**	0.50±0.18 0.11±0.26**	0.39±0.21 -0.00±0.19***	0.34±0.18 -0.05±0.12***	0.29±0.11 ^ -0.11±0.10	0.20±0.06 ^* -0.20±0.08*	0.20±0.09* -0.20±0.09*
The danazol group	0.05	0.39 ± difference	Small dose group	2.5	0.42 ± difference	0.58±0.26 0.16±0.29*	0.49±0.22 0.10±0.26**	0.50±0.18 0.11±0.26**	0.39±0.21 -0.00±0.19***	0.34±0.18 -0.05±0.12***	0.63±0.28 ^* 0.22±0.31*	0.65±0.24 ^* 0.23±0.30***	0.58±0.23 0.17±0.27**
Middle dosage group	5	0.45 ± difference	Small dose group	2.5	0.42 ± difference	0.58±0.26 0.16±0.29*	0.63±0.21 ^* 0.19±0.27*	0.56±0.16 0.11±0.17***	0.49±0.13 0.05±0.17***	0.45±0.17 0.01±0.21***	0.63±0.28 ^* 0.22±0.31*	0.65±0.24 ^* 0.23±0.30***	0.58±0.23 0.17±0.27**
Middle dosage group	5	0.45 ± difference	Heavy dose of group	10	0.40 ± difference	0.42±0.22 0.02±0.21**	0.63±0.21 ^* 0.19±0.27*	0.56±0.16 0.11±0.17***	0.49±0.13 0.05±0.17***	0.45±0.17 0.01±0.21***	0.39±0.15 -0.01±0.1**	0.30±0.11 -0.05±0.23***	0.27±0.11 ^ -0.13±0.10*

Annotate: compare with matched group: ^{△ △}P＜0.01.Compare with model group: ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001.

Self compares: ^#P＜0.05; , ^##P＜0.01, ^###＜0.001.

The agent of this medicament composition granule is little, in, all have obvious inhibition oxytocin to cause the effect of uterine motility 30,40,50,60 minutes the time after big three dosage groups, indometacin group and the administration of danazol group, with model group (oxytocin group) relatively (P＜0.05～P＜0.001).

11 medicament composition granule agent of experimental example are to the microcirculatory influence experiment of mice mesentery

60 random packet of healthy mice, every day, gastric infusion was 1 time, successive administration 7 days.Anisodamine group (65-42 group) administered intramuscular 1 time, after the administration 15 minutes, after other each administration group last administration 30 minutes with pentobarbital sodium intraperitoneal anesthesia (50mg/kg), open the abdominal cavity, pull out one section intestinal loop, be tiled on the plate, drip warm saline on the mesentery, with anti-drying, partial insulation, tail vein injection 15% macromolecule right rotary glycoside (10ml/kg), injection back 10,20, in the time of 30 minutes at body surface microcirculation visualizer, the blood fluidised form of different time after the observation administration, statistical procedures employing reference differential technique compares between organizing.

Model group tail vein award the injection macromolecule right rotary glycoside after the blood flow fluidised form by+level be changed to +++level, flow velocity causes circulatory disorders by slowing down gradually soon, stops until blood flow.Behind this medicament composition granule agent small dose group tail vein injection macromolecule right rotary glycoside during 10min and model group notable difference (u＜0.05) is relatively arranged, in, behind heavy dose of group and the danazol group tail vein injection macromolecule right rotary glycoside 10,20, during 30min, the microcirculatory fluidised form of mice mesentery is obviously improved, the blood flow fluidised form mostly is ++ below the level, significant difference (u＜0.05～u＜0.01) is respectively relatively arranged with model group.

The influence (n=10) of table 11. pair mice mesentery microcirculation disturbance blood fluidised form

Group	Dosage g/kg	Different time blood fluidised form after the administration
																	10(min)					20(min)					30(min)
		-	+	++	+++		-	+	++	+++		-	+	++	+++		10(min)					20(min)					30(min)
		-	+	++	+++		-	+	++	+++		-	+	++	+++		Model group		0	6	4	0		0	5	4	1		0	4	3	3
The 65-42 group	5.2mg	9	1	0	0	**	7	3	0	0	**	5	4	1	0	**	Model group		0	6	4	0		0	5	4	1		0	4	3	3
The 65-42 group	5.2mg	9	1	0	0	**	7	3	0	0	**	5	4	1	0	**	The danazol group	0.1	3	7	0	0	*	2	8	0	0	*	1	9	0	0	*
This medicament composition granule agent group	3.75	2	8	0	0	*	1	3	6	0		1	3	4	2		The danazol group	0.1	3	7	0	0	*	2	8	0	0	*	1	9	0	0	*
This medicament composition granule agent group	3.75	2	8	0	0	*	1	3	6	0		1	3	4	2		This medicament composition granule agent group	7	2	8	0	0	*	2	7	1	0	*	2	6	2	0	*
This medicament composition granule agent group	15	5	5	0	0	**	4	4	2	0	*	2	5	3	0	*	This medicament composition granule agent group	7	2	8	0	0	*	2	7	1	0	*	2	6	2	0	*

Annotate: compare with model group ^*U＜0.05. ^*U＜0.01

The result shows, in the agent of this medicament composition granule, heavy dose of group has certain protective role to local disturbance of blood circulation.

12 medicament composition granule agent of experimental example antiinflammatory action---to the granulomatous influence experiment of rat

Animal is divided into 5 groups at random, rat is with 1% pentobarbital sodium intraperitoneal anesthesia during experiment, the routine disinfection skin of axillary fossa, open otch, to weigh cotton balls (the heavy 30mg of each cotton balls) behind autoclaving, every cotton balls adds Duropenin 1mg/0.1ml, and it is subcutaneous to implant left and right side axillary region respectively after 50 ℃ of oven for drying.Performing the operation began gastric infusion the same day, administration every day 1 time, and continuous 7 days, the cervical vertebra dislocation was put to death after 7 days, took out cotton balls and claimed weight in wet base with electronic balance, claimed dry weight in 60 degree baking ovens placements again after 12 hours then.Respectively organize granulomatous average weight in wet base and dry weight (all deducting the raw cotton ball weight) after calculating relatively causes inflammation, compare between organizing with the t check.

In the agent of this medicament composition granule, the dry weight and the weight in wet base of heavy dose of group and aspirin group granuloma cotton balls all be starkly lower than model group (P＜0.05～0.01), the agent of this medicament composition granule has the effect of obvious inhibition granuloma induced by implantation of cotton pellets.Dry weight, the weight in wet base of small dose group granuloma cotton balls are also light than model group, but compare no significant difference with model group.

The agent of this medicament composition granule of table 12. is to (the X ± SD) of the granulomatous influence of rat due to the cotton balls

Group	Dosage (g/kg)	Number of animals (n)	Cause scorching back cotton balls weight (mg)
			Cause scorching back cotton balls weight (mg)		Weight in wet base	Dry weight
			Model group		Weight in wet base	Dry weight	10	895.60±194.83	151.30±32.08
The aspirin group	0.15	10	Model group		688.10±55.30 ^**	115.80±11.12 ^**	10	895.60±194.83	151.30±32.08
The aspirin group	0.15	10	This medicament composition granule agent group	2.5	688.10±55.30 ^**	115.80±11.12 ^**	10	765.60±72.33	131.30±14.79
This medicament composition granule agent group	5	10	This medicament composition granule agent group	2.5	745.40±81.14 ^*	120.60±20.15 ^*	10	765.60±72.33	131.30±14.79
This medicament composition granule agent group	5	10	This medicament composition granule agent group	10	745.40±81.14 ^*	120.60±20.15 ^*	10	701.60±90.78 ^*	119.90±18.77 ^*

Annotate: compare with model group: ^*P＜0.05, ^*P＜0.01.

13 medicament composition granule agent of experimental example analgesic activity---Dichlorodiphenyl Acetate brings out the influence test of pain

Mice is divided into 5 groups, every group 10, each treated animal gastric infusion every day 1 time, continuous 7 days, pethidine group gastric infusion 1 time, model group are given with the volume normal saline, and each group is 30 minutes lumbar injection 0.6% acetic acid (0.2ml/ only) after the last administration all, observe each treated animal in 20 minutes and turn round the body number of times, between organizing relatively (t check) by what acetic acid brought out.

Can cause the more persistent pain stimulation of mice behind the lumbar injection acetic acid, writhing response appears in mice.In the agent of this medicament composition granule, heavy dose of all have the effect that reduces pain in various degree, animal to turn round the body number of times obviously to be less than model group (P＜0.05～0.01).

Table 13: to the influence of mice pain (turning round body) effect (X ± SD)

Grouping	Dosage (g/kg)	Number of animals (only)	Turn round the body number of times
Grouping	Dosage (g/kg)	Number of animals (only)	Turn round the body number of times	Model group		10	74550±8.14
The dolantin group	0.05	10	34.80±9.76 ^***	Model group		10	74550±8.14
The dolantin group	0.05	10	34.80±9.76 ^***	This medicament composition granule agent group	3.75	10	70.80±14.93
This medicament composition granule agent group	7.5	10	58.10±20.07 ^*	This medicament composition granule agent group	3.75	10	70.80±14.93
This medicament composition granule agent group	7.5	10	58.10±20.07 ^*	This medicament composition granule agent group	15	10	51.10±21.16 ^**

Annotate: compare with matched group ^*P＜0.05; ^*P＜0.01; ^{* *}P＜0.001.

14 medicament composition granule agent of experimental example are to the influence test of immunologic hypofunction mice serum agglutinin

Get 66 of mices, be divided into 6 groups at random, every group 11, administration every day 1 time, successive administration 7 days, administration is administration the 3rd day after 48 hours, and every mouse peritoneal is injected 5% sheep red blood cell (SRBC) 0.25ml and carried out immunity, and the while is subcutaneous injection cyclophosphamide 100mg/kg 1 time (except the blank group) again.Got blood by the eyeball rear vein beard in the 6th day after the immunity, centrifugal, separation of serum is measured anti-SRBC antibody titer in the serum with micro-agglutination.

The antibody product obviously reduces in the cyclophosphamide group mice serum, with blank group comparing difference remarkable (P＜0.01); In the agent of this medicament composition granule, heavy dose of group all improved the level that cyclophosphamide causes anti-sheep red blood cell antibody in the immunologic hypofunction mice serum significantly, with cyclophosphamide group comparing difference highly significant (P＜0.01～0.001); The positive control drug levamisole has also significantly improved the antibody response (P＜0.01) of mice to sheep red blood cell.Illustrate that the agent of this medicament composition granule can improve the humoral immune function that cyclophosphamide causes the immunologic hypofunction mice.

Table 14: this medicament composition granule agent

To the effect of immunologic hypofunction mice serum agglutinin (n=11 X ± SD)

Group	Dosage (g/kg)	The antibody product	t	P
Group	Dosage (g/kg)	The antibody product	t	P	The blank group		21.261±4.741	3.6662	＜0.01
The cyclophosphamide group	100mg	14.786±3.440			The blank group		21.261±4.741	3.6662	＜0.01
The cyclophosphamide group	100mg	14.786±3.440			The levamisole group	30mg	20.287±3.927	3.4947	＜0.01
This medicament composition granule agent group	3.75	14.841±5.184	0.0293	＞0.05	The levamisole group	30mg	20.287±3.927	3.4947	＜0.01
This medicament composition granule agent group	3.75	14.841±5.184	0.0293	＞0.05	This medicament composition granule agent group	7.5	20.824±5.788	2.9742	＜0.01
This medicament composition granule agent group	15	22.427±4.367	4.5586	＜0.001	This medicament composition granule agent group	7.5	20.824±5.788	2.9742	＜0.01

Annotate: compare with the cyclophosphamide group.

15 medicament composition granule agent of experimental example are to the influence experiment of immunologic hypofunction mice phagocytic function

Get 60 of mices, be divided into 6 groups at random, every day, gastric infusion was 1 time, successive administration 7 days, administration is that mouse subcutaneous injection cyclophosphamide 100mg/kg (except the blank group) is given in administration the 3rd day after 48 hours, and mouse tail vein injection is only given with 10% india ink 0.2ml/ in the 8th day of administration.The eyeball rear vein beard was got blood 0.02ml respectively back 2 minutes of injection and 10 minutes, join lysed erythrocyte in the test tube that the 4ml distilled water is housed, be UV-120-02 type spectrophotometric determination optical density (OD) value of 600nm then at wavelength, calculate in the unit interval speed K of carbon clearance in the serum.OD ₂And OD ₁₀Injected behind the india ink 2 minutes and the OD value of 10 minutes institute's blood samplings t respectively ₂And t ₁₀For getting the blood time.Result of the test t check.

Table 15: this medicament composition granule agent

To the influence of immunologic hypofunction mice phagocytic function (n=10 X ± SD)

Group	Dosage (g/kg)	Carbon clearance index (K)	t	P
Group	Dosage (g/kg)	Carbon clearance index (K)	t	P	Matched group		0.0210±0.0050	2.3027	＜0.05
The cyclophosphamide group	100mg	0.0152±0.0062			Matched group		0.0210±0.0050	2.3027	＜0.05
The cyclophosphamide group	100mg	0.0152±0.0062			The levamisole group	30mg	0.0302±0.0098	4.0903	＜0.001
This medicament composition granule agent group	3.75	0.0210±0.0078	1.8407	＞0.05	The levamisole group	30mg	0.0302±0.0098	4.0903	＜0.001
This medicament composition granule agent group	3.75	0.0210±0.0078	1.8407	＞0.05	This medicament composition granule agent group	7.5	0.0250±0.0124	2.2353	＜0.05
This medicament composition granule agent group	15	0.0289±0.0109	3.4548	＜0.01	This medicament composition granule agent group	7.5	0.0250±0.0124	2.2353	＜0.05

Annotate: compare with the cyclophosphamide group.

It is as shown in the table, and the cyclophosphamide group has reduced the monocytic phagocytic function of normal mouse significantly, with blank group comparing difference remarkable (P＜0.05); The carbon clearance index of positive control drug levamisole is significantly higher than cyclophosphamide group (P＜0.001), in the agent of this medicament composition granule, heavy dose of group can obviously improve cyclophosphamide and cause immunologic hypofunction mice carbon clearance index, significantly distinguish (P＜0.05～0.01) with cyclophosphamide group comparing difference.This medicine causes immunologic hypofunction mice non-specific immunity to cyclophosphamide potentiation.

16 medicament composition granule agent of experimental example are to the influence experiment of the tardy paraphilia reaction of immunologic hypofunction mice

Get 70 of mices, be divided into 7 groups at random, 10 every group, administration every day 1 time, successive administration 8 days, except that the blank treated animal, each was organized mice and evenly is applied in 1 sensitization of abdominal part depilation place with 1%DNFB acetone olive oil solution 50 μ l in administration the 3rd day.Simultaneously except that blank group and model group animal, all animal subcutaneous injection cyclophosphamide 100mg/kg 1 time.The 5th day whole mices after sensitization evenly are applied in every mouse right ear two sides with the same antigen 20 μ l and attack.Attack back 24h, the dislocation of mice cervical vertebra is put to death, cut left and right sides auricular concha, take off the auricle of diameter 8mm, weigh with card punch.Swelling degree result carries out the t check.

Table 16: this medicament composition granule agent

To the effect of immunologic hypofunction mice FPR (n=10 X ± SD)

Group	Dosage (g/kg)	Swelling degree FPR (mg)	t	P
Group	Dosage (g/kg)	Swelling degree FPR (mg)	t	P	The blank group		5.60±0.84
Model group		11.00±2.35△	6.8425	＜0.001	The blank group		5.60±0.84
Model group		11.00±2.35△	6.8425	＜0.001	The cyclophosphamide group	100mg	3.90±0.74#	9.1129	＜0.001
The levamisole group	30mg	10.10±1.10 ^*	14.7887	＜0.001	The cyclophosphamide group	100mg	3.90±0.74#	9.1129	＜0.001
The levamisole group	30mg	10.10±1.10 ^*	14.7887	＜0.001	This medicament composition granule agent group	3.75	5.30±0.67 ^*	4.4349	＜0.001
This medicament composition granule agent group	7.5	6.20±0.78 ^*	6.7647	＜0.001	This medicament composition granule agent group	3.75	5.30±0.67 ^*	4.4349	＜0.001
This medicament composition granule agent group	7.5	6.20±0.78 ^*	6.7647	＜0.001	This medicament composition granule agent group	15	8.20±0.91 ^*	11.5932	＜0.001

Annotate: △ and blank group are relatively; # and model group are relatively; ^*Compare with the cyclophosphamide group.

It is as shown in the table, and model control group mice ear degree obviously increases, and compares P＜0.001 with the blank group, illustrates that tardy paraphilia reaction forms; Cyclophosphamide group mice ear degree is obviously light than model group, and with the remarkable P of model group comparing difference＜0.001, the immunologic hypofunction mouse model forms; Positive control drug levamisole group and cyclophosphamide group immunologic function P＜0.001 that relatively is significantly improved, be subjected to the agent of this medicament composition granule of reagent little, in, heavy dose of group all causes immunologic hypofunction mice ear degree to cyclophosphamide that significant potentiation is arranged, with the very remarkable equal P of cyclophosphamide group comparing difference＜0.001, this medicine has potentiation to the cellular immune function that cyclophosphamide causes the immunologic hypofunction mice.

Embodiment 1: medicament composition granule agent preparation

Radix Paeoniae Rubra 313g Radix Bupleuri 208g Pollen Typhae 208g Radix Astragali 417g

Radix Angelicae Sinensis 250g Rhizoma Chuanxiong 208g Fructus Psoraleae 208g Rhizoma Corydalis 313g

Ramulus Cinnamomi 125g Herba Asari 62g Myrrha (processed) 125g

More than ten simply, Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis extract 3 times with the alcohol heating reflux of 6 times of amounts 60%, each 2 hours, the merging ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 6 times of water gagings, soak 2 hours, and water vapour distillation 8 hours is collected volatile oil, and aqueous solution and medicinal residues are standby; Get volatile oil and carry out inclusion with the betacyclodextrin of 6 times of amounts and the water of 60 times of amounts, 40 ℃ were stirred 1 hour, and cold preservation 12 hours filters, and 40 ℃ of cold drying of inclusion complex are ground into fine powder, and are standby; Radix Bupleuri, Radix Paeoniae Rubra, Pollen Typhae and the Radix Astragali add 8 times of water gagings, extract 3 times, each 1 hour, close with medicinal residues such as above-mentioned Ramulus Cinnamomi for the third time and fry in shallow oil, collecting decoction filters, filtrate and above-mentioned aqueous solution merge, survey relative density 1.10-1.15 when amalgamation liquid is concentrated into 50 ℃, add ethanol and make and contain the alcohol amount and reach 70%, cold preservation is spent the night, filter, survey relative density 1.15-1.20 when medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis reclaims ethanol to 50 ℃ respectively, drying is with above-mentioned volatile oil betacyclodextrin inclusion complex fine powder and an amount of (about 500g) mixing of dextrin, make granule, drying is made granule 1000g, promptly.

Embodiment 2: the preparation of medicament composition capsule agent

Radix Paeoniae Rubra 7kg Radix Angelicae Sinensis 7kg Pollen Tyjphae 4kg

Rhizoma Chuanxiong 6kg Ramulus Cinnamomi 2kg Rhizoma Corydalis 8kg

Radix Bupleuri 4kg Radix Astragali 11kg Fructus Psoraleae 4kg

Herba Asari 2kg Myrrha (processed) 2kg

Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis are measured 70% alcohol heating reflux with 5 times, extract 2 times, and each 3 hours, merge ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 5 times of water gagings, soak 3 hours, and water vapour distillation 7 hours is collected volatile oil, and aqueous solution and medicinal residues are standby; Get volatile oil and carry out inclusion with the betacyclodextrin of 8 times of amounts and the water of 60 times of amounts, 60 ℃ were stirred 0.5 hour, and cold preservation 12 hours filters, and 35 ℃ of cold drying of inclusion complex are ground into fine powder, and are standby; Radix Bupleuri, Radix Paeoniae Rubra, Pollen Typhae and the Radix Astragali add 9 times of water gagings, extract 2 times, each 1.5 hours, medicinal residues such as the general's extracting solution second time and above-mentioned Ramulus Cinnamomi close and fry in shallow oil again, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, when being concentrated into 50 ℃, surveys amalgamation liquid relative density 1.10, add ethanol and make and contain alcohol amount and reach 80%, cold preservation is spent the night, and filters, when reclaiming ethanol to 50 ℃ respectively, surveys medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis relative density 1.15, drying, with above-mentioned volatile oil beta cyclodextrin inclusion complex fine powder and dextrin by weight 1: 0.5 mixing, make granule, drying is made capsule.

Embodiment 3: the preparation of pharmaceutical composition tablet

Radix Paeoniae Rubra 8kg Radix Angelicae Sinensis 5kg Pollen Tyjphae 6kg

Rhizoma Chuanxiong 4kg Ramulus Cinnamomi 4kg Rhizoma Corydalis 7kg

Radix Bupleuri 6kg Radix Astragali 9kg Fructus Psoraleae 6kg

Herba Asari 1kg Myrrha (processed) 4kg

Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis are measured 50% alcohol heating reflux with 7 times, extract 2 times, and each 1.5 hours, merge ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 7 times of water gagings, soak 1.5 hours, and water vapour distillation 9 hours is collected volatile oil, and aqueous solution and medicinal residues are standby; Get volatile oil and carry out inclusion with the betacyclodextrin of 4 times of amounts and the water of 80 times of amounts, 40 ℃ were stirred 1.5 hours, and cold preservation 12-18 hour, filter, 35 ℃ of cold drying of inclusion complex are ground into fine powder, and are standby; Radix Bupleuri, Radix Paeoniae Rubra, Pollen Typhae and the Radix Astragali add 7 times of water gagings, extract 2 times, each 0.5 hour, medicinal residues such as the general's extracting solution second time and above-mentioned Ramulus Cinnamomi close and fry in shallow oil again, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, when being concentrated into 50 ℃, surveys amalgamation liquid relative density 1.15, add ethanol and make and contain alcohol amount and reach 80%, cold preservation is spent the night, and filters, when reclaiming ethanol to 50 ℃ respectively, surveys medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis relative density 1.20, drying, with above-mentioned volatile oil beta cyclodextrin inclusion complex fine powder and dextrin by weight 1: 0.5 mixing, make granule, drying is made tablet.

Embodiment 4: the preparation of granule

Radix Paeoniae Rubra 313g Radix Bupleuri 208g Pollen Typhae 208g Radix Astragali 417g Radix Angelicae Sinensis 250g

Rhizoma Chuanxiong 208g Fructus Psoraleae 208g Rhizoma Corydalis 313g Ramulus Cinnamomi 125g Herba Asari 62g

Myrrha (processed) 125g

Make granule 1000g altogether, endometriosis patient takes, each 8g, every day 3 times.

Embodiment 5: the preparation of capsule

Radix Paeoniae Rubra 7kg Radix Angelicae Sinensis 7kg Pollen Tyjphae 4kg Rhizoma Chuanxiong 6kg Ramulus Cinnamomi 2kg

Rhizoma Corydalis 8kg Radix Bupleuri 4kg Radix Astragali 11kg Fructus Psoraleae 4kg Herba Asari 2kg

Myrrha (processed) 2kg

Make capsule 5750g altogether, endometriosis patient takes, each 2g, every day 3 times.

Embodiment 6: the preparation of tablet

Radix Paeoniae Rubra 8kg Radix Angelicae Sinensis 5kg Pollen Tyjphae 6kg Rhizoma Chuanxiong 4kg Ramulus Cinnamomi 4kg

Rhizoma Corydalis 7kg Radix Bupleuri 6kg Radix Astragali 9kg Fructus Psoraleae 6kg Herba Asari 1kg

Myrrha (processed) 4kg

Make tablet 6200g altogether, endometriosis patient takes, each 2g, every day 3 times.

Embodiment 7: the preparation of oral agents

Myrrha (processed) 4kg

Extract by above-mentioned preparation technology, add adjuvant, make oral liquid, every bottle of 100ml, totally 3100 bottles, endometriosis patient takes, each 10ml, every day 3 times.

Embodiment 8: the preparation of drop pill

Myrrha (processed) 2kg

Extract by above-mentioned preparation technology, add adjuvant, make drop pill, every 0.05g.Endometriosis patient takes, one time 20,3 times on the one.

Embodiment 9: the discrimination method in the method for quality control of pharmaceutical composition

A. get this medicament composition granule agent agent 4g, porphyrize adds methanol 20ml, and supersound process 20 minutes filters, and filtrate is as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 12 μ l, reference substance solution 10 μ l, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 40: 5: 10: 0.2 chloroform-ethyl acetate-methanol-formic acid was developing solvent, launch, take out, dry, spray is with 50% phosphoric acid ethanol solution of 5% vanillin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

B. get psoralen and isopsoralen reference substance, add methanol respectively and make the solution that every 1ml contains 0.3mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 12 μ l, reference substance solution 8 μ l under the above-mentioned discriminating A item, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 8: 2 normal hexane-ethyl acetates was developing solvent, launch, take out, dry, spray is put under the 365nm ultra-violet lamp and is inspected with 10% sodium hydrate methanol solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

C. get this medicament composition granule agent agent 5g, porphyrize adds ethanol 15ml, and supersound process 1 hour filters, and filtrate is as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 2g of Rhizoma Chuanxiong control medicinal material, and each adds ethanol 10ml, and supersound process is 1 hour respectively, filters, and filtrate is medical material solution in contrast; According to thin layer chromatography test, draw each 10 μ l of above-mentioned need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 9: 1 cyclohexane extraction-ethyl acetates was developing solvent, launched, and took out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

D. get this medicament composition granule agent agent 12g, porphyrize adds ammonia solution 2ml and chloroform 30ml, and supersound process 40 minutes filters, and filtrate is reclaimed chloroform to doing, and residue adds ethanol 5ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw each 12 μ l of above-mentioned two kinds of solution, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 5: 4: 0.4 normal hexane-ethyl acetate-ammonia was developing solvent, launch, take out, dry, smoked clear with iodine vapor to the speckle colour developing, inspect under the daylight; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

E. get this medicament composition granule agent agent 24g, porphyrize is put in the round-bottomed flask of 250ml, adds entry 50ml, connects volatile oil extractor, adds water from the determinator upper end and makes and be full of scale, and 60～90 ℃ add petroleum ether 1.5ml, connect reflux condensing tube then; Flask contents is heated to boils, and keep little and boiled 2 hours, stop heating, get petroleum ether layer as need testing solution; Other gets Herba Asari control medicinal material 10g and extracts volatile oil, adds chloroform 30ml and makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 15 μ l, reference substance solution 3 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 95: 5 benzene-ethyl acetates was developing solvent, launch, take out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

F. get this medicament composition granule agent agent 6g, porphyrize adds kieselguhr 2g, mixing, the methanol solution 20ml that adds 2% sodium hydroxide, heating and refluxing extraction 1 hour filters, the filtrate evaporate to dryness adds water 30ml and makes dissolving, uses the water saturated n-butanol extraction of 40ml at every turn, extract 3 times, merge n-butyl alcohol liquid, with water washing 2 times, discard water liquid, with 1% potassium dihydrogen phosphate washing 1 time, discard water liquid again, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.3mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 12 μ l, reference substance solution 6 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution with 20: 7: 2 chloroform-methanol-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be baked to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Embodiment 10: the content assaying method in the pharmaceutical composition method of quality control

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 70: 430 acetonitrile-water is mobile phase; The detection wavelength is 230nm; Theoretical cam curve is calculated by peoniflorin should be not less than 3000;

It is an amount of that the preparation of reference substance solution takes by weighing the peoniflorin reference substance, adds methanol and make the solution that every 1ml contains 0.5mg, promptly;

Content under this medicament composition granule agent agent content uniformity item is got in the preparation of need testing solution, and porphyrize is got 1.2g, put in the tool plug conical flask, it is fixed to claim, adds methanol 20ml, claim to decide weight, supersound process 25 minutes is taken out, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter with the 0.45um microporous filter membrane, get subsequent filtrate, promptly;

Algoscopy is drawn above-mentioned reference substance solution 10 μ l, need testing solution 15 μ l respectively, injects chromatograph of liquid, measures, promptly;

This product every bag (8g dress) contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 23.0mg; Embodiment 11: the method for quality control of pharmaceutical composition

This product every bag (8g dress) contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 23.0mg.

Claims

1, the endometriotic pharmaceutical composition of a kind of treatment is characterized in that this pharmaceutical composition is to be made by following raw material medicaments in part by weight:

Radix Paeoniae Rubra 7-8 weight portion Radix Angelicae Sinensis 5-7 weight portion Pollen Tyjphae 4-6 weight portion

Herba Asari 1-2 weight portion Myrrha (processed) 2-4 weight portion.

2, pharmaceutical composition as claimed in claim 1 is characterized in that this pharmaceutical composition is to be made by following raw material medicaments in part by weight:

Herba Asari 1.5 weight portion Myrrha (processed)s 3 weight portions.

3, pharmaceutical composition as claimed in claim 1 is characterized in that this pharmaceutical composition is to be made by following raw material medicaments in part by weight:

Herba Asari 2 weight portion Myrrha (processed)s 2 weight portions.

4, pharmaceutical composition as claimed in claim 1 is characterized in that this pharmaceutical composition is to be made by following raw material medicaments in part by weight:

Herba Asari 1 weight portion Myrrha (processed) 4 weight portions.

5,, it is characterized in that said composition can make dosage form clinically a kind of or that pharmaceutically accept: tablet, pill, capsule, granule, drop pill, soft capsule, suppository, suspensoid, oral liquid or enema as claim 1,2,3 or 4 described pharmaceutical compositions.

6, as claim 1,2,3 or 4 described preparation of drug combination methods, it is characterized in that this method may further comprise the steps:

Get it filled compositions crude drug ten simply by proportioning dosage, and Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis are doubly measured the alcohol heating reflux of 50-70% with 5-8, extract 2-4 time, and each 1.5-3 hour, merge ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 3-7 times of water gaging, soaked 0.5-3 hour, and water vapour distillation 2-8 hour, collect volatile oil, aqueous solution and medicinal residues are standby; Get the water that beta-schardinger dextrin-that volatile oil doubly measures with 4-8 and 40-80 doubly measure and carry out inclusion, 30-60 ℃ was stirred 0.5-1.5 hour, and cold preservation 10-18 hour, filter, inclusion complex 30-55 ℃ cold drying is ground into fine powder, and is standby; Radix Bupleuri, Radix Paeoniae Rubra, the Pollen Typhae and the Radix Astragali add 7-10 times of water gaging, extract 2-4 time, each 0.5-1.5 hour, medicinal residues such as last extracting solution and above-mentioned Ramulus Cinnamomi are closed fry in shallow oil, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, when being concentrated into 50 ℃, surveys amalgamation liquid relative density 1.10-1.15, add ethanol and make and contain alcohol amount and reach 60-80%, cold preservation is spent the night, and filters, when reclaiming ethanol to 50 ℃ respectively, surveys medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis relative density 1.15-1.20, drying, with above-mentioned volatile oil beta-cyclodextrin inclusion complex fine powder and dextrin by weight 1: the 0.5-1.5 mixing, make granule, drying is made capsule, tablet or granule.

7, preparation of drug combination method as claimed in claim 6 is characterized in that this method may further comprise the steps:

Get it filled compositions crude drug ten simply by proportioning dosage, and Radix Angelicae Sinensis, Rhizoma Chuanxiong, Fructus Psoraleae, Rhizoma Corydalis are measured 60% alcohol heating reflux with 6 times, extract 3 times, and each 2 hours, merge ethanol extract, medicinal liquid is standby; Ramulus Cinnamomi, Herba Asari, Myrrha add 6 times of water gagings, soak 2 hours, and water vapour distillation 8 hours is collected volatile oil, and aqueous solution and medicinal residues are standby; Get volatile oil and carry out inclusion with the beta-schardinger dextrin-of 6 times of amounts and the water of 60 times of amounts, 40 ℃ were stirred 1 hour, and cold preservation 12 hours filters, and 40 ℃ of cold drying of inclusion complex are ground into fine powder, and are standby; Radix Bupleuri, Radix Paeoniae Rubra, the Pollen Typhae and the Radix Astragali add 8 times of water gagings, extract 3 times, each 1 hour, again medicinal residues such as extracting solution and above-mentioned Ramulus Cinnamomi are closed and fry in shallow oil, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, when being concentrated into 50 ℃, surveys amalgamation liquid relative density 1.10-1.15, add ethanol and make and contain alcohol amount and reach 70%, cold preservation is spent the night, and filters, when reclaiming ethanol to 50 ℃ respectively, surveys medicinal liquid such as filtrate and above-mentioned Radix Angelicae Sinensis relative density 1.15-1.20, drying, with above-mentioned volatile oil beta-cyclodextrin inclusion complex fine powder and dextrin by weight 1: 1 mixing, make granule, drying is made capsule, tablet or granule.

8,, it is characterized in that discrimination method in this method comprises a kind of and/or several in the following discriminating as the method for quality control of claim 1,2,3 or 4 described pharmaceutical compositions:

A. get the drug combination preparation of 1/6 times of phase taking dose on the same day, porphyrize adds methanol 15-25ml, and supersound process 10-30 minute, filter, filtrate is as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 0.3-0.6mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 5-15 μ l, reference substance solution 5-15 μ l, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, chloroform-ethyl acetate-methanol-formic acid with 35-45: 4-6: 8-12: 0.15-0.25 is developing solvent, launch, take out, dry, spray is with the 45-55% phosphoric acid ethanol solution of 3-6% vanillin, and it is clear to be heated to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get the drug combination preparation of 5/24 times of phase taking dose on the same day, porphyrize adds ethanol 10-20ml, and supersound process 0.5-1.5 hour, filter, filtrate is as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 1.5-2.5g of Rhizoma Chuanxiong control medicinal material, and each adds ethanol 8-12ml, and supersound process is 0.5-1.5 hour respectively, filters, and filtrate is medical material solution in contrast; Test according to thin layer chromatography, draw each 5-15 μ l of above-mentioned need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 8-10: 0.8-1.2 cyclohexane extraction-ethyl acetate is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

D. get the drug combination preparation of 1/2 times of phase taking dose on the same day, porphyrize adds ammonia solution 1.5-2.5ml and chloroform 25-35ml, and supersound process 30-50 minute, filter, filtrate is reclaimed chloroform to doing, and residue adds ethanol 4-6ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methanol and makes the solution that every 1ml contains 0.15-0.25mg, in contrast product solution; Test according to thin layer chromatography, draw each 10-15 μ l of above-mentioned two kinds of solution, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 4-6: 3-5: 0.3-0.5 normal hexane-ethyl acetate-ammonia is developing solvent, launch, take out, dry, smoked clear with iodine vapor to the speckle colour developing, inspect under the daylight; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

E. get the drug combination preparation of 1 times of phase taking dose on the same day, porphyrize is put in the round-bottomed flask of 200-300ml, add entry 40-60ml, connect volatile oil extractor, add water from the determinator upper end and make and be full of scale, 60～90 ℃ add petroleum ether 1.0-2.0ml, connect reflux condensing tube then; Flask contents is heated to boils, and kept 1.5-2.5 hour, stop heating, get petroleum ether layer as need testing solution; Other gets Herba Asari control medicinal material 8-12g and extracts volatile oil, adds chloroform 25-35ml and makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 10-20 μ l, reference substance solution 2-5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 92-98: 4-6 benzene-ethyl acetate is developing solvent, launch, take out, dry, spray is with 0.8-0.12% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

F. get the drug combination preparation of 1/4 times of phase taking dose on the same day, porphyrize, add kieselguhr 1.5-2.5g, mixing adds the methanol solution 15-25ml of 2% sodium hydroxide, heating and refluxing extraction 0.5-1.5 hour, filter, the filtrate evaporate to dryness adds water 25-35ml and makes dissolving, each with the water saturated n-butanol extraction of 35-45ml, extract 2-4 time, merge n-butyl alcohol liquid, with water washing 1-3 time, discard water liquid, with 0.8-1.2% potassium dihydrogen phosphate washing 1-2 time, discard water liquid, n-butyl alcohol liquid evaporate to dryness again, residue adds methanol 1.5-2.5ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.2-0.4mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 10-15 μ l, reference substance solution 5-7 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 18-22: 6-8: lower floor's solution of 1.5-2.5 chloroform-methanol-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be baked to the speckle colour developing at 100-110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

9, the method for quality control of pharmaceutical composition as claimed in claim 8 is characterized in that discrimination method in this method comprises a kind of and/or several in the following discriminating:

A. get it filled 1/6 times of compositions preparation day taking dose, porphyrize adds methanol 20ml, and supersound process 20 minutes filters, and filtrate is as need testing solution; Other gets the peoniflorin reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 12 μ l, reference substance solution 10 μ l, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 40: 5: 10: 0.2 chloroform-ethyl acetate-methanol-formic acid was developing solvent, launch, take out, dry, spray is with 50% phosphoric acid ethanol solution of 5% vanillin, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get it filled 5/24 times of compositions preparation day taking dose, porphyrize adds ethanol 15ml, and supersound process 1 hour filters, and filtrate is as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material, each 2g of Rhizoma Chuanxiong control medicinal material, and each adds ethanol 10ml, and supersound process is 1 hour respectively, filters, and filtrate is medical material solution in contrast; According to thin layer chromatography test, draw each 10 μ l of above-mentioned need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 9: 1 cyclohexane extraction-ethyl acetates was developing solvent, launched, and took out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

D. get it filled 1/2 times of compositions preparation day taking dose, porphyrize adds ammonia solution 2ml and chloroform 30ml, and supersound process 40 minutes filters, and filtrate is reclaimed chloroform to doing, and residue adds ethanol 5ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography, draw each 12 μ l of above-mentioned two kinds of solution, put respectively in same be on the prefabricated lamellae of silica gel G of adhesive with the sodium carboxymethyl cellulose, with 5: 4: 0.4 normal hexane-ethyl acetate-ammonia was developing solvent, launch, take out, dry, smoked clear with iodine vapor to the speckle colour developing, inspect under the daylight; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

E. the compositions preparation of getting it filled is equivalent to a day taking dose, and porphyrize is put in the round-bottomed flask of 250ml, adds entry 50ml, connects volatile oil extractor, adds water from the determinator upper end and makes and be full of scale, and 60～90 ℃ add petroleum ether 1.5ml, connect reflux condensing tube then; Flask contents is heated to boils, and keep little and boiled 2 hours, stop heating, get petroleum ether layer as need testing solution; Other gets Herba Asari control medicinal material 10g and extracts volatile oil, adds chloroform 30ml and makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 15 μ l, reference substance solution 3 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with 95: 5 benzene-ethyl acetates was developing solvent, launch, take out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

F. get it filled 1/4 times of compositions preparation day taking dose, porphyrize adds kieselguhr 2g, mixing, the methanol solution 20ml that adds 2% sodium hydroxide, heating and refluxing extraction 1 hour filters, the filtrate evaporate to dryness adds water 30ml and makes dissolving, uses the water saturated n-butanol extraction of 40ml at every turn, extract 3 times, merge n-butyl alcohol liquid, with water washing 2 times, discard water liquid, with 1% potassium dihydrogen phosphate washing 1 time, discard water liquid again, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 0.3mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned need testing solution 12 μ l, reference substance solution 6 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, lower floor's solution with 20: 7: 2 chloroform-methanol-water is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be baked to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

10,, it is characterized in that the assay in this method comprises following assay method as the method for quality control of claim 1,2,3 or 4 described pharmaceutical compositions:

Content under this drug combination preparation content uniformity item is got in the preparation of need testing solution, and porphyrize is got 1.0-1.5g, put in the tool plug conical flask, it is fixed to claim, adds methanol 15-25ml, claim to decide weight, supersound process 20-30 minute, take out, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter with the 0.40-0.50um microporous filter membrane, get subsequent filtrate, promptly;

Algoscopy is drawn above-mentioned reference substance solution 5-15 μ l, need testing solution 14-20 μ l respectively, injects chromatograph of liquid, measures, promptly;

11, require the method for quality control of 10 described pharmaceutical compositions as profit, it is characterized in that assay in this method is for comprising following assay method:

The preparation of the need testing solution content under the compositions preparation content uniformity item of getting it filled, porphyrize is got 1.2g, put in the tool plug conical flask, it is fixed to claim, adds methanol 20ml, claim to decide weight, supersound process 25 minutes is taken out, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter with the 0.45um microporous filter membrane, get subsequent filtrate, promptly;

Pharmaceutical composition per diem taking dose contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 69.0mg.

12, as claim 1,2, the application of 3 or 4 described pharmaceutical compositions in the endometriotic medicine of preparation treatment.

13, the application of pharmaceutical composition as claimed in claim 12 is characterized in that described treatment endometriosis is meant the content that reduces follicule-stimulating hormone (FSH) FSH, lutropin LH, prolactin antagonist PRL or estradiol E2, dwindles the Ectopic Endometrium volume, alleviates Ectopic Endometrium weight, alleviates the endometriosis lesion degree, improves the rheol trend of animal blood, reduces the uterine activity amplitude and suppress uterine motility, improves the microcirculatory effect of mesentery, the protective effect to local disturbance of blood circulation, antiinflammatory action, analgesic activity or raise immunity effect.