CN103893676A

CN103893676A - Preparation technology of traditional Chinese medicinal composition for treating primary dysmenorrhea

Info

Publication number: CN103893676A
Application number: CN201210576101.4A
Authority: CN
Inventors: 唐了平; 郭银汉; 金倩; 和芳; 周春莺; 唐全红
Original assignee: BEIJING CHUANGLI-KECHUANG PHARMACY TECHNOLOGY DEVELOPMENT Co Ltd
Current assignee: BEIJING CHUANGLI-KECHUANG PHARMACY TECHNOLOGY DEVELOPMENT Co Ltd
Priority date: 2012-12-27
Filing date: 2012-12-27
Publication date: 2014-07-02
Anticipated expiration: 2032-12-27
Also published as: CN103893676B

Abstract

The invention discloses a preparation technology of a traditional Chinese medicinal composition for treating primary dysmenorrheal. The technology comprises the following steps: 1, carrying out reflux extraction on Ligusticum wallichii, Corydalis tuber and (processed) Combined Spicebush Root in ethanol, collecting the above obtained filtrate, and collecting filter residues in a container; 2, percolating Chinese angelica with ethanol, collecting the obtained percolated liquid, and collecting filter residues in a container; 3, distilling Rhizoma Cyperi and Rhizoma Curcumae with water to extract volatile oil, collecting the volatile oil, collecting an aqueous solution obtained after distillation for later use, and collecting medicinal residues in a container for later use; and 4, mixing the filter residues obtained in step 1 with the filter residues obtained in step 2 and Medicinal Cyathula Root, decocting, mixing the obtained decoction with the solution obtained in step 3, concentrating, adding the volatile oil, and uniformly mixing. The technology improves original traditional Chinese medicinal composition preparation technologies, reduces the waste of raw materials, and improves the extraction degree of effective components in the traditional Chinese medicinal composition, and products obtained in the invention have the advantages of easy drying, stability and good curative effect.

Description

A kind of preparation technology of Chinese medicine composition for the treatment of primary dysmenorrhea

Technical field

The present invention relates to a kind of preparation technology of Chinese medicine composition for the treatment of primary dysmenorrhea, particularly relate to a kind of preparation technology of Chinese medicine composition for the treatment of qi stagnation and blood stasis type primary dysmenorrhea.

Background technology

Dysmenorrhea refers to that women periodically occurs the discomforts such as the spastic pain of obvious hypogastric region, falling inflation or soreness of waist pain on the occasion of menstrual period and before and after passing through, and severe patient can be with nausea and vomiting, dripping cold sweat, deadly cold hand and foot, and even severe pain is to the person of fainting.Dysmenorrhea is divided into two kinds of constitutional and Secondary cases, and primary dysmenorrhea refers to the dysmenorrhea of organ without organic disease; Secondary dysmenorrhea mostly is genitals the dysmenorrhea of organic disease.

Epidemiological study shows that primary dysmenorrhea is one of modal disease in current gynaecopathia, shows according to domestic sampling survey, and China's dysmenorrhes incidence rate is 33.19%, and wherein primary dysmenorrhea incidence rate is 36.06%.In dysmenorrhea generating process, the age is key factor.Primary dysmenorrhea often occurs in blue or green few phase, is after menophania, in 1-2, to start morbidity mostly, and after 30 years old, incidence rate starts to decline; Pain starts mostly after menstrual onset, appears at the earliest premenstrual 12 hours, pass through first day the most violent, remission after 2-3 days; It is spastic that pain is often, and with nausea,vomiting,diarrhea, dizziness, the symptom such as weak, severe patient complexion is turned white, is in a cold sweat.Dysmenorrhea presents periodical attack and vicious cycle, causes patient neurasthenia toward contact, makes patient feel painful, and prolonged and repeated outbreak also can affect mental health, such as agitation, irritability etc.The pathogenic factor complexity of dysmenorrhea, repeatability is large, treats comparatively thornyly, brings huge inconvenience to women's physical and mental health and work and study.

Doctor trained in Western medicine scholar thinks that primary dysmenorrhea generally starts to occur in menophania, mostly be uterine contraction and ischemia and cause neuropsychiatric pain, it is generally acknowledged relevant with come off (membranous dysmenorrhea), uterine hypoplasia, metrocampsis, narrow, the bad body appearance of neck tube, physical factors, allergy state and Nervous and Mental Factors etc. of inner membrance cast.Mainly increase because prostaglandin is synthetic, abnormal uterine shrink cause that hypoxic-ischemic, sensory nerve are subject to that multiple stimulation, the threshold of pain reduce, other cause as effects such as heredity campaigns.Primary dysmenorrhea has multiple therapy methods at present, conventional medicine has: NSAID (non-steroidal anti-inflammatory drug), oral contraceptive, calcium ion channel blocker, spasmolytic tranquilizer, receptor stimulating agent, vitamin E etc., these are all short-term good analgesic effect, but can not solve problem in essence, and easily recurrence, side effect are large, and its application is clinically very limited.

NSAID (non-steroidal anti-inflammatory drug) is the most frequently used first-line treatment medicine, such medicine is PGSI, to reduce the synthetic of prostaglandin, uterus tension force and contractility is declined by blocking-up Cycloxygenase, thereby alleviate the hysterospasm contraction that prostaglandin causes, dysmenorrhea is alleviated.Many studies have shown that after taking medicine truly has reduction through blood PG content.According to difference report, after nearly 64%-95% patient applies this type of medicine, subjective symptom alleviates.As aspirin, ibuprofen, naproxen, fluorine mefenamic acid, indometacin etc., effective percentage 30%-80%, but untoward reaction is more, take gastrointestinal tract and hepatorenal damage as main.

Oral contraceptive can reach more than 90% the effective percentage of primary dysmenorrhea treatment.This type of medical instrument has dual function, can suppress on the one hand inner membrance growth, reduces amount of bleeding, and the prostaglandin that generates and discharge is reduced; On the other hand can ovulation inhibition, reduce estrogenic content in blood, prostaglandin in blood, vasopressin and oxytocin level are reduced, thereby play the effect that suppresses uterine activity.Moreover, owing to having suppressed hypothalamic-pituitary-ovarian axis function, body inner estrogen was on close level follicular phase morning, and now inner membrance Prostaglandin is also minimum, has been applicable to contraception claimer.Because need are taken medicine continuously, therefore oral contraceptive is still Second line Drug for Most patients.This class medicine also has obvious impact to organism metabolism simultaneously, thereby has more untoward reaction.

Its mechanism of action of calcium ion channel blocker is to disturb calcium ion to pass through cell membrane, and stop calcium ion to discharge in cell inner membrance, suppressing calcium ion flows into outward in cell through Uterine Smooth Cell film, thereby inhibition smooth muscle contraction, removing hysterospasm shrinks, blood vessel dilating, improves uterus blood supply, therefore can treat dysmenorrhea.Can only maintain 5 hours but this type of medicine alleviates pain, easily recur and there is certain side effect.

The traditional Chinese medical science think dysmenorrhoea disease position in uterus, punching appoints, take " stagnation of QI and blood may bring about pain " or " not Rong Ze pain " as main mechanism.Its pathogeny is mainly the impact that between menstrual period, health is subject to various paathogenic factors, causes punching to appoint stasis blocking or cold accumulating in meridian, makes QI-blood circulation not smooth, and the circulation of uterus menses is obstructed, consequently " stagnation of QI and blood may bring about pain "; Or punching is appointed, uterus lose in moistening foster, consequently " not Rong Ze pain ".Can be divided into by pathogeny: qi stagnation and blood stasis type, cold blood stasis type, deficiency of qi and blood type etc., traditional Chinese medical herbal treatment dysmenorrhea is with a long history, determined curative effect.According to theory of Chinese medical science, a lot of effective methods are developed, as acupuncture, massage etc., also have much Chinese medicine be no matter single medicinal material or compound preparation all to improving few abdomen pain in premenstrual or menstrual period, distending pain of the breast, the dysmenorrhea symptom such as do not relax uncomfortable in chest has remarkable effect, and curative effect is lasting, after drug withdrawal, do not recur, be deeply subject to the great kindness of extensive patients.Qi stagnation and blood stasis type patient is more and more in recent years, but most of Chinese patent medicine is mostly from cold blood stasis, stagnation of QI anemia developing drugs kind, therefore cannot meet patient's needs.Street drug mainly contains TONGJINGBAO KELI, Tongjinglin Granules, HUAHONG PIAN, the precious sheet of woman, WUJI BAIFENG WAN, YUANHU ZHITONG PIAN etc.TONGJINGBAO KELI, mainly in cold-condensing type dysmenorrhea, adopts a large amount of removing blood stasis medicines in side, easily causes patient's blood deficiency symptom in treatment.Tongjinglin Granules is mainly in caused by energy stagnation and blood stasis dysmenorrhea, but square taste of Chinese medicine is various, and dose is large.HUAHONG PIAN is mainly applicable to secondary dysmenorrhea, can not meet primary dysmenorrhea patient's demand.The precious sheet of woman is mainly for the amenorrhea causing in blood stasis, dysmenorrhea, puerperal abdonimal pain.WUJI BAIFENG WAN is for the menoxenia of QI and blood deficiency, asthenic body.YUANHU ZHITONG PIAN is mainly used in hypochondriac pain, headache and the menstruation stomachache of caused by energy stagnation and blood stasis, has nauseating, dizzy, weakly, uncomfortable in chestly waits performance.

Application number is to disclose a kind of Chinese medicine composition for the treatment of primary dysmenorrhea in 200910079396.2 patent application, and this medicine is mainly used in treating qi depression to blood stasis primary dysmenorrhea.

Its prescription is as follows:

Radix Angelicae Sinensis 10-20 part Rhizoma Chuanxiong 5-15 part Rhizoma Cyperi 5-15 part Radix Linderae 5-10 part Rhizoma Curcumae 5-15 part Rhizoma Corydalis 5-10 part Radix Cyathulae 5-15 part

Its preparation technology is as follows:

1) get Radix Angelicae Sinensis, Rhizoma Chuanxiong, Rhizoma Cyperi, the Radix Linderae and Rhizoma Curcumae and be ground into coarse powder, the 6-10 that adds water doubly measures, and soaks 0.5-3h, extraction by steam distillation volatile oil, and the another device of aqueous solution after distillation is for subsequent use; Medicinal residues decoct with water, and filter, and filtrate and above-mentioned aqueous solution merge, and filtrate is concentrated;

2) Radix Cyathulae, Rhizoma Corydalis add alcohol reflux three times, filter merging filtrate, reclaim ethanol, merge concentratedly with above-mentioned concentrated solution, and drying and crushing, adds above-mentioned volatile oil, make peroral dosage form with common process.

Up to now, have no other documents openly report adopt Different Preparation prepare this medicine.But in the preparation technology that this patent provides, most medicines are all that the approach that adopts water extraction to be dried again obtains effective ingredient, the Chinese medicine composition that adopts this method for making to make, its active constituent content is low, in output process, waste of raw materials is large, the pharmaceutical preparations obtaining is difficult to be dried, and the stability of medicine is low.

Therefore, be badly in need of providing a kind of prepare treat primary dysmenorrhea Chinese medicine composition new technology, to provide, curative effect is better, good stability, safe drugs.

Summary of the invention

The object of the present invention is to provide a kind of preparation technology of Chinese medicine composition for the treatment of primary dysmenorrhea, compared with the compositions that compositions prepared by this technique is prepared with the method for making that provides of contrast patent, its drug absorption Du Gengjia, bioavailability are higher, better efficacy.

The preparation technology of the oral preparation of Chinese traditional medicinal for the treatment of primary dysmenorrhea provided by the invention, specific as follows:

Prescription:

Preparation technology:

1) get Radix Linderae 5-10 part, add the stirring of rice vinegar 1-2 part, moistening 0.5-2h, after vinegar liquid is sucked, with 60% microwave heating power, heating 2-10min, takes out, and cools, for subsequent use.

2) get Rhizoma Chuanxiong 5-15 part, Rhizoma Corydalis 5-10 part, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 60-90% ethanol as solvent, soaked overnight, add 6-9 and doubly measure 60-90% alcohol reflux 1-3 time, each 0.5-2h, filter merging filtrate, decompression recycling ethanol, collect filtrate for later use, another device is collected filtering residue.

3) get Radix Angelicae Sinensis 10-20 part and be ground into coarse powder, make solvent with 60-90% ethanol, dipping 24-48h, slowly percolation, collects the liquid of just filtering, and another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collects the continuous liquid of filtering, and merging filtrate is for subsequent use.Another device is collected filtering residue.

4) get Rhizoma Cyperi 5-15 part, Rhizoma Curcumae 5-15 part and be ground into coarse powder, add the water that 6-9 doubly measures, soak 1-3h, hydrodistillation extracts volatile oil, and extraction time is 2-5h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use.

5) get Radix Cyathulae 5-15 part, with above-mentioned 2), 3), 4) gained medicinal residues add the decocting that 6-9 doubly measures and boil 1-2h in step, filter, filtrate and above-mentioned 2), 3), 4) gained solution merges in step, at 70 ℃, being evaporated to relative density is 1.1-1.5, dry, add above-mentioned volatile oil, mix and get final product.

Said method also comprises that the product obtaining to step 5) adds appropriate adjuvant, makes various dosage forms.Further preferably, described dosage form includes but not limited to granule, tablet, pill, powder, capsule.

Preferably:

2) get Rhizoma Chuanxiong 5-15 part, Rhizoma Corydalis 5-10 part, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 70-80% ethanol as solvent, soaked overnight, add 8-9 and doubly measure 70-80% alcohol reflux 2 times, each 1-1.5h, filter merging filtrate, decompression recycling ethanol, collect filtrate for later use, another device is collected filtering residue.

3) get Radix Angelicae Sinensis 10-20 part and be ground into coarse powder, make solvent with 70-80% ethanol, dipping 24-36h, slowly percolation, collects the liquid of just filtering, and another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collects the continuous liquid of filtering, and merging filtrate is for subsequent use.Another device is collected filtering residue.

4) get Rhizoma Cyperi 5-15 part, Rhizoma Curcumae 5-15 part and be ground into coarse powder, add the water that 7-9 doubly measures, soak 1-1.5h, hydrodistillation extracts volatile oil, and extraction time is 3-4h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use.

5) get Radix Cyathulae 5-15 part, with above-mentioned 2), 3), 4) gained medicinal residues add the decocting that 7-9 doubly measures and boil 1-1.5h in step, filter, filtrate and above-mentioned 2), 3), 4) gained solution merges in step, at 70 ℃, being evaporated to relative density is 1.2-1.5, dry, add above-mentioned volatile oil, mix and get final product.

More preferably:

1) get 10 parts of the Radixs Linderae, add 2 parts of stirrings of rice vinegar, moistening 1h, after vinegar liquid is sucked, with 60% microwave heating power, heating 5min, takes out, and cools, for subsequent use.

2) get 15 parts of Rhizoma Chuanxiongs, 10 parts of Rhizoma Corydalis, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 70% ethanol as solvent, soaked overnight, adds 8 times of amount 70% alcohol reflux 2 times, each 1.5h is excellent, filters merging filtrate, decompression recycling ethanol, collects filtrate for later use, and another device is collected filtering residue.

3) get 20 parts of Radix Angelicae Sinensis and be ground into coarse powder, make solvent with 70% ethanol, dipping 24h, slowly percolation, collects the liquid of just filtering, and another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collects the continuous liquid of filtering, and merging filtrate is for subsequent use.Another device is collected filtering residue.

4) 15 parts of 15 parts of Rhizoma Cyperis, Rhizoma Curcumae are ground into coarse powder, add the water of 8 times of amounts, soak 2.5h, hydrodistillation extracts volatile oil, and extraction time is 4h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use.

5) get 15 parts of Radix Cyathulaes, with above-mentioned 2), 3), 4) gained medicinal residues add 8 times of amounts in step decocting boils 1.5h, filter, filtrate and above-mentioned 2), 3), 4) gained solution merges in step, under 70 ℃ of conditions, being evaporated to relative density is 1.2, dry, add above-mentioned volatile oil, mix and get final product.

More preferably, said method also comprises to step 5) products therefrom and adds appropriate adjuvant, makes various dosage forms.Further preferably, described dosage form includes but not limited to granule, tablet, pill, powder, capsule.

For example: by obtained pharmaceutical preparations, add 10 parts of dried cream powders, 10 parts of sucrose, 5 parts, dextrin, adds appropriate 70% ethanol granulation, dry, to obtain final product.

For example: by obtained pharmaceutical preparations, add 10 parts of dried cream powders, 10 parts of sucrose, 5 parts, dextrin, adds appropriate 70% ethanol granulation, dry, granulate tabletting, film coating or sugar-coat, to obtain final product.

For example: by obtained pharmaceutical preparations, every 10 parts of fine powders add 10 parts of refined honeys and make concentrated honeyed pill, to obtain final product.

For example: by obtained pharmaceutical preparations, grind, mix, pack and get final product.

For example: by obtained pharmaceutical preparations, add 10 parts of starch, 10 parts of magnesium stearate, add 70% appropriate amount of ethanol granulation, dry, granulate, encapsulated and get final product.

The preparation technology that preparation technology provided by the present invention and patent (application number is 200910079396.2) provide has significant difference, specific as follows:

1. Radix Angelicae Sinensis mixes distillation extraction and has made independent percolation into and extract from former preparation technology: concrete shows as by original first water soaking 3h, then distills 6h, and medicinal residues decoct 2h and made soak with ethanol 2.5h repercolation 24h into, and medicinal residues decoct 1.5h.

Pass through By consulting literatures, conventionally the extraction process adopting for medical material Radix Angelicae Sinensis is water distillation or alcohol reflux, the present invention adopts percolation to extract, percolation technological operation is relatively simple, and compared with water distillation or alcohol reflux Radix Angelicae Sinensis, its active constituent content of product that percolation extraction Radix Angelicae Sinensis obtains is higher; Ethanol used herein can be recycled simultaneously, has reduced production cost.

2. Rhizoma Chuanxiong, the Radix Linderae two taste medicines in prescription, mixing distillation extraction from former preparation technology has made alcohol reflux into: specific as follows: former technique is first by two taste medical material water soaking 3h, then distill 6h, medicinal residues decoct 2h, preparation technology of the present invention changes soak with ethanol into and spends the night, each 1.5h that refluxes 2 times, filtering residue decocts 1.5h.

3. in prescription, the mode of the extraction of Rhizoma Cyperi, Rhizoma Curcumae two taste medical materials is constant, but the time become 2.5h from original water soaking 3h, distillation 6h becomes 4h, filtering residue decocts 2h and becomes 1.5h; Entirety reduction extraction time is 3h.

4. prescription Chinese crude drug Rhizoma Corydalis is by original ethanol extraction 3 times, and each 3h becomes first soaked overnight, then 2 each 1.5h of alcohol reflux, and filtering residue decocts 1.5h; Entirety reduction extraction time 4.5h, the medicinal residues after refluxing in addition are also fully used, and have reduced the waste of crude drug.

5. prescription Chinese crude drug Radix Cyathulae is become and is decocted 1.5h from 3 each 3h of original alcohol reflux, has reduced preparation technology's required time, has saved production cost simultaneously.

specific embodiment mode

Below in conjunction with specific embodiment, the invention will be further described; and the not restriction to invention; according to prior art well known in the art; embodiments of the present invention are not limited to this; therefore all this areas of making according to present disclosure be equal to replacement, all belong to protection scope of the present invention.

embodiment 1: different method for makings are prepared pharmaceutical preparations

1. prescription

Radix Angelicae Sinensis 20kg(20 part) Rhizoma Chuanxiong 15kg(15 part) Rhizoma Cyperi 15kg(15 part) Radix Linderae 10kg(10 part) Rhizoma Curcumae 15kg(15 part) Rhizoma Corydalis 10kg(10 part) Radix Cyathulae 15kg(15 part)

2. adopt method for making provided by the invention to prepare pharmaceutical preparations

specific as follows:

1) get Radix Linderae 10kg, add the rice vinegar of 2 deals to stir, moistening 1h, after vinegar liquid is sucked, with 60% microwave heating power, heating 5 min, take out, and cool, for subsequent use.

2) get Rhizoma Chuanxiong 15kg, Rhizoma Corydalis 10kg, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 70% ethanol as solvent, soaked overnight, adds 8 times of amount 70% alcohol reflux 2 times, each 1.5h, filters merging filtrate, decompression recycling ethanol, collects filtrate for later use, and another device is collected filtering residue.

3) get 20kg Radix Angelicae Sinensis powder and be broken into coarse powder, make solvent with 70% ethanol, dipping 24h, slowly percolation, collects the liquid of just filtering, and another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collects the continuous liquid of filtering, and merging filtrate is for subsequent use.Another device is collected filtering residue.

4) 15kg Rhizoma Cyperi, 15kg Rhizoma Curcumae are ground into coarse powder, add the water of 8 times of amounts, soak 2.5h, hydrodistillation extracts volatile oil, and extraction time is 4h, collects volatile oil, and distillation rear solution is collected for subsequent use, and the another device of medicinal residues is collected for subsequent use.

5) get Radix Cyathulae 15kg, with above-mentioned 2), 3), 4) gained medicinal residues add 8 times of amounts in step decocting boils 1.5h, filters filtrate and above-mentioned 2), 3), 4) in solution merge, under 70 ℃ of conditions, being evaporated to relative density is 1.2, is dried.Add above-mentioned volatile oil, mix and obtain pharmaceutical preparations A.

adopt the preparation technology of contrast patent to prepare Radix Angelicae Sinensis dysmenorrhea pharmaceutical preparations

1) get Radix Angelicae Sinensis 20kg, Rhizoma Chuanxiong 15kg, Rhizoma Cyperi 15kg, Radix Linderae 10kg and Rhizoma Curcumae 15kg and be ground into coarse powder, add 10 times of amounts, soak 3h, extraction by steam distillation volatile oil, extraction time is 6h, collects volatile oil, the another device of aqueous solution after distillation is collected for subsequent use;

2) medicinal residues add 10 times of water gagings and decoct 2h, filter, filtrate and the merging of above-mentioned aqueous solution, under 70 ℃ of conditions filtrate to be concentrated into relative density be 1.05-1.2; Radix Cyathulae 15kg, Rhizoma Corydalis 10kg add 10 times of amount 70% alcohol reflux three times, and each 2h filters merging filtrate, reclaims ethanol, merge with above-mentioned concentrated solution, under 70 ℃ of conditions, being concentrated into relative density is 1.15-1.40, drying and crushing, add above-mentioned volatile oil, mix and obtain pharmaceutical preparations B.

embodiment 2: the active constituent content measuring test of Different Preparation gained prepared product

Medicine described in this patent, its prescription comprises Radix Angelicae Sinensis, Radix Cyathulae, Rhizoma Chuanxiong, the Radix Linderae, Rhizoma Cyperi, Rhizoma Corydalis, Rhizoma Curcumae seven flavor medicine material, Radix Angelicae Sinensis, Rhizoma Chuanxiong are the monarch drug of prescription described in this patent, Radix Cyathulae is adjuvant drug, from version pharmacopeia First in 2010 and pertinent literature, the main effective ingredient of Radix Angelicae Sinensis, Radix Cyathulae, Rhizoma Chuanxiong is ferulic acid; Data in literature shows that ferulic acid has the pharmacological actions such as the prostaglandin of enhancing activity, analgesia, alleviating vascular spasm, in conjunction with primary dysmenorrhea pathogeny and primary dysmenorrhea person's clinical manifestation, therefore select the determination object that ferulic acid is this test.

ferulaic acid content determination test

specific as follows:

1, instrument and reagent

1) instrument: instrument Agilent-1100 type high performance liquid chromatograph (Anjelen Sci. & Tech. Inc), Agilent1100 chromatographic work station, ultraviolet spectra detector, Hypersil C18 chromatographic column (4.6mm × 250mm, 5 μ m), Agirlent sample injection bottle, analytical balance (precision ten thousand/), TU-1900 ultraviolet-uisible spectrophotometer.

2) reagent: methanol (Beijing North fine chemistry Co., Ltd, lot number: 20091114, chromatographically pure), water is tri-distilled water, other reagent are analytical pure, ferulic acid reference substance (Nat'l Pharmaceutical & Biological Products Control Institute, product batch number: 20081123), pharmaceutical preparations A, the self-control of pharmaceutical preparations B(our company).

2, method and result

1) chromatographic condition

(4.6mm × 250mm, m), mobile phase is that methanol-1.0% glacial acetic acid solution is 25: 75 to 5 μ, flow velocity: 1.0ml/min, detection wavelength: 321nm, column temperature: 25 ℃, sample size is 10.0 μ l to adopt Hypersil C18 chromatographic column.Theoretical cam curve is calculated and is greater than 3000 with ferulic acid peak value.

2) solution preparation

Reference substance solution preparation: accurately weighed ferulic acid reference substance 0.1mg, put in 5ml volumetric flask, add methanol constant volume to scale, stand for standby use.

Need testing solution preparation: analytical balance takes pharmaceutical preparations A, the each 10g of pharmaceutical preparations B, add respectively 10g kieselguhr, porphyrize after mix homogeneously, be placed in respectively cable type extractor according, add respectively 70% methanol solution 200ml, respectively extract 5h, extracting solution is evaporated to about 4ml, residue is transferred to respectively in 20ml volumetric flask after dissolving by mobile phase, adds mobile phase to be settled to scale, for subsequent use after leaving standstill.

3) preparation of standard curve

Accurate ferulic acid reference substance solution that concentration is 20ug/ml 10,20,40,80,160, the 320ul of drawing, respectively to 2ml volumetric flask, add mobile phase to be settled to scale, draw respectively 10ul and inject high performance liquid chromatograph, record its peak area, take sample size as abscissa, peak area is vertical coordinate, drawing standard curve, obtaining regression equation is Y=3.84 × 10 ⁴x+0.05 × 10 ⁴(r=0.9998).Result shows that ferulaic acid content is good in 0.025-1.200mg scope internal linear relation.

4) precision test

The same need testing solution 100ul of accurate absorption, repeats sample introduction 6 times, measures its average peak area, and its RSD is 0.97%, and result shows that this law precision is good.

5) study on the stability

Respectively accurately draw same test sample 10.0ul, 0,4,8,12,16,20, the each sample introduction of 24h 1 time, the RSD of its results peaks area is 0.76%, shows that ferulic acid is basicly stable in 24h.

6) replica test

Get 5 parts, the sample of same lot number, prepare sample solution and measure the content of ferulic acid according to the preparation method of ferulic acid need testing solution, according to linear relationship measurement result, in sample, the average content of ferulic acid is 0.35mg/g, and RSD is 1.62%.Result shows that this law repeatability is good.

7) application of sample recovery experiment

Take the about 2.0g of sample (medicine prepared product A) of known content, totally 6 parts.Accurately weighed, precision adds ferulic acid reference substance solution (0.05mg/g) 24ml respectively, measures content according to the method under assay item, and the while 99.86%, RSD is 1.28%, and application of sample reclaims good.

8) assay

Get pharmaceutical preparations A, pharmaceutical preparations B, prepare solution according to the preparation method of need testing solution, precision takes reference substance solution and the each 10ul of need testing solution, injects high performance liquid chromatograph.Impurity peaks in chromatographic peak and the sample of ferulic acid can reach baseline separation, and theoretical cam curve is calculated the content that is greater than 3000. calculating ferulic acids according to ferulic acid peak, the results are shown in Table 1.Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

± S represents; Between many groups, relatively, variance is used variance analysis together, and heterogeneity of variance is used non parametric tests, has significant difference when P < 0.05.

Ferulaic acid content measurement result in table 1 Different Preparation gained prepared product

Note: ^★represent compared with the ferulaic acid content of technique group of the present invention and the ferulaic acid content of contrast patent technique group P < 0.05.

From table 1, the ferulaic acid content of technique group of the present invention, compared with contrasting the ferulaic acid content of patent technique group, has significant difference (P < 0.05).Ferulic acid is the effective ingredient of medicine of the present invention, the above results explanation, adopt content that method for making provided by the invention prepares its effective ingredient ferulic acid of the obtained product content apparently higher than ferulic acid in the preparation technology's products therefrom adopting described in contrast patent, therefore, illustrate further, preparation technology provided by the present invention is better than contrasting the preparation technology described in patent.

embodiment 3: the impact of the pharmaceutical preparations of Different Preparation gained on the mice thermostimulation threshold of pain

1, test material: pharmaceutical preparations A, the B of preparation in embodiment 1 of the present invention, content is 9g/ bag (every bag of about 15.6g of symphysis medicine).

2, experimental animal: Kunming mouse, body weight 20 ± 2g.

3, trial drug dosage: this experiment adopts gastric infusion, and pharmaceutical preparations A administration group, pharmaceutical preparations B administration group are all established high, medium and low dosage group, and dosage is respectively 4g/kg, 2g/kg, 1g/kg, according to the administration of 20ml/kg body weight; Matched group: gavage gives isopyknic water.

4, test method: get some of Kunming kind female mices, under room temperature environment, put into the 500ml beaker that thermostat water bath top is heated to 0.5 ℃ of 55 scholar, each one, record mice with stopwatch and certainly drop into beaker to occurring licking the time (s) of metapedes as the pain threshold of this Mus, so measure 2 times, interval 30min, gets average and is the front pain threshold of medicine (meansigma methods is no more than 30s person for qualified).

Get 70 of mices that meet above-mentioned test requirements document, be divided at random 7 groups by body weight: matched group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.After administration 1 time, after 30min, measure pain threshold by front method, its pain threshold of mice that pain threshold exceedes 60s calculates by 60s, calculates by 60s, and pain threshold improves percentage rate (%)=(pain threshold-Basic Pain Threshold value after administration)/Basic Pain Threshold value × 100%.Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

± S represents; Between many groups, relatively, variance is used variance analysis together, heterogeneity of variance non parametric tests, and P < 0.05 has significant difference.

The impact of table 2 Different Preparation gained pharmaceutical preparations on mice thermostimulation pain threshold

Group	Dosage (g/kg)	The pain threshold (s) that after administration, 30min measures	Percentage rate (%) is improved in the threshold of pain
				Matched group	—	17.58±1.73	—
B low dosage	1	23.64±2.41 ^★	34.47
				A low dosage	1	25.22±2.05 ^★	43.5
Dosage in B	2	24.08±1.89 ^★	36.9
				Dosage in A	2	25.91±1.92 ^★●	47.4
B high dose	4	24.60±2.43 ^★	39.9
				A high dose	4	26.61±2.20 ^★?	51.4

Note: ^★represent that each administration group is compared with matched group, P < 0.05, ^●represent compared with the middle dosage group of pharmaceutical preparations A and the middle dosage group of pharmaceutical preparations B, P < 0.05, ^?represent compared with the high dose group of pharmaceutical preparations A and the high dose group of pharmaceutical preparations B P < 0.05.

From table 2 result, after administration 30min, the pain threshold of the measured mice of high, medium and low dosage group of pharmaceutical preparations A and pharmaceutical preparations B, compared with matched group, all have a significant difference (P < 0.05), this shows that pharmaceutical preparations A and pharmaceutical preparations can reduce the pain threshold of mice; Under same dosage level, the pain threshold of pharmaceutical preparations A group is greater than pharmaceutical preparations B group, and high, under two dosage levels, the pain threshold of pharmaceutical preparations A group, compared with pharmaceutical preparations B group, all has significant difference (P < 0.05).

The above results shows, adopts the pharmaceutical preparations of two kinds of method for making gained all can improve the pain threshold of mice thermostimulation; The pharmaceutical preparations A that simultaneously adopts preparation technology's gained provided by the invention is more remarkable effect for the preparation technology's gained pharmaceutical preparations B described in adopting contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 4: the impact of the pharmaceutical preparations of Different Preparation gained on the reaction of mice acetic acid twisting

2, experimental animal: Kunming mouse, body weight 20 ± 2g.

4, test method: get 70 male and female half and half of Kunming mouse, be divided at random 7 groups by sex, body weight, 10 every group, be respectively matched group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.Successive administration 3 days, once a day, after last administration 30min, each Mus lumbar injection 0.6% glacial acetic acid 0.1ml/10g, start to record mouse writhing number of times and the inferior time (incubation period) that occurs writhing of each mouse's head in 20min, analgesia rate (%)=(writhing number of times after basic writhing number of times-administration)/basic writhing number of times × 100%.Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

The impact of table 3 Different Preparation gained pharmaceutical preparations on the reaction of mice acetic acid twisting

Group	Dosage (g/kg)	Incubation period (s)	Writhing number of times/20min	Analgesia rate (%)
					Matched group	—	3.3±1.0	38.6±2.6	—
B low dosage	1	6.9±1.5 ^★	26.1±1.8 ^★	32.4
					A low dosage	1	7.4±1.8 ^★	20.9±2.1 ^★▲	45.9
Dosage in B	2	7.2±1.6 ^★	25.4±1.8 ^★	34.2
					Dosage in A	2	9.9±1.1 ^★●	19.6±2.2 ^★●	49.2
B high dose	4	7.7±1.3 ^★	24.9±1.9 ^★	35.5
					A high dose	4	10.6±1.2 ^★?	18.9±1.7 ^★?	51.0

Note: ^★represent that each administration group is compared with matched group, P < 0.05, ^▲represent that pharmaceutical preparations A low dose group is compared with pharmaceutical preparations B low dose group, P < 0.05, ^●represent in pharmaceutical preparations A that dosage group is compared with dosage group in pharmaceutical preparations B, P < 0.05, ^?represent that pharmaceutical preparations A high dose group is compared with pharmaceutical preparations B high dose group, P < 0.05.

From table 3 result, with matched group comparison, writhing incidence rate, the writhing of each administration group all have significant difference (P < 0.05) incubation period, illustrate that pharmaceutical preparations A and pharmaceutical preparations B can reduce writhing number of times, increase writhing incubation period.Obtained by data in table, under same dosage level, pharmaceutical preparations A group compares with pharmaceutical preparations B group, incubation period is longer, writhing number of times still less, and high, under two dosage levels, the incubation period of pharmaceutical preparations A group and pharmaceutical preparations B group more all have significant difference (P < 0.05), in addition, under high, medium and low each dosage level, the writhing number of times of pharmaceutical preparations A group and pharmaceutical preparations B group relatively, all have significant difference (P < 0.05).

The above results shows, adopts the mouse writhing reaction due to the prepared pharmaceutical preparations Dichlorodiphenyl Acetate of two kinds of method for makings all to have good therapeutic effect, extended to a certain extent incubation period, reduced writhing number of times; Simultaneously, the pharmaceutical preparations A that adopts preparation technology's gained provided by the invention is more remarkable effect for the preparation technology's gained pharmaceutical preparations B described in adopting contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 5: the pharmaceutical preparations of Different Preparation gained brings out the impact of dysmenorrhea model in mice writhing response on oxytocin

2, experimental animal: Kunming mouse, body weight 20 ± 2g.

3, trial drug dosage: this experiment adopts gastric infusion, and pharmaceutical preparations A administration group, pharmaceutical preparations B administration group are all established high, medium and low dosage group, and dosage is respectively 4g/kg, 2g/kg, 1g/kg, according to the administration of 20ml/kg body weight; Matched group and model group: gavage gives isopyknic water.

4, test method: get 80 of Kunming kind female mices, be divided at random 8 groups by body weight, 10 every group, be respectively matched group, model group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.Except matched group, all the other respectively organize all according to dosage 0.2mg/ subcutaneous injection diethylstilbestrol 0.2mg/, and continuous 10 days, to improve uterus sensitivity.Injecting diethylstilbestrol the 11st day, starting administration, for three days on end, after last administration lh, according to dosage 2u/ all lumbar injection oxytocin 2u/ of each mice, records mouse writhing and reacts the incubation period and the interior writhing number of times of 30min that occur.Analgesia rate (%)=(writhing number of times after administration-basic writhing number of times)/basic writhing number of times × 100%.

Observe rear 30min, put to death rat, peeled off rat uterus, taken 0.5g with precision instrument, added 1ml normal saline, homogenate, centrifugal, get supernatant, to be measured.Press rat prostate element F _2aenzyme-linked immunologic detecting kit, rat prostate element PGE ₂enzyme-linked immunologic detecting kit requires to carry out PGF _2a, PGE ₂the detection of content.

Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

Table 4 Different Preparation gained pharmaceutical preparations brings out the impact of rat dysmenorrhea model writhing response on oxytocin

Group	Dosage (g/kg)	Writhing incubation period (min)	Writhing number of times (30min)	Pain suppression ratio
					Matched group	—	28.6±2.99 ^u	0.5±1.04 ^u	—
Model group	—	3.6±1.1	39.2±2.6	—
					B low dosage	1	6.4±1.7 ^★	27.0±2.1 ^★	31.12
A low dosage	1	7.4±1.8 ^★	21.5±2.5 ^★▲	45.15
					Dosage in B	2	6.9±1.6 ^★	26.0±1.9 ^★	33.67
Dosage in A	2	8.8±1.8 ^★●	19.9±2.5 ^★●	49.23
					B high dose	4	7.4±1.1 ^★	25.5±1.8 ^★	34.95
A high dose	4	9.8±0.9 ^★?	19.3±2.3 ^★?	50.77

Note: ^urepresent that matched group is compared with model group, P < 0.05, ^★represent that each administration group is compared with model group, P < 0.05, ^▲represent that pharmaceutical preparations A low dose group is compared with pharmaceutical preparations B low dose group, P < 0.05, ^●represent in pharmaceutical preparations A that dosage group is compared with dosage group in pharmaceutical preparations B, P < 0.05, ^?represent that pharmaceutical preparations A high dose group is compared with pharmaceutical preparations B high dose group, P < 0.05.

From table 4, with matched group comparison, the writhing incidence rate of model group gained, writhing all have significant difference (P < 0.05) incubation period, show this test modeling success.Each administration group is compared with model group, mouse writhing number of times reduces, writhing prolongation of latency, and each group all have significant difference (P < 0.05), illustrates that each administration group is directed to oxytocin and brings out rat dysmenorrhea model writhing response and all have good effect.

The above results shows, under same dosage level, pharmaceutical preparations A group is compared with pharmaceutical preparations B group, and still less, writhing response time of occurrence is longer for mouse writhing number of times; High, under two dosage levels, compared with pharmaceutical preparations B group, all have significant difference (P < 0.05) incubation period of pharmaceutical preparations A group; Under each dosage group, the writhing number of times of pharmaceutical preparations A group, compared with pharmaceutical preparations B group, all has significant difference (P < 0.05).

The above results shows, the rat dysmenorrhea model writhing response that adopts two kinds of prepared pharmaceutical preparations of different method for makings to bring out for oxytocin all has good therapeutic effect, can reduce mouse writhing number of times, extends writhing incubation period; And more remarkable effect for the gained pharmaceutical preparations B that the pharmaceutical preparations A that simultaneously adopts preparation technology's gained provided by the invention is prepared with respect to the preparation technology described in adopting contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

Table 5 Different Preparation gained pharmaceutical preparations brings out rat uterus tissue in rat dysmenorrhea model to oxytocin

PGF _2a, PGE ₂impact

Group	Dosage (g/kg)	PGF _2ａ(ng/ml)	PGE ₂(ng/ml)
				Matched group	——	41.30±2.52	5.55±0.44
Model group	——	76.40±2.45 ^u	2.67±0.39 ^u
				B low dosage	1	69.18±2.79 ^★	3.15±0.38 ^★
A low dosage	1	62.00±3.25 ^★▲	3.99±0.27 ^★▲
				Dosage in B	2	56.37±2.38 ^★	3.87±0.21 ^★
Dosage in A	2	48.60±2.63 ^★●	4.87±0.31 ^★●
				B high dose	4	52.58±2.62 ^★	4.16±0.24 ^★
A high dose	4	41.13±2.62 ^★?	5.15±0.23 ^★?

From table 5, with matched group comparison, model group is at the measured PGF of uterine cancer cell _2acontent is higher, has significant difference (P < 0.05), measured PGE ₂content reduces, and has significant difference (P < 0.05), and the above results and analysis show this test modeling success.

Each administration group compared with model group, the PGF in uterine cancer cell _2acontent is higher, has significant difference (P < 0.05), PGE ₂content reduces, and has significant difference (P < 0.05), shows that each administration group brings out rat uterus tissue PGF in rat dysmenorrhea model to oxytocin _2athe increase of content and PGE ₂the reduction of content all has remarkable result.

Under same dosage level, compared with pharmaceutical preparations B group, pharmaceutical preparations A group is at the measured PGF of uterine cancer cell _2acontent is higher, has significant difference (P < 0.05), measured PGE ₂content reduces, and has significant difference (P < 0.05).

The above results shows, the rat dysmenorrhea model that adopts two kinds of prepared pharmaceutical preparations of different method for makings to bring out for oxytocin all has good therapeutic effect, can increase PGF in uterine cancer cell _2acontent, reduces PGE ₂content; And the pharmaceutical preparations A that adopts preparation technology's gained provided by the invention is more remarkable effect for the preparation technology's gained pharmaceutical preparations B described in adopting contrast patent, further illustrate preparation technology provided by the present invention and be better than contrasting the preparation technology described in patent.

embodiment 6: the impact of the pharmaceutical preparations of Different Preparation gained on rat uterus ligament blood capillary site crossing number

2, experimental animal: Kunming kind rat 210 ± 10g.

3, trial drug dosage: pharmaceutical preparations A administration group, pharmaceutical preparations B administration group are all established high, medium and low dosage group, and dosage is respectively 4g/kg, 2g/kg, 1g/kg.

4, test method: get 70 male and female half and half of Kunming kind rat, be divided at random 7 groups by sex, body weight, 10 every group, be respectively matched group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.Successive administration 3 days.Each group rat is pressed after the anesthesia of 45mg/kg dosage lumbar injection pentobarbital sodium, cut abdominal cavity and pull out Aconitum carmichaeli Debx. palace and a ligament, be fixed in 37 ℃ of 10ml Rockwell physiological balance liquids, stablize after 20min, observe blood capillary before counting administration at 1mm with 4 × 10 times of inverted microscopes ²intersection in scope is counted, and ligation pylorus is pressed 20mL/kg administration for each group in duodenum, and Normal group gives isopyknic distilled water, after administration 30min, observes blood capillary site crossing number with method.Number × 100% is opened in increase percentage rate (%)=(the open number in open number-matched group blood capillary intersection site, blood capillary intersection site after administration)/matched group blood capillary intersection site.Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used ± S represents; Between many groups, relatively, variance is used variance analysis together, and heterogeneity of variance is used non parametric tests, has significant difference when P < 0.05.

The impact of table 6 Different Preparation gained pharmaceutical preparations on the open number in rat capillary intersecting blood vessels site

Group	Dosage (g/kg)	The blood capillary intersection open number in site (individual)	Increase percentage rate (%)
				Matched group	—	4.2±1.3	—
B low dosage	1	5.9±1.3 ^★	40.5
				A low dosage	1	7.1±1.6 ^★	69.0
Dosage in B	2	6.1±1.52 ^★	45.2
				Dosage in A	2	7.2±1.2 ^★	71.4
B high dose	4	6.4±1.5 ^★	52.3
				A high dose	4	7.9±1.4 ^★?	88.1

Note: ^★represent that each administration group is compared with matched group, P < 0.05, ^?represent that pharmaceutical preparations A high dose group is compared with pharmaceutical preparations B high dose group, P < 0.05.

As shown in Table 6, compared with matched group, the measured blood capillary of each administration group intersects the open number in site and has significant difference (P < 0.05), illustrates that this tests related pharmaceutical preparations and all have the effect of invigorating blood circulation; Under same dosage level, compare with pharmaceutical preparations B group, under each dosage level, pharmaceutical preparations A organizes the measured open number in blood capillary intersection site all increase, and high dose has significant difference (P < 0.05), show that pharmaceutical preparations A is better than the pharmaceutical preparations B effect of invigorating blood circulation.

The above results shows, adopts the effect of all invigorating blood circulation of two kinds of prepared pharmaceutical preparations of different method for makings; The pharmaceutical preparations A that simultaneously the adopts preparation technology's gained provided by the invention more remarkable effect of invigorating blood circulation for the preparation technology's gained pharmaceutical preparations B described in adopting contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 7: the impact of the pharmaceutical preparations of Different Preparation gained on blood stasis type hemorheology of rat

2, experimental animal: Kunming kind rat 210 ± 10g.

4, test method: get 80 male and female half and half of Kunming kind rat, be divided at random 8 groups by sex, body weight, 10 every group, be respectively matched group, model group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.Successive administration 7 days.After last administration 30min, except matched group, all the other respectively organize according to dosage 0.08ml/100g subcutaneous injection (sc) 0.1g/Bl adrenalin hydrochloride injection, control rats subcutaneous injection equivalent normal saline, after 2h, except matched group, all the other each group rats are all immersed in 0-4 ℃ of frozen water and carry out cold stimulation 5min, subcutaneous injection equivalent adrenalin hydrochloride 0.4ml/kg again after 2h, fasting 16h after disposing.According to dosage 25% urethane 0.4ml/100g intraperitoneal injection of anesthesia, ventral aorta blood sampling, heparin sodium anticoagulant, the full-automatic blood flow of the centrifugal rear employing side instrument that accelerates detects the indexs such as whole blood viscosity, plasma viscosity, erythrocyte electrophoretic time and packed cell volume.Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

As seen from the results in Table 7, with matched group comparison, whole blood viscosity, plasma viscosity and the packed cell volume of model group has significant difference (P < 0.05), shows this test blood stasis type modeling success; Under high, medium and low dosage level, whole blood viscosity, plasma viscosity and the packed cell volume of each dosage group all has significant difference (P < 0.05) compared with model group, under low dosage level, the plasma viscosity of each dosage group, packed cell volume all have significant difference (P < 0.05) compared with model group, show that pharmaceutical preparations A, pharmaceutical preparations B can improve dense, the sticky state of stasis syndrome rat model blood, have the effect of certain blood circulation promoting and blood stasis dispelling;

Compare with pharmaceutical preparations B group, under height, middle dosage level, whole blood viscosity, plasma viscosity and the packed cell volume of pharmaceutical preparations A group all have significant difference (P < 0.05), and compare with pharmaceutical preparations B group, under low dosage level, pharmaceutical preparations A group plasma viscosity and packed cell volume all have significant difference (P < 0.05), show that pharmaceutical preparations A improves the more remarkable effect of dense, the sticky state of stasis syndrome rat model blood than pharmaceutical preparations B.

The above results shows, adopts two kinds of prepared pharmaceutical preparations of method for making all can improve dense, the sticky state of stasis syndrome rat model blood; Adopt the pharmaceutical preparations A of preparation technology's gained provided by the invention for the preparation technology's gained pharmaceutical preparations B described in adopting contrast patent simultaneously, impact effect to blood stasis type hemorheology of rat is more remarkable, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 8: the pharmaceutical preparations of Different Preparation gained on rat due to oxytocin in the uterotonic impact of body

2, experimental animal: Kunming kind rat 210 ± 10g.

3, trial drug dosage: pharmaceutical preparations A administration group, pharmaceutical preparations B administration group are all established high, medium and low dosage group, and dosage is respectively 4g/kg, 2g/kg, 1g/kg, according to the administration of 20ml/kg body weight.

4, test method: get 80 male and female half and half of Kunming kind rat, be divided at random 8 groups by sex, body weight, 10 every group, be respectively matched group, model group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.2mgkg-1 subcutaneous injection diethylstilbestrol according to dosage before experiment, every day 1 time, for three days on end.According to dosage 50mgkg-1 subcutaneous injection pentobarbital sodium anesthetized rat, cut open the belly, find out a side cornua uteri, peel off gently uterus fatty tissue around, stitching a cotton thread at cornua uteri mid point is connected with tonotransducer, the vagina end of cornua uteri and ovary end are sewn to respectively on the fulcrum at special two ends, glass infuser bottom, then stomach wall are wrapped up to glass infuser bottom surrounding and sew up, add nutritional solution.Apart from adding one of heat-insulating lamp, make temperature maintain 37 ℃ ± 0.15 ℃ suitably, after curve shrinkage balance, record the mean tension (g) in its curve movement 10min.According to dosage 0.1U/kg gives oxytocin by femoral vein, and contraction is stablized backward nutritional solution and splashed into medicine, and matched group, model group splash into normal saline to nutritional solution.After curve shrinkage balance, record the mean tension (g) in its curve movement 10min, and calculate its suppression ratio (suppression ratio=(model group muscular tension-model group muscular tension)/model group muscular tension × 100%).Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

Table 8 Different Preparation gained pharmaceutical preparations on rat due to oxytocin in the uterotonic impact of body

Group	Dosage (g/kg)	Muscular tension after administration (g)	Suppression ratio (%)
				Matched group	—	0.69±0.03	—
Model group	—	2.62±0.37 ^u	—
				B low dosage	1	1.99±0.14 ^★	24.05
A low dosage	1	1.83±0.17 ^★	30.15
				Dosage in B	2	1.96±0.22 ^★	25.19
Dosage in A	2	1.74±0.15 ^★	33.59
				B high dose	3	1.84±0.21 ^★	29.77
A high dose	3	1.57±0.23 ^★?	40.07

Note: ^urepresent that matched group is compared with model group, P < 0.05, ^★represent that each administration group is compared with model group, P < 0.05, ^?represent that pharmaceutical preparations A high dose group is compared with pharmaceutical preparations B high dose group, P < 0.05.

Shown by table 8 result, for muscular tension after administration, model group and matched group relatively have significant difference (P < 0.05), show this test modeling success; After the administration of each administration group, muscular tension and model group more all have significant difference (P < 0.05), show that pharmaceutical preparations A, pharmaceutical preparations B all can reduce uterus muscle tension force.

Under high dose level, for reducing muscular tension, pharmaceutical preparations A is than pharmaceutical preparations B more remarkable effect, and both have significant difference (P < 0.05).Above-mentioned analytic explanation, the pharmaceutical preparations A that adopts preparation technology's gained provided by the invention is than adopting preparation technology's gained pharmaceutical preparations B described in contrast patent to reducing the uterus muscle tension force more remarkable effect of the rat model due to oxytocin, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 9: the impact that the pharmaceutical preparations of Different Preparation gained shrinks Isolated Rat Uterus due to oxytocin

2, experimental animal: Kunming kind rat 210 ± 10g.

4, test method: get 80 of Kunming kind rats and be divided at random 8 groups without pregnant history female rats by body weight, 10 every group, be respectively matched group, model group, the high, medium and low dosed administration group of pharmaceutical preparations A, the high, medium and low dosed administration group of pharmaceutical preparations B.Rat is 2mgkg according to dosage before experiment ^-1subcutaneous injection diethylstilbestrol, cause artificial rutting period to improve the sensitivity of uterus to medicine, every day 1 time, for three days on end, suck ether to mice, after anesthesia, cut open rapidly and get uterus, clip uterus 2cm, be placed in the sulculus that fills 30ml Rockwell liquid, be connected on sensor, bath temperature remains on 36 ℃ ± 0.5 ℃, in groove, pass into oxygen by air drain, a 1-2 per second minute bubbles, in order to guarantee that uterus is in more stable active state, to the in addition heavy heavy burden of 1g of uterus, after uterine contraction is stable, according to dosage 300UL-1 adds oxytocin inj, uterine smooth muscle is shunk strongly.After contraction is stable, in Rockwell liquid, add medicine, matched group adds normal saline to Rockwell liquid, then observe the muscular tension after number of contractions and the administration of every 10min after administration, and calculate suppression ratio (suppression ratio=(model group muscular tension-administration group muscular tension)/model group muscular tension × 100%).Experimental result adopts SPSS18.0 statistical software to carry out statistical procedures, and data are used

The impact that table 9 Different Preparation gained pharmaceutical preparations shrinks Isolated Rat Uterus due to oxytocin

Group	Dosage (g/kg)	Frequency (inferior/10min)	Muscular tension after administration (g)	Suppression ratio (%)
					Matched group	—	7.39±1.04	5.64±1.24	—
Model group	—	12.72±1.67 ^u	56.18±1.96 ^u	—
					B low dosage	1	10.33±1.42 ^★	40.57±3.03 ^★	27.79
A low dosage	1	8.47±1.28 ^★▲	34.39±1.97 ^★▲	38.79
					Dosage in B	2	9.57±1.41 ^★	35.52±1.49 ^★	36.77
Dosage in A	2	7.41±1.09 ^★●	20.39±1.86 ^★●	63.7
					B high dose	3	7.88±1.23 ^★	31.49±1.94 ^★	43.95
A high dose	3	6.37±1.05 ^★?	16.93±1.53 ^★?	69.86

As shown in Table 9, after the isolated uterine contraction frequency that model group is surveyed and administration, muscular tension has significant difference (P < 0.05) with matched group than all, and this test modeling success is described; After the isolated uterine contraction frequency that each administration group is surveyed and administration, muscular tension is with model group comparison, all there is significant difference (P < 0.05), illustrate that pharmaceutical preparations A, pharmaceutical preparations B all can significantly reduce isolated uterine contraction frequency and muscular tension; Under high, medium and low dosage level, compare with pharmaceutical preparations B group, after pharmaceutical preparations A organizes the isolated uterine contraction frequency surveyed and administration, muscular tension all has significant difference (P < 0.01), shows that pharmaceutical preparations A is to reducing isolated uterine contraction frequency and uterus muscle tension force more remarkable effect.

The above results shows, adopts two kinds of prepared pharmaceutical preparations of method for making all to have considerable influence for the rat model isolated uterine due to oxytocin, can reduce isolated uterine contraction frequency and uterus muscle tension force; The pharmaceutical preparations A that simultaneously adopts preparation technology's gained provided by the invention is than the preparation technology's gained pharmaceutical preparations B more remarkable effect adopting described in contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

embodiment 10: the clinical comparison test of the pharmaceutical preparations of Different Preparation gained

1, grouping situation: the 100 routine patients that meet diagnostic criteria are divided into test group and matched group at random, i.e. pharmaceutical preparations A test group and pharmaceutical preparations B matched group, wherein test group 50 examples, matched group 50 examples.

2, treatment medication and the course for the treatment of

Test group: oral drugs prepared product A9 gram, day three times, serve on 7 days.Matched group: oral drugs prepared product B9 gram, every day three times.Two groups all started to take medicine in premenstrual 4 days, serve on 7 days, and treating continuously 3 menstrual cycle is a course for the treatment of.After the course for the treatment of, add up curative effect.

3, criterion of therapeutical effect

Clinical efficacy standard is carried out with reference to " guideline of clinical investigations for the treatment of by Chinese herbs dysmenorrhea ".

Dysmenorrhea curative effect determinate standard is as follows:

Clinical recovery: after taking medicine, integration returns to 0 point, stomachache and other transference cures, 3 not recidivists of menstrual cycle after drug withdrawal.

Effective: the rear integration for the treatment of is reduced to front below 1/2 of integration for the treatment of, and stomachache obviously alleviates, and all the other symptoms take a turn for the better, and disobedience analgesic can be adhered to work.

Effective: after treatment, integration is reduced to the 1/2-3/4 of the front integration for the treatment of, and stomachache alleviates, all the other symptoms take a turn for the better, and take analgesic and can adhere to work.

Invalid: stomachache and other symptoms are without changer.

Traditional Chinese medical science disease curative effect determinate standard formula is: integration × 100% before therapeutic index (n)=(the rear integration of integration-treatment before treatment)/treatment

Traditional Chinese medical science disease curative effect determinate standard is as follows:

Short Term Clinical is cured: n >=90%

Effective: 90% > n >=70%

Effective: 70% > n >=30%

Invalid: n < 30%

4 and data analysis

Measurement data adopts t check or variance analysis, and enumeration data adopts χ ²check, clinical-grade data relatively adopts RiDit to analyze.Measurement data used is all used

± S represents, all data acquisitions are analyzed with SPSS18.0 statistical software.Determine that P < 0.05 is for there being significant difference.

5, physical data

1) two groups of patient ages distribute

A table 10 liang group patient age distributes

Group	N	15-20	21-25	26-30	30-35	Mean age
							Test group	50	9	17	18	6	25±4.85
Matched group	50	8	20	17	5	25.18±4.81

Shown in upper table: minimum 15 years old of test group age, maximum 35 years old; Minimum 15 years old of matched group age, maximum 35 years old, through χ ²check, two groups of patient age there was no significant differences (P > 0.05), have comparability.Through t check, two groups of patient age there was no significant differences (P > 0.05), have comparability.

2) two groups of patient's course of disease situations comparison

Table 11 liang group patient course of disease situation comparison

As shown above: through χ ²check, two groups of patient's course of disease there was no significant differences (P > 0.05), have comparability.

3) dysmenorrhea integration situation comparison before two groups of patient treatments

Dysmenorrhea integration situation comparison before table 12 liang group patient treatment

Group	N	Symptom integral before treatment
			Test group	50	11.58±4.26
Matched group	50	11.83±4.04

Shown in upper table: through χ ²check, dysmenorrhea integration situation there was no significant difference (P > 0.05) before two groups of patient treatments, has comparability.Through t check, severity extent there was no significant difference (P > 0.05) before two groups of patient treatments, has comparability.

4) severity extent comparison before two groups of treatments

Severity extent comparison before table 13 liang group patient treatment

As shown above: through χ ²check, severity extent there was no significant difference (P > 0.05) before two groups of patient treatments, has comparability.

6, result

Through 1 the course for the treatment of analysis and observation to efficacy result be summarized as follows:

Dysmenorrhea curative effect comparison after table 14 liang group treatment:

Group	Number of cases	Recovery from illness	Effective	Effectively	Invalid	Total effective rate
							Test group	50	31	17	10	2	96%
Matched group	50	20	16	5	9	82%

Analyze through RiDit, two groups are compared P < 0.05, show that the therapeutic effect of test group is better than matched group.The pharmaceutical preparations A that shows to adopt preparation technology's gained provided by the invention is more obvious than adopting the preparation technology's gained pharmaceutical preparations B therapeutic effect described in contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

The comparison of table 15 pain scores

After two groups of patient treatments, pain scores is learned processing by statistics, and two groups of pain scores P < 0.05, have significant difference, demonstrate test group analgesic effect better.The pharmaceutical preparations A that shows to adopt preparation technology's gained provided by the invention is more obvious than adopting the preparation technology's gained pharmaceutical preparations B analgesic effect described in contrast patent, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

Durante dolors comparison after table 16 liang group patient treatment

After two groups of patient treatments, durante dolors is learned processing by statistics, and two groups of durante dolors P < 0.05, have significant difference, and test group durante dolors is shorter simultaneously.Show that patient takes after the pharmaceutical preparations A of preparation technology's gained provided by the invention, durante dolors is shorter, further illustrates preparation technology provided by the present invention and is better than contrasting the preparation technology described in patent.

Traditional Chinese medical science disease curative effect comparison after table 17 liang group treatment ( ± S)

After two groups of patient treatments, the scoring of traditional Chinese medical science disease is learned and is processed by statistics, and two groups of traditional Chinese medical science disease scoring P < 0.05, have significant difference.

embodiment 11: the pharmaceutical preparations stability test contrast of Different Preparation gained

Influence factor's test: the pharmaceutical preparations A, the pharmaceutical preparations B that get gained in embodiment 1 carry out hot test (60 ℃), high wet test (relative humidity 92.5%, 25 ℃) and exposure experiments to light (4500lx), respectively at sampling in the 0th, 5,10 days, check according to every test method, and with 0D comparison, the results are shown in following table 18.

The pharmaceutical preparations influence factor result of the test of table 18 Different Preparation gained

The demonstration of table 18 result, under hot conditions, there is respectively the phenomenons such as layering, granularity increase, mastic water outlet in Radix Angelicae Sinensis dysmenorrhea pharmaceutical preparations A, B, illustrates that pharmaceutical preparations A is more stable under hot conditions than pharmaceutical preparations B at the 10th day, the 5th day; Under super-humid conditions, there is the phenomenons such as layering, granularity increase, mastic water outlet in DO prepared product B, shows that pharmaceutical preparations A is more stable under super-humid conditions than pharmaceutical preparations B at the 10th day; Under illumination, there is the phenomenons such as layering, granularity increase, mastic water outlet in pharmaceutical preparations A, B, both stability there was no significant differences are described under illumination at the 10th day.In sum, under above-mentioned influence factor's condition, adopt the pharmaceutical preparations A of preparation technology's gained provided by the invention can show that than the preparation technology's gained pharmaceutical preparations B described in employing contrast patent is more stable preparation technology provided by the invention is better than contrasting the preparation technology of patent.

In sum, in active constituent content measuring test, adopt the present invention its active constituent content of medicine of preparing of described production technology apparently higher than the prepared medicine of preparation technology adopting described in contrast patent; Meanwhile, aspect pharmacodynamics and clinical efficacy, adopt the present invention the medicine prepared of described production technology, be better than equally adopting the prepared medicine of preparation technology described in contrast patent; The medicine that simultaneously adopts preparation technology of the present invention to prepare is more stable.In sum, can obtain preparation technology provided by the present invention and be better than contrasting the preparation technology described in patent.

embodiment 12: the preparation of the pharmaceutical preparations granule of preparation technology's gained that employing this patent provides

By the pharmaceutical preparations A of gained in embodiment 1, add appropriate dried cream powder, sucrose, dextrin, appropriate amount of ethanol granulation, dry, mix and get final product.

embodiment 13: the preparation of the pharmaceutical preparations tablet of preparation technology's gained that employing this patent provides

By the pharmaceutical preparations A of gained in embodiment 1, add appropriate dried cream powder, sucrose, dextrin, appropriate amount of ethanol granulation, dry, granulate, tabletting, film coating or sugar-coat, to obtain final product.

embodiment 14: the preparation of the pharmaceutical preparations pill of preparation technology's gained that employing this patent provides

By the pharmaceutical preparations A of gained in embodiment 1, add appropriate refined honey and make concentrated honeyed pill, to obtain final product.

embodiment 15: the preparation of the pharmaceutical preparations powder of preparation technology's gained that employing this patent provides

By the pharmaceutical preparations A of gained in embodiment 1, grind, mix, pack and get final product.

embodiment 16: the preparation of the pharmaceutical preparations capsule of preparation technology's gained that employing this patent provides

By the pharmaceutical preparations A of gained in embodiment 1, add appropriate amount of starch and magnesium stearate, with the ethanol granulation of debita spissitudo, dry, granulate, encapsulated and get final product.

Claims

1. treat a preparation technology for the Chinese medicine composition of primary dysmenorrhea, described technique is specific as follows:

Prescription:

Preparation technology:

1) get Radix Linderae 5-10 part, add the stirring of rice vinegar 1-2 part, moistening 0.5-2h, after vinegar liquid is sucked, with 60% microwave heating power, heating 2-10min, takes out, and cools, for subsequent use;

2) get Rhizoma Chuanxiong 5-15 part, Rhizoma Corydalis 5-10 part, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 60-90% ethanol as solvent, soaked overnight, add 6-9 and doubly measure 60-90% alcohol reflux 1-3 time, each 0.5-2h, filter merging filtrate, decompression recycling ethanol, collect filtrate for later use, another device is collected filtering residue;

3) get Radix Angelicae Sinensis 10-20 part and be ground into coarse powder, make solvent, dipping 24-48h, slowly percolation with 60-90% ethanol, collect the liquid of just filtering, another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collect the continuous liquid of filtering, merging filtrate is for subsequent use, and another device is collected filtering residue;

4) get Rhizoma Cyperi 5-15 part, Rhizoma Curcumae 5-15 part and be ground into coarse powder, add the water that 6-9 doubly measures, soak 1-3h, hydrodistillation extracts volatile oil, and extraction time is 2-5h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use;

2. the preparation technology who treats according to claim 1 the Chinese medicine composition of primary dysmenorrhea, is characterized in that:

2) get Rhizoma Chuanxiong 5-15 part, Rhizoma Corydalis 5-10 part, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 70-80% ethanol as solvent, soaked overnight, add 8-9 and doubly measure 70-80% alcohol reflux 2 times, each 1-1.5h, filter merging filtrate, decompression recycling ethanol, collect filtrate for later use, another device is collected filtering residue;

3) get Radix Angelicae Sinensis 10-20 part and be ground into coarse powder, make solvent, dipping 24-36h, slowly percolation with 70-80% ethanol, collect the liquid of just filtering, another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collect the continuous liquid of filtering, merging filtrate is for subsequent use, and another device is collected filtering residue;

4) get Rhizoma Cyperi 5-15 part, Rhizoma Curcumae 5-15 part and be ground into coarse powder, add the water that 7-9 doubly measures, soak 1-1.5h, hydrodistillation extracts volatile oil, and extraction time is 3-4h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use;

3. according to the preparation technology who treats the Chinese medicine composition of primary dysmenorrhea described in claim 1-2, it is characterized in that:

1) get 10 parts of the Radixs Linderae, add 2 parts of stirrings of rice vinegar, moistening 1h, after vinegar liquid is sucked, with 60% microwave heating power, heating 5min, takes out, and cools, for subsequent use;

2) get 15 parts of Rhizoma Chuanxiongs, 10 parts of Rhizoma Corydalis, the Radix Linderae (process of preparing Chinese medicine) and be ground into coarse powder take 70% ethanol as solvent, soaked overnight, adds 8 times of amount 70% alcohol reflux 2 times, each 1.5h is excellent, filters merging filtrate, decompression recycling ethanol, collects filtrate for later use, and another device is collected filtering residue;

3) get 20 parts of Radix Angelicae Sinensis and be ground into coarse powder, make solvent with 70% ethanol, dipping 24h, slowly percolation, collects the liquid of just filtering, and another device is preserved, and continues percolation, till the nearly colourless or micro-yellow of percolate, collects the continuous liquid of filtering, and merging filtrate is for subsequent use, and another device is collected filtering residue;

4) 15 parts of 15 parts of Rhizoma Cyperis, Rhizoma Curcumae are ground into coarse powder, add the water of 8 times of amounts, soak 2.5h, hydrodistillation extracts volatile oil, and extraction time is 4h, collects volatile oil, and it is for subsequent use that distillation rear solution is collected, and the another device of medicinal residues is collected for subsequent use;

4. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-3, it is characterized in that, described technique also comprises to the pharmaceutical preparations obtaining and adds appropriate amount of auxiliary materials, makes various dosage forms.

5. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-4, it is characterized in that, described technique also comprises to the pharmaceutical preparations obtaining and adds appropriate dried cream powder, sucrose, dextrin, add again appropriate 70% ethanol, dry, granulation agent.

6. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-4, it is characterized in that, described technique also comprises to the pharmaceutical preparations obtaining, and adds appropriate dried cream powder, sucrose, dextrin, add again appropriate 70% ethanol granulation, dry, granulate tabletting, film coating or sugar-coat, make tablet.

7. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-4, it is characterized in that, described technique also comprises to obtained pharmaceutical preparations, adds appropriate refined honey, the concentrated pill of making.

8. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-4, it is characterized in that, described technique also comprises obtained pharmaceutical preparations, grinds, and mixes, and packing is made powder.

9. according to the preparation technology of the Chinese medicine composition of the treatment primary dysmenorrhea described in claim 1-4, it is characterized in that, described technique also comprises to obtained pharmaceutical preparations, add appropriate amount of starch, magnesium stearate, then add appropriate 70% ethanol granulation, dry, granulate, the encapsulated capsule of making.

10. the preparation technology of the Chinese medicine composition for the treatment of primary dysmenorrhea according to claim 1, is characterized in that, described dosage form includes but not limited to granule, tablet, pill, powder and capsule.