CN104161806A

CN104161806A - Traditional Chinese medicine composition for treating toothache and preparation method thereof

Info

Publication number: CN104161806A
Application number: CN201410407324.7A
Authority: CN
Inventors: 鲍家科; 茅向军; 许乾丽; 左鼎; 王尉; 王德甫; 王晓春; 周宁; 冯泽熹; 曾香兰
Original assignee: Guizhou Tongjitang Pharmaceutical Co Ltd
Current assignee: Sinopharm Group Tongjitang Guizhou Pharmaceutical Co Ltd
Priority date: 2014-08-19
Filing date: 2014-08-19
Publication date: 2014-11-26
Anticipated expiration: 2034-08-19
Also published as: CN104161806B

Abstract

The invention relates to a traditional Chinese medicine composition for treating toothache and a preparation method thereof. The traditional Chinese medicine composition is prepared from the following medicinal materials in parts by weight by virtue of the following preparation steps: taking 50-400 parts of valeriana officinalis, 20-100 parts of safflower, 50-300 parts of garden balsam and 5-50 parts of camphorwood, performing distillation extraction on the valeriana officinalis, soaking the dreg in ethanol before percolation, soaking the garden balsam, the safflower and the camphorwood in the ethanol before percolation, adding a pharmaceutically acceptable carrier or a diluting agent or not, and preparing into a pharmaceutically acceptable form according to a conventional preparation process. The traditional Chinese medicine composition is high in active ingredient content, and free from toxic and side effects, and the toothache treating effect of the composition is superior to that of the prior art.

Description

A kind of Chinese medicine composition for the treatment of toothache and preparation method thereof

Technical field

The present invention relates to Medicinal invention field, be specifically related to a kind of Chinese medicine composition for the treatment of toothache and preparation method thereof.

Background technology

Toothache is a kind of common disease, and it shows as: red swelling of gingiva, chance caloric stimulation pain, buccal swelling etc.Toothache causes dental pulp (dental nerve) to infect caused by gingivitis and periodontitis, dental caries (decayed tooth) or jackknifing tooth mostly; The traditional Chinese medical science thinks that toothache is due to due to the reasons such as diseases caused by exogenous pathogenic factor ailment said due to cold or exposure, excessiveness of stomach-fire, the hyperactivity of fire of suffering from a deficiency of the kidney, worm-eaten tooth.Toothache is the cardinal symptom of primary disease, early stage, and gingiva is itched, uncomfortable, halitosis, and then red swelling of gingiva, soft, easily hemorrhage, pain, repeatedly outbreak; With the passing of time the periodontal membrane of gingiva and tooth root is destroyed, forms a sack, is periodontal pocket, in bag, often have pus to overflow, inflammation continues to expand, and can become periodontal abscess, aggravation, local pain, swelling, be just rigid, after become softly, have fluctuation, can wear out voluntarily, flow out pus, after coming to a head, pain can alleviate, or outbreak repeatedly, very painful.

The medicine for external use that is used for the treatment of at present toothache mainly contains: tincture, powder, gargarism etc.These drug uses inconvenience of getting up, and action time is not long yet, can not bring into play for a long time drug effect; Oral medicine mainly be take antibiotic antiinflammatory as main, but side effect is larger; Acute pulpitis, emergency processing is often carried out to open marrow operation in oral cavity, but much patient old and that have a nervous feared state of mind is often difficult to accept.

The disclosed Yatongxiao for curing toothache of Chinese patent 95100457.3 and preparation technology, have obvious analgesia, detumescence, teeth consolidating effect to dental disease, adopts steam distillation to obtain Valeriana officinalis var. latifolia volatile oil component, and petroleum ether soaks 5 days to obtain paste; Flos Impatientis, Flos Carthami and Lignum cinnamomi camphorae obtain extracting solution for 7 days by soak with ethanol; Clearly, this process cycle time is long, and production efficiency is low, in addition, contains abundant volatile oil component in Lignum cinnamomi camphorae, only by soaking, can not effectively extract, and this invention active constituent content is low, and impurity is many.

The method for making of disclosed Toothache tinctures in the < < of National Drug Administration national drug standards > > [WS-10589-(ZD-0589)-2002] compound recipe Toothache tinctures quality standard, is specially: it is standby that Valeriana officinalis var. latifolia is collected distillate through vapor distillation; It is standby that water extraction liquid is condensed into thick paste; After medicinal residues are dry, add 75% ethanol, soak 7 days, extract and thick paste mix, and after standing 24 hours, filter, and solution decompression reclaims ethanol and obtains extractum; Flos Impatientis, Lignum cinnamomi camphorae, Flos Carthami add the airtight immersion of 60% ethanol 7 days, filter, and get filtrate and merge with above-mentioned extractum, mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol be 45%, obtain.But do not mention Valeriana officinalis var. latifolia medicinal residues are carried out to the technique of percolation and the technique that Flos Impatientis, Lignum cinnamomi camphorae, Flos Carthami are soaked to rear percolation.

The said method production cycle is long, and the difficult proposition of effective ingredient, and content is on the low side, affects the curative effect of medicine, carries inconvenience.

Inventor, in order to overcome the above problems, has researched and developed a kind of new method.

Summary of the invention

The object of this invention is to provide a kind of Chinese medicine composition for the treatment of toothache.

Another object of the present invention is to provide a kind of Chinese medicine composition preparation method for the treatment of toothache.

The present invention also provides the application of Chinese medicine composition in the medicine of preparation treatment gingivitis, toothache that dental caries causes or gingival swelling and pain.

The Chinese medicine composition for the treatment of toothache provided by the invention, this Chinese medicine composition is made by being prepared as follows technique by the medicine of following weight parts: Valeriana officinalis var. latifolia 50-400 part, Flos Carthami 20-100 part, Flos Impatientis 50-300 part, Lignum cinnamomi camphorae 5-50 part

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 1-8 times of water gaging distillation extraction 2-6h;

2) collect in volatile oil refrigerator and save backup, it is standby that water extraction liquid is condensed into thick paste;

3) after medicinal residues are dried, add 8-20 and doubly measure after 50%-85% soak with ethanol 1-3h, the speed percolation with 2-5Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 20-30h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add 10-20 and doubly measure after 50%-70% soak with ethanol 4-24h, and with the speed percolation of 0.5-3Bv/h, percolate and above-mentioned thick paste mix, and filter, and add volatile oil, are adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 5%-50%;

The Chinese medicine composition for the treatment of toothache provided by the invention, this Chinese medicine composition is preferably made by being prepared as follows technique by the medicine of following weight parts: Valeriana officinalis var. latifolia 50-400 part, Flos Carthami 20-100 part, Flos Impatientis 50-300 part, Lignum cinnamomi camphorae 5-50 part

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 4 times of water gaging distillation extraction 5h;

3) after medicinal residues are dried, add after 10 times of amount 75% soak with ethanol 2h, the speed percolation with 3Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 24h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 15 times of amount 60% soak with ethanol 6h, and with the speed percolation of 1Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 45%;

6) add or do not add pharmaceutically acceptable carrier or diluent, preparation process is made acceptable dosage form on pharmaceutics routinely.

Chinese medicine composition weight proportion of the present invention is preferred: Valeriana officinalis var. latifolia 80-350 part, Flos Carthami 30-90 part, Flos Impatientis 80-260 part, Lignum cinnamomi camphorae 10-45 part.

Chinese medicine composition weight proportion of the present invention is preferred: Valeriana officinalis var. latifolia 100-300 part, Flos Carthami 40-80 part, Flos Impatientis 90-240 part, Lignum cinnamomi camphorae 15-40 part.

Chinese medicine composition weight proportion of the present invention is preferred: Valeriana officinalis var. latifolia 130-260 part, Flos Carthami 50-75 part, Flos Impatientis 100-180 part, Lignum cinnamomi camphorae 20-35 part.

Chinese medicine composition weight proportion of the present invention is preferred: 200 parts of Valeriana officinalis var. latifolia, 60 parts, Flos Carthami, 110 parts of Flos Impatientiss, 30 parts of Lignum cinnamomi camphoraes.

Chinese medicine composition of the present invention is solid preparation or liquid preparation or membranous patch.

Solid preparation of the present invention is sheet, capsule, granule or pill; Described liquid preparation is tincture, oral liquid, injection.

Pharmaceutically acceptable carrier of the present invention or diluent are selected from one or more in film former, wetting agent, filler, binding agent, disintegrating agent, lubricant, surfactant, diluent or correctives.

The application of Chinese medicine composition of the present invention in the medicine of preparation treatment gingivitis, toothache that dental caries causes or gingival swelling and pain.

Described filler is selected from dextrin, starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline Cellulose or glucose etc.;

Described film former is selected from hydroxypropyl emthylcellulose, polyvinyl alcohol, carbomer.

Described binding agent be selected from cellulose derivative, alginate, gelatin or polyvinylpyrrolidone, etc.;

Described disintegrating agent is selected from microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose or cross-linking sodium carboxymethyl cellulose;

Described lubricant is selected from stearic acid, Polyethylene Glycol, calcium carbonate, sodium bicarbonate, micropowder silica gel, Pulvis Talci or magnesium stearate;

Described surfactant is selected from dodecylbenzene sodium sulfonate, stearic acid, Pluronic F68, fatty acid Pyrusussuriensis is smooth or Polysorbate (tween) etc.;

Described correctives is selected from aspartame, Sucralose or saccharin sodium, lactose.

Described wetting agent is glycerol.

Described diluent is water etc.

The Chinese medicine composition for the treatment of toothache provided by the invention has the following advantages:

1, compare with prior art:

1) the disclosed Yatongxiao for curing toothache of Chinese patent application 95100457.3 of the prior art and preparation technology, this process cycle time is long, production efficiency is low, active constituent content is low, impurity is many, the content of S-A Hydroxysafflor yellow A is 3.998mg/g after measured, Quercetin 0.521mg/g, kaempferide 5.312mg/g.

2) method for making of disclosed Toothache tinctures in the < < of National Drug Administration national drug standards > > of the prior art [WS-10589-(ZD-0589)-2002] compound recipe Toothache tinctures quality standard, its shortcoming is: effective component content is on the low side, the content of S-A Hydroxysafflor yellow A is 4.115mg/g after measured, Quercetin 0.553mg/g, kaempferide 5.528mg/g, and quality is unstable, action time is not long.

2, the present invention soaks rear percolation by the extraction of Valeriana officinalis var. latifolia vapor distillation, medicinal residues; By percolation after Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae soak with ethanol, this technique has been saved ethanol consumption, has reduced extraction time, reduced production cost, effective component content is high, and the high-load of S-A Hydroxysafflor yellow A has reached 4.668mg/g after measured, Quercetin 0.688mg/g, kaempferide 6.119mg/g.

3, the present invention take Valeriana officinalis var. latifolia expelling wind and removing dampness, invigorate blood circulation that to dredge through, regulating QI to relieve pain be monarch drug; Flos Carthami promoting blood circulation to restore menstrual flow, eliminating stasis to stop pain, Rhodiola dumulosa (Franch.) S.H.Fu promoting blood circulation to remove obstruction in the collateral, wind-expelling pain-stopping, Lignum cinnamomi camphorae expelling wind and cold, vital energy regualting and blood circulation-promoting pain relieving, three is adjuvant altogether.All medicines share, and have promoting blood circulation to remove blood stasis, the effects such as reducing swelling and alleviating pain, and the clinical toothache causing for gingivitis, pericoronitis, dental caries or gingival swelling and pain, determined curative effect, without obvious toxic-side effects.

The specific embodiment

Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.

Described times of amount is weight multiple, and Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add 15 times of amount 60% ethanol, for adding 60% ethanol of 15 times of Flos Impatientiss, Flos Carthami, Lignum cinnamomi camphorae weight.

Embodiment 1: compound recipe toothache pad pasting

Compound recipe forms: Valeriana officinalis var. latifolia 200g, Flos Carthami 60g, Flos Impatientis 110g, Lignum cinnamomi camphorae 30g

Preparation method:

6) add 1.5% hydroxypropyl emthylcellulose and 1% glycerol, swelling 24h, is laid on the glass plate that scribbles a small amount of liquid paraffin, dries, and obtains.

Embodiment 2: compound recipe toothache pad pasting

Compound recipe forms: Valeriana officinalis var. latifolia 50g, Flos Carthami 20g, Flos Impatientis 50g, Lignum cinnamomi camphorae 5g

Preparation method:

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 1 times of water gaging distillation extraction 2h;

3) after medicinal residues are dried, add after 8 times of amount 50% soak with ethanol 1, the speed percolation with 2Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 20h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 10 times of amount 50% soak with ethanol 4h, and with the speed percolation of 0.5Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 5%;

Embodiment 3: compound recipe toothache pad pasting

Compound recipe forms: 50 parts of Valeriana officinalis var. latifolia 400g, Flos Carthami 100g, Flos Impatientis 300g, Lignum cinnamomi camphoraes

Preparation method:

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 8 times of water gaging distillation extraction 6h;

3) after medicinal residues are dried, add after 20 times of amount 85% soak with ethanol 3h, the speed percolation with 5Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 30h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 20 times of amount 70% soak with ethanol 24h, and with the speed percolation of 3Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 50%;

Embodiment 4 compound recipe Toothache tinctures

Preparation method:

6) pack in bottle, obtain tincture.

Embodiment 5 compound recipe Toothache tinctures

Compound recipe forms: Valeriana officinalis var. latifolia 80g, Flos Carthami 30g, Flos Impatientis 80g, Lignum cinnamomi camphorae 10g

Preparation method:

6) pack in bottle, obtain tincture.

Embodiment 6 compound recipe Toothache tinctures

Compound recipe forms: Valeriana officinalis var. latifolia 350g, Flos Carthami 90g, Flos Impatientis 260g, Lignum cinnamomi camphorae 45g

Preparation method:

6) pack in bottle, obtain tincture.

Embodiment 7 compound recipe toothache capsules

Compound recipe forms: Valeriana officinalis var. latifolia 100g, Flos Carthami 40g, Flos Impatientis 90g, Lignum cinnamomi camphorae 15g

Preparation method:

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 3 times of water gaging distillation extraction 3h;

3) after medicinal residues are dried, add after 12 times of amount 60% soak with ethanol 2h, the speed percolation with 4Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 22h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 12 times of amount 60% soak with ethanol 5h, and with the speed percolation of 1Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 10%;

6) add dextrin and the lactose of 3 times, mix, granulation, dry, pack in Capsules, obtain capsule.

Embodiment 8 compound recipe toothache sprays

Compound recipe forms: Valeriana officinalis var. latifolia 300g, Flos Carthami 80g, Flos Impatientis 240g, Lignum cinnamomi camphorae 40g.

Preparation method:

1) by proportioning, take each taste Chinese medicine, Valeriana officinalis var. latifolia is added to 6 times of water gaging distillation extraction 5h;

3) after medicinal residues are dried, add after 18 times of amount 85% soak with ethanol 2.5h, the speed percolation with 4Bv/h, obtains percolate;

4) percolate and the concentrated thick paste of water extraction liquid are mixed, after standing 28h, filter, solution decompression reclaims ethanol, and to obtain thick paste standby;

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 18 times of amount 65% soak with ethanol 20h, and with the speed percolation of 2Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 50%;

6) pack in aerosol container, obtain spray.

Embodiment 9 compound recipe toothache tablets

Compound recipe forms: Valeriana officinalis var. latifolia 130g, Flos Carthami 50g, Flos Impatientis 100g, Lignum cinnamomi camphorae 20g.

Preparation method:

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 15 times of amount 60% soak with ethanol 6h, and with the speed percolation of 1Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 5%;

6) add dextrin and the lactose of 5 times, mix, tabletting, obtains tablet.

Embodiment 10 compound recipe toothache granules

Compound recipe forms: Valeriana officinalis var. latifolia 260g, Flos Carthami 75g, Flos Impatientis 180g, Lignum cinnamomi camphorae 35g.

Preparation method:

5) Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add after 15 times of amount 60% soak with ethanol 6h, and with the speed percolation of 1Bv/h, percolate and above-mentioned thick paste mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol in medicinal liquid be 10%;

6) add dextrin and the lactose of 3 times, mix, granulate, dry, granulate, obtains granule.

Embodiment 11 compound recipe toothache oral liquids

Preparation method:

6) add the pure water of 3 times and the sweeting agent of 0.2 times, mix, perfusion, gland, sterilizing, obtains oral liquid.

Embodiment 12 compound recipe toothache injection

Compound recipe forms: Valeriana officinalis var. latifolia 200g, Flos Carthami 60g, Flos Impatientis 110g, Lignum cinnamomi camphorae 30g.

Preparation method:

6) add 1000 liters of waters for injection, with the filtering with microporous membrane of 0.45 μ m, stir and make mix homogeneously in 30 minutes, embedding, sterilizing, obtains injection.

Embodiment 13 compound recipe toothache pills

Preparation method:

6) add the dextrin of 5 times, mix, pill, dry, obtain pill.

Test example 1 contrast test

1, sample: product in the middle of embodiment of the present invention 1-13 and S-A Hydroxysafflor yellow A, Quercetin, kaempferide content in comparative example 1, comparative example 2 centre product are contrasted.

2, technique: the middle product of embodiment 1-13 are prepared from by the technique of embodiment of the present invention 1-13, comparative example 1 is prepared from by following technique with comparative example 2:

Formula and the method for making of comparative example 1 Chinese patent application 95100457.3

Form: Valeriana officinalis var. latifolia 35.7g Flos Impatientis 37.5g Flos Carthami 15g Lignum cinnamomi camphorae 10g

Method for making: Valeriana officinalis var. latifolia is cleaned in rearmounted steamer, carries out steam distillation, fresh of every 100kg, collects distillate 50kg standby; (2) Valeriana officinalis var. latifolia root is cleaned and dried, add petroleum ether and flooded powder, airtight, to soak and filter afterwards for 5 days, filtrate obtains faint yellow paste after reclaim under reduced pressure petroleum ether, continues to be incubated and to evaporate into without till petroleum ether on water-bath, standby; (3) get Flos Impatientis, Flos Carthami, rod wood, after preparing, add up 1.2 times of 60% airtight immersion of ethanol of dose by 8:1.5:0.5, whisk every day once, within 7 days, filter afterwards, the 10%-12% ethanol that residue adds former dose again embathes filtration, and merging filtrate is standby; (4) get (1), (2), (3) each group respectively in 10%, 2.5%, 87.5% ratio mixing, standing 24 hours, filtration, standby after the assay was approved; (5) subpackage, every bottle of 30ml, capping, pastes label, packing, through sleeve sample after the assay was approved, warehouse-in, puts the storage of shady and cool place.

Comparative example 2: formula and the method for making of disclosed compound recipe Toothache tinctures in the < < of National Drug Administration national drug standards > > [WS-10589-(ZD-0589)-2002] compound recipe Toothache tinctures quality standard

Form: Valeriana officinalis var. latifolia 200g Flos Carthami 60g Flos Impatientis 110g Lignum cinnamomi camphorae 30g

Method for making: above four tastes, it is standby that Valeriana officinalis var. latifolia is collected distillate through vapor distillation; It is standby that water extraction liquid is condensed into thick paste; After medicinal residues are dry, add 75% ethanol, soak 7 days, extract and thick paste mix, and after standing 24 hours, filter, and solution decompression reclaims ethanol and obtains extractum; Flos Impatientis, Lignum cinnamomi camphorae, Flos Carthami add the airtight immersion of 60% ethanol 7 days, filter, and get filtrate and merge with above-mentioned extractum, mix, and filtration, adds volatile oil, is adjusted to ormal weight, and to make amount of alcohol be 45%, obtain.

3, content assaying method

[assay] measured according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2010).

(1) S-A Hydroxysafflor yellow A assay

Chromatographic condition and system suitability experiment are filler with octadecylsilane chemically bonded silica; Methanol-0.4% phosphoric acid solution (26:74) is mobile phase; Detection wavelength is 403nm.Theoretical cam curve is calculated and should be not less than 3000 by S-A Hydroxysafflor yellow A peak.

Reference substance solution preparation

It is appropriate that precision takes S-A Hydroxysafflor yellow A reference substance, adds 25% methanol and make the solution that contains 0.1mg in every 1ml, obtains.

The preparation of need testing solution

Sample thief, precision takes 0.8g, puts in tool plug conical flask, and precision adds 25% methanol 25ml, weighed weight.Supersound process (power 300W, frequency 50kHz) 30 minutes, lets cool. weighed weight again.The weight of supplying less loss with 25% methanol, shakes up, and filters, and gets subsequent filtrate, obtains.

Algoscopy

Precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and injection liquid chromatography, measures, and obtains.

(2) kaempferide and quercetin content are measured

Chromatographic condition and system suitability experiment

Octadecylsilane chemically bonded silica is filler; Methanol-0.4% phosphoric acid solution (42:58) is mobile phase; Detection wavelength is 360nm.Number of theoretical plate calculates and is not less than 3000 by emodin.

Reference substance solution preparation

Precision takes kaempferide and Quercetin reference substance is appropriate, adds methanol and makes respectively kaempferide (C ₁=104 μ g/ml) and Quercetin (C ₂=9 μ g/ml) mix reference substance solution, obtain.

The preparation of need testing solution

Precision takes sample 1.5g, puts in tool plug conical flask, and precision adds 70% ethanol 50ml, weigh, on water-bath, reflux is 1 hour, takes off, let cool, weigh, with 70% ethanol, supply the weight of loss, filter, get subsequent filtrate 25ml, put in tool plug conical flask, in water-bath, volatilize, add methanol-25% hydrochloric acid (4:1) mixed liquor 25ml, reflux 30min in water-bath, take off, let cool, be transferred in 50ml measuring bottle, with methanol gradation, wash conical flask, washing liquid is incorporated in measuring bottle, add methanol to scale, shake up, filter, get subsequent filtrate, obtain.

Algoscopy

4, result: the S-A Hydroxysafflor yellow A in embodiment of the present invention 1-13 sample, Quercetin, kaempferide are apparently higher than comparative example 1, comparative example 2 (in Table 1).

Table 1: assay result

Test example 2 preparation technology parameters preferably

1 Valeriana officinalis var. latifolia vapor distillation technique orthogonal test

The investigation of 1.1 extraction times

Consult related documents data and learn, the factor that affects vapor distillation mainly contains: distillation time, amount of water and medical material soak time.Select three factors, with volatile oil, extract and be entirely index, the volatile oil yield of different time sections is investigated.Press the first method in appendix X. determination of volatile oil method of < < Chinese Pharmacopoeia > > (version in 2010), take 3 parts of each 100g of Valeriana officinalis var. latifolia, cut into and be about thick section of 1cm, add 6 times of water gagings, with volatile oil extractor, extract, the volume of record volatile oil of different extraction time, as shown in table 2.

The receipts amount of table 2 different time volatile oil

Note: in table, data are volatile oil volume (ml)

As can be seen from the table, when being distilled to 6 hours, volatile oil extracts completely substantially, and the receipts amount of first three hour volatile oil is below 55%, therefore selects 4,5,6h is as the investigation of Valeriana officinalis var. latifolia vapor distillation time.

1.2 quadrature factor level designs

On the basis of trial test, choose extraction time (A), amount of water (B), soak time (C) three factors.Above-mentioned three factors, respectively get 3 levels, carry out L ₉(3 ⁴) orthogonal experiment, using the fingers such as dry cream yield, volatile oil volume, borneol acetate class content as investigating index, carry out the screening of extraction process condition.Orthogonal Experiment and Design factor level table, in Table 3.

Table 3 factor level table

The preparation of 1.3 samples

Take Valeriana officinalis var. latifolia 100g, totally 9 parts, moisture 66.8%.By orthogonal test condition, carry out vapor distillation, collect in volatile oil refrigerator and preserve, the centrifugal 10min of 3000r/min after water extraction liquid filters, standby, medicinal residues dry rear standby.

1.4 dry cream yields

By above-mentioned water extraction liquid water bath method after centrifugal, then move in 105 ℃ of baking ovens dry 3 hours, take out, put immediately in exsiccator and place 30 minutes, precise weighing, calculates dry cream yield, the results are shown in Table 4.

1.5 take chemical composition content as index optimization extraction process

1.5.1 the assay chromatographic condition of borneol acetate

Phenyl post; The methanol-water (70:30) of take is mobile phase; Detection wavelength is 203nm.

1.5.2 the preparation of reference substance solution

It is appropriate that precision takes borneol acetate reference substance, adds dehydrated alcohol and make the solution that contains borneol acetate 1.623mg in every 1ml dehydrated alcohol, obtains.

1.5.3 the preparation of need testing solution

Get the resulting volatile oil of Valeriana officinalis var. latifolia vapor distillation orthogonal experiment, put in separatory funnel, add ethyl acetate extraction three times, each 15ml, combined ethyl acetate liquid, low temperature is waved to about 2ml, adds methanol and is transferred to 50ml measuring bottle, as storing solution.Get storing solution 10ml, in 25ml measuring bottle, add methanol to scale, shake up, filter, get subsequent filtrate, obtain.

1.5.4 algoscopy

1.5.5 experimental result

Take dry cream yield, volatile oil volume, borneol acetate class content is evaluation index, carries out comprehensive grading, result of the test and variance analysis in Table 4, table 5.

Table 4 orthogonal experiment plan is taken into account result table

Y＝30×(X ₁/X ₁max)+20×(X ₂/X ₂max)+50×(X ₃/X ₃max)

Table 5 variance analysis

* show P < 0.05, * * shows P < 0.01

Intuitive analysis and variance analysis from comprehensive grading result, the factor size that affects vapor distillation is followed successively by A > B > C, it is distillation time > amount of water > soak time, and factor A has significance to the impact of Valeriana officinalis var. latifolia vapor distillation, therefore select A ₂, by intuitive analysis, determine that extracting optimised process is A ₂b ₃c ₃.

Determining of 1.6 Valeriana officinalis var. latifolia vapor distillation optimum extraction processes

By above analysis result, can reach a conclusion: the comprehensive grading to chemical constituents determination results such as dry cream yield, volatile oil volume, borneol acetate content carries out variance analysis, can find out, factor A compares with factor B, C, there is significant difference, that is to say that factor A has a significant impact Valeriana officinalis var. latifolia vapor distillation extraction effect, according to the result of intuitive analysis, select A ₂, optimum extraction process is A ₂b ₃c ₃.Because factor B, C do not have a significant impact extraction effect, the consideration of enhancing productivity from saving time, selects time good technique A ₂b ₁c ₁.Finally reach a conclusion: for vapor distillation, extract volatile oil in Valeriana officinalis var. latifolia, preferably extraction process is A ₂b ₁c ₁, be: add 4 times of water gagings, vapor distillation extracts 5h.For further confirming the reasonability of this technique, it is carried out to demonstration test, the results are shown in Table 6.

Table 6 demonstration test result

As shown in Table 6, the dry cream yield of this extraction process yield and orthogonal test, volatile oil volume, borneol acetate content are basically identical, but this selection process is saved amount of water, reduced extraction time, reduce production cost, this stable process conditions, reasonable have been described, be applicable to large production.Therefore, determine that extraction process is: add 4 times of water gagings, vapor distillation extracts 5h.

2 medicinal residues percolation technique orthogonal tests

2.1 quadrature factor level designs

Comparative example 2 techniques are: add 75% soak with ethanol 7 days.From the enhance productivity that saves time, comparative example 2 is soaked and changed percolation into, and percolation technological parameter is studied.According to preliminary experiment and documents and materials, the factor that affects percolation mainly contains: concentration of alcohol (A), alcohol adding amount (B), flow velocity (C) and soak time (D).Above-mentioned four factors, respectively get 3 levels, carry out L ₉(3 ⁴) orthogonal experiment, using dry cream yield, general flavone content as investigating index, carry out the screening of extraction process condition.Orthogonal test factor level table is in Table 7.

Table 7 factor level table

The preparation of 2.2 samples

Take Valeriana officinalis var. latifolia 100.0g, 10 parts.By definite process conditions, carry out vapor distillation, collect in volatile oil refrigerator and save backup, it is standby that water extraction liquid is condensed into thick paste.After medicinal residues dry, a by comparative example 2 techniques, extract, add 75% soak with ethanol 7 days, filter and obtain filtrate No. 0.All the other nine parts mix, and pulverize, then are divided into nine parts, by orthogonal test condition, carry out percolation, collect the liquid of filtering, and are settled to 250ml 1,4, No. 7, and all the other are settled to 500ml, obtain.

2.3 dry cream yields

Accurate each 25ml of above-mentioned filtrate that draws, puts in the evaporating dish of own constant weight, water bath method, then move in 105 ℃ of baking ovens dry 3 hours, and take out, to put immediately in exsiccator and place 30 minutes, precise weighing, calculates dry cream yield, the results are shown in Table 8.

2.4 take chemical composition content as index optimization extraction process

2.4.1 the preparation of sample reserve liquid

Take the medicinal residues 30g (moisture content 12% of Valeriana officinalis var. latifolia after vapor distillation, be equivalent to dry medical material 26.4g), according to Valeriana officinalis var. latifolia medicinal residues percolation orthogonal experiment condition, carry out percolation, sample 1,4,7 sample liquid standardize solution are to 250ml, other standardize solution, to 500ml, obtain.

2.4.2 the preparation of reference substance solution

Get 120 ℃ of control substance of Rutin that are dried to constant weight appropriate, add 70% appropriate amount of ethanol, make every 1ml70% ethanol containing the reference substance solution of anhydrous rutin 0.1989mg.

2.4.3 the preparation of standard curve

Precision measures reference substance solution 1ml, 2ml, 4ml, 6ml, 8ml, 10ml, puts respectively in 25ml measuring bottle, respectively adds 70% ethanol to 10ml, add 5% sodium nitrite solution 1ml, shake up, place 6min, add 10% aluminum nitrate solution 1ml, shake up, place 6min, add NaoH test solution 10ml, then add 70% ethanol to scale, shake up, place 15 minutes.Take corresponding solution as blank, according to ultraviolet visible spectrophotometry, at the wavelength place of 510nm, measure absorbance, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.

2.4.4 algoscopy

The sample reserve liquid 3ml that precision measures Valeriana officinalis var. latifolia medicinal residues percolation orthogonal experiment, puts in 25ml measuring bottle, adds 70% ethanol to scale, shakes up, as blank.Another precision measures 3ml, puts in 25ml measuring bottle, and the method under sighting target directrix curve preparation from " adding 70% ethanol to 10ml ", is measured absorbance in accordance with the law immediately, from standard curve, reads the amount of anhydrous rutin in percolate, and calculating, obtains.

2.4.5 experimental result

Take extractum yield, general flavone content is evaluation index, carries out comprehensive grading, result of the test and variance analysis in Table 8, table 9.

Table 8 orthogonal experiment plan is taken into account result table

Y＝30×(X ₁/X ₁max)+70×(X ₂/X ₂max)

Table 9 variance analysis

* show P < 0.05, * * shows P < 0.01

Intuitive analysis and variance analysis from comprehensive grading result, the factor size that affects percolation extraction effect is followed successively by D > A > B > C, it is soak time > concentration of alcohol > alcohol adding amount > flow velocity, and factor A, D have significance to the impact of Valeriana officinalis var. latifolia medicinal residues percolation effect, therefore select A ₂d ₃, by intuitive analysis, determine that percolation technique is A ₂b ₃c ₁d ₃.

Determining of 2.5 Valeriana officinalis var. latifolia medicinal residues optimum extraction processes

Comprehensive grading to chemical constituents determination results such as extractum yield, general flavone contents carries out variance analysis, can find out, factor A, D compare with factor B, C, there is significant difference, that is to say that factor A, D have a significant impact Valeriana officinalis var. latifolia medicinal residues percolation effect, according to the result of intuitive analysis, select A ₂d ₃, optimum extraction process is A ₂b ₃c ₁d ₃.Because factor B, C do not have a significant impact percolation effect, from the cost-saving consideration of enhancing productivity, select B ₁c ₃, inferior good extraction process is A ₂b ₁c ₃d ₃.Finally reach a conclusion: for the percolation experiment of Valeriana officinalis var. latifolia medicinal residues, preferably extraction process is A ₂b ₁c ₃d ₃, be: add after the soak with ethanol 2h of 10 times of amounts 75%, with 3Bv/h, carry out percolation.For further confirming the reasonability of this technique, it is carried out to demonstration test, the result of the test of demonstration test result and comparative example 2 soaking technologies is compared to analysis simultaneously.The results are shown in Table 10,11.

Table 10 demonstration test result

Table 11 soaking technology result of the test

By table 10,11 is known, and the dry cream yield of this extraction process yield and orthogonal test, general flavone content approach, and obviously high than comparative example 2 extraction process yields.This selection process has been saved ethanol consumption simultaneously, has reduced extraction time, has reduced production cost, and this stable process conditions, reasonable are described, is applicable to large production.Therefore, determine that extraction process is: add after the soak with ethanol 2h of 10 times of amounts 75%, with the speed of 3Bv/h, carry out percolation.

3 Flos Impatientiss, Lignum cinnamomi camphorae, Percolation Processes of Carthamus Tinctorius L orthogonal test

3.1 quadrature factor level designs

Comparative example 2 techniques are: add 60% soak with ethanol 7 days.From the enhance productivity that saves time, comparative example 2 is soaked and changed percolation into, and percolation technological parameter is studied.According to preliminary experiment and documents and materials, the factor that affects percolation mainly contains: concentration of alcohol (A), alcohol adding amount (B), flow velocity (C) and soak time.Above-mentioned four factors, respectively get 3 levels, carry out L ₉(3 ⁴) orthogonal experiment, using chemical index (extractum yield, S-A Hydroxysafflor yellow A, Quercetin, kaempferide content) as investigating index, carry out the screening of extraction process condition.Orthogonal test factor level table is in Table 12.

Table 12 factor level table

The preparation of 3.2 samples

According to prescription ratio, take Flos Impatientis 55g, Flos Carthami 30g, Lignum cinnamomi camphorae 15g, each 10 parts, wherein 1 part is extracted by comparative example 2 techniques, adds 60% soak with ethanol 7 days, filters and obtains filtrate No. 0.All the other 9 parts are carried out percolation by orthogonal test condition, collect the liquid of filtering, and add water to 1000ml 1,4, No. 7; 2, add water to 1500ml 5, No. 8; 3, add water to 2000ml 6, No. 9, obtain.

3.3 dry cream yields

Accurate each 25ml of above-mentioned filtrate that draws, puts in the evaporating dish of own constant weight, water bath method, then move in 105 ℃ of baking ovens dry 3 hours, and take out, to put immediately in exsiccator and place 30 minutes, precise weighing, calculates dry cream yield, the results are shown in Table 13.

3.4 take chemical composition content as index optimization extraction process

3.4.1 sample reserve liquid

By orthogonal experiment condition, carry out percolation, 1,4, No. 7 sample liquid standardize solution of sample are to 1000ml, and 2,5, No. 8 standardize solution are to 1500ml, and 3,6, No. 9 standardize solution, to 2000ml, obtain.

3.4.2HPLC method is measured S-A Hydroxysafflor yellow A content in percolate

3.4.2.1 chromatographic condition and system suitability

Take octadecylsilane chemically bonded silica as filler; Methanol-0.4% phosphoric acid solution (26:74) of take is mobile phase; Detection wavelength is 403nm.

3.4.2.2 the preparation of reference substance solution

It is appropriate that precision takes S-A Hydroxysafflor yellow A reference substance, adds 25% methanol and make the solution that contains 0.1339mg in every 1ml, obtains.

3.4.2.3 the preparation of need testing solution

Precision measures percolate 5ml, puts in 25ml measuring bottle, adds 25% methanol to scale, shakes up, and filters, and gets subsequent filtrate, obtains.

3.4.2.4 assay

Precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and injection liquid chromatography, measures, and calculate S-A Hydroxysafflor yellow A, the results are shown in Table 13.

3.4.3HPLC method is measured the content of kaempferide and Quercetin in percolate simultaneously

3.4.3.1 chromatographic condition and system suitability

Take octadecylsilane chemically bonded silica as filler; Methanol-0.4% phosphoric acid solution (40:60) of take is mobile phase; Detection wavelength is 360nm.

3.4.3.2 the preparation of reference substance solution

Precision takes kaempferide, Quercetin reference substance is appropriate, adds methanol and makes the mixed solution that contains respectively kaempferide, Quercetin 104.4 μ g, 8.9 μ g in every 1ml methanol, obtains.

3.4.3.3 the preparation of need testing solution

Precision measures percolate 10ml, puts in tool plug conical flask, and water-bath volatilizes, take off, add methanol-25% hydrochloric acid (4:1) mixed liquor 25ml, reflux 40min in water-bath, take off, let cool, be transferred in 50ml measuring bottle, with methanol gradation, wash conical flask, washing liquid is incorporated in measuring bottle, adds methanol to scale, shake up, filter, get subsequent filtrate, obtain.

3.4.3.4 assay

Precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and injection liquid chromatography, measures, and calculates the content of kaempferide, Quercetin, the results are shown in Table 13.

3.4.4 experimental result

Extractum yield, S-A Hydroxysafflor yellow A, Quercetin, kaempferide content are carried out to comprehensive grading, result of the test and variance analysis in Table 13, table 14.

Table 13 orthogonal experiment plan is taken into account result table

Y＝20×(X ₁/X ₁max)+20×(X ₂/X ₂max)+30×(X ₃/X ₃max)+30×(X ₄/X ₄max)

Table 14 variance analysis

* show P < 0.05, * * shows P < 0.01

Intuitive analysis and variance analysis from comprehensive grading result, the factor size that affects percolation extraction effect is followed successively by A > B > C > D, it is concentration of alcohol > alcohol adding amount > flow velocity > soak time, and factor A, B, C have significance to the impact of the percolation effect of Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae, therefore select A ₂b ₂c ₁, by intuitive analysis, determine that percolation technique is A ₂b ₂c ₁d ₃.

Determining of 3.5 Flos Impatientiss, Lignum cinnamomi camphorae, three medical material optimum extraction processes of Flos Carthami

Comprehensive grading to chemical constituents determination results such as dry cream yield, S-A Hydroxysafflor yellow A, Quercetin, kaempferide content carries out variance analysis, can find out, factor A, B, C compare with factor D, there is significant difference, that is to say that factor A, B, C have a significant impact three medical material percolation effects such as Flos Impatientiss, according to the result of intuitive analysis, select A ₂b ₂c ₁, optimum extraction process is A ₂b ₂c ₁d ₃.Because factor D does not have a significant impact percolation extraction effect, from the consideration of enhancing productivity, select D ₁, inferior good extraction process is A ₂b ₂c ₁d ₁.Finally reach a conclusion: for the percolation experiment of Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae, preferably extraction process is A ₂b ₂c ₁d ₁, be: add after the soak with ethanol 6h of 15 times of amounts 60%, with 1Bv/h, carry out percolation.For further confirming the reasonability of this technique, it is carried out to demonstration test, the result of the test of demonstration test result and comparative example 2 soaking technologies is compared to analysis simultaneously.The results are shown in Table 15,16.

Table 15 demonstration test result

Table 16 soaking technology result of the test

By table 15,16 is known, and the dry cream yield of this extraction process yield and orthogonal test, S-A Hydroxysafflor yellow A, Quercetin, kaempferide content approach, and obviously high than comparative example 2 extraction process yields.This selection process has reduced extraction time simultaneously, has reduced production cost, and this stable process conditions, reasonable are described, is applicable to large production.Therefore, determine that optimum extraction process is: add after the soak with ethanol 6h of 15 times of amounts 60%, with the speed of 1Bv/h, carry out percolation.

Compound recipe toothache sticking film forming technical study in test example 3 embodiment

Before preparation extraction process is studied, determine that optimum extraction process is: Valeriana officinalis var. latifolia adds 4 times of water gagings, vapor distillation extracts 5h; After medicinal residues dry, add after 10 times of amount 75% soak with ethanol 2h, with the speed percolation of 3Bv/h; Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add 60% soak with ethanol 6h, 15 times of amount 1Bv/h percolation.By this technique, prepare medicinal liquid, add appropriate substrate, after abundant swelling, stir, paving, to the glass plate of certain area, is dried and get final product.This membrane can be directly released into painful area by medicine, plays the effect of quickly alleviating pain, and there is transportation, carry, feature easy to use.

The selection of 1 substrate

The key of preparing membranous patch is to select substrate (adhesion material), desirable adhesion material should be nontoxic, without absorbing, stable performance, have good biocompatibility, adhesion is suitable, effect rapidly, easily mixes with medicine but chemical reaction does not occur and do not affect its release, cheap and easy to get.According to documents and materials, conventional have chitosan, cellulosic derivant, a polyacrylic.Chitosan has good bioadhesive, biocompatibility, safety, and can strengthen the absorption of mucosa to medicine, but that its pharmacokinetic property is affected by deacetylation degree is larger; Hypromellose is most widely used cellulose derivative.Have good film property, its formed film is transparent, tough and tensile, is difficult for adhesion during production, can greatly improve medicine stability.Have bibliographical information, take 50% ethanol as solvent, the filming performance of the hydroxypropyl emthylcellulose liquid containing 10% is good; Carbomer is the high molecular polymer being cross-linked to form by acrylic acid and poly-alkyl sucrose or poly-alkyl tetramethylolmethane, main as thickening agent, suspending agent and binding agent in medicament.This product home made article divides basic, normal, high three viscosity specifications, is equivalent to respectively the external products such as CP-941, CP-934, CP-940, and wherein CP-934 is known as in can doing to take substrate now.Bibliographical information once used carbomer940 (P represents this product pharmacopeia level) to make a kind of oral preparation substrate, its oral administration is done to the experiments such as acute toxicity, allergy and mucous membrane irritation, result LD50 value is consistent with bibliographical information, illustrates that carbomer is as the substrate safety of oral formulations.According to the splendid film property of HPMC and the desirable adhesiveness of CP, select HPMC and HPMC and CP-934 to mix the substrate that is used as this membrane.

1.1 test method

According to documents and materials, after preliminary experiment, the HPMC of selection 2% and 1% CP.2% HPMC is added separately into the liquid, after abundant swelling, stir, be taped against on glass plate and dry, make medicine film No. 1.The CP of 2% HPMC and 1% is added into the liquid simultaneously, after abundant swelling, stir, be taped against on glass plate and dry, make medicine film No. 2.

1.2 result of the test

No. 1 medicine film energy swelling is even, the easy film forming of good fluidity, and the membrane forming is even.No. 2 medicine films can not complete swelling, adularescent flocculent deposit, and the membrane drying is very inhomogeneous.Therefore select HPMC as this product substrate.

The proportioning of 2 substrate and medicine is selected

For further improving outward appearance, the quality of membrane, increase the plasticity of membrane, add a small amount of glycerol].And the consumption of HPMC is further investigated.

2.1 test method

Adopt orthogonal design, take HPMC, glycerol is two factors, and concentration is 3 levels, by orthogonal table L ₉(3 ⁴) operation, take adhesion time as main screening foundation, carry out the screening of formulation and technology.Orthogonal test factor level table is in Table 17.

Table 17 factor level table

2.2 adhesion times are measured

Reference literature data, the fresh pig large intestine of choosing decontamination, opposed flattened is some, cuts open and shakeouts, and pastes respectively each 6 of each tested number prescription films on mucosa, in the water bath with thermostatic control that to hang over 37 ℃, pH be 6.2, records the adhesion time of each membrane.The results are shown in Table 18,19.

Table 18 orthogonal experiment plan is taken into account result table

Table 19 variance analysis

Result of the test is carried out to intuitive analysis and variance analysis discovery, and the factor that affects adhesion time is followed successively by B>A, i.e. glycerol concentration >HPMC concentration, and glycerol concentration has significance to the impact of adhesion time, therefore select B ₁.The result of film forming situation shows that film forming situation ideal when HPMC concentration is 1.5% considers and thinks that prescription 4 meets designing requirement.

3 prepare membranous patch

3.1 experimental technique

According to prescription ratio, take every medical material in prescription, Valeriana officinalis var. latifolia adds 4 times of water gagings, and vapor distillation extracts 5h, collects in volatile oil refrigerator and saves backup, and it is standby that water extraction liquid is condensed into thick paste.After medicinal residues dry, add after 10 times of amount 75% soak with ethanol 2h, with the speed percolation of 3Bv/h, the concentrated thick paste of percolate and water extraction liquid mixes, and after standing 24h, filters, and solution decompression reclaims ethanol and obtains extractum; Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae add 60% soak with ethanol 6h, 15 times of amount 1Bv/h percolation, and filter liquid and extractum mix, and filter, and add volatile oil, and being adjusted to ormal weight and making amount of alcohol is 45%.Add 1.5%HPMC and 1% glycerol, after abundant swelling, stir evenly, paving is to scribbling on the glass plate of a small amount of liquid paraffin respectively, and room temperature is dried and be get final product.

Result

Preparation process, from analyzing character and the enhance productivity that saves time of the relevant chemical composition of each medical material, is divided into three parts.Valeriana officinalis var. latifolia vapor distillation orthogonal test, Valeriana officinalis var. latifolia medicinal residues percolation orthogonal test and Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae percolation orthogonal test.Orthogonal Experiment and Design all adopts L ₉(3 ⁴) orthogonal table, the dynamic change of chemical index, as investigating index, is carried out the screening of extraction process condition.

Determined the extraction process condition of Valeriana officinalis var. latifolia vapor distillation, Valeriana officinalis var. latifolia medicinal residues, Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae medical material.Valeriana officinalis var. latifolia optimum extraction process is: add 4 times of water gagings, vapor distillation extracts 5h; Valeriana officinalis var. latifolia medicinal residues optimum extraction process is: add after the soak with ethanol 2h of 10 times of amounts 75%, with the speed of 3Bv/h, carry out percolation; Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae medical material optimum extraction process are: add after the soak with ethanol 6h of 15 times of amounts 60%, with the speed of 1Bv/h, carry out percolation.

The prescription substrate composition of membranous patch and preparations shaping technique have been carried out to screening test, preparations shaping process conditions have been determined: by definite technique, prepare medicinal liquid, add 1.5%HPMC and 1% glycerol, after abundant swelling, stir evenly, paving is to scribbling on the glass plate of a small amount of liquid paraffin respectively, and room temperature is dried and be get final product.

Test example 4 pharmacodynamic experiments

Experiment material

1. laboratory animal

Kunming mouse: male and female half and half, body weight (20 ± 2) g, by Changsha Kaifu District Dong Chuan Animal Science service department (quality certification number: doctor moves word 090012) and the Guiyang Medical College animal center (quality certification number: SCXK Guizhou Province 2002-0001) provide.Normal diet is raised, clean level, 18～24 ℃ of laboratory temperatures, laboratory relative humidity: 65%～75%.

Wistar rat: male and female half and half, body weight (200 ± 20) g, is provided by Guiyang Medical College animal center.The quality certification number: SCXK (Guizhou Province) 2002-0001.Normal diet is raised, clean level, 18～25 ℃ of laboratory temperatures, laboratory relative humidity: 65%～75%.

SD rat: male and female half and half, body weight (200 ± 20) g, by Chongqing, Teng Xin Bioisystech Co., Ltd provides, the quality certification number: SCXK (Chongqing) 2007-0003.Normal diet is raised, clean level, 18～24 ℃ of laboratory temperatures, laboratory relative humidity: 65%～75%.

2. main medicine and reagent

Good Huangpu pharmaceutical Co. Ltd that transports by sea on spasmolysis and analgesia tincture provides, lot number: 20100903

Sodium sulfide China Chengdu Kingsoft chemical reagent company limited, lot number: 20080106

Dong great chemical plant, glacial acetic acid Tianjin, lot number: 20090902

Chuanjiang River, dimethylbenzene Chongqing chemical reagent factory, lot number: 901201

Chloral hydrate Chemical Reagent Co., Ltd., Sinopharm Group, lot number: 080314

Solution on Chemical Reagents in Shanghai company of azovan blue Chinese Medicine group import subpackage, F20020913

Methanamide, Beijing Chemical Plant, 20080814

Acetone Chemical Reagent Co., Ltd., Sinopharm Group, lot number: F 20090802

Sodium chloride injection Guizhou Kelun Pharmaceutical Co., Ltd., lot number: B090527

Ether China Chengdu Kingsoft chemical reagent company limited, lot number: 20090106

Technological experiment medical material used is provided by Guizhou Tongjitang Pharmaceutical Co., Ltd, and medical material kind is identified through Chen Deyuan researcher.Four Chinese medicine material is tested by its statutory standards respectively, all complies with relevant regulations.

Compound recipe toothache pad pasting is made by the formula of embodiment 1 and preparation method.

3. key instrument

Hot plate dolorimeter Kunming Electronic Instruments Plant

Ultraviolet-visible spectrophotometer UV-1750 Shimadzu Instrument Ltd.

Centrifuge TGL-16G Anting Scientific Instrument Factory, Shanghai

XS205DU-analytical balance Mei Teletuo benefit (METTLER TOLEDO)

TC-10K type electronic balance Changshu Shuan Jie test instrunment factory

1ml syringe Wuxi De Fan Instrument Ltd.

One, the embodiment of the present invention 1 compound recipe toothache pad pasting pharmacodynamics checking

1. hot-plate instrument is brought out to the impact of mice vola pain model

1.1 animal groupings

By 50 of female mices, abdominal part depilation, is divided into 5 groups at random, 10 every group, is respectively model group, high (the about 0.11g crude drug/㎝ of compound recipe toothache pad pasting ²), in (about 0.05g crude drug/㎝ ²), low (about 0.03g crude drug/㎝ ²) dosage group, positive drug control group (spasmolysis and analgesia tincture, 0.24g/ml).

1.2 experimental technique

Mice adaptability is fed two days later, and model group is smeared administration with normal saline abdominal part; Spasmolysis and analgesia tincture group is smeared administration by 10ml/kg in abdominal part; Membrane high dose, middle dosage, low dose group are all by 1 ㎝ ²/ be only affixed on depilation place skin, stick 2h every day, continuous use 2 days.Before medication, mice is placed on hot plate box, controls temperature in (55.0 ± 0.5) ℃, record after mice is put into hot plate box and lick metapedes required time to it, within the 2nd day, survey again 1 time, using the meansigma methods of the continuous two days Basic Pain Threshold value before as administration.After medication in the 3rd day 30,60,90,120min measures respectively pain threshold by above-mentioned Basic Pain Threshold pH-value determination pH method.

1.3 detect index

The time that occurs first foot phenomenon after adding after the administration of observed and recorded mice when 30min, 60min, 90min, 120min in hot-plate instrument.

2. glacial acetic acid is caused to the impact of mouse web portion writhing pain reaction

2.1 animal groupings

By 50 of male and female half and half mices, abdominal part depilation, is divided into 5 groups at random, 10 every group, is respectively model group, high (the about 0.11g crude drug/㎝ of compound recipe toothache pad pasting ²), in (about 0.05g crude drug/㎝ ²), low (about 0.03g crude drug/㎝ ²) dosage group, positive drug control group (spasmolysis and analgesia tincture, 0.24g/ml).

2.2 experimental technique ^]

Mice adaptability is fed two days later, and model group is smeared administration with normal saline abdominal part; Spasmolysis and analgesia tincture group is smeared administration by 10ml/kg in abdominal part; Membrane high dose, middle dosage, low dose group are all by 1 ㎝ ²/ be only affixed on depilation place skin, stick 2h every day, continuous use 2 days.After the 3rd day medication 1h, the glacial acetic acid 0.2ml of every mouse peritoneal injection 0.8%, records respectively the writhing number of times of mice in 10min and 20min and respectively organizes mouse writhing suppression ratio.

Writhing suppression ratio: (model group writhing number of times-administration group writhing number of times)/model group writhing number of times * 100%

2.3 detect index

Observed and recorded is respectively organized the writhing number of times of mice appearance within the time of writhing response (incubation period) and 10min, 20min first appear after glacial acetic acid in injection.

3. xylol causes the impact of mice auricle swelling and capillary permeability variation model

3.1 animal groupings

By 50 of male and female half and half mices, abdominal part depilation, is divided into 5 groups at random, 10 every group, is respectively model group, high (the about 0.11g crude drug/㎝ of compound recipe toothache pad pasting film ²), in (about 0.05g crude drug/㎝ ²), low (about 0.03g crude drug/㎝ ²) dosage group, positive drug control group (spasmolysis and analgesia tincture, 0.24g/ml)

3.2 experimental technique

Mice adaptability is fed two days later, and model group is smeared administration with normal saline abdominal part; Spasmolysis and analgesia tincture group is smeared administration by 10ml/kg in abdominal part; Membrane high dose, middle dosage, low dose group are all by 1 ㎝ ²/ be only affixed on depilation place skin, stick 2h every day, continuous use 2 days.In administration in the 3rd day simultaneously in mouse tail vein injection 1% azovan blue 0.2ml.After last administration 1h, each is organized mouse right ear two sides and drips 10 μ l caused by dimethylbenzene xylene inflammation, puts to death mice after 30min, with 9mm card punch, in the same position of left and right ear, lays circular auricle.Weigh left and right auricle weight record.Auris dextra sheet is shredded and puts into the EP pipe that fills 4ml35% acetone soln.After 38 ℃ of immersion 48h of water-bath, take out leachate, with the centrifugal 10min of 1500r/min.Get supernatant and in 590nm wavelength place, survey absorbance OD value.

Swelling=auris dextra sheet weight-left auricle weight

Ear swelling suppression ratio: (model group swelling average-administration group swelling average)/model group swelling average * 100%

3.3 detect index

Mice left and right auricle weight, auris dextra sheet leachate OD value.

4. the impact on rat hindleg acute soft tissue injury

4.1 animal groupings

50 of SD rats are divided into 5 groups at random, 10 every group, are respectively model group, high (the about 0.11g crude drug/㎝ of compound recipe toothache pad pasting film ²), in (about 0.05g crude drug/㎝ ²), low (about 0.03g crude drug/㎝ ²) (spasmolysis and analgesia tincture 0.24g/ml), is respectively organized equal male and female half and half above for dosage group, positive drug control group.

4.2 experimental technique

24h before experiment,, loses hair or feathers after 24h to the left back huckle depilation of rat with sodium sulfide solution, etherization rat, and with beating device, the left back leg outer side of rat is caused to acute soft tissue injury, depending on subcutaneous hemorrhage, be that within several minutes, having touched obvious tumefaction sense is modeling success.Model group is smeared administration with normal saline in redness place; Spasmolysis and analgesia tincture group is smeared administration by 10ml/kg in redness place, and it is degree that application area be take over impaired area 1/2; Membrane high dose, middle dosage, low dose group are all by 2 ㎝ ²/ being only affixed on impaired place, wiping is once seen in medication every day afterwards, successive administration 6 days.Rat is carried out to active state scoring, outward appearance scoring.

4.3 detect index

5. Data Processing in Experiment

Experimental data all with represent, adopt SPSS13.0 statistical software to analyze.Adopt one factor analysis of variance (One-Way ANOVA) to carry out significance,statistical analysis.Group difference significance adopts LSD (Least-significant difference) method in one factor analysis of variance to carry out multiple comparisons check, and P ﹤ 0.05 is for there being significant difference.

6. the impact of compound recipe toothache pad pasting on mice vola pain

Spasmolysis and analgesia tincture group demonstrates stronger analgesic activity after administration 1h, but effect declines to some extent when 2h.The high, medium and low dosage group of membrane acts on obviously after administration 1h, and to after administration during 2h effect continue strongly, when administration 1h, analgesic activity is weaker than spasmolysis and analgesia tincture group, after administration 1.5h, analgesic activity is better than spasmolysis and analgesia tincture group.The high, medium and low dosage group of membrane strengthens along with dosage increases analgesic activity, and high, medium and low dosage all has stronger analgesic activity.In Table 20.

Table 20 is respectively organized the comparison of hot plate method in mice pain experiment pain threshold

With comparison before administration, ^*p<0.05, ^*p<0.01, ^{* *}p<0.001; With the comparison of feminine gender group, ^▲p<0.05,

^{▲ ▲}p<0.01, ^{▲ ▲ ▲}p<0.001; With the comparison of spasmolysis and analgesia tincture group, ^#p<0.05, ^##p<0.01; N=10

7. the effect of compound recipe toothache pad pasting to the experiment of mouse writhing pain

High, the middle dosage group of spasmolysis and analgesia tincture group and membrane all shows stronger analgesic activity.Three dosage of membrane and its analgesic activity are proportionate, membrane high dose group analgesia intensity and spasmolysis and analgesia tincture category seemingly, in membrane dosage group ease pain intensity compared with spasmolysis and analgesia tincture a little less than, membrane low dose group is without obvious analgesic activity.Preclinical relatively in, spasmolysis and analgesia tincture and negative group have very significant difference, and that other respectively organize difference is not obvious.In Table 21.

Table 21 is respectively organized the comparison of mouse writhing number of times

With model group comparison, ^*p<0.05, ^*p<0.01, ^{* *}p<0.001; With the comparison of spasmolysis and analgesia tincture group, ^▲p<0.05, ^{▲ ▲}p<0.01, ^{▲ ▲} ^▲p<0.001; N=10

8. the impact of compound recipe toothache pad pasting xylol induced mice auricle edema

The mice auricle swelling that each medication group dimethylbenzene causes is all obviously suppressed.With model group comparison, P<0.05, P<0.001.Wherein the effect of membrane high dose group is the strongest, and inhibitory rate of intumesce is 56.52%, higher than spasmolysis and analgesia tincture group (45.65%); In membrane, dosage group suppression ratio is 36.96%; Membrane low dose group suppression ratio is 13.04%.Show that compound recipe toothache pad pasting increases antiinflammatory action with dosage and strengthens.In Table 22.

Table 22 is respectively organized the comparison of auricle edema due to mice dimethylbenzene

9. the impact of compound recipe toothache pad pasting xylol induced mice inflammation auricle capillary permeability

The absorbance comparison that each medication group and model group Mice Auricle Evans blue dyestuff ooze out, difference all has very significantly meaning (P<0.001).Capillary permeability that due to each medication group xylol, inflammation causes increases and dyestuff oozes out and all has very strong inhibitory action, and membrane senior middle school low dose group is similar to positive control medicine spasmolysis and analgesia tincture activity, all has stronger antiinflammatory action.In Table 23.

Table 23 is respectively organized the absorbance comparison that Mice Auricle dyestuff oozes out

With model group comparison, ^*p<0.05, ^*p<0.01, ^{* *}p<0.001; N=10

10. the impact of compound recipe toothache pad pasting on rat acute soft tissue injury

After modeling success, each is organized rat and hinders limb and all occur obvious tumefaction, skin cyan ecchymosis and subcutaneous extvavasated blood, hemorrhage or edema, and be limping state, after modeling the same day the most serious, the condition of the injury alleviates gradually subsequently, kinestate is normal gradually.After the administration of compound recipe toothache pad pasting, 3d, 4d, 5d, 6d are to the effect of being significantly improved of rat hindlimb active state, and after administration, 4d, 5d, 6d rat soft tissue injury local appearance are significantly improved.Appraisal result in Table 24, table 25

The impact of table 24 compound recipe toothache pad pasting on rat soft tissue injury moving obstacle

With model group comparison, ^*p<0.05, ^*p<0.01, ^{* *}p<0.001

The impact of table 25 compound recipe toothache pad pasting on rat soft tissue injury local appearance

With model group comparison, ^*p<0.05, ^*p<0.01, ^{* *}p<0.001

Two, the embodiment of the present invention 4 compound recipe Toothache tinctures pharmacodynamic experiments

1. hot-plate instrument is brought out to the impact of mice vola pain model

1.1 animal groupings

50 of female mices, are divided into 5 groups at random, 10 every group, be respectively matched group, compound recipe Toothache tinctures high (0.08g/kg), in (0.04g/kg), low (0.02g/kg) dosage group, spasmolysis and analgesia tincture group (0.1g/kg).

1.2 experimental technique

70 of Kunming kind mices, female, body weight 18-22g.The qualified mice of screening before test, is chosen at and is preheated to Basic Pain Threshold value that the constant temperature hot-plate instrument of 55 ± 0.5 ℃ the records mice between 5-30s.Replication twice, get twice threshold of pain meansigma methods as administration before Basic Pain Threshold value.By qualified mice random packet, give every mice 0.2ml and smear two metapedes.After administration 15,30,60,90,120min measures respectively mice pain threshold, surpass 60s and still without reflector, take out and calculate with 60s.

1.3 detect index

The time that occurs first foot phenomenon after adding after the administration of observed and recorded mice when 15min, 30min, 60min, 90min, 120min in hot-plate instrument.

2.1 animal groupings

50 of male and female half and half mices, are divided into 5 groups at random, 10 every group, be respectively matched group, compound recipe Toothache tinctures high (0.08g/kg), in (0.04g/kg), low (0.02g/kg) dosage group, spasmolysis and analgesia tincture group (0.1g/kg).

2.2 experimental technique

Mice adaptability is fed two days later, and after the glacial acetic acid solution of 10ml/kg lumbar injection 0.6%, abdominal part is smeared and respectively organized medicine immediately, records respectively the writhing number of times of mice in 10min and 20min and respectively organizes mouse writhing suppression ratio.Writhing suppression ratio: (model group writhing number of times-administration group writhing number of times)/model group writhing number of times * 100%

2.3 detect index

3.1 animal groupings

3.2 experimental technique

Mice adaptability is fed two days later, and matched group is smeared administration with normal saline; All the other each groups are smeared after administration 5min, dimethylbenzene is applied to two sides, mouse right ear front and back with 20 μ l, make its auricle two sides cause inflammation, tail vein injection 1% Evans blue 10ml/kg after administration 20min, left auricle, for contrast, is cut ears after 20min, with diameter 6mm card punch, lay two auricles, in electronic balance, take respectively two ear weight, with the difference of two ears, represent swelling index.Auris dextra sheet is soaked in Methanamide, and 38 ℃ of water-bath 48h ooze out dyestuff, in the centrifugal 10min of 1500rpm, get supernatant in 722 spectrophotometer 590nm place's colorimetric determination absorbance A values.

Swelling=auris dextra sheet weight-left auricle weight

3.3 detect index

Mice left and right auricle weight, absorbance.

4. the impact on rat hindleg acute soft tissue injury

4.1 animal groupings

Half and half 50 of wistar rat male and female are divided into 5 groups at random, every group 10, be respectively matched group, compound recipe Toothache tinctures high (0.08g/kg), in (0.04g/kg), low (0.02g/kg) dosage group, spasmolysis and analgesia tincture group (0.1g/kg).

4.2 experimental technique

24h before experiment,, loses hair or feathers after 24h to the left back huckle depilation of rat with sodium sulfide solution, chloral hydrate anesthesia rat, and with beating device, the left back leg outer side of rat is caused to acute soft tissue injury, depending on subcutaneous hemorrhage, be that within several minutes, having touched obvious tumefaction sense is modeling success.The successful wist ar of modeling rat respectively organized to smear every day be administered once, application area take surpass impaired area more than 1/2 for spending, common administration 9 days.Observe the mobility after Damage of Rats, local appearance, and give score every day relatively.In 6d, 9d, put to death each 5 of rats, get damage location 4% formaldehyde and fix, carry out histopathologic examination.Micro-Microscopic observation is respectively organized muscular tissue, according to striated muscle tissue pathological observation standard recording.

4.3 detect index

5. hot plate is caused to the impact of mice pain threshold

Spasmolysis and analgesia tincture and the high, medium and low dosage group of compound recipe Toothache tinctures mice after smearing administration 15mi n to 90min pain threshold all apparently higher than matched group, there is statistical significance (P<0.01, P<0.05).Prompting compound recipe Toothache tinctures has certain analgesic activity.In Table 26.

The impact of table 26 compound recipe Toothache tinctures on mice hot plate induced pain

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

6. glacial acetic acid is caused to the impact of mouse writhing reaction

The incubation period that compound recipe Toothache tinctures 0.08g/kg, 0.04g/kg, tri-dosage group energy significant prolongation writhings of 0.02g/kg occur, 0.08g/kg dosage group significantly reduces the writhing number of times of mice.Prompting compound recipe Toothache tinctures has certain inhibitory action to chemical stimulation induced pain.In Table 27.

The impact of table 27 compound recipe Toothache tinctures on mouse writhing pain

Note: with matched group comparison, * * P<0.01*P<0.05

7. the impact of xylol induced mice auricle edema

Experimental result prompting, compound recipe Toothache tinctures 0.08g/kg, 0.04g/kg dosage group xylol induced mice auricle edema have significant inhibitory action, and its suppression ratio is respectively 30.6%, 28.6%; Prompting compound recipe Toothache tinctures has certain inhibitory action to acute inflammation.In Table 28.

The impact of table 28 compound recipe Toothache tinctures xylol induced mice auricle edema

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

8. impact capillary permeability due to auricle inflammation being changed

Result is as follows.Compound recipe Toothache tinctures 0.08g/kg, the increase of 0.04g/kg dosage group xylol induced mice auricle capillary permeability have obvious inhibitory action (P<0.05).In Table 29.

The impact that table 29 compound recipe Toothache tinctures increases capillary permeability due to inflammation auricle

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

9. the impact on rat soft tissue injury

After modeling success, each is organized rat and hinders limb and all occur obvious tumefaction, skin cyan ecchymosis and subcutaneous extvavasated blood, hemorrhage or edema, and be limping state, after modeling the same day the most serious, the condition of the injury alleviates gradually subsequently, kinestate is normal gradually.After the administration of compound recipe Toothache tinctures, 2d, 3d, 5d, 6d, 7d, 8d, 9d are significantly improved rat hindlimb moving obstacle and outward appearance, the results are shown in Table 30-33.

Table 30 compound recipe Toothache tinctures is on the impact of the moving obstacle of rat soft tissue injury (1～6d)

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

Table 31 compound recipe Toothache tinctures is on the impact of the moving obstacle of rat soft tissue injury (7～9d)

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

Table 32 compound recipe Toothache tinctures is on the impact of the local appearance of rat soft tissue injury (1～6d)

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

Table 33 compound recipe Toothache tinctures is on the impact of the local appearance of rat soft tissue injury (7～9d)

Note: with matched group comparison, ^*p<0.01 ^*p<0.05

The pharmacodynamic experiment of test example 5 screening extraction processes

4 taste medical materials in the compound recipe Toothache tinctures prescription of the disclosed Guizhou Tongjitang Pharmaceutical Co., Ltd of the < < of National Drug Administration national drug standards > > [WS-10589-(ZD-0589)-2002], except Valeriana officinalis var. latifolia extracts by former technique, Valeriana officinalis var. latifolia medicinal residues and Flos Impatientis etc. change percolation into by former immersion, adopt Orthogonal Experiment and Design to Valeriana officinalis var. latifolia vapor distillation, percolation technique is carried out preferably, with mice plantalgia threshold value, writhing number of times, ear swelling test result refers to as investigating index, optimize rational extraction process route.Factor level table sees the following form:

Valeriana officinalis var. latifolia steam distillation technique orthogonal test

Medicinal residues percolation technique orthogonal test

Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae percolation technique orthogonal test

1. adopt hot-plate instrument to bring out mice vola pain model

1.1 mice vola pain experiments

1.1.1 animal grouping

110 of mices are divided into 11 groups at random, 10 every group, be respectively model group, spasmolysis and analgesia tincture group (about 0.24g crude drug/ml), nine kinds of middle product (about 0.4g crude drug/kg) totally nine groups of experiments, above each group is female mice.

1.1.2 experimental technique

Mice adaptability is fed after 24h, and hot-plate instrument is preheated to 55 ℃, and every mice is placed in hot-plate instrument and tests the front pain threshold of its administration record, and happiness leaper rejects, and take between 10-50s is qualified mice.Administration is smeared with 1ml syringe in metapedes bottom, experiment every mice left and right on the same day, and each medication group dosage is 10ml/kg, and model group is given equivalent normal saline.Pain threshold after observed and recorded administration 30,60,90, after 120min, and calculate the threshold of pain and improve percentage rate.

1.1.3 detect index

The 1.2 pain experimental result intuitive analysis of hot plate method vola and variance analyses.

2. adopt glacial acetic acid to cause mouse writhing pain model

2.1 mouse writhing pain experiments

2.1.1 animal grouping

110 of mices are divided into 11 groups at random, 10 every group, be respectively product (about 0.4g crude drug/kg) totally nine groups in the middle of model group, spasmolysis and analgesia tincture group (about 0.24g crude drug/ml), nine kinds of experiments, respectively organize equal male and female half and half above.

2.1.2 experimental technique

Mice adaptability is fed after 24h, and glacial acetic acid successively decreases and injects standby Mus by 0.8%-0.6% Concentraton gradient, and the mice of take produces writhing but be not dead is glacial acetic acid concentration standard, is identified for the glacial acetic acid concentration of modeling.Experiment gave every whole abdominal part of mice and smeared corresponding medicine the same day, and dosage is 10ml/kg, gives normal saline for negative group, after administration 20min, to mice lower-left lumbar injection 0.2ml glacial acetic acid solution, and starts timing observational record.

2.1.3 detect index

2.2 intuitive analysis of writhing experimental result and variance analyses.

3. adopt and prepare inflammatory animal model with dimethylbenzene and Evans blue

3.1 mice auricle swellings and capillary permeability experiment

3.1.1 animal grouping

3.1.2 experimental technique

Mice adaptability is fed after 24h, by the dosage of 10min/kg, each group mice left and right ear is coated with to corresponding medicine, negative group is coated with equivalent normal saline, and in mouse tail vein injection 1% azovan blue 0.2ml, after administration 20min, with liquid-transfering gun, in mouse right ear tow sides, respectively drip 10 μ l dimethylbenzene to scorching.After administration 1h, the dislocation of mice cervical vertebra is put to death, and along auricle base portion, cuts ears, with 9mm card punch, in the same position of left and right ear, lays circular auricle, on electronic balance, claim same mice left and right auricle weight record, auris dextra sheet is shredded and puts into the EP pipe that fills 4ml35% acetone soln.After 38 ℃ of immersion 48h of water-bath, take out leachate, with the centrifugal 10min of 1500r/min.Get supernatant and in 590nm wavelength place, survey absorbance A, the suppression ratio that experiment with computing group Mice Auricle dyestuff oozes out.

3.1.3 detect index

3.1.3.1 observation ear swelling degree, the ear swelling suppression ratio of the swelling of animal model auricle inflammation

3.1.3.2 leachate absorbance.

3.2 inflammation test visual result is analyzed and variance analysis.

The pharmacodynamic experiment result of screening extraction process

1. Valeriana officinalis var. latifolia vapor distillation craft screening

The effect of product to the experiment of mice vola pain in the middle of 1.1 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice vola pain experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that extends mice pain threshold aspect, but in the middle of most of product aspect extending mice pain threshold with before matched group and administration, compared significant difference (P<0.05 or 0.01); The above results explanation, different Extraction techniques have a direct impact the relevant drug action of mice vola pain.(in Table 34)

Table 34 mice hot plate vola pain experimental result

Before administration group and administration, compare, ^*represent significant difference (P<0.05), ^*represent difference highly significant (P<0.01).. administration group and negative control group comparison, ^▲represent significant difference (P<0.05), ^{▲ ▲}represent difference highly significant (P<0.01).

1.2 vola pain experimental result intuitive analysis and variance analyses

The 90min pain threshold after mice vola pain experiment administration of take carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 35, table 36)

Table 35 orthogonal experiment plan is taken into account result

Table 36 variance analysis

* show P<0.05

The 90min pain threshold after mice vola pain experiment administration of take is evaluation index, by carrying out intuitive analysis and variance analysis, learn, the factor size that affects extraction effect is followed successively by A>B>C, be distillation time > amount of water > soak time, by intuitive analysis, determine that extraction process is A ₁b ₃c ₃.

The impact of product on mouse writhing pain in the middle of 1.3 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mouse writhing experiment grade demonstration: in the middle of different process experiment, product have certain difference for the effect that reduces mouse writhing number of times, but in the middle of most of, product are being compared significant difference (P<0.05 or 0.01) aspect minimizing writhing number of times with negative group.Positive group spasmolysis and analgesia tincture has been compared utmost point significant difference (P<0.001) in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.(in Table 37)

Table 37 mouse writhing experimental result

Administration group and negative control group comparison, ^*represent significant difference (P<0.05), ^*represent difference highly significant (P<0.01), ^{* *}represent difference very significantly (P<0.001).

1.4 intuitive analysis of writhing experimental result and variance analyses

In the mouse writhing pain of take experiment 20min, writhing number of times carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 38, table 39)

Table 38 orthogonal experiment plan is taken into account result

Table 39 variance analysis

* show P<0.05

In the mouse writhing of take experiment 20min, writhing number of times is evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by C>A>B, be soak time > distillation time > amount of water, by intuitive analysis, determine that extraction process is A ₂b ₁c ₃.

The effect of product to mice auricle swelling in the middle of 1.5 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice auricle swelling experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that suppresses mice inflammation auricle edema, and in the middle of major part, product are suppressing to have compared significant difference (P<0.05 or 0.01) with negative group aspect auricle edema.Positive group medicine has been compared utmost point significant difference (P<0.001) with product in the middle of part in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.(in Table 40)

Table 40 mice auricle swelling experimental result

1.6 ear swelling test visual results are analyzed and variance analysis

The mice auricle swelling of take experiment auricle swelling degree carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 41, table 42)

Table 41 orthogonal experiment plan is taken into account result

Table 42 variance analysis

* show P<0.05

The RBC index of testing with Mouse Blood deficiency and excess is changed into evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by A>B>C, be distillation time > amount of water > soak time, by intuitive analysis, determine that extraction process is A ₂b ₃c ₁.

The effect of product to mice inflammation auricle capillary permeability in the middle of 1.7 9 groups of technological experiments

Mice capillary permeability experiment grade shows: positive drug spasmolysis and analgesia tincture with all in the middle of product cause comparing with matched group and all having significant difference (P<0.05) aspect capillary permeability increase because of auricle inflammation suppressing mice; The above results explanation, the relevant drug action that different Extraction techniques cause mice capillary permeability to increase to inflammation-inhibiting all has a direct impact.(in Table 43)

The A value of table 43 test group and control group mice auricle dyestuff transudate

Administration group and negative control group comparison, ^*represent significant difference (P<0.05), ^*represent difference highly significant (P<0.01), ^{* *}represent difference very significantly (P<0.001)

1.8 intuitive analysis of capillary permeability experimental result and variance analyses

The mice inflammation auricle of take oozes out dyestuff absorbance and carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 44, table 45)

Table 44 orthogonal experiment plan is taken into account result

Table 45 variance analysis

* show P<0.05

The auricle of the mice capillary permeability of take experiment oozes out dyestuff absorbance as evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by C>A>B, be distillation time > amount of water > soak time, by intuitive analysis, determine that extraction process is A ₃b ₁c ₃.

The optimum extraction condition that 1.9 pharmacodynamic experiments filter out

Comprehensive above 4 pharmacodynamic experiment acquired results can optimize Valeriana officinalis var. latifolia vapor distillation optimum extraction process: A ₂b ₃c ₃, be: add 8 times of water gagings, soak 2h, vapor distillation extracts 5h.

2. the screening of medicinal residues percolation technique

The effect of product to the experiment of mice vola pain in the middle of 2.1 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice vola pain experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that extends mice pain threshold aspect, but in the middle of most of product aspect extending mice pain threshold with before matched group and administration, compared significant difference (P<0.05 or 0.01); The above results explanation, different Extraction techniques have a direct impact the relevant drug action of mice vola pain.(in Table 46)

Table 46 mice hot plate vola pain experimental result

2.2 vola pain experimental result intuitive analysis and variance analyses

The 90min pain threshold after mice vola pain experiment administration of take carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 47, table 48)

Table 47 orthogonal experiment plan is taken into account result

Table 48 variance analysis

* show P<0.05

In the pain experiment of vola, 90min pain threshold is evaluation index, by carrying out intuitive analysis and variance analysis, learn, the factor size that affects extraction effect is followed successively by B>C>D>A, be alcohol adding amount > flow velocity > soak time > concentration of alcohol, by intuitive analysis, determine that extraction process is A ₃b ₃c ₃d ₂.

The impact of product on mouse writhing pain in the middle of 2.3 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mouse writhing experiment grade demonstration: in the middle of different process experiment, product have certain difference for the effect that reduces mouse writhing number of times, but in the middle of most of, product are being compared significant difference (P<0.05 or 0.01) aspect minimizing writhing number of times with negative group.Positive group spasmolysis and analgesia tincture has been compared utmost point significant difference (P<0.001) with product in the middle of part in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.

(in Table 49)

Table 49 mouse writhing experimental result

2.4 intuitive analysis of writhing experimental result and variance analyses

In the mouse writhing pain of take experiment 20min, writhing number of times carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 50, table 51)

Table 50 orthogonal experiment plan is taken into account result

Table 51 variance analysis

* show P<0.05

The writhing number of times of the mouse writhing of take experiment is evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by A>B>D>C, it is concentration of alcohol > alcohol adding amount > soak time > flow velocity, and factor A has significance to the impact of writhing number of times, therefore select A ₂, by intuitive analysis, determine that extraction process is A ₂b ₃c ₁d ₃.

The effect of product to mice auricle swelling in the middle of 2.5 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice auricle swelling experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that suppresses mice inflammation auricle edema, and in the middle of major part, product are suppressing to have compared significant difference (P<0.05 or 0.01) with negative group aspect auricle edema.Positive group medicine has been compared utmost point significant difference (P<0.001) with product in the middle of part in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.(in Table 52)

Table 52 mice auricle swelling experimental result

2.6 ear swelling test visual results are analyzed and variance analysis

The mice auricle swelling of take experiment auricle swelling degree carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 53, table 54)

Table 53 orthogonal experiment plan is taken into account result

Table 54 variance analysis

* show P<0.05

The swelling of the mice auricle swelling of take experiment is evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by A>B>C>D, it is concentration of alcohol > alcohol adding amount > flow velocity > soak time, and factor A, B, C have significance to the impact of swelling, therefore select A ₂, B ₂, C ₁, by intuitive analysis, determine that extraction process is A ₂b ₂c ₁d ₂.

The effect of product to mice inflammation auricle capillary permeability in the middle of 2.7 9 groups of technological experiments

Mice capillary permeability experiment grade shows: positive drug spasmolysis and analgesia tincture with most of in the middle of product cause comparing with matched group and all having significant difference or utmost point significant difference (P<0.05, P<0.01 or P<0.001) aspect capillary permeability increase because of auricle inflammation suppressing mice; The above results explanation, the relevant drug action that different Extraction techniques cause mice capillary permeability to increase to inflammation-inhibiting all has a direct impact.(in Table 55)

The A value of table 55 test group and control group mice auricle dyestuff transudate

2.8 intuitive analysis of capillary permeability experimental result and variance analyses

The mice inflammation auricle of take oozes out dyestuff absorbance and carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 56, table 57)

Table 56 orthogonal experiment plan is taken into account result

Table 57 variance analysis

* show P<0.05

What the mice capillary permeability of take was tested oozes out dyestuff absorbance as evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by A>D>B>C, be concentration of alcohol > soak time > alcohol adding amount > flow velocity, by intuitive analysis, determine that extraction process is A ₂b ₂c ₃d ₂.

The optimum extraction condition that 2.9 pharmacodynamic experiments filter out

Comprehensive above 4 pharmacodynamic experiment acquired results can optimize Valeriana officinalis var. latifolia medicinal residues optimum extraction process: A ₂b ₂c ₁d ₂, be: add after 15 times of amount 75% soak with ethanol 1h, with the flow velocity of 1Bv/h, carry out percolation.

3. the screening of Flos Impatientis, Flos Carthami, Lignum cinnamomi camphorae percolation technique

The effect of product to the experiment of mice vola pain in the middle of 3.1 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice vola pain experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that extends mice pain threshold aspect, but in the middle of most of product aspect extending mice pain threshold with before matched group and administration, compared significant difference (P<0.05 or 0.01); The above results explanation, different Extraction techniques have a direct impact the relevant drug action of mice vola pain.(in Table 58)

Table 58 mice hot plate vola pain experimental result

3.2 vola pain experimental result intuitive analysis and variance analyses

The 90min pain threshold after mice vola pain experiment administration of take carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 59, table 60)

Table 59 orthogonal experiment plan is taken into account result

Table 60 variance analysis

* show P<0.05

The 90min pain threshold in the experiment of mice vola pain of take is evaluation index, by carrying out intuitive analysis and variance analysis, learn, the factor size that affects extraction effect is followed successively by D>B>A>C, it is soak time > alcohol adding amount > concentration of alcohol > flow velocity, and factor D has significance to the impact of pain threshold, therefore select D ₂, by intuitive analysis, determine that extraction process is A ₃b ₂c ₂d ₂.

The impact of product on mouse writhing pain in the middle of 3.3 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mouse writhing experiment grade demonstration: in the middle of different process experiment, product have certain difference for the effect that reduces mouse writhing number of times, but in the middle of most of, product are being compared significant difference (P<0.05 or 0.01) aspect minimizing writhing number of times with negative group.Positive group spasmolysis and analgesia tincture has been compared utmost point significant difference (P<0.001) with product in the middle of part in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.(in Table 61)

Table 61 mouse writhing experimental result

3.4 intuitive analysis of writhing experimental result and variance analyses

In the mouse writhing pain of take experiment 20min, writhing number of times carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 62, table 63)

Table 62 orthogonal experiment plan is taken into account result

Table 63 variance analysis

* show P<0.05

The writhing number of times of the mouse writhing of take experiment is evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by D>C>B>A, it is soak time > flow velocity > alcohol adding amount > concentration of alcohol, and factor B, C, D have significance to the impact of writhing number of times, therefore select B ₂, C ₂, D ₂, by intuitive analysis, determine that extraction process is A ₃b ₂c ₂d ₂.

The effect of product to mice auricle swelling in the middle of 3.5 9 groups of technological experiments

After nine groups of middle product administrations of technological experiments, mice auricle swelling experiment grade shows: in the middle of different process experiment, product have certain difference for the effect that suppresses mice inflammation auricle edema, but in the middle of all, product are suppressing to have compared significant difference (P<0.05 or 0.01) with negative group aspect auricle edema.Positive group medicine has been compared utmost point significant difference (P<0.001) in the effect aspect minimizing mouse writhing number of times with negative group: the above results explanation, different Extraction techniques have a direct impact the relevant drug action of mouse writhing pain.(in Table 64)

Table 64 mice auricle swelling experimental result

3.6 ear swelling test visual results are analyzed and variance analysis

The mice auricle swelling of take experiment auricle swelling degree carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 65, table 66)

Table 65 orthogonal experiment plan is taken into account result

Table 66 variance analysis

* show P<0.05

The swelling of the mice auricle swelling of take experiment is evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by C>B>A>D, be flow velocity > alcohol adding amount > concentration of alcohol > soak time, by intuitive analysis, determine that extraction process is A ₃b ₂c ₁d ₁.

The effect of product to mice inflammation auricle capillary permeability in the middle of 3.7 9 groups of technological experiments

Mice capillary permeability experiment grade shows: positive drug spasmolysis and analgesia tincture with all in the middle of product cause comparing with matched group and all having significance or utmost point significant difference (P<0.05, P<0.01 or P<0.001) aspect capillary permeability increase because of auricle inflammation suppressing mice; The above results explanation, the relevant drug action that different Extraction techniques cause mice capillary permeability to increase to inflammation-inhibiting all has a direct impact.

(in Table 67)

The A value of table 67 test group and control group mice auricle dyestuff transudate

Group	Size of animal (n)	A value
			Negative group	10	0.0144±0.0039
Spasmolysis and analgesia tincture group	10	0.0061±0.0013***
			No. 1 medicine	10	0.0102±0.0049*
No. 2 medicines	10	0.0086±0.0050**
			No. 3 medicines	10	0.0089±0.0064*
No. 4 medicines	10	0.0100±0.0044*
			No. 5 medicines	10	0.0075±0.0059**
No. 6 medicines	10	0.0103±0.0042*
			No. 7 medicines	10	0.0078±0.0049**
No. 8 medicines	10	0.0078±0.0042**
			No. 9 medicines	10	0.0092±0.0061*

Administration group and negative control group comparison, ^*represent significant difference (P<0.05), ^*represent difference highly significant (P<0.01), * * * represents difference very significantly (P<0.001)

3.8 intuitive analysis of capillary permeability experimental result and variance analyses

The mice inflammation auricle of take oozes out dyestuff absorbance and carries out intuitive analysis and variance analysis as index, and interpretation of result is as follows: (in Table 68, table 69)

Table 68 orthogonal experiment plan is taken into account result

Table 69 variance analysis

* show P<0.05

What the mice capillary permeability of take was tested oozes out dyestuff absorbance as evaluation index, by carrying out intuitive analysis and variance analysis, obtain, the factor size that affects extraction effect is followed successively by B>C>A>D, it is alcohol adding amount > flow velocity > concentration of alcohol > soak time, and factor A, B, C have significance to the impact of absorbance, therefore select A ₃, B ₂, C ₂, by intuitive analysis, determine that extraction process is A ₃b ₂c ₂d ₂.

The optimum extraction condition that 3.9 pharmacodynamic experiments filter out

Comprehensive above 4 pharmacodynamic experiment acquired results can optimize three taste medical material optimum extraction process: the A such as Flos Impatientis ₃b ₂c ₂d ₂, be: add after 15 times of amount 70% soak with ethanol 12h, with the flow velocity of 2Bv/h, carry out percolation.

Although, above used general explanation, the specific embodiment and test, the present invention is described in detail, on basis of the present invention, can make some modifications or improvements it, and this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.

Claims

1. treat a Chinese medicine composition for toothache, it is characterized in that this Chinese medicine composition made by being prepared as follows technique by the medicine of following weight parts: Valeriana officinalis var. latifolia 50-400 part, Flos Carthami 20-100 part, Flos Impatientis 50-300 part, Lignum cinnamomi camphorae 5-50 part

2. Chinese medicine composition according to claim 1, is characterized in that this Chinese medicine composition made by being prepared as follows technique by the medicine of following weight parts: Valeriana officinalis var. latifolia 50-400 part, Flos Carthami 20-100 part, Flos Impatientis 50-300 part, Lignum cinnamomi camphorae 5-50 part

3. Chinese medicine composition according to claim 1 and 2, is characterized in that Chinese medicine weight proportion is wherein: Valeriana officinalis var. latifolia 80-350 part, Flos Carthami 30-90 part, Flos Impatientis 80-260 part, Lignum cinnamomi camphorae 10-45 part.

4. Chinese medicine composition according to claim 1 and 2, is characterized in that Chinese medicine weight proportion is wherein: Valeriana officinalis var. latifolia 100-300 part, Flos Carthami 40-80 part, Flos Impatientis 90-240 part, Lignum cinnamomi camphorae 15-40 part.

5. Chinese medicine composition according to claim 1 and 2, is characterized in that Chinese medicine weight proportion is wherein: Valeriana officinalis var. latifolia 130-260 part, Flos Carthami 50-75 part, Flos Impatientis 100-180 part, Lignum cinnamomi camphorae 20-35 part.

6. Chinese medicine composition according to claim 1 and 2, is characterized in that Chinese medicine weight proportion is wherein: 200 parts of Valeriana officinalis var. latifolia, 60 parts, Flos Carthami, 110 parts of Flos Impatientiss, 30 parts of Lignum cinnamomi camphoraes.

7. Chinese medicine composition according to claim 1 and 2, is characterized in that, described Chinese medicine composition is solid preparation or liquid preparation or membranous patch.

8. Chinese medicine composition according to claim 7, is characterized in that: described solid preparation is sheet, capsule, granule or pill; Described liquid preparation is tincture, oral liquid, injection.

9. Chinese medicine composition according to claim 1 and 2, it is characterized in that, described pharmaceutically acceptable carrier or diluent are selected from one or more in film former, wetting agent, filler, diluent, binding agent, disintegrating agent, lubricant, surfactant or correctives.

10. the application of Chinese medicine composition according to claim 1 and 2 in the medicine of preparation treatment gingivitis, toothache that dental caries causes or gingival swelling and pain.