CN102293948A

CN102293948A - Preparation for treating children dyspepsia endogenous heat, and preparation method and detection method of preparation

Info

Publication number: CN102293948A
Application number: CN2010102055903A
Authority: CN
Inventors: 汤明昌; 蒋平
Original assignee: Individual
Current assignee: Individual
Priority date: 2010-06-22
Filing date: 2010-06-22
Publication date: 2011-12-28

Abstract

The invention discloses a preparation for treating children dyspepsia endogenous heat, and a preparation method and a quality control method of the preparation. The preparation consists of the following raw materials in part by weight: 3 to 9 parts of cultured calculus bovis (or in vitro cultured calculus bovis ), 0.75 to 2.25 parts of artificial musk, 3 to 9 parts of borneol, 60 to 180 parts of ramulus et uncus uncariae, 60 to 180 parts of gastrodia elata, 15 to 45 parts of stiff silkworm heated with bran, 15 to 45 parts of scorpion, 15 to 45 parts of coptis chinensis, 15 to 45 parts of tabasheer, 5 to 15 parts of cinnabar, 15 to 45 parts of rhubarb, 15 to 45 parts of arisaema cum bile roasted with rice wine, 15 to 45 parts of thunberg fritillary bulb, 15 to 45 parts of processed pinellia ternata, 60 to 180 parts of tangerine root, and 60 to 180 parts of talc. The preparation is strict in a formula, has the effects of clearing heat and relieving convulsion, and dispelling wind and reducing phlegm, and is used for children dyspepsia endogenous heat; and the preparation process and the quality control method are scientific and reasonable; and by the quality control method, the quality of a product can be effectively controlled.

Description

Internally heated preparation of a kind of treatment children's's dyspepsia and preparation method thereof and detection method

Technical field

The present invention relates to internally heated preparation of a kind of treatment children's's dyspepsia and preparation method thereof and method of quality control, belong to technical field of Chinese medicine.

Background technology

Children's's physiological feature shows that the digestion aspect is a little less than the strong spleen of stomach, promptly to be easy to take in too much the heavy food of high heat, and can't in time to digest.Food rests on the digestive tract overlong time and promptly forms dyspepsia, and the dyspepsia dyspepsia is the inducement of numerous disease, the early symptom of infantile dyspepsia as seen: thick fur is greasy, abnormal smells from the patient is deep in mouthful, it is not normal to defecate, and it is not stable to sleep, i.e. the traditional Chinese medical science said " disorder of the stomach leading to insomnia with restlessness "; Dyspepsia causes the miopragia of children's's the intestines and stomach, develops into then: a warm abdomen heat, anorexia, belch are made full, vomiting sour, wheezing sound in the throat, feverish sensation over the palm, stool a few days 1 time, oliguria with reddish urine muddiness, agitation is cried angry not peaceful, have with thirsty disease, the milk regurgitation that has, food, the heat-phlegm etc. of waiting of low grade fever.

The food that is difficult to digest time of staying in digestive tract has been grown, and will ferment and produces heat, and instant stagnation-derived fever causes that children's's body burns, the vomiting sputum, and cough is fidgetyly got impatient, and is restless and sleepless, constipation due to dry stool etc.

Children's's (particularly infant of 6 months to 3 years old) is because its distinctive physiological reason and onset speed is exceedingly fast, (1～2 day) can the hyperpyrexia symptom occur because of the dyspepsia interior-heat at short notice, hyperpyrexia reaches 39～40 ℃ and occurs convulsions, obnubilation, the i.e. traditional Chinese medical science said " convulsion with spasms, coma " often.Reason is: the dyspepsia fermentation produces toxin, and dyspepsia causes alteration of intestinal flora, and pathogenic bacterium breed the generation toxin in a large number; Toxin enters blood circulation, causes hyperpyrexia; When infantile hyperpyrexia, the metabolism of neurocyte, zmount of oxygen consumption and blood flow can change, simultaneously, children's's central nervous system also can be in over-drastic excitatory state, this " excitement " can have influence on one of children's does not have the fully-developed cerebral tissue---and " thalamus ", make it to produce intensive discharge, and pass to other position of brain, that is: convulsive attack.Hyperpyrexia convulsion is very serious to children's's harm, and faint from fear continuing more than 10 minutes may be because of the cerebral anoxia overlong time causes the neurocyte infringement, continues more than 30 minutes half then and leaves in various degree delayed ischemic neurological deficits.

According to the etiology and pathogenesis and the characteristics of incidence of heat syndrome in children's's dyspepsia, answer heat clearing away, relieving convulsion, diffusing wind on the method for treatment, reduce phlegm.At present, though the Chinese medicine and western medicine of commercially available such disease of treatment is various, have the few of definite curative effect.For more effectively preventing and treating such disease, the applicant develops a kind of safe and effective new drug.

Artificial Moschus, Borneolum Syntheticum, Calculus Bovis three flavors match in the prescription, with the refreshment of having one's ideas straightened out, heat-clearing and toxic substances removing, are used for the epidemic febrile disease unconsciousness due to high fever and faint from fear; Ramulus Uncariae Cum Uncis, Rhizoma Gastrodiae, Scorpio, Bombyx Batryticatus match, and with relieving spasm by subduing liver-wind, are used for intenseness of heat and send out convulsion, convulsion with spasms; Rhizoma Coptidis, Concretio Silicea Bambusae, Cinnabaris, Radix Et Rhizoma Rhei match, with clear away heart-fire, calm the nerves, pathogenic fire purging, be used for irritability, palpitation with fear insomnia, irritated, constipation; The Rhizoma Pinelliae, Arisaema Cum Bile, Bulbus Fritillariae Thunbergii, dried tangerine peel, Talcum match, with wind arresting convulsion, clearing away heat to stop vomiting, circulation of qi promoting expectorant, the expelling summer-heat of loosing; Full side's heat clearing away, relieving convulsion, the wind that looses, reduce phlegm and usefulness, prescription is rigorous, determined curative effect.

On January 31st, 2004, State Food and Drug Administration sends " about the notice of Calculus Bovis and succedaneum use problem thereof ": for the new drug that contains Calculus Bovis of clinical anxious severe disease disease medicine variety that contains Calculus Bovis in the national drug standards prescription and the approval of national drug supervisory and management department, Calculus Bovis in the prescription can be substituted uses that feed intake of Calculus Bovis equivalent with Calculus Bovis cultivating, In vitro cultured Calculus Bovis, but it is alternative not to be able to the artificial Calculus Bovis; Other kinds that contain Calculus Bovis can substitute the use that feeds intake of Calculus Bovis equivalent with Calculus Bovis cultivating, In vitro cultured Calculus Bovis or artificial Calculus Bovis with the Calculus Bovis in the prescription.The scope of application to the artificial Calculus Bovis of this document limits, and can be equal to use with natural Calculus Bovis to Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), also is the affirming of quality, drug effect, clinical efficacy to Calculus Bovis cultivating (or In vitro cultured Calculus Bovis).

The natural Calculus Bovis shortage of resources can not satisfy clinical application, and the Calculus Bovis cultivating that using character of the present invention is close (or In vitro cultured Calculus Bovis) replaces natural Calculus Bovis, has scientific basis and policy support.With regard to active ingredient, in existing statutory standards, the bilirubin (active ingredient) that natural Calculus Bovis and Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) contain must not be less than 35.0%.

In addition, in order to control drug quality fully and effectively,, must formulate scientific and reasonable method of quality control to guarantee its clinical efficacy.

Summary of the invention

The objective of the invention is to: internally heated preparation of a kind of treatment children's's dyspepsia and preparation method thereof and method of quality control are provided, the preparation that is provided is used for children's's dyspepsia interior-heat and causes that the cough body burns, and the vomiting sputum is fidgetyly got impatient, restless and sleepless, convulsion with spasms, coma, constipation due to dry stool, its determined curative effect, have no side effect, and preparation method is scientific and reasonable, product quality is easily controlled.

The present invention constitutes like this: component calculates by weight: it is to be by 3～9 parts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) by it; 0.75～2.25 part of artificial Moschus; 3～9 parts of Borneolum Syntheticums; 60～180 parts of Ramulus Uncariae Cum Uncis; 60～180 parts in Rhizoma Gastrodiae; 15～45 parts of Bombyx Batryticatus (parched with bran); 15～45 parts of Scorpios; 15～45 parts of Rhizoma Coptidis; 15～45 parts of Concretio Silicea Bambusaes; 5～15 parts in Cinnabaris; 15～45 parts of Radix Et Rhizoma Rhei; 15～45 parts of Arisaema Cum Bile (wine is processed); 15～45 parts of Bulbus Fritillariae Thunbergiis; 15～45 parts of Rhizoma Pinelliae (processed); tangerine 60～180 parts; 60～180 parts in Talcum is made.

Specifically, component calculates by weight: it is made for 120 parts by 6 parts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), 1.5 parts of artificial Moschuss, 6 parts of Borneolum Syntheticums, 120 parts of Ramulus Uncariae Cum Uncis, 120 parts in Rhizoma Gastrodiae, 30 parts of Bombyx Batryticatus (parched with bran), 30 parts of Scorpios, 30 parts of Rhizoma Coptidis, 30 parts of Concretio Silicea Bambusaes, 10 parts in Cinnabaris, 30 parts of Radix Et Rhizoma Rhei, 30 parts of Arisaema Cum Bile (wine is processed), 30 parts of Bulbus Fritillariae Thunbergiis, 30 parts of Rhizoma Pinelliae (processed), 120 parts of dried tangerine peels, Talcum.

Described preparation is powder, granule, capsule.

The preparation method of the internally heated preparation of a kind of children's's of treatment dyspepsia of the present invention is:

The first step: get Talcum, Concretio Silicea Bambusae, two flavor medical materials are ground into impalpable powder with high speed disintegrator, 80 ℃ of dryings are crossed 150 mesh sieves after 2 hours, standby.

Second step: get Cinnabaris, add purified water with grinder and adopt elutriation to be ground into impalpable powder, cross 120 mesh sieves after 2 hours, add the first step gained medicated powder of 2 times of amounts of Cinnabaris again, ground 30 minutes with 40 ℃ of dryings of electric heating constant temperature oven, standby; Get Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), be ground into impalpable powder, cross 120 mesh sieves after 2 hours, add the first step gained medicated powder of 2 times of amounts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) again, ground 30 minutes with 60 ℃ of dryings of electric heating constant temperature oven with grinder, standby; Get the artificial Moschus, put into mortar, add a small amount of purified water (component meter by weight, artificial Moschus: water=3: 1), be ground into impalpable powder, cross 120 mesh sieves after 2 hours with 40 ℃ of dryings of electric heating constant temperature oven, the first step gained medicated powder that adds 2 times of amounts of artificial Moschus again ground 30 minutes, and is standby; Get Borneolum Syntheticum, add the first step gained medicated powder of 2 times of amounts, be ground into impalpable powder, cross 120 mesh sieves with high speed disintegrator, standby.

The 3rd step: get 5 flavor medical materials such as Bombyx Batryticatus (parched with bran), Scorpio, Arisaema Cum Bile (wine is processed), Bulbus Fritillariae Thunbergii, Rhizoma Pinelliae (processed), after 2 hours, mixing is ground into impalpable powder with high speed disintegrator respectively at 80 ℃ of dryings, crosses 120 mesh sieves, and is standby.

The 4th step: Ramulus Uncariae Cum Uncis cuts into the section that length is 10mm, and Rhizoma Gastrodiae, Rhizoma Coptidis, Radix Et Rhizoma Rhei cut into the thin slice that thickness is 2～3mm, and dried tangerine peel cuts into the filament that width is 2～3mm.More than 5 flavor medical materials respectively at 80 ℃ of dryings after 2 hours, mixing is ground into impalpable powder with high speed disintegrator, crosses 120 mesh sieves, and is standby.

The 5th step: in multidirectional movement mixer, second four kinds of medicated powder that go on foot gained were mixed 30 minutes with remaining first step gained medicated powder; The 3rd step of property adding gained medicated powder continues to mix 30 minutes again; Last the 4th step of property adding gained medicated powder continues to mix 30 minutes.

The 6th step: final mixed medicated powder is made different preparations according to a conventional method, as powder, granule, capsule etc.

Described powder is preparation like this:

The 6th step: final mixed medicated powder is carried out packing by certain loading amount specification, promptly.

Described granule is preparation like this:

The 6th step: final mixed medicated powder is made granule with the spray of boiling granulating machine with purified water or alcoholic solution, carry out packing by certain loading amount specification again, promptly.

Described capsule is preparation like this:

The 6th step: will final mixed medicated powder spray and make granule, recharge and be capsule, or directly be filled to capsule, promptly with final mixed medicated powder with purified water or alcoholic solution with the boiling granulating machine.

The method of quality control of the internally heated preparation of a kind of children's's of treatment dyspepsia of the present invention is: described method of quality control mainly comprise in character, the discriminating project partly or entirely, inspection and assay; Wherein differentiate and comprise that the thin layer chromatography that preparation is carried out microscopical identification and Rhizoma Coptidis, Borneolum Syntheticum differentiates that assay is the assay to Calculus Bovis cultivating in the preparation (or In vitro cultured Calculus Bovis).

Described method of quality control comprises:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

Differentiate: it is an amount of that (1) gets this preparation, and the rearmounted microscopically of load is observed: prism of calcium oxalate is present in the parenchyma cell; Calcium oxalate sand crystal exists in the parenchyma cell; The wall of the thin cell of wood is beaded and thickens, and pit is obvious; Contain gelatinizing polysaccharide fragment of tissue chance iodine liquid and be brown or light brown brown, dissolve when meeting the heating of chloral hydrate liquid; Irregular sheet is colourless, has and peels off vestige layer by layer; Calcium oxalate cluster crystal is big, diameter 60～140 μ m; Reticulate vessel is bigger, non-lignify; The stone cell foresythia is similar round polygon or spindle, and the hole ditch is very thin, and the laminated striation that has is obvious; The body wall fragment is faint yellow to yellow, tool reticular texture and circular trichopore, visible sometimes sepia bristle; The body wall fragment is colourless, and there is superfine mycelium on the surface; The starch grain oval, diameter 35～48 μ m, the omphalion point-like, herringbone or horse-hof shape are positioned at a less end, and laminated striation is fine and closely woven and obvious; Starch grain pentagon or polygon, omphalion point-like or starlike often is stained with granular substance, is golden yellow or pale brown color; The dark brownish red of irregular fine particle, glossy, the edge dark yellow; Amorphous agglomerate is fallow, is embedded with tiny prism of calcium oxalate.

(2) get this preparation 1g, add methanol 10ml, supersound process 15 minutes, centrifugal, get the supernatant evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 10ml, shines medical material solution in pairs with legal system; Get the berberine hydrochloride reference substance again, add methanol and make the solution that 1ml contains 0.5mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1) is developing solvent, put in the chromatography cylinder of ammonia saturated with vapor, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle.

(3) get this preparation 1g, add ethyl acetate 10ml, supersound process 15 minutes, centrifugal, get supernatant as need testing solution; Other gets the Borneolum Syntheticum reference substance, adds ethyl acetate and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-acetone (9: 1) is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Check: granularity is measured according to an appendix XI of Chinese Pharmacopoeia version in 2010 B, should be all by No. six sieves.

The outward appearance uniformity should meet under an appendix IB of Chinese Pharmacopoeia version in 2010 the powder item regulation to the outward appearance uniformity.

Moisture is measured according to an appendix IX of Chinese Pharmacopoeia version in 2010 H aquametry, must not cross 9.0%.

Content uniformity should meet under an appendix IB of Chinese Pharmacopoeia version in 2010 the powder item regulation to content uniformity.

Microbial limit should be up to specification according to the inspection of an appendix X of Chinese Pharmacopoeia version in 2010 III C microbial limit test.

Assay: shine an appendix VI of Chinese Pharmacopoeia version in 2010 D high effective liquid chromatography for measuring:

The system suitability test is a filler with the octadecylsilane chemically bonded silica, dimethyl sulfoxide-acetonitrile-0.5mol/L Spirit of Mindererus. (regulating pH value with glacial acetic acid is 5.3) (6: 6: 7) is a mobile phase, the detection wavelength is 452nm, and theoretical cam curve is pressed the bilirubin peak and calculated, and should be not less than 1000.

The preparation precision of reference substance solution takes by weighing the bilirubin reference substance 10.0mg that is dried to constant weight through phosphorus pentoxide, puts in the brown volumetric flask of 100ml, adds dimethyl sulfoxide-acetonitrile (7: 3) and makes dissolving, and be diluted to scale, shakes up, in contrast product solution.

The preparation precision of need testing solution takes by weighing this preparation 1g, put in the brown volumetric flask of 25mL, accurate dimethyl sulfoxide-the acetonitrile (7: 3) that adds is an amount of, supersound process 15min, the above-mentioned solvent of reuse is settled to scale, shakes up, and filters with microporous filter membrane (0.45 μ m), discard filtrate just, get subsequent filtrate as need testing solution.

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

The every 1g of this preparation contains Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) with bilirubinometer, must not be less than 2.5mg.

Side of the present invention separates: we are made up of ten six medicaments such as Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), artificial Moschus, Borneolum Syntheticum, Ramulus Uncariae Cum Uncis, Rhizoma Gastrodiae, Bombyx Batryticatus (parched with bran), Scorpio, Rhizoma Coptidis, Concretio Silicea Bambusae, Cinnabaris, Radix Et Rhizoma Rhei, Arisaema Cum Bile (wine is processed), Bulbus Fritillariae Thunbergii, Rhizoma Pinelliae (processed), dried tangerine peel, Talcums.Artificial Moschus, Borneolum Syntheticum, Calculus Bovis are the key medicine of calentura in the side, and the three matches, and with the refreshment of having one's ideas straightened out, heat-clearing and toxic substances removing, are used for the epidemic febrile disease unconsciousness due to high fever and faint from fear, with the critical symptom of rapid alleviation convulsions; Ramulus Uncariae Cum Uncis, Rhizoma Gastrodiae, Scorpio, Bombyx Batryticatus four flavors are good at controlling wind, and four match, and with relieving spasm by subduing liver-wind, are used for intenseness of heat and send out convulsion, convulsion with spasms; Rhizoma Coptidis, Concretio Silicea Bambusae, Cinnabaris, Radix Et Rhizoma Rhei match, and can clear away heart-fire well, calm the nerves, pathogenic fire purging, are used for irritability, palpitation with fear insomnia, irritated, constipation; The Rhizoma Pinelliae, Arisaema Cum Bile, Bulbus Fritillariae Thunbergii, tangerine four flavors are all arrogated to oneself removing heat-phlegm, are equipped with the Talcum of expelling summer-heat again, with wind arresting convulsion, clearing away heat to stop vomiting, circulation of qi promoting expectorant, the expelling summer-heat of loosing; Full side's heat clearing away, relieving convulsion, the wind that looses, reduce phlegm and usefulness, so the multiple critical symptom of children's's dyspepsia can be controlled and eliminate to compatibility rapidly, and eliminate the cause of disease.

Innovative point of the present invention is:

1. preparation heat clearing away of the present invention, relieving convulsion, the wind that looses, reduce phlegm and usefulness, be used for heat syndrome in children's's dyspepsia.

2. the preparation method of preparation of the present invention is provided.

3. the method for quality control of preparation of the present invention is provided.

For the preparation technology who verifies preparation of the present invention is easy to large-scale production, have good quality controllability and stability, and have excellent curative and higher safety, the applicant has carried out a series of experimental study, and is specific as follows:

One, preparation prescription screening:

Ramulus Uncariae Cum Uncis, Bombyx Batryticatus, Rhizoma Gastrodiae, Scorpio are come from " the Ramulus Uncariae Cum Uncis drink " of " medical science main story " volume eight that the Ming Dynasty positive moral second last of the twelve Earthly Branches (1515) Yu Tuan writes, cure mainly a little less than the spleen and stomach in children gas due to infantile convulsion; Scorpio, Bombyx Batryticatus, Cinnabaris, Calculus Bovis, Borneolum Syntheticum, Rhizoma Coptidis, Rhizoma Gastrodiae, Arisaema Cum Bile come from " Wanbing Huichun, Curative Measures for All Diseases " that 15 years Wanli year of the Ming Dynasty, (1587) Gong Tingxian was write and roll up seven " a thousand pieces of gold is diffusing ", cure mainly all phlegm dyspneas of children's, anxious chronic infantile convulsion; Radix Et Rhizoma Rhei, Cinnabaris come from that " Chinese pharmacopoeia version " children's indigestion powder " in 2000 cures mainly children's and stops breast and stop eating abdominal distention constipation, dyspnea with cough due to overabundant phlegm; The present invention is " Ramulus Uncariae Cum Uncis drink ", " a thousand pieces of gold looses ", " children's indigestion powder " is harmonious; on flavour of a drug, make suitably plus-minus according to the etiology and pathogenesis of heat syndrome in children's's dyspepsia, and the key medicine that is equipped with treatment epidemic febrile disease such as Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), artificial Moschus, Borneolum Syntheticum forms, relieving convulsion to reach heat clearing away; the effect of diffusing air slaking expectorant; be used for children's's dyspepsia interior-heat and cause that the cough body burns, the vomiting sputum is fidgetyly got impatient; restless and sleepless; convulsion with spasms, coma, constipation due to dry stool.

Two, Study on Preparation

1. process route:

Talcum, Concretio Silicea Bambusae → pulverizing → drying → sieve → medicated powder are 1.

Cinnabaris → water flies → dry → sieve → add medicated powder 1. → grinding → medicated powder 2.

Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) → grinding → drying → sieve → add medicated powder 1. → grinding → medicated powder 3.

Artificial Moschus → add water grinding → drying → sieve → add medicated powder 1. → grinding → medicated powder 4.

Borneolum Syntheticum → adding medicated powder 1. → pulverize → sieve → medicated powder 5.

Bombyx Batryticatus (parched with bran), Scorpio, Bulbus Fritillariae Thunbergii

→ drying → mixing → pulverize → sieve → medicated powder is 6. respectively

Arisaema Cum Bile (wine is processed), Rhizoma Pinelliae (processed)

The Ramulus Uncariae Cum Uncis cutting

Drying → the mixing of Rhizoma Gastrodiae, Rhizoma Coptidis, Radix Et Rhizoma Rhei shave → respectively → pulverize → sieve → medicated powder 7.

Dried tangerine peel chops up silk

1. 5. 4. 3. 2. medicated powder mix → add medicated powder with residue medicated powder and 6. mix → add medicated powder and 7. mix → finished product medicated powder

2. process chart

See Fig. 1

3. process rationality research

The same or similar medical material of character is pulverized simultaneously, can be improved crush efficiency, can play blended effect again; Adopt specific process to pulverize separately by its characteristic expensive thin medical material, and mix with Pulvis Talci India bloom again after grinding dilution with the Pulvis Talci India bloom of 2 times of amounts, help the homodisperse of expensive thin medical material; Various medicated powder mix step by step by the character principle identical or close with part by weight, can guarantee blended uniformity, and avoid the simple total dispersion non-uniform phenomenon that causes that mixes of each medicated powder.

3.1 the pulverizing of Talcum and Concretio Silicea Bambusae: Talcum is a silicates mineral talc family Talcum, be flat, rhomboid or irregular bulk, crystal is six sides and rhombus plate, and matter is softer and real, hardness is 1, proportion 2.7～2.8 can scrape white lead with fingernail, and that touches has a lubricious sense, no hygroscopicity, put do not collapse in the water diffusing, odorless, tasteless; Mainly contain the hydration magnesium silicate, also can be used as adjuvant after Talcum is pulverized and use; To be grass family Chinese textile bamboo or schizostachyum chinense Rendle stung behind the hole and the bleeding sap of storing up between ring by the Xylocopa dissimilis (Lep.) of parasitism Concretio Silicea Bambusae, and drying condenses into block, main aluminium hydroxide, tripoli, aluminium sesquioxide, iron sesquioxide; The two quality is close, preferentially the two mixed powder is broken into impalpable powder (medicated powder 1.), and can be used as the diluent of the thin medical material of Fang Zhonggui.

3.2 the pulverizing of Cinnabaris, Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), artificial Moschus, Borneolum Syntheticum:

3.2.1 Cinnabaris is the mineral drug of main Containing Sulfur hydrargyrum, and often is mingled with broken end such as small amounts of sandstone, water flies into impalpable powder, and after the drying, 1. " 3.1 " the gained medicated powder that adds with 2 times of amounts dilute, and grinds evenly, obtains medicated powder 2.; So operation can pure medicine and is avoided Cinnabar to rise, lump, fly upward.(water flies: Cinnabaris is water-fast mineral drug, adds proper amount of clear water in grinder, and Cinnabaris is ground to form pasty state, add water gaging again and stir, coarse powder promptly sinks, and in time inclines to suspension, the coarse grain that sinks row again grinds, like this repeatable operation, till porphyrize, impurity that at last can not suspendible discards, the suspension merging that anteversion and retroversion go out is left standstill, treat post precipitation, the top clear water that inclines, the gained precipitate grinds to form superfine powder.) the water Cinnabaris can make it purer, be convenient to preparation, selecting cinnabar content is index, carry out assay relatively with the direct comminuting method of high speed disintegrator: water intaking Cinnabaris powder, high speed disintegrator is pulverized each 0.3g of Cinnabaris powder, totally 6 parts, the accurate title, decide, put in the 250ml flask, add sulphuric acid 10ml and potassium nitrate 1.5g heating and make dissolving, put cold, add water 50ml, and add potassium permanganate solution to showing pink, and drip 2% copperas solution again to red the disappearance, add ammonium ferric sulfate indicator solution 2ml, with ammonium thiocyanate liquid (0.1mol/L) titration, promptly get (the 0.1mol/L ammonium thiocyanate liquid of every 1ml is equivalent to the 11.63mg cinnabar).

Table 1 elutriation and directly pulverizing gained Cinnabaris fine powder purity comparison

The result shows: elutriation can obviously improve cinnabar content, can remove most of impurity Cinnabaris is played purified effect (RSD=0.6669%), and make content reach pharmacopeia requirement (this product Containing Sulfur hydrargyrum must not be less than 96.0%); Directly pulverizing then can not be removed the impurity in the Cinnabaris, and content difference is bigger, and medicine is not had pure effect.

3.2.2 Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) body is light; matter is crisp; be ground into impalpable powder with grinder; cross 120 mesh sieves after 2 hours with 60 ℃ of dryings of electric heating constant temperature oven; 1. " 3.1 " the gained medicated powder that adds with 2 times of amounts dilute again; grind evenly, obtain medicated powder 3., the Calculus Bovis the avoided powder after dilution is ground rises, lumps, flies upward.

3.2.3 the artificial Moschus mainly contains muscone, steroidal compounds etc., its effective ingredient can be dissolved in organic solvents such as dehydrated alcohol, chloroform, is soluble in ether, but water insoluble, stable in properties under the room temperature; So use mortar, add a small amount of purified water (component meter by weight, artificial Moschus: water=3: 1), be ground into impalpable powder, cross 120 mesh sieves after 2 hours with 40 ℃ of dryings of electric heating constant temperature oven, 1. " 3.1 " the gained medicated powder that adds with 2 times of amounts dilute again, grind evenly, obtain medicated powder 4..

3.2.4 Borneolum Syntheticum very easily distils, easily oxidation becomes Camphora in air, therefore adopts " 3.1 " the gained medicated powder that mixes 2 times of amounts in the Borneolum Syntheticum 1. to be ground into impalpable powder with high speed disintegrator, crosses 120 mesh sieves, obtains medicated powder 5..1. medicated powder be Pulvis Talci and India bloom, is mainly mineralogical composition, and no hygroscopicity, thus can play good dispersion, Stabilization to Borneolum Syntheticum, and make crushing operation be easy to carry out.See Table 2.

The effect that pulverizing was pulverized, mixed to table 2 Borneolum Syntheticum separately compares

Remarks: the investigation amount is 2000g.

The result shows: Borneolum Syntheticum is pulverized easy adhering device separately, pulverizes the back powder and easily bonds, and is mobile poor; Mix and to pulverize that then Borneolum Syntheticum can by Pulvis Talci and the India bloom disperses and dilution, avoid adhering device, and powder is loose, good fluidity helps follow-up married operation.

3.3 the pulverizing of Bombyx Batryticatus (parched with bran), Scorpio, Arisaema Cum Bile (wine is processed), Bulbus Fritillariae Thunbergii, Rhizoma Pinelliae (processed):

5 flavor medical materials such as Bombyx Batryticatus (parched with bran), Scorpio, Arisaema Cum Bile (wine is processed), Bulbus Fritillariae Thunbergii, Rhizoma Pinelliae (processed), after 2 hours, mixing is ground into impalpable powder with high speed disintegrator respectively at 80 ℃ of dryings, crosses 120 mesh sieves, obtains medicated powder 6..

Arisaema Cum Bile is the fine powder of Rhizoma Arisaematis (processed) (tuber of aroid Rhizoma Arisaematis) and the square block that cattle and sheep, Fel Sus domestica process from strand, and matter is crisp frangible; Bulbus Fritillariae Thunbergii is the underground bulb of liliaceous plant Bulbus Fritillariae Thunbergii, and quality is solid; Rhizoma Pinelliae (processed) is that the tuber of the aroid Rhizoma Pinelliae is concocted and processed through Calx, Radix Glycyrrhizae, and quality is loose and crisp; Above-mentioned three kinds of medicines all contain a large amount of starch.Bombyx Batryticatus (parched with bran) forms through parched with bran for the lethal rigid polypide of larva infection muscardine of silkworm section insecticide silkworm, and matter is hard and crisp, is all the insects medicine with Scorpio.

The insects medicine is pulverized separately, and its fragment of tissue very easily adheres to machine and screen cloth can't carry out crushing screening, so starch-containing many Arisaema Cum Bile, Bulbus Fritillariae Thunbergii, the Rhizoma Pinelliae mix pulverizing with it in the general side, then can in time absorb and disperses the insects medicine, avoids adhesion.See Table 3.

Crushing effect is pulverized, mixed to table 3 Bombyx Batryticatus Scorpio separately relatively

Remarks: investigation amount 3000g, mix in the pulverizing and carry out proportioning in the prescription ratio.

The result shows: Bombyx Batryticatus, two kinds of insect medicines of Scorpio are pulverized easy adhering device separately, pulverize the back powder and easily bond, and mobile difference can't be sieved; Mix and to pulverize then that the insect medicine fragment can be disperseed and dilution by starch-containing many medicines such as Arisaema Cum Bile, Bulbus Fritillariae Thunbergii, the Rhizoma Pinelliae, avoid adhering device, and powder is loose, good fluidity helps sieving and follow-up married operation.

3.4 the pulverizing of Ramulus Uncariae Cum Uncis, Rhizoma Gastrodiae, Rhizoma Coptidis, Radix Et Rhizoma Rhei, dried tangerine peel:

Ramulus Uncariae Cum Uncis cuts into the section that length is 10mm, and Rhizoma Gastrodiae, Rhizoma Coptidis, Radix Et Rhizoma Rhei cut into the thin slice that thickness is 2～3mm, and dried tangerine peel cuts into the filament that width is 2～3mm.More than 5 flavor medical materials respectively at 80 ℃ of dryings after 2 hours, mixing is ground into impalpable powder with high speed disintegrator, crosses 120 mesh sieves, obtains medicated powder 7..

Ramulus Uncariae Cum Uncis, Rhizoma Gastrodiae, Rhizoma Coptidis, Radix Et Rhizoma Rhei, dried tangerine peel are fibrous medical material, if this type of medicine is mixed pulverizing with starch-containing many medicine or mineral drug, then can reduce the crushing effect of various medicines simultaneously, so this class medical material carries out suitable cutting and drying after sorting out separately in the general side, and select the stronger high speed disintegrator (omnipotent flour mill) of grindability to pulverize.Pulverizing is divided into 2 times to be carried out, and disposes 40 mesh sieve sheets for the first time and is ground into coarse powder, disposes 80 mesh sieve sheets for the second time coarse powder is pulverized once more, and can obtain can be by the impalpable powder of 120 mesh sieves.

3.5 the research of medicated powder hybrid technique

Table 4 is operated the ratio (component calculating by weight) of the various medicated powder that obtain in above-mentioned breaking method:

As seen: medicated powder ratio 1.～7. is=150: 30: 18: 4.5: 18: 150: 420

3.5.1 hybrid plan one (each medicated powder is mixed step by step)

If above-mentioned 7 kinds of medicated powder are mixed in the lump, then can because of medicated powder character, proportional difference too greatly can't mix homogeneously, so mix after will medicated powder classifying.

(1) medicated powder total amount 2.～5. is=70.5; Remaining medicated powder 1.=150-20-12-3-12=103, this and medicated powder total amount 2.～5. is close, and above-mentioned medicated powder is all based on Pulvis Talci, thus can be with above-mentioned medicated powder prior to mixing 30 minutes in the multidirectional movement mixer, mixing the back gross weight is 173.5.

(2) total amount was 173.5 after 1.～5. medicated powder mixed, and this and medicated powder amount 150 6. is the most approaching, so it can be mixed 30 minutes with multidirectional movement mixer again, mixing the back gross weight is 323.5.

(3) total amount was 323.5 after 1.～6. medicated powder mixed, and this and medicated powder amount 420 7. is approaching, thus it can be mixed 30 minutes with multidirectional movement mixer, the medicated powder that gets product after the mixing, gross weight is 743.5.

3.5.2 hybrid plan two (with the direct disposable mixing of all medicated powder)

1.～7. medicated powder all placed multidirectional movement mixer, mixed 30 minutes, the medicated powder that gets product, component meter by weight, gross weight is 743.5.

3.5.3 the mixed effect of two kinds of hybrid plans is relatively:

(1) percentile mensuration of moisture absorption and comparison:

Screening index: 72 hours moisture absorption percentage rate.The result: 72 hours moisture absorption percentage rate that two kinds of hybrid plans make finished product medicated powder see Table 5

72 hours moisture absorption percentage rate of two kinds of obtained medicated powder of hybrid plan of table 5

Remarks: investigation amount 30g

As shown in Figure 2, the medicated powder moisture absorption percentage rate that hybrid plan one makes is less than hybrid plan two, so scheme one is better than scheme two.(2) mensuration of mixing uniformity and comparison: the finished product medicated powder to scheme one, scheme two is respectively got 9 duplicate samples by the principle of Stratified Sampling respectively, the selection bilirubin is an index, measure content, according to the requirement of " pharmaceutical production checking guide " version (Chemical Industry Press) in 2003 to the medicine mixing uniformity, assay result's relative standard deviation (RSD) should≤2%.

Method: according to an appendix VI of Chinese Pharmacopoeia version in 2010 D high effective liquid chromatography for measuring.

The preparation precision of need testing solution takes by weighing medicated powder 1g, put in the brown volumetric flask of 25mL, accurate dimethyl sulfoxide-the acetonitrile (7: 3) that adds is an amount of, supersound process 15min, the above-mentioned solvent of reuse is settled to scale, shakes up, and filters with microporous filter membrane (0.45 μ m), discard filtrate just, get subsequent filtrate as need testing solution.

Two kinds of hybrid plans of table 6 make finished medicines powder content uniformity relatively

The result shows: even two scheme content averages are very approaching, but the content results intensity of hybrid plan one is apparently higher than hybrid plan two.Illustrate that the finished product medicated powder uniformity that hybrid plan one makes is better than hybrid plan two, so select the hybrid technique of hybrid plan one as preparation of the present invention.

3.6 the research of moulding process

3.6.1 finished product medicated powder bulk density is measured: take by weighing medicated powder and pack in the graduated cylinder, with weight divided by its average bulk density of volume calculations.Measurement result sees Table 7.

The measurement result of table 7 bulk density

We recipe quantity can be total to the 743.5g powder, dress up 2500 capsules, the powder charge thing is 0.2974g in then every capsules, its bulk density is 0.54g/ml, and shared volume is 0.5507mL, according to the capacity of all size capsule shells, No. 0 capsule approx. volume is 0.75mL, No. 1 capsule approx. volume is 0.55mL, selects No. 1, No. 0 capsule shells all can load the medicated powder of preparation of the present invention, so the capsule of preparation of the present invention is easy to realize.

3.6.2 the mensuration of finished product medicated powder angle of repose: adopt the fixed funnel method to measure the finished product medicated powder angle of repose that hybrid plan one makes, 5 times measurement result sees Table 8.

The measurement result of table 8 angle of repose

As seen from the above table, 5 times measurement result is all spent less than 35 angle of repose, shows this product good fluidity, is easy to carry out the packing of powder, is easy to direct filled capsules, and is suitable for particulate preparation.So preparation of the present invention is easy to be prepared into dosage forms such as powder, capsule, granule.

4 pilot scales: 10 times according to recipe quantity feed intake, and the detailed data of three batches of pilot products sees Table 9.

The detailed data of three batches of pilot products of table 9

The result shows: determined preparation technology is suitable for suitability for industrialized production.On conventional art, expensive thin medical materials such as Cinnabaris, Calculus Bovis, artificial Moschus are confined to the processing of loosing of small lot ball more, are complete manual the manufacturing, and quality can't standardization, limited (output≤1kg/ criticizes) in batches; Preparation technology provided by the present invention can realize the big production of scale, can will be amplified in batches more than 100kg/ criticizes, and minimum retail packaging unit＞100,000/batch.

Three, the research of method of quality control

The method of quality control of the internally heated preparation of a kind of children's's of treatment dyspepsia of the present invention is: described method of quality control mainly comprise in character, the discriminating project partly or entirely, inspection and assay; Wherein differentiate and comprise that the thin layer chromatography that preparation is carried out microscopical identification and Rhizoma Coptidis, Borneolum Syntheticum differentiates that assay is the assay to Calculus Bovis cultivating in the preparation (or In vitro cultured Calculus Bovis).Can realize the inspection of all main medicines among the other side by this method of quality control, thereby control the quality of preparation of the present invention effectively, all sidedly.Existing division is as follows:

1. character: determine that according to 3 batches of finished products of pilot scale the powder character of preparation of the present invention is: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

2. differentiate: (1) is the discriminating of microscopic features.It is an amount of to get this preparation, and the rearmounted microscopically of load is observed: prism of calcium oxalate is present in the parenchyma cell; Calcium oxalate sand crystal exists in the parenchyma cell; The wall of the thin cell of wood is beaded and thickens, and pit is obvious; Contain gelatinizing polysaccharide fragment of tissue chance iodine liquid and be brown or light brown brown, dissolve when meeting the heating of chloral hydrate liquid; Irregular sheet is colourless, has and peels off vestige layer by layer; Calcium oxalate cluster crystal is big, diameter 60～140 μ m; Reticulate vessel is bigger, non-lignify; The stone cell foresythia is similar round polygon or spindle, and the hole ditch is very thin, and the laminated striation that has is obvious; The body wall fragment is faint yellow to yellow, tool reticular texture and circular trichopore, visible sometimes sepia bristle; The body wall fragment is colourless, and there is superfine mycelium on the surface; The starch grain oval, diameter 35～48 μ m, the omphalion point-like, herringbone or horse-hof shape are positioned at a less end, and laminated striation is fine and closely woven and obvious; Starch grain pentagon or polygon, omphalion point-like or starlike often is stained with granular substance, is golden yellow or pale brown color; The dark brownish red of irregular fine particle, glossy, the edge dark yellow; Amorphous agglomerate is fallow, is embedded with tiny prism of calcium oxalate.

The powder of each medicine has its distinctive microscopic features in the side, can detect the existence of most medicines one by one according to these features, and is significant to control of product quality.Corresponding relation such as following table:

Microscopic features	Corresponding medicine
		Prism of calcium oxalate is present in the parenchyma cell; Calcium oxalate sand crystal exists in the parenchyma cell; The wall of the thin cell of wood is beaded and thickens, and pit is obvious; Contain gelatinizing polysaccharide fragment of tissue chance iodine liquid and be brown or light brown brown, dissolve when meeting the heating of chloral hydrate liquid.	Rhizoma Gastrodiae, Ramulus Uncariae Cum Uncis, dried tangerine peel
Calcium oxalate cluster crystal is big, diameter 60～140 μ m; Reticulate vessel is bigger, non-lignify; The stone cell foresythia is similar round polygon or spindle, and the hole ditch is very thin, and the laminated striation that has is obvious.	Radix Et Rhizoma Rhei, Rhizoma Coptidis
		The body wall fragment is colourless, and there is superfine mycelium on the surface; The body wall fragment is faint yellow to yellow, tool reticular texture and circular trichopore, visible sometimes sepia bristle.	Bombyx Batryticatus, Scorpio
The starch grain oval, diameter 35～48 μ m, the omphalion point-like, herringbone or horse-hof shape are positioned at a less end, and laminated striation is fine and closely woven and obvious; Starch grain pentagon or polygon, omphalion point-like or starlike often is stained with granular substance, is golden yellow or pale brown color.	Bulbus Fritillariae Thunbergii, the Rhizoma Pinelliae, Arisaema Cum Bile
		Irregular sheet is colourless, has and peels off vestige layer by layer; Amorphous agglomerate is fallow, is embedded with tiny prism of calcium oxalate.	The artificial Moschus
The dark brownish red of irregular fine particle, glossy, the edge dark yellow.	Cinnabaris, Concretio Silicea Bambusae

(2) differentiate for the thin layer chromatography of Rhizoma Coptidis in the side: get this preparation 1g, add methanol 10ml, supersound process 15 minutes, centrifugal, get the supernatant evaporate to dryness, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 10ml, shines medical material solution in pairs with legal system; Get the berberine hydrochloride reference substance again, add methanol and make the solution that 1ml contains 0.5mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1) is developing solvent, put in the chromatography cylinder of ammonia saturated with vapor, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle.

With Rhizoma Coptidis control medicinal material, berberine hydrochloride reference substance is contrast, carries out thin layer chromatography and differentiates that control of product quality is had important differential significance.

1. the preparation of need testing solution is with reference to " Rhizoma Coptidis of Chinese pharmacopoeia version in 2010 is differentiated the thin layer discriminating under the item, and adopting methanol is that solvent supersonic extracts, and the gained background is noiseless, the gained clear spot.

2. the selection of chromatographic condition is with reference to " developing solvent in the thin layer discriminating under a Rhizoma Coptidis discriminating of Chinese pharmacopoeia version in 2010 item, in the gained dot set, separating degree is good, and R _fValue is suitable.

3. experiment condition: lamellae silica gel G F ₂₅₄Lamellae, self-control, 110 ℃ of activation 30min;

Developing solvent benzene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1)

Launch 27 ℃ of temperature, relative humidity 60%, exhibition is apart from 12cm;

Rhizoma Coptidis control medicinal material, berberine hydrochloride reference substance are all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute;

Inspect under the inspection knowledge ultra-violet lamp (365nm);

4. as a result in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle; In the negative control chromatograph, noiseless with reference substance chromatograph relevant position.This discriminating detects effective, and method is feasible.

(3) differentiate for the thin layer chromatography of Borneolum Syntheticum in the side: get this preparation 1g, add ethyl acetate 10ml, supersound process 15 minutes, centrifugal, get supernatant as need testing solution; Other gets the Borneolum Syntheticum reference substance, adds ethyl acetate and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-acetone (9: 1) is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Ministerial drug in the Borneolum Syntheticum side of being, its powder does not have the microscopical identification feature, thus the thin layer chromatography discriminating of selecting for use the Borneolum Syntheticum reference substance to set up Borneolum Syntheticum for contrast, to control its quality.

1. the preparation of need testing solution employing ethyl acetate is that solvent supersonic extracts.

2. the selection toluene-acetone (9: 1) of chromatographic condition is developing solvent, sprays 5% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 105 ℃, and in the dot set, separating degree is good, and Rf value is about 0.5.

3. experiment condition: lamellae silica gel g thin-layer plate, self-control, 110 ℃ of activation 30min;

Developing solvent: toluene-acetone (9: 1) launches temperature: 24 ℃, and relative humidity: 72%, exhibition distance: 12cm;

Borneolum Syntheticum reference substance: available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute;

Inspection is known spray with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.

4. the result is in the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; In the negative control chromatograph, noiseless with reference substance chromatograph corresponding position.This discriminating detects effective, and method is feasible.

3. check: by " regulation under (appendix I B) powder item of Chinese pharmacopoeia version in 2010 is carried out conventional quality control to the powder of preparation of the present invention.

3.1 moisture: (lot number: 081101,081102,081103) the moisture check result is respectively 5.3%, 6.1%, 4.9% to three batches of powder samples of the present invention, and is all up to specification.

3.2 the inspection of content uniformity: (lot number: content uniformity check result 081101,081102,081103) is all up to specification for three batches of powder samples of the present invention.

3.3 granularity: (lot number: granularity 081101,081102,081103) checks that the result all can all meet the regulation of department of pediatrics medication to granularity by No. six sieves to three batches of powder samples of the present invention.

3.4 the outward appearance uniformity: three batches of powder samples of the present invention (lot number: 081101,081102,081103) all meet under an appendix IB of Chinese Pharmacopoeia version in 2010 the powder item regulation to the outward appearance uniformity.

3.5 microbial limit checks that according to an appendix X of Chinese Pharmacopoeia version in 2010 III C microbial limit test the result is all up to specification.

4. assay: the mensuration that is Calculus Bovis cultivating among the other side (or In vitro cultured Calculus Bovis).

The quality control of Calculus Bovis adopts the content (recording in one one of Chinese Pharmacopoeia version in 2010) of spectrophotometry Calculus Bovis mesobilirubin in the existing quality standard; The present invention adopts reversed phase high-performance liquid chromatography; set up the bilirubinic content assaying method of Calculus Bovis in the preparation of the present invention; reversed phase high-performance liquid chromatography has the separation efficiency height than spectrophotography; specificity is strong; advantages such as sensitivity height can be controlled the quality of Calculus Bovis cultivating in the preparation of the present invention (or In vitro cultured Calculus Bovis) better.This content assaying method and correlational study process thereof are as follows:

4.1 content assaying method:

4.2 chromatographic condition and specificity are investigated

4.2.1 instrument and reagent: instrument Baseline 810 System high performance liquid chromatographs (U.S. Waters company) comprise 510 type infusion pump (double pump), 7725i injector, 486 type UV-detector; KQ-50 type supersonic generator (city of Kunshan Dianshan Lake detecting instrument factory); Tianjin, island AUW 220D type double-range analytical balance (day island proper Tianjin company); Phs-2C type acidometer (Shanghai thunder magnetic instrument plant).Acetonitrile is chromatographically pure (U.S. Tedia company), and all the other reagent are analytical pure, and water is the new system purified water; Sample (lot number: 081101,081102,081103), bilirubin reference substance (lot number: 10077-0301 is for assay usefulness, available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute).

4.2.2 chromatographic condition: chromatographic column: Hypersil C18 (4.6mm * 250mm, 10 μ m) (Dalian Inst of Chemicophysics, Chinese Academy of Sciences); Mobile phase: dimethyl sulfoxide-acetonitrile-0.5mol/L Spirit of Mindererus. (regulating pH value with glacial acetic acid is 5.3) (6: 6: 7); Detect wavelength: 452nm; Column temperature: room temperature, flow velocity 1.0ml/min.With this understanding, bilirubin and other components all can reach baseline separation, separating degree (Rf)＞1.5.Calculate post with the bilirubin peak and imitate its theoretical cam curve 〉=1000.

4.3 the preparation of reference substance solution: precision takes by weighing the bilirubin reference substance 10.0mg that is dried to constant weight through phosphorus pentoxide, puts in the brown volumetric flask of 100ml, adds dimethyl sulfoxide-acetonitrile (7: 3) and makes dissolving, and be diluted to scale, shake up, product solution in contrast, concentration is 0.1mg/ml.

The preparation of need testing solution: precision takes by weighing this preparation 1g, put in the brown volumetric flask of 25mL, accurate dimethyl sulfoxide-the acetonitrile (7: 3) that adds is an amount of, supersound process 15min, the above-mentioned solvent of reuse is settled to scale, shakes up, filter with microporous filter membrane (0.45 μ m), discard filtrate just, get subsequent filtrate as need testing solution, its concentration and reference substance solution concentration basically identical.

4.4 the investigation of linear relationship: precision takes by weighing bilirubin reference substance 11.3mg, puts in the brown volumetric flask of 25ml, adds dimethyl sulfoxide-acetonitrile (7: 3), dissolves and is diluted to scale, shakes up.The above-mentioned solution 10ml of accurate absorption, do to wait doubly dilution successively with dimethyl sulfoxide-acetonitrile (7: 3), concentration is respectively 0.452,0.226,0.113,0.056 0.028mg/ml is by " 4.2 " described chromatographic condition sample introduction 10 μ L, measure peak area (A), with A concentration (C) is made linear regression, get regression equation: A=6253731C-11736, r=0.9999.As seen in 0.028～0.452mg/ml scope, the linear relationship of peak area and concentration is good.See Table 1.

The investigation result of table 1 linear relationship

4.5 precision experiment: No. 3 solution (concentration 0.113mg/ml) under the line taking sexual relationship item, continuous sample introduction 6 times, each 10 μ L measure peak area, calculate.The result: the RSD of peak area is 0.98%.

4.6 stability experiment: the powder sample (lot number: 081101) of getting preparation of the present invention, prepare need testing solution by " 4.3 " method, place down in room temperature (20～25 ℃), respectively 0,1,2, repeat sample introduction 3 times in the 4h different time sections, each sample introduction 10 μ L, measure peak area, the result: need testing solution is placed in the 4h basicly stable.

4.7 repeated experiment: precision take by weighing preparation of the present invention the powder sample (lot number: 081101), totally 6 parts, measure as stated above, the calculating average content is 3.0mg/g, RSD is 1.70%.

4.8 response rate experiment: precision takes by weighing bilirubin reference substance 14.5mg, puts in the brown volumetric flask of 50ml, dissolves and is diluted to scale, product storing solution in contrast with dimethyl sulfoxide-acetonitrile (7: 3).Precision takes by weighing the powder sample of the preparation of the present invention of known content of bilirubin (3.0mg/g), and (lot number: 081101) 0.5g is totally 9 parts, add reference substance storing solution 4.0 respectively, 5.0,6.0ml, then by the preparation method operation of need testing solution down in " 4.3 ", calculate the amount of recording by external standard method, the results are shown in Table 2.Bilirubin average recovery rate (n=3) is respectively 97.69%, 98.16%, 97.51%; RSD is respectively 1.34%, 1.07%, and 0.68%.

Table 2 bilirubin response rate experimental result mg, n=9

4.9 the formulation of sample determination and content limit: the powder sample by 7 batches of preparations of the present invention of " 4.1 " method mensuration the results are shown in Table 3.

Sample content of bilirubin (mg/g)=reference substance concentration * Ai ÷ Ar * diluted sample multiple ÷ sample sample weighting amount

Ai: test sample peak area integrated value; Ar: reference substance peak area integrated value.

Table 3 seven batch sample content of bilirubin measurement results

The result that seven batch samples record, its average content is 2.94mg/g, so determine content limit be: the every 1g of this preparation contains Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) with bilirubinometer, must not be less than 2.5mg.

5. stability test

For investigating this product time dependent rule under the influence of temperature, humidity, packing, the effect duration of setting up this product by stability test.According to relevant stability techniques requirement in " provisions for new drugs approval ", and in conjunction with " requirement of new Chinese medicine stability test ", test agent in three batches of this product is carried out stability study.

5.1 test method: testing three batch sample lot numbers is 081101,081102,081103; Content of the test is 6 months accelerated tests (40 ℃ ± 2 ℃, RH75% ± 5%), 30 months long term tests (25 ℃ ± 2 ℃, RH60% ± 10%); The investigation project is character, discriminating, inspection and assay.

5.2 accelerated test: 3 batches in sample, the Aluminum-plastic composite bag packing is 40 ℃ ± 2 ℃ in temperature, places under the condition of relative humidity 75% ± 5%.Take a sample respectively once the 0th, 1,2,3 and 6 the end of month at duration of test, press the study on the stability project and detect.

5.3 long term test: 3 batches in sample, Aluminum-plastic composite bag packing is 25 ℃ ± 2 ℃ in temperature, relative humidity 60% ± 10% condition under place.Take a sample respectively once the 0th, 6,12,18,24,30 the end of month at duration of test, press the study on the stability project and detect.

5.4 result: this product is after accelerated test (6 months) and long term test (30 months), its character, discriminating, inspection, assay are not all seen significant change, illustrate that this product has good stability in this packing (Aluminum-plastic composite bag packing), its effect duration is defined as 24 months.

6. in sum: method of quality control of the present invention has realized that all play inspection, discriminating and the assay of main therapeutical effect medicine among the other side, and comprised to preparation by " inspection routinely that Chinese pharmacopoeia is carried out, thus controlled the quality of preparation effectively, all sidedly.

Four, main pharmacodynamics data

1. the pharmacological research of single medicine in the preparation of the present invention

Preparation of the present invention is the most important with the effect of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), artificial Moschus, Borneolum Syntheticum, Cinnabaris; its influence to the full side of preparation of the present invention also is maximum; so this four Chinese medicine is wherein carried out pharmacology's analysis, therefrom excavates the mechanism of action of preparation of the present invention.

1.1 Calculus Bovis cultivating (or In vitro cultured Calculus Bovis): the bitter cold of the property of medicine is rushed down clearly, and causing resuscitation with aromatic drugs is gone into the heart, Liver Channel, both has been apt to cool liver, has cleared away heart-fire and relieving spasm by subduing liver-wind, and arresting convulsion is calmed the nerves, and clears away heart-fire, eliminates the phlegm, the refreshment of having one's ideas straightened out.Its main component contains bilirubin and cholic acid.Modern pharmacological research shows: Calculus Bovis has calmness, convulsion and refrigeration function, and the energy blood vessel dilating makes blood pressure drops, and the oral Calculus Bovis of rabbit can significantly increase the erythrocyte in the blood, and Detoxication is arranged, and Calculus Bovis also has antiinflammatory, anti-infective effect simultaneously.

1.2 artificial Moschus: the hot wet of the property of medicine is extremely fragrant, goes into heart spleen channel, and the property of walking to scurry is very strong, and both kind logical the closing of having one's ideas straightened out is the refreshment analepsia, controls the key medicine of closing the disease coma, and kind again promoting blood circulation to restore menstrual flow is opened the heap soil or fertilizer over and around the roots of meridians and held back.Modern pharmacological research shows: the artificial Moschus is dual regulation to the central nervous system, can alleviate cerebral edema significantly, strengthens the toleration of central nervous system to ischemia, improves cerebral circulation; The effect that its main component muscone has boosting and increases respiratory frequency anesthetized cat; In addition, the artificial Moschus also has antiinflammatory, effect such as antibacterial.

1.3 Borneolum Syntheticum: the property of medicine is arduous cool, goes into the heart, spleen, lung meridian, and its refreshment of having one's ideas straightened out is like the artificial Moschus and power is inferior.Modern pharmacological research shows: Borneolum Syntheticum topical application energy sensation nerve, can improve blood-brain barrier permeability rapidly, and, amount height long in the brain savings time, this causing resuscitation with aromatic drugs effect for Borneolum Syntheticum provides foundation; Borneolum Syntheticum still has antibacterial, antiinflammatory action, can prolong the time of mice anoxia enduring.

1.4 Cinnabaris: the property of medicine is sweet, is slightly cold, and goes into heart channel, tranquillization with heavy prescription, detoxifcation.Be used for the mind uneasiness, severe palpitation, insomnia, diseases such as infantile convulsion.The Cinnabaris mind of calming is applicable to the disease of various mind uneasinesses.As hyperactivity of heart-fire, dysphoria and insomnia, the medicines such as Rhizoma Coptidis, Magnetitum that can cooperate clearing away heart-fire for tranquillization are with using; As unconsciousness due to high fever, can join heat clearing away, the Calculus Bovis of having one's ideas straightened out, artificial Moschus and be equal to usefulness; As the hot infantile convulsion of expectorant, Concretio Silicea Bambusae, the Arisaema Cum Bile that can join the eliminating phlegm arresting convulsion are equal to usefulness.Modern pharmacological research shows: Cinnabaris can obviously reduce the awakening time of insomnia rat, faint from fear the excitation of antagonism amfetamine, pentetrazole class medicine due to the prolongation S sleep, postponement caffeine and sodium benzoate, promote the chloral hydrate hypnosis, bring into play angst resistance effect by reducing the synthetic of 5-HT or discharging.

2. the pharmaceutical research of the full side of preparation of the present invention

Cure mainly according to this side's function, in laboratory research the pharmacologically active of preparation of the present invention (powder), now be summarized as follows:

2.1 be subjected to the reagent thing: preparation of the present invention (powder), lot number 081101 is equivalent to crude drug 1g/g (listed dosage is the crude drug amount in the literary composition).

2.2 laboratory animal: Kunming mouse, the NIH mice, the SD rat is provided by Sichuan medical courses in general institute Experimental Animal Center, all meets healthy primary standard.Japan large ear rabbit is provided by Chengdu biologics factory of the Ministry of Agriculture.

2.3 method and result

2.3.1 refrigeration function

(1) vaccine is caused the rabbit that influences of fever in rabbits, male and female all have, body weight 2.4 ± 0.2kg, prohibit water 8h before the experiment, 1h measures normal body temperature (anus temperature) 2 times at interval, choose 2 temperature difference and be no more than 0.3 ℃, mean body temperature is 38.3～39.5 ℃ rabbit, and auricular vein is annotated Typhoid And Paratyphoid A, B Vaccine 0.5ml/kg, surveys body temperature behind the 1h again, get the fever in rabbit that heats up more than 0.9 ℃, evenly be divided into 4 groups by the height that heats up, 10 every group, two administration groups gavage preparation 6g/kg of the present invention respectively, 2g/kg, positive controls gavages 1.5% aspirin 10ml/kg (dosage 0.15g/kg), blank gavages the 10ml normal saline, and each group is all in 1,2,3,4, each take temperature of 5h 1 time is surveyed 5 times altogether, calculate and respectively organize each time point body temperature variation, the results are shown in Table 1.

Table 1 preparation of the present invention causes the influence of fever in rabbits to vaccine

With the same period blank group relatively: a:P＜0.05; B:P＜0.01; C:P＜0.001 (following table together)

Table 1 is the result show, 1h body temperature obviously raises behind the rabbit injection vaccine, preparation 5g/kg dosage group of the present invention body temperature behind administration 1h increases significantly decrease (P＜0.01), 2g/kg dosage group also behind administration 2h body temperature increase significantly decrease (P＜0.05), illustrate that preparation of the present invention causes fever in rabbits to vaccine significant refrigeration function is arranged.

(2) on Carrageenan causes the male rat that influences of rat fever, and body weight 155 ± 25g prohibits water 12h before the experiment, interval 1h measures normal body temperature (anus temperature) 2 times with the temperature indication, select 38～39 ℃ of mean body temperatures, the temperature difference is no more than 0.2 ℃ 36 of rats, is divided into 4 groups at random by normal body temperature.In the subcutaneous 1% carrageenin 0.1ml pyrogenicity of injecting respectively of every Mus two hind legs foot sole of the foot portion, press the gastric infusion of dosage shown in the table 2 subsequently, 4～8h after pyrogenicity measures each Mus body temperature 1 time every 1h with quadrat method, surveys altogether 5 times.

Table 2 on Carrageenan causes the influence of rat fever

With the same period blank group relatively: a:P＜0.05; B:P＜0.01

Table 2 is the result show, 4h body temperature obviously raises behind the rat injection carrageenin, and preparation 4g/kg dosage group of the present invention, 2g/kg dosage group all are lower than matched group at each fervescence of generating heat in period, have the trend of bringing down a fever significantly (P＜0.01, P＜0.05).

2.3.2 anti-inflammatory and analgesic effect

What (1) xylol caused mice ear influences 75 of male mices, is divided into 5 groups at random, and gastric infusion 3d continuously is that proinflammatory agent is measured and respectively organized the ear swelling degree with dimethylbenzene.The result: matched group, preparation high dose group of the present invention (6g/kg), middle dosage group (4g/kg), low dose group (2g/kg), positive control administration prednisone 20mg/kg ear swelling degree are respectively 10.9 ± 1.8,7.3 ± 2.6,7.8 ± 2.5,8.9 ± 2.1 and 7.6 ± 2.4mg.High, medium and low dosage of preparation of the present invention and 20mg/kg prednisone have remarkable antiinflammatory action, relatively are respectively P＜0.001, P＜0.01, P＜0.05 and P＜0.001 with matched group.

(2) influence 75 of male mices to what the mouse skin capillary permeability increased, be divided into 5 groups at random, continuous gastric infusion 3d, 1h after the last administration, every caudal vein is injected 1% azovan blue 0.1ml/10g, inject 0.1% histamine phosphate 0.1ml in the hypogastric region Intradermal immediately, epigastrium depilation district melted paraxylene 20 μ l, put to death mice behind the 20min, cut the scorching position of caused by dimethylbenzene xylene locus coeruleus skin, shred the back and immerse in the 2.5ml acetone normal saline, room temperature is placed 24h, centrifugal, get supernatant and put 721 spectrophotometer 610nm colorimetrics, directly compare with the OD value; Indigo plant is dyed degree (the indigo plant degree of dying is divided into 4 grades) around the hypogastric region skin of peeling off and overturn, each Mus histamine skin mound of staging comparison.The results are shown in Table 3.

The table 3 pair influence that the mouse skin capillary permeability increases

The result shows: increase for the caused mouse skin capillary permeability of dimethylbenzene, the high, medium and low dosage of preparation of the present invention all has remarkable inhibitory action, increases for capillary of skin permeability due to the histamine, and high, middle dosage has obvious inhibition effect.

(3) 50 of 18～22g mices are got in the Dichlorodiphenyl Acetate influence that causes pain mouse writhing reaction, male and female half and half, and grouping and administration see Table 4.Each treated animal every day according to dosage gastric infusion once, 4d continuously.Behind last administration 45min, the glacial acetic acid 0.2ml/ of every Mus lumbar injection 0.6% only, in the observed and recorded 15min mice pain turn round the body number of times, and calculate the analgesia suppression ratio.The results are shown in Table 4.

Table 4 Dichlorodiphenyl Acetate causes the influence of pain mouse writhing reaction

Result of the test shows: compare with the blank group, positive controls, preparation 6g/kg of the present invention, 4g/kg, 2g/kg group Dichlorodiphenyl Acetate induced mice painful writhing response have significant inhibitory effect, and results suggest preparation of the present invention has significant analgesic activity.

2.3.3 antibacterial action

(1) the in-vitro antibacterial test adopts " test tube doubling dilution " to measure the minimum inhibitory concentration (MIC) of preparation of the present invention to 8 kinds of common pathogens.The result shows that preparation of the present invention all has certain bacteriostasis to golden Portugal bacterium, first type and group B streptococcus, streptococcus pneumoniae, escherichia coli, dysentery bacterium, Bacillus proteus, bacillus pyocyaneus, and MIC is respectively 0.042,0.042,0.028,0.021,0.037,0.037,0.085,0.025g/ml.

(2) the protective effect mice that mice gold Portugal bacterium is infected is 75, and male and female half and half are divided into 5 groups at random by sex, body weight, continuously gastric infusion 3d.1h after the last administration, the above-mentioned golden Portugal of every Mus lumbar injection bacteria suspension (the prerun fatality rate is 90%) 0.5ml, 2h administration 1 time (dosage reduces by half) again after the microbiological contamination is observed in the 24h and is respectively organized the dead mouse situation.The result compares with the blank group, preparation 6,4 of the present invention, 2g/kg group, acetylspiramycin 1g/kg group (positive control) animal dead number is respectively 14/15,4/15,6/15,7/15,6/15, the mortality rate (comparing P＜0.01) of pointing out preparation of the present invention can significantly reduce mouse infection gold Portugal bacterium with the blank group.

2.3.4 antitussive action: 75 of male mices, be divided into 5 groups at random, 1h experimentizes after adopting ammonia to draw to cough the administration of method mouse stomach, accepts to take out the cough number of times in the record 2min after ammonia stimulates 1min.Result: matched group, preparation 6g/kg group of the present invention, the average cough number of times of 4g/kg group, 2g/kg group and codeine 30mg/kg group mice is respectively 44.1 ± 20.3,21.8 ± 22.4,31.3 ± 21.7,41.2 ± 23.6 and 19.8 ± 20.6 (P＜0.01) show that preparation high dose group of the present invention (6g/kg), middle dosage group (4g/kg) stimulate the mouse cough that causes that obvious antitussive action is arranged to ammonia.

2.3.5 phlegm-dispelling functions: 60 of mices, male and female all have, and are divided into 4 groups at random, and 30min behind the gastric infusion measures the phenol red concentration (with the OD value representation) of respectively organizing mouse breathing road irrigating solution with phenol red method.Matched group is 0.037 ± 0.019 as a result; Preparation 6g/kg group of the present invention is respectively 0.064 ± 0.019 (P＜0.01) and 0.051 ± 0.014 (P＜0.05) with the 4g/kg group; Ammonium chloride 2g/kg group is 0.060 ± 0.021 (P＜0.01), shows that preparation of the present invention can make the secretory volume of mouse breathing road mucosa obviously increase, and has significant phlegm-dispelling functions.

2.3.6 anti-hyperpyrexia convulsion effect: 100 male and female half and half of rat, body weight (50 ± 20) g is by inducing the hyperpyrexia convulsion rat model, randomly draw 15 groups in contrast, all the other 85 are screened, and convulsions person is taken place in the 5min, when fainting from fear beginning, take out, enter experimental group; Have 61 Mus in 85 Mus and enter experimental group; Experimental group is divided into A group, B, C, D group once more at random; B, C, D organize the normal saline solution that gives 6g/kg, 4g/kg, 2g/kg preparation of the present invention after each convulsions immediately; A group and blank group give with the volume normal saline solution; The next day, induces and faints from fear coinduction 10 times 1 time; Blank group is put 5min in 37 ℃ of water-baths, totally 10 times; The record rat is fainted from fear incubation period, convulsions persistent period.Rat convulsions result is induced in hot bath: rats in normal control group is moved about freely in 37.0 ℃ of water-baths, no abnormal variation before and after the water outlet; Hyperpyrexia convulsion group rat with afterbody and lower limb body support, moves about in 44.0 ℃ of hot water once in a while; As time goes on, rat begins irritated going up and jumps in hot water, blindly seeks, and is adherent once more upright after the several seconds, occurs nodding, forelimb clonic spasm, hind leg clonic spasm and develop into complete tetanus-grand mal; After the convulsive attack 6 times, behavior change appears in the part rat, shows as irritability, the strong or bradykinesia of aggressivity, the movable minimizing; Phenomenons such as screaming can appear in rat when convulsions intensity was high, respiratory rhythm is irregular, extremity cyanosis; A group, B group, C group, D group all induce febrile convulsion, and 3 grades of convulsions accounts for 31%, 4 grade of convulsions and accounts for 48%, 5 grade of convulsions and account for 21%; 6 rats are long because of the convulsions time, and mouth and nose secretions is more and die from convulsions respectively the 6th, 8,9 time; The results are shown in Table 5.

Table 5 is respectively organized the convulsions incubation period of rat and is fainted from fear the persistent period

The result shows: blank group and test group (6g/kg, 4g/kg, 2g/kg) rat relatively: fainting from fear has significant difference incubation period (P＜0.05, the persistent period difference of fainting from fear has significance (P＜0.05).The prolongation of latency of fainting from fear after the administration is described, the persistent period shortens, and shows that this medicine tool has control to faint from fear to take place to a certain extent and the effect of outbreak.

Above experimental result shows, preparation of the present invention has remarkable antipyretic effect to the caused fever in rabbits of TAB, the rat fever that on Carrageenan causes has remarkable antipyretic effect, rat paw edema and dimethylbenzene induced mice ear swelling due to the obvious inhibition carrageenin, the mouse skin capillary permeability that reduction histamine or dimethylbenzene cause increases, in addition, phenolsulfonphthalein test shows that this product can increase the secretion of respiratory mucosa, the performance phlegm-dispelling functions induces rat convulsions tool that the effect of generation of control convulsions and outbreak is arranged to a certain extent to hot bath.

Five, toxicological information

1. acute toxicity test

1.1 material: formulation samples lot number 081101 of the present invention, every 1g is equivalent to crude drug in whole 1g); Kunming kind white mice, male and female half and half, body weight 20 ± 2g, Sichuan University's West China medical college Experimental Animal Center provides, and meets healthy one grade standard.

1.2 experiment condition: in the experimentation, animal freely drinks water, feed, and room temperature keeps 18～21 ℃, relative humidity 70～80%, natural lighting, ventilation fan ventilates, and feedstuff is the mice particulate material, and Sichuan Province's medical courses in general institute Experimental Animal Center provides.

1.3 method and result: get 60 of mices, be divided into six groups at random, 10 every group by sex, body weight, high dose group gavages preparation of the present invention, and all the other are respectively organized medication and adopt the dilution of proportional diluted method, and dose ratio is 1: 0.85, administration, the toxic reaction and the death condition of observing animal in the week.Movable minimizing of mice or crawl volt are motionless after the administration, the ptosis, the perpendicular hair of part mice, dyspnea, increase toxic reaction with dosage and increase the weight of severe patient death, the postmortem immediately of dead mice, perusal main organs, no abnormality seen, survival mice are put to death after 7 days and are observed main organs, also no abnormality seen.Calculate the LD of mice oral administration ₅₀And 95% fiducial limit, the results are shown in Table 1:

Table 1 studies on acute toxicity result

Conclusion: preparation of the present invention is to the LD of mice oral administration ₅₀Be 197.6 ± 17.96 crude drugs/kg (for 109 times of a clinical consumption per day)

2. long term toxicity test

2.1 material: preparation of the present invention, the 0.9g/ bag, 1 0.9g, 2 times/d, promptly clinical consumption per day is a 1.8g/20kg body constitution amount, lot number: 081102; Laboratory animal is the healthy SD rat, male and female half and half, and body weight 150 ± 20g meets healthy primary standard.

2.2 method: get 104 of rats, be divided into 4 groups at random by the body constitution amount, male and female half and half, every group 25, be respectively high, medium and low dosage group and blank group, dosage is respectively 4.5g crude drug/(kgd), the 2.7g crude drug/(kgd), 0.9g crude drug/(kgd) (is equivalent to 50 times of clinical consumption per day respectively, 30 times, 10 times), the blank group gives normal saline solution 5ml, 1 time/d, gavage 60 days continuously.Per 2 weeks claim body constitution amounts 1 time before the administration and during the administration, and adjust dosage that experimental session is observed rat general state, sign, food ration, drinking-water and feces situation every day by the average body quality.Discovery has the animal of toxic reaction should take out single cage raising, primary part observation.Find dead or the postmortem in time of dying animal.Finish back 24 hours in the last administration, all rat is got blood from the eyelid vascular plexus and does routine blood test and hepatic and renal function detection, and each is organized 15 rats of drive and does the pathology inspection; Drug withdrawal 20 days, matched group and heavy dose of each remaining rat of group are got blood equally, put to death and make routine blood test, hepatic and renal function and pathologic finding.

2.3 the results are shown in Table 1,2,3.

The influence that table 1 pair rat body weight increases

Table 2 pair rat blood is learned the influence of index

The influence of table 3 pair rats'liver, renal function

The result shows by table 1: during the administration of high, medium and low dosage group and after the drug withdrawal 20 days, body weight gain was normal, with matched group there was no significant difference (P＞0.05) relatively.

The result shows by table 2: after 60 days, compare with matched group by erythrocyte, leukocyte, platelet count and content of hemoglobin in administration for high, medium and low dosage group, difference that there are no significant (P＞0.05).Drug withdrawal 20 days, the above index of heavy dose of group and matched group be no significant difference (P＞0.05) relatively.

Table 3 is the result show: high, medium and low dosage group was administration 60 days, and glutamate pyruvate transaminase, alkali phosphatase, blood urea nitrogen, creatine concentration and matched group compare in the blood, difference that there are no significant (P＞0.05).Drug withdrawal 20 days, the above index of heavy dose of group and matched group be no significant difference (P＞0.05) relatively.

Check pathological section, each administration group and matched group are not relatively seen obvious pathological change; Gastrointestinal mucosa is complete, no congestion and edema and damage; The lobules of liver structure is clear, and hepatocyte does not have the cloudy swelling degeneration; Renal cortex internal glomerulus and renal tubules do not have the cloudy swelling degeneration yet; Meninges is complete, no congestion and edema, and the arrangement and the cellular morphology of various types of cells are normal; Adrenal gland, spleen, testis and ovary are all no abnormal; Each group all has the lung of individual animal to have deposition blood stasis and inflammatory cell group fast, but not statistically significant, may be that medicine is choked due to the lung when irritating stomach.

2.4 conclusion: preparation of the present invention is with 4.5g crude drug/(kgd), 2.7g crude drug/(kgd), 0.9g crude drug/(kgd) (be equivalent to 50 times of clinical consumption per day respectively, 30 times, 10 times) gavage 60 days continuously for rat, to body weight gain, behavioral activity, routine blood test and hepatic and renal function all do not have obvious influence, and main organs does not also have obvious histopathology damage, 20 days no slowness toxic reactions illustrate this clinical drug trial dosage safety after the drug withdrawal.

Six, clinical data

Preparation of the present invention has carried out clinical research through 4 tame hospitals of domestic clinical pharmacology the Al-Qaeda terrorist organization, and the result is as follows:

1. the observation of curative effect that preparation for treating infantile hyperpyrexia of the present invention is fainted from fear

1.1 data and method

1.1.1 hyperpyrexia convulsion infant 72 examples (male 36 examples, women 36 examples), 7 months～8 years old age, diagnostic criteria: mainly according to FC (hyperpyrexia convulsion) diagnosis and the typing standard that propose in the neural academic conference of nineteen eighty-three whole nation children's.Include standard in: 1. clinical manifestation meets above-mentioned CFC diagnostic criteria; 2. had 1 outbreak in average per 3 months at least; 3. can adhere to following up a case by regular visits at least 6 months.Exclusion standard: 1. infant has liver, renal function and hematological abnormalities person; 2. to drug allergy person; 3. family members disagree with therapist.68 routine hyperpyrexia convulsion infants are divided into two groups at random: 38 examples (male 19 examples, women 19 examples) are organized in treatment, 1～8 years old age, The median age 2.8 years old, auxillary temperature (39.8 ± 0.5) ℃.Matched group 34 examples (male 17 examples, women 17 examples), 1.1～7.8 years old age, The median age 2.7 years old, auxillary temperature (39.7 ± 0.4) ℃.Aspect difference not statistically significants (P＞0.05) such as two groups of patients' sex, age, the degree of heat.

1.1.2 Therapeutic Method treatment group infant is taken preparation 0.09g/kg of the present invention, matched group gives antondin injection (dosage:＜2 years old 0.7mL, 2～7 years old 1～1.5mL, 1.5～2.0mL) intramuscular injection in 7～13 years old, diazepam (dosage: 0.5kg/mg, maximal dose is no more than 20mg) intramuscular injection.All treatment is 1 medication, and 1h, 2h observe the routine number of spasmolytic after administration, and 1h, 2h observe 1～2 ℃ of body temperature decline or reduce to normal routine number.

1.1.3 curative effect judging standard produce effects: medication 1h twitches and to stop, and 1h body temperature descends 1～2 ℃ or reduce to normal; Effectively: medication 2g twitches and to stop, and 2h endosome relaxing the bowels with purgatives of warm nature falls 1～2 ℃ or reduce to normal; Invalid: twitching behind the medication 1h does not alleviate even worsens, and body temperature does not have decline and rises on the contrary behind the 2h.

1.1.4 relatively adopt x between the statistical procedures group ²Check, there is statistical significance P＜0.05 for difference.

1.2 result

1.2.1 the whole infants of lab testing are all made conventional electroencephalogram behind the normal 7～10d of body temperature, unusual 8 examples (22.2%), and wherein 6 examples are the slow wave outburst, spike/sharp wave appears in 2 examples, basilic rhythm slowing down 1 example.CT/MRI checks that 12 examples are all no abnormal.

1.2.2 spasmolytic and bring down a fever relatively treatment group of effect after medication 1, the spasmolytic effect and 1 of 2h, the effect of bringing down a fever in the 2h be apparently higher than matched group, difference has statistical significance (P＜0.01).See Table 1.

The table 1 liang group spasmolytic curative effect relatively [example (%)] of bringing down a fever

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

1.3 discuss: hyperpyrexia convulsion is meant that infant shows effect period for the first time, and the prevalence of FC is reported as 2.2%～5% abroad, and China is 3.9%, in whole children, about 5%～6% FC took place, and was one of modal convulsions type of children's therefore.From this group result as can be seen, preparation administration of the present invention and the intramuscular injection antondin curative effect difference of bringing down a fever has statistical significance (P＜0.01), shows that preparation of the present invention has tangible anti-hyperpyrexia convulsion effect.

2. the observation of curative effect of preparation for treating children's dyspepsia of the present invention

2.1 physical data: 210 routine infants.Wherein male 110 examples, women 100 examples, 8 months～12 years old age.Case all meets the diagnostic criteria of child's dyspepsia.The course of disease is in 2d, and body temperature is more than 37.5 ℃.Be divided into two groups at random, each organizes 105 examples.

2.2 Therapeutic Method: treatment group (preparation of the present invention) adopted below 1 years old, and is oral, 2 times/d, and each 0.5g; 1～5 year old, oral, 2 times/d, each 0.9g; 6～10 years old, oral, 2 times/d, each 1.2g, 11～12 years old, oral, 2 times/d, each 1.5 administrated method.Matched group (valiant fighter's digestive liquid) adopted below 1 years old, and is oral, and 2 times/d, each 5ml; 1～5 year old, oral, 2 times/d, each 10ml; 6～10 years old, oral, 2 times/d, each 15ml; 11～12 years old, oral, 2 times/d, the method for administration of each 15ml.3d is 1 course of treatment.

2.3 criterion of therapeutical effect: observe medication 72h to judge curative effect, recovery from illness and produce effects, improvement are added up in the lump to effectively total.

2.4 sings and symptoms: observe and each group of record take medicine before and the variation of clinical symptoms such as abdominal distention of the 3d infant of taking medicine, vomiting sputum, lassitude, appetite decline, poor sleep, constipation due to dry stool, and between the effective percentage of each group symptom organized relatively.

2.5 result

2.5.1 two groups of curative effects relatively: the clinical symptoms change that infant is taken medicine behind the 3d sees Table 1,2, and comprehensive therapeutic effect relatively sees Table 3.

Table 1 liang group infant abdominal distention, vomiting sputum, listless clinical symptoms change be [n (%)] relatively

Annotate: compare with matched group: a:P＜0.05, b:P＜0.01.

Table 2 liang group infant appetite, sleep, stool clinical symptoms change be [n (%)] relatively

Annotate: compare with matched group: a:P＜0.05, b:P＜0.01.

The table 3 liang group clinical comprehensive therapeutic effect of infant [n (%)]

Annotate: compare with matched group: a:P＜0.05

2.5.2 adverse effect: treatment group, matched group have 1,3 infant when empty stomach is oral gastrointestinal reactions such as stomach upset and diarrhoea to be arranged respectively, do not influence and continue treatment, do not have irritated the reaction and take place.

The clinical symptoms of two groups of infants all obtains to a certain degree improvement behind the 3d 2.5.3 take medicine, but that the effective percentage of all indexs is organized in treatment is the highest, with matched group vomiting sputum effective percentage relatively, P＜0.01; Improvement effect listless to coma, sleep obviously is better than matched group, P＜0.01; Improvement effect treatment group to constipation due to dry stool is better than matched group, P＜0.05; Comparison to comprehensive therapeutic effect; The treatment group obviously is better than matched group, P＜0.05;

Clinical observation shows: utilize that heat syndrome has heat clearing away relieving convulsion in the preparation for treating children's dyspepsia of the present invention, the wind expectorant effect of loosing obviously is better than matched group improving on the comprehensive therapeutic effects such as clinical sign and total effective rate, is worth promoting the use in clinical.

The specific embodiment:

Embodiments of the invention 1: Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) 6g, artificial Moschus 1.5g, Borneolum Syntheticum 6g, Ramulus Uncariae Cum Uncis 120g, Rhizoma Gastrodiae 120g, Bombyx Batryticatus (parched with bran) 30g, Scorpio 30g, Rhizoma Coptidis 30g, Concretio Silicea Bambusae 30g, Cinnabaris 10g, Radix Et Rhizoma Rhei 30g, Arisaema Cum Bile (wine is processed) 30g, Bulbus Fritillariae Thunbergii 30g, Rhizoma Pinelliae (processed) 30g, tangerine 120g, Talcum 120g

The 6th step: final mixed medicated powder is carried out packing by certain loading amount specification, promptly get powder.

Embodiments of the invention 2: Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) 6g, artificial Moschus 1.5g, Borneolum Syntheticum 6g, Ramulus Uncariae Cum Uncis 120g, Rhizoma Gastrodiae 120g, Bombyx Batryticatus (parched with bran) 30g, Scorpio 30g, Rhizoma Coptidis 30g, Concretio Silicea Bambusae 30g, Cinnabaris 10g, Radix Et Rhizoma Rhei 30g, Arisaema Cum Bile (wine is processed) 30g, Bulbus Fritillariae Thunbergii 30g, Rhizoma Pinelliae (processed) 30g, tangerine 120g, Talcum 120g

The 6th step: final mixed medicated powder is made granule with the spray of boiling granulating machine with purified water or alcoholic solution, carry out packing by certain loading amount specification again, promptly get granule.

The embodiment of the invention 3: Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) 6g, artificial Moschus 1.5g, Borneolum Syntheticum 6g, Ramulus Uncariae Cum Uncis 120g, Rhizoma Gastrodiae 120g, Bombyx Batryticatus (parched with bran) 30g, Scorpio 30g, Rhizoma Coptidis 30g, Concretio Silicea Bambusae 30g, Cinnabaris 10g, Radix Et Rhizoma Rhei 30g, Arisaema Cum Bile (wine is processed) 30g, Bulbus Fritillariae Thunbergii 30g, Rhizoma Pinelliae (processed) 30g, tangerine 120g, Talcum 120g

The 6th step: will final mixed medicated powder spray and make granule, recharge and be capsule, or directly be filled to capsule, promptly get capsule with final mixed medicated powder with purified water or alcoholic solution with the boiling granulating machine.

The embodiment of the invention 4: described method of quality control comprises:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

The embodiment of the invention 5: described method of quality control can comprise:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

Differentiate: it is an amount of to get this preparation, and the rearmounted microscopically of load is observed: prism of calcium oxalate is present in the parenchyma cell; Calcium oxalate sand crystal exists in the parenchyma cell; The wall of the thin cell of wood is beaded and thickens, and pit is obvious; Contain gelatinizing polysaccharide fragment of tissue chance iodine liquid and be brown or light brown brown, dissolve when meeting the heating of chloral hydrate liquid; Irregular sheet is colourless, has and peels off vestige layer by layer; Calcium oxalate cluster crystal is big, diameter 60～140 μ m; Reticulate vessel is bigger, non-lignify; The stone cell foresythia is similar round polygon or spindle, and the hole ditch is very thin, and the laminated striation that has is obvious; The body wall fragment is faint yellow to yellow, tool reticular texture and circular trichopore, visible sometimes sepia bristle; The body wall fragment is colourless, and there is superfine mycelium on the surface; The starch grain oval, diameter 35～48 μ m, the omphalion point-like, herringbone or horse-hof shape are positioned at a less end, and laminated striation is fine and closely woven and obvious; Starch grain pentagon or polygon, omphalion point-like or starlike often is stained with granular substance, is golden yellow or pale brown color; The dark brownish red of irregular fine particle, glossy, the edge dark yellow; Amorphous agglomerate is fallow, is embedded with tiny prism of calcium oxalate.

The embodiment of the invention 6: described method of quality control can comprise:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

Differentiate: (1) gets this preparation 1g, adds methanol 10ml, and supersound process 15 minutes is centrifugal, gets the supernatant evaporate to dryness, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 10ml, shines medical material solution in pairs with legal system; Get the berberine hydrochloride reference substance again, add methanol and make the solution that 1ml contains 0.5mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1) is developing solvent, put in the chromatography cylinder of ammonia saturated with vapor, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle.

(2) get this preparation 1g, add ethyl acetate 10ml, supersound process 15 minutes, centrifugal, get supernatant as need testing solution; Other gets the Borneolum Syntheticum reference substance, adds ethyl acetate and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to an appendix VI of Chinese Pharmacopoeia version in 2010 B thin layer chromatography, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-acetone (9: 1) is developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing at 105 ℃.In the examination chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The embodiment of the invention 7: described method of quality control can comprise:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

The embodiment of the invention 8: described method of quality control can comprise:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.

Description of drawings: Fig. 1 process chart

Fig. 2 sucting wet curve figure

Fig. 3 bilirubin canonical plotting.

Claims

1. treat the internally heated preparation of children's's dyspepsia for one kind, it is characterized in that: calculate according to composition by weight: it is by 3～9 parts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis); 0.75～2.25 part of artificial Moschus; 3～9 parts of Borneolum Syntheticums; 60～180 parts of Ramulus Uncariae Cum Uncis; 60～180 parts in Rhizoma Gastrodiae; 15～45 parts of Bombyx Batryticatus (parched with bran); 15～45 parts of Scorpios; 15～45 parts of Rhizoma Coptidis; 15～45 parts of Concretio Silicea Bambusaes; 5～15 parts in Cinnabaris; 15～45 parts of Radix Et Rhizoma Rhei; 15～45 parts of Arisaema Cum Bile (wine is processed); 15～45 parts of Bulbus Fritillariae Thunbergiis; 15～45 parts of Rhizoma Pinelliae (processed); tangerine 60～180 parts; Talcum is prepared from for 60～180 parts.

2. according to the internally heated preparation of the described treatment of claim 1 children's's dyspepsia, it is characterized in that: calculate according to composition by weight: it is to be prepared from for 120 parts by 6 parts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), 1.5 parts of artificial Moschuss, 6 parts of Borneolum Syntheticums, 120 parts of Ramulus Uncariae Cum Uncis, 120 parts in Rhizoma Gastrodiae, 30 parts of Bombyx Batryticatus (parched with bran), 30 parts of Scorpios, 30 parts of Rhizoma Coptidis, 30 parts of Concretio Silicea Bambusaes, 10 parts in Cinnabaris, 30 parts of Radix Et Rhizoma Rhei, 30 parts of Arisaema Cum Bile (wine is processed), 30 parts of Bulbus Fritillariae Thunbergiis, 30 parts of Rhizoma Pinelliae (processed), 120 parts of dried tangerine peels, Talcum.

3. according to claim 1 or the internally heated preparation of 2 described treatment children's dyspepsia, it is characterized in that: described preparation is powder, granule, capsule.

4. according to the preparation method of heating agent in the arbitrary described treatment children's dyspepsia of claim 1～3, it is characterized in that:

The 6th step: final mixed medicated powder is made different preparations according to a conventional method.

5. according to the preparation method of heating agent in the described treatment of the claim 4 children's dyspepsia, it is characterized in that: powder is preparation like this:

6. according to the preparation method of heating agent in the described treatment of the claim 4 children's dyspepsia, it is characterized in that: granule is preparation like this:

Second step; Get Cinnabaris, add purified water with grinder and adopt elutriation to be ground into impalpable powder, cross 120 mesh sieves after 2 hours, add the first step gained medicated powder of 2 times of amounts of Cinnabaris again, ground 30 minutes with 40 ℃ of dryings of electric heating constant temperature oven, standby; Get Calculus Bovis cultivating (or In vitro cultured Calculus Bovis), be ground into impalpable powder, cross 120 mesh sieves after 2 hours, add the first step gained medicated powder of 2 times of amounts of Calculus Bovis cultivating (or In vitro cultured Calculus Bovis) again, ground 30 minutes with 60 ℃ of dryings of electric heating constant temperature oven with grinder, standby; Get the artificial Moschus, put into mortar, add a small amount of purified water (component meter by weight, artificial Moschus: water=3: 1), be ground into impalpable powder, cross 120 mesh sieves after 2 hours with 40 ℃ of dryings of electric heating constant temperature oven, the first step gained medicated powder that adds 2 times of amounts of artificial Moschus again ground 30 minutes, and is standby; Get Borneolum Syntheticum, add the first step gained medicated powder of 2 times of amounts, be ground into impalpable powder, cross 120 mesh sieves with high speed disintegrator, standby.

7. according to the preparation method of heating agent in the described treatment of the claim 4 children's dyspepsia, it is characterized in that: capsule is preparation like this:

The 3rd step; Get 5 flavor medical materials such as Bombyx Batryticatus (parched with bran), Scorpio, Arisaema Cum Bile (wine is processed), Bulbus Fritillariae Thunbergii, Rhizoma Pinelliae (processed), after 2 hours, mixing is ground into impalpable powder with high speed disintegrator respectively at 80 ℃ of dryings, crosses 120 mesh sieves, and is standby.

8. according to the detection method of heating agent in the described treatment of the claim 1～3 children's dyspepsia, described preparation is a powder, it is characterized in that: described detection method comprises character, discriminating, inspection and assay project; Wherein differentiate and comprise that the thin layer chromatography that preparation is carried out microscopical identification and Rhizoma Coptidis, Borneolum Syntheticum differentiates that assay is the assay to Calculus Bovis cultivating in the preparation (or In vitro cultured Calculus Bovis); This detection method specifically comprises following project:

Character: product is lurid powder; Gas is fragrant cool, bitter in the mouth.