CN102008704A

CN102008704A - Detection method for composition having middle-warming stomach harmonizing function

Info

Publication number: CN102008704A
Application number: CN2010105936503A
Authority: CN
Inventors: 付立家; 付建家
Original assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Current assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Priority date: 2007-05-23
Filing date: 2007-05-23
Publication date: 2011-04-13
Anticipated expiration: 2027-05-23
Also published as: CN102008704B

Abstract

The invention discloses a pharmaceutical composition detection method, material drugs of the pharmaceutical composition are as follows: elecampane, fructus amomi, white atractylodes rhizome, pericarpium citri reticulatae, Poria cocos, Pinellia ternate(prepared), rhizoma cyperi, immature bitter orange, Amomum kravanh, bark official magnolia, Pogostemon cablin and liquorice, and the detection method detects content of Costundide by high performance liquid chromatography.

Description

A kind of detection method with compositions of warming middle-JIAO for easing the stomach effect

The present invention is for dividing an application, and the original bill application number is 200710099505.8, and the original bill applying date is on 05 23rd, 2007, and the original bill name is called composition and method of making the same and the method for quality control with warming middle-JIAO for easing the stomach effect.

Technical field

The present invention relates to a kind of Chinese medicine composition and preparation method thereof and method of quality control, particularly relate to a kind of Chinese medicine composition and preparation method thereof and method of quality control with warming middle-JIAO for easing the stomach effect.

Background technology

Taste are ill, certainly will influence the source of nutrition of human body, also are directly connected to the generation of disease, development and prognosis.So-called " the normal gas of normal person is reported in stomach, the normal gas of stomach person normal person also, people's extreme hypofunction of the stomach day is contrary, and is dead against the person." gastropathy should cause that people's attention is taken precautions against early.

Motherland's medical science is thought: diseases caused by exogenous pathogenic factor cold-evil, not only the cloudy gas of easy damaged taste influence spleen fortuneization ability, also easily making cold stagnate in, hinder the operation of gas, cold is invaded between the stomach lung, makes the blood unable to walk, gas can not lead to, solid and generation is trembled with fear and is ached.Pathology: abdominal pain due to accumulation of cold, meet cold rnning and weigh, must warm up then and relax the abdominal distention discomfort.Modern scholar thinks, people's stomach some near stomach wall, cold air is if directly invade and epigastrium, but reflexive ground causes stomach and vasoconstriction thereof, the stomach motor function gets muddled, thereby produces spasmodic colic, stomach feeling of repletion, poor appetite, even feel sick vomiting.

Gastropathy is caused by multiple reason, hyperchlorhydria, and pepsinia increases, nerve and endocrine dysfunction, the gastrin secretion increases, drinking and eating irregularly, smoking, reasons such as excessive drinking and gastrointestinal antibacterial and unusual procreation.General following a few class medicines (1) antacid: relief of symptoms.(2) histamine's receptor blocking agent generally has side effect to kidney.(3) anticholinergic agents, price is expensive, and side effect is big, should not take for a long time.(4) gastric mucosal protection medicine and improve medicine for stomach dynamic.General antacid, medication is adhered in protection gastric mucosa medicine and antimicrobial drug collocation; can not frequently change dressings the short time, medication after meal and medication ante cibum will the difference effects midway, and anticholinergic should not share by the medicine for stomach dynamic opposite with effect; due to illness and different and, constantly grope curative effect.

Therefore inventing a kind of medicine with warming middle-JIAO for easing the stomach effect is necessary.

Summary of the invention

The object of the invention is to provide a kind of Chinese medicine composition with warming middle-JIAO for easing the stomach effect;

The object of the invention also is to provide a kind of Chinese medicine composition preparation method with warming middle-JIAO for easing the stomach effect;

The object of the invention also is to provide a kind of method of quality control with Chinese medicine composition of warming middle-JIAO for easing the stomach effect.

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

Chinese medicine composition with warming middle-JIAO for easing the stomach effect of the present invention is to be made by the crude drug of following weight ratio:

Radix Aucklandiae 30-100g Massa Medicata Fermentata 150-300g Rhizoma Atractylodis Macrocephalae 50-200g

Pericarpium Citri Reticulatae 50-200g Poria 50-200g Rhizoma Pinelliae (processed) 50-200g

Rhizoma Cyperi (vinegar system) 30-100g Fructus Aurantii Immaturus (stir-fry) 30-100g Fructus Amomi Rotundus (shelling) 30-100g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 30-100g Fructus Chaenomelis 30-100g Rhizoma Zingiberis Recens 10-60g

Fructus Jujubae 20-100g;

The above-mentioned raw materials optimum ratio is:

Radix Aucklandiae 70g Massa Medicata Fermentata 200g Rhizoma Atractylodis Macrocephalae 100g

Pericarpium Citri Reticulatae 110g Poria 110g Rhizoma Pinelliae (processed) 110g

Rhizoma Cyperi (vinegar system) 60g Fructus Aurantii Immaturus (stir-fry) 60g Fructus Amomi Rotundus (shelling) 60g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 60g Fructus Chaenomelis 60g Rhizoma Zingiberis Recens 30g

Fructus Jujubae 50g;

Chinese medicine composition with warming middle-JIAO for easing the stomach effect of the present invention can be made by the crude drug of following weight ratio:

Radix Aucklandiae 30-100g Fructus Amomi 30-100g Rhizoma Atractylodis Macrocephalae 50-200g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 30-100g Herba Pogostemonis 30-100g Radix Glycyrrhizae 10-90g

Rhizoma Zingiberis Recens 10-60g Fructus Jujubae 20-100g;

The above-mentioned raw materials optimum ratio is:

Radix Aucklandiae 30-50g Fructus Amomi 30-50g Rhizoma Atractylodis Macrocephalae 120-200g

Pericarpium Citri Reticulatae 120-200g Poria 120-200g Rhizoma Pinelliae (processed) 120-200g

Rhizoma Cyperi (vinegar system) 30-60g Fructus Aurantii Immaturus (stir-fry) 30-60g Fructus Amomi Rotundus (shelling) 30-60g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 30-60g Herba Pogostemonis 30-60g Radix Glycyrrhizae 50-90g

Rhizoma Zingiberis Recens 20-40g Fructus Jujubae 40-80g;

Compositions of the present invention technology adding adjuvant is routinely made clinical acceptable forms such as tablet, capsule, oral liquid, drop pill, spray, granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.

The preparation method of Chinese medicinal composition granules of the present invention is:

The Rhizoma Pinelliae and Rhizoma Zingiberis Recens are made solvent with the 60-80% ethanol that medical material 5-7 doubly measures, and flood after 20-30 hour, and with the speed percolation of per minute 1～6ml, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubaes such as medicinal residues and all the other Poria decoct with water each 1-2.5 hour 2-3 time, collecting decoction, filter, filtrate and above-mentioned aqueous solution merge, and concentrate relative density 1.10 (50～55 ℃), put cold, add equivalent ethanol, left standstill 20-30 hour, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, and being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, cane sugar powder 1-3 part, dextrin 1-3 part, ethanol are an amount of, make granule, drying adds above-mentioned volatile oil, mixing, promptly.

Pharmaceutical composition method of quality control of the present invention comprises one or more in following discrimination method and/or the assay:

Differentiate:

(1) get the quite described composition material medicine of preparation 28-32g, add ethanol-strong ammonia solution (1-2: 1-2) 5ml, moistening was placed 15-30 minute, add chloroform 18-25ml, supersound process 15-30 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (10-15: 4-9: 2-6: 2-5: be developing solvent 1), put in the chromatography cylinder of ammonia saturated with vapor, launch, take out, dry, spray is with the 0.4-0.7% ethanol solution of ninhydrin, 105 ℃ of bakings about 4-7 minute; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical purple dot;

(2) get the quite described composition material medicine of preparation 10g,, porphyrize adds water 20ml, chloroform 20ml, hydrochloric acid 3ml, and reflux 1-2 hour, divide and get chloroform layer, wash with water to neutrality, volatilize, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other removes hydrogen constuslactone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw control medicinal material solution 1 μ l, sample solution 2～4 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-toluene-ethyl acetate-glacial acetic acid (8-12: 18-22: 5-8: 0.4-0.7) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) get the quite described composition material medicine of preparation 28-32g, porphyrize, add water 50ml dissolving, add diethyl ether and 2-4 time (20-30ml that at every turn adds diethyl ether, saturated nacl aqueous solution 5-15ml) of saturated nacl aqueous solution extraction, divide and get ether solution, room temperature volatilizes, residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1-2 hour, filter, the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (30～60 ℃)-acetone-ethyl acetate (80-90: 12-18: 1) be developing solvent, presaturation 10-20 minute, launch, take out, dry, uviol lamp is observed (365nm) down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

(4) get the quite described composition material medicine of preparation 10g,, porphyrize, the 30ml that adds diethyl ether, reflux 25-35 minute, put coldly, filter, filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets magnolol, honokiol reference substance, adds methanol respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw the about 10 μ l of need testing solution, reference substance solution 8 μ l, put respectively on same silica gel g thin-layer plate, with toluene-methanol (25-30: 1) be developing solvent, launch, take out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

(1) according to high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler (4.6 * 150mm, 5 μ m) with octadecylsilane chemically bonded silica; Methanol-water (10-16: 6-9) be mobile phase; The detection wavelength is 225nm, and theoretical cam curve is calculated by costunolide should be not less than 3000;

The preparation of reference substance solution: the accurate Rhizoma Beesiae Calthaefoliae hydrocarbon lactone reference substance 1mg that claims, put in the 25ml measuring bottle, add the ethyl acetate dissolving, and be diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), promptly;

The preparation of need testing solution: get the quite described composition material medicine of preparation 20g,, put in the tool plug Erlenmeyer flask, the accurate chloroform 100ml that adds, close plug shakes up, and claims to decide weight, and placement is spent the night, supersound process (power 200-400W, frequency 40KH _Z) 20-40 minute, take out, put cold, close plug claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shake up, filter, precision is measured subsequent filtrate 20ml, put in the evaporating dish, volatilize, residue adds an amount of methanol, slight fever makes dissolving, is transferred in the 5ml measuring bottle, and is diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), promptly;

Algoscopy: accurate respectively reference substance liquid, each 10 μ l injection chromatograph of liquid of test sample liquid drawn, measure, promptly.

(2) according to high effective liquid chromatography for measuring

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With acetonitrile-water-phosphoric acid (18: 82: 0.1) is mobile phase; The detection wavelength is 283nm, and number of theoretical plate is pressed naringin and calculated not the end in 2500;

It is an amount of that the preparation of reference substance solution is taken at 110 ℃ of naringin reference substances that are dried to constant weight, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 80 μ g, promptly;

The preparation of need testing solution: get the quite described composition material medicine of preparation 10g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, supersound process (power 250W, frequency 40kHz) 45 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5ml, put in the 10ml measuring bottle, add methanol and be diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly;

Algoscopy: accurate respectively reference substance solution and the need testing solution 10 μ l of drawing, inject chromatograph of liquid, measure, promptly.

Pharmaceutical composition method of quality control of the present invention is preferably as follows one or more in discrimination method and/or the assay:

Differentiate:

(1) get present composition granule 15g, add ethanol-strong ammonia solution (1: 1) 5ml, moistening was placed 20 minutes, added chloroform 20ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1) is developing solvent, puts in the chromatography cylinder of ammonia saturated with vapor, launches, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, about 5 minutes of 105 ℃ of bakings; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical purple dot;

(2) get present composition granule 5g, porphyrize adds water 20ml, chloroform 20ml, hydrochloric acid 3ml, and reflux 1 hour is divided and got chloroform layer, washes with water to neutrality, volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other removes hydrogen constuslactone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw control medicinal material solution 1 μ l, sample solution 2～4 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-toluene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) get present composition granule 15g, porphyrize adds water 50ml dissolving, add diethyl ether and three times (25ml that at every turn adds diethyl ether, saturated nacl aqueous solution 10ml) of saturated nacl aqueous solution extraction, divide and get ether solution, room temperature volatilizes, residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1 hour filters, and the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (30～60 ℃)-acetone-ethyl acetate (84: 15: 1) is developing solvent, presaturation 15 minutes launches, and takes out, dry, uviol lamp is observed (365nm) down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

(4) get present composition granule 5g, porphyrize, the 30ml that adds diethyl ether, reflux 30 minutes is put coldly, filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets magnolol, honokiol reference substance, adds methanol respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw the about 10 μ l of need testing solution, reference substance solution 8 μ l, put respectively on same silica gel g thin-layer plate, with toluene-methanol (27: 1) is developing solvent, launches, and takes out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

(1) according to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test are filler (4.6 * 150mm, 5 μ m) with octadecylsilane chemically bonded silica; Methanol-water (13: 7) is a mobile phase; The detection wavelength is 225nm, and theoretical cam curve is calculated by costunolide should be not less than 3000;

The accurate Rhizoma Beesiae Calthaefoliae hydrocarbon lactone reference substance 1mg that claims of the preparation of reference substance solution puts in the 25ml measuring bottle, adds the ethyl acetate dissolving, and is diluted to scale, shakes up, and filters with microporous filter membrane (0.45 μ m), promptly;

It is an amount of that granule is got under the present composition granule content uniformity in the preparation of need testing solution, porphyrize, and precision takes by weighing about 10g, puts in the tool plug Erlenmeyer flask, the accurate chloroform 100ml that adds, close plug shakes up, and claims to decide weight, placement is spent the night, supersound process (power 250W, frequency 40KH _Z) 30 minutes, take out, put cold, close plug claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shake up, filter, precision is measured subsequent filtrate 20ml, put in the evaporating dish, volatilize, residue adds an amount of methanol, slight fever makes dissolving, is transferred in the 5ml measuring bottle, and is diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), promptly;

(2) according to high effective liquid chromatography for measuring

The preparation of need testing solution: get this product under the content uniformity item, porphyrize is got 5g, and accurate the title decides, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, supersound process (power 250W, frequency 40kHz) 45 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5ml, put in the 10ml measuring bottle, add methanol and be diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly;

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

Described composition quality control method can be applied to the various dosage forms of compositions, as clinical acceptable forms such as tablet, capsule, oral liquid, drop pill, spray, granules, each dosage form is when carrying out quality control, the all unified conversion of selected sample size is the amount of crude drug, as " get preparation be equivalent to as described in composition material medicine 10g ", as granule promptly quite be: composition granule 5g.

The present composition has good drug effect, compares existing preparation and shows good drug effect.The method of quality control of Chinese medicine composition provided by the present invention, be by obtaining behind the creative experiment sieving of big measuring, pass through screening in the discrimination method to sample treatment, the selection of developing solvent, make and differentiate that specificity is fine, and method is economic and practical, the result is quick, and can both use different lamellaes.Pass through screening in the content assaying method to sample, test sample processing method, the selection of developing solvent, make content assaying method effectivelyly to carry out quality control, and will compare more stable that product that additive method measures shows on pharmacological effect with the product that this method is measured to product.

The specific embodiment

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

Choose medicine group of the present invention and carry out pharmacodynamic experiment research: medicine group I of the present invention (embodiment 1 preparation); Medicine group II of the present invention (embodiment 7 preparations)

The influence of 1 pair of rat gastric juice of experimental example secretory function

70 of rats are divided into 7 groups at random, and first, second, third group are irritated stomach medicine group of the present invention I suspension 2.0,3.0,4.0g/kg respectively, fourth, penta, oneself group are irritated pharmaceutical preparation 2.0,3.0, the 4.0g/kg of stomach medicine group of the present invention II, heptan, group was irritated stomach with the volume normal saline, successive administration 7 days, fasting 24 hours (can't help water) after the last administration, etherization, open the abdominal cavity, the ligation pylorus injects above-mentioned various medicine 1.8ml (0.9g) through duodenum, sew up the abdominal cavity then, fasting was prohibited water 5 hours.The reuse etherization is opened the abdominal cavity after 5 hours, and the ligation cardia takes off stomach, filter gastric juice with two layers of cloth respectively, to the scale test tube, with the centrifugal 15min of 3000r/min, record supernatant liquid measure is a gastric juice, uses acidometer xylometric measurement, the special capillary test method of wheat respectively, measures free acidity.Total acidity and pepsin activity the results are shown in following table:

Influence to rat gastric juice secretory function

Annotate: ^*P＜0.01

The result shows: invention medicine group I and medicine group II of the present invention all can promote in Mus gastric secretion raising free acidity and total acidity output, compare with the normal saline matched group, pepsin activity is not made significant difference, and the effect of invention medicine group I same dose group is better than medicine group II of the present invention.

The influence of 2 pairs of mouse small intestine ahead runnings of experimental example

70 of mices, be divided into 7 groups at random, first, second, third group are irritated stomach invention medicine group I suspension 4.0,6.0,8.0g/kg respectively, fourth, penta, oneself group are irritated stomach embodiment 7 suspension 6.0g/kg, heptan, group was irritated stomach with the volume normal saline, successive administration 7 days, fasting 24 hours (can't help water) after the last administration, irritating the Weishang respectively states medicine and is made into physiological water and contains each suspension 0.2ml/10g body weight of 10% of carbon powder and arabic gum, with the cervical vertebra dislocation method mice is put to death after 15 minutes, cut the abdominal cavity open, take out gastrointestinal tract.Cut off attached to the mesentery on the intestinal tube, intestinal tube is not added traction ground be tiled in (a little saline on the glass plate) on the glass plate lightly.With the pylorus is starting point, measures the displacement of charcoal end in intestinal tube and the total length of small intestinal (from pylorus to ileocecus), and the displacement of calculating every mice charcoal end accounts for small intestinal total length percentage rate, i.e. intestinal propulsion rate the results are shown in Table:

Group	Dosage (g/kg)	Intestinal propulsion rate (%)
			N.S		60.54±5.21
Invention medicine group I	4.0	81.18±4.73 ^**
			Invention medicine group I	6.0	82.27±5.48 ^**
Invention medicine group I	8.0	84.23±4.15 ^**
			Medicine group II of the present invention	4.0	72.23±4.29 ^*
Medicine group II of the present invention	6.0	73.87±5.09 ^*
			Medicine group II of the present invention	8.0	76.47±4.89 ^*

Annotate: ^*P＜0.01; ^*P＜0.05

The result shows: invention medicine group I can promote the motion of mouse small intestine, with the normal saline matched group significant difference P＜0.01 is arranged relatively, and medicine group II of the present invention also has the mouse small intestine of promotion motion effect, but invention medicine group I effect is not strong.

The influence of experimental example 3 Dichlorodiphenyl Acetate type gastric ulcers

70 of rats, be divided into 7 groups at random, fasting is 24 hours then, can't help water, under etherization, cut abdomen open by the sterile working and draw stomach, behind the 2 dipping glacial acetic acid of the Lu scraps of paper with diameter 5mm, be posted on the 30Second of serous coat place, greater gastric curvature both sides, remove the Lu scraps of paper rapidly, and dry with rayon balls, the reduction body of stomach is sewed up stomach wall.Postoperative begins feed and gastric infusion next day, grouping, dosage, through medicine with experiment 1, fasting is 24 hours after the last administration, dislocation is put to death, cut open the belly immediately and get stomach and use 1% formalin fixed, write down gastric pathological changes situation, with ulcer length summation (mm) as ulcer index, carry out statistical procedures, the results are shown in Table:

The influence of Dichlorodiphenyl Acetate type gastric ulcer

Group	Dosage (g/kg)	Ulcer index (mm)
			N.S		140±2.83
Invention medicine group I	2.0	5.6±0.12 ^***
			Invention medicine group I	3.0	2.9±0.10 ^***
Invention medicine group I	4.0	3.2±0.11 ^***
			Medicine group II of the present invention	2.0	5.4±0.12 ^***
Medicine group II of the present invention	3.0	3.0±0.13 ^***
			Medicine group II of the present invention	4.0	3.9±0.13 ^***

Annotate: ^{* *}P＜0.001

The result shows: medicine group of the present invention has the obvious suppression effect to rat acetic acid type gastric ulcer, with the normal saline matched group significant differences P＜0.001 is arranged relatively, and it does in order to the invention medicine group I effect of 3.0g/kg best.

Experimental example 4 is differentiated screening experiment

We get the different preparations of medicine of the present invention, carry out microscopical identification respectively, prove that this discrimination method is stable, are applicable to that the thin layer of all preparations under this its preparation process is differentiated.

The thin layer discrimination method of 1 Radix Aucklandiae:

1) selection of different need testing solution preparation methoies:

Need testing solution one: get pharmaceutical preparation 5g of the present invention, porphyrize adds water 20ml, chloroform 20ml, hydrochloric acid 3ml, and reflux 1 hour is divided and got chloroform layer, washes with water to neutrality, volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution;

Need testing solution two: get pharmaceutical preparation 15g of the present invention, porphyrize adds 60-90 ℃ of petroleum ether 50ml, flooded 25-35 minute, and supersound process 25-35 minute, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution.

2) different developing solvents and ratio:

Developing solvent one: petroleum ether (60-90 ℃)-benzene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5); Developing solvent two: cyclohexane extraction-acetone (10: 1).

3) selection of different contrast solutions

Contrast solution one: the dehydrocostuslactone reference substance adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution;

Contrast solution two: Radix Aucklandiae control medicinal material 0.5g, add ethyl acetate 10ml, supersound process 25-35 minute, filter, filtrate is medical material solution in contrast.

According to the thin layer chromatography test, put above-mentioned two kinds of need testing solutions, two kinds of each 5 μ l of contrast solution respectively at two silica gel g thin-layer plates, with above-mentioned two kinds of different developing solvents, launch, take out, to dry, spray is with 1% vanillin ethanol solution of sulfuric acid, and hot blast blows to clear spot.Compare the color developing effect of the need testing solution of two kinds of extracting method preparations, the results are shown in following table:

As can be seen from the above table, adopt the color developing effect of need testing solution one, developing solvent one good, the Pass Test requirement.And reference substance solution is good than control medicinal material solution color developing effect, so two kinds of methods are all feasible.

2) developing solvent proportioning preferred in the discrimination method of the above-mentioned Radix Aucklandiae: draw the need testing solution 10 μ l of preparation as stated above, reference substance solution one, two each 5 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 ℃)-toluene-ethyl acetate-glacial acetic acid proportioning is (5: 30: 2: 0.5) (5: 25: 7: 0.5) (10: 20: 7: 0.5) (15: 15: 7: 0.5) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, it is clear that hot blast blows to the speckle colour developing, puts respectively under daylight and the ultra-violet lamp (365nm) and inspect.Observe the unfolded effect of test sample on each lamellae, the results are shown in following table:

The developing solvent proportioning	5∶30∶2∶0.5	5∶25∶7∶0.5	10∶20∶7∶0.5	15∶15∶7∶0.5
					Rf value	0.38	0.40	0.45	0.78

Developing solvent proportioning as can be seen from the above table is 10: 20: 7: 0.5 o'clock, it is best that need testing solution launches effect, and appearance hangover, speckle separate phenomenons such as bad.

3) mensuration of dehydrocostuslactone detectability in the discrimination method of the above-mentioned Radix Aucklandiae contains 0.03mg, 0.05mg, 0.1mg, 0.2mg in every 1ml reference substance solution.Reference substance solution one, two each 1 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 ℃)-toluene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, it is clear that hot blast blows to the speckle colour developing, puts respectively under daylight and the ultra-violet lamp (365nm) and inspect.Observe the effect of test sample principal spot colour developing on the lamellae, the results are shown in following table:

Test sample concentration is when 0.1mg/ml as can be seen from the above table, and it is clear to develop the color on lamellae, is fit to test requirements document.So with 10 ^-6G is decided to be the detectability of dehydrocostuslactone.

4) sample solution concentration preferred in the discrimination method of the above-mentioned Radix Aucklandiae contains this product 1g, 3g, 0.5g, 0.7g in every 1ml need testing solution.Reference substance solution one, two each 1 μ l need testing solution, 3 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60-90 ℃)-toluene-ethyl acetate-glacial acetic acid (27: 53: 19: 1) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, it is clear that hot blast blows to the speckle colour developing, puts respectively under daylight and the ultra-violet lamp (365nm) and inspect.Observe the effect of test sample principal spot colour developing on the lamellae, the results are shown in following table:

Test sample concentration is when 0.5g/ml as can be seen from the above table, and it is clear to develop the color on lamellae, is fit to test requirements document.So 5g/ml is decided to be the test sample concentration of dehydrocostuslactone.

4) negative control test

Get the negative sample that lacks the Radix Aucklandiae, prepare negative control solution, launch the back and on reference substance solution one corresponding position, corresponding speckle do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method (2).And corresponding light speckle is arranged on reference substance solution two correspondence positions, illustrate that selected identification experiment does not have specificity, so get rid of contrast two.

2, the thin layer discrimination method of Poria:

1) selection of different need testing solution preparation methoies:

Need testing solution one: get pharmaceutical preparation 15g of the present invention, porphyrize adds water 50ml dissolving, add diethyl ether and three times (25ml that at every turn adds diethyl ether, saturated nacl aqueous solution 1ml) of saturated nacl aqueous solution extraction, divide and get ether solution, room temperature volatilizes, residue adds normal hexane 0.5ml makes dissolving, as need testing solution;

Need testing solution two: get pharmaceutical preparation 5-15g of the present invention, the 40-60ml that adds diethyl ether, reflux, extract, 20-40 minute, filter, filtrate volatilizes, and residue adds the 0.5-2ml normal hexane makes dissolving, as need testing solution.

Need testing solution three: get pharmaceutical preparation 5-15g of the present invention, the 25-75ml that adds diethyl ether, reflux, extract, 15-45 minute, filter, filtrate volatilizes, and residue adds the 0.5-1.5ml normal hexane makes dissolving, as need testing solution.

2) selection of different contrast solutions

Contrast solution one: the Poria control medicinal material, the 50ml that adds diethyl ether, reflux 1 hour was considered, and the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml makes dissolving, in contrast medical material solution;

Contrast solution two: the Poria control medicinal material, the 40-60ml that adds diethyl ether, reflux 20-40 minute, filter, filtrate volatilizes, and residue adds normal hexane 0.5-2ml makes dissolving, in contrast medical material solution.

Contrast solution three: the Poria control medicinal material, the 25-75ml that adds diethyl ether, reflux 15-45 minute, filter, filtrate volatilizes, and residue adds normal hexane 0.5-1.5ml makes dissolving, in contrast medical material solution.

Test according to thin layer chromatography, put above-mentioned two kinds of need testing solutions, two kinds of each 5 μ l of contrast solution respectively at two silica gel g thin-layer plates, with petroleum ether (30-60 ℃)-acetone-ethyl acetate (84: 15: 1) is developing solvent, presaturation 15 minutes, launch, take out, dry, put and observe (365nm) under the uviol lamp.Compare the color developing effect of the need testing solution of two kinds of extracting method preparations, the results are shown in following table:

As can be seen from the above table, adopt the color developing effect of need testing solution one, developing solvent one good, the Pass Test requirement.

2) the middle developing solvent proportioning of above-mentioned discrimination method (2) is preferred: draw above-mentioned need testing solution one and reference substance solution one each 5 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-acetone-ethyl acetate (84: 15: 1) is developing solvent, presaturation 15 minutes, launch, take out, dry, put and observe (365nm) under the uviol lamp.Observe the unfolded effect of test sample on each lamellae, the results are shown in following table:

The developing solvent proportioning	84∶15∶1	60∶5∶0.5	70∶5∶0.5	100∶30∶20
					Launch effect	Good	Better	Better	Difference

Developing solvent proportioning as can be seen from the above table is 84: 15: 1 o'clock, and it is best that need testing solution launches effect, and appearance hangover, speckle separate phenomenons such as bad, unintelligible.

3) the middle sample solution point sample amount of above-mentioned discrimination method (2) is preferred, get need testing solution 1 μ l, 2 μ l, 3 μ l, 5 μ l, control medicinal material solution one 5 μ l, putting respectively on same silica gel g thin-layer plate, is developing solvent with petroleum ether (30-60 ℃)-acetone-ethyl acetate (84: 15: 1), presaturation 15 minutes, launch, take out, dry, put and observe (365nm) under the uviol lamp.Observe the effect of test sample principal spot colour developing on the lamellae, the results are shown in following table:

Test sample point sample amount is when 5 μ l as can be seen from the above table, and color developing effect is good on lamellae, is fit to test requirements document.

4) negative control test

Get the negative sample that lacks Poria, prepare negative control solution, launch the back and on control medicinal material solution correspondence position, corresponding speckle do not occur, illustrate that selected identification experiment specificity is strong according to above-mentioned need testing solution preparation method.

3, the thin layer discrimination method of Cortex Magnoliae Officinalis:

The selection of different need testing solution preparation methoies:

Method one: get pharmaceutical preparation 5g of the present invention, 30ml adds diethyl ether; Reflux 0.5 hour is taken out, and puts coldly, filters, and filtrate evaporate to dryness, residue add ethyl acetate 2ml makes dissolving, as need testing solution.

Method two: get pharmaceutical preparation 5g of the present invention, add methanol 50ml, close plug, jolting 30 minutes filters, and filtrate is as need testing solution.

Drawing each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene one methanol (96.5: 3.5), launches, and takes out, and dries, and spray is with 1% vanillin sulfuric acid solution, 100 ℃ be heated to speckle develop the color clear.Compare the color developing effect of the need testing solution of two kinds of extracting method preparations, the results are shown in following table:

Extracting method	Method one	Method two
			Color developing effect	Color developing effect is good	It is clear to develop the color, and interference is arranged.

As can be seen from the above table, the color developing effect of two method need testing solutions is all good, but the method two negative test has interference, so method one Pass Test requirement.

Experimental example 5 assay screening tests

1. test apparatus

Detecting instrument (room temperature detection): Agilent 1100 type high performance liquid chromatographs;

Octadecylsilane chemically bonded silica (4.6 * 150mm, 5 μ m)

Producer: Agilent Technologies Anjelen Sci. ﹠ Tech. Inc (China)

Mobile phase: methanol-water (13: 7)

Detect wavelength: 225nm

Flow velocity: 1.0ml/min

Column temperature: room temperature

The reference substance source: costunolide is purchased lot number: the 1512-200001 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

Assay method: the preparation method of getting by need testing solution under [assay] item prepares sample liquid; And by preparing the blank sample that lacks the Radix Aucklandiae under [method for making] item, the preparation negative controls.Filter with microporous filter membrane (0.45 μ m).Accurate respectively each the 10 μ l of negative controls, reference substance liquid and need testing solution that draw inject chromatograph of liquid, measure, promptly.

2. the investigation of test sample condition determination:

(1) investigation of extraction solvent amount:

By method under the assay item need testing solution is detected.Take by weighing three parts of every part of 5g of this product by the method precision under the preparation of need testing solution, put respectively in the tool plug conical flask, accurate respectively chloroform 15ml, 50ml, the 100ml of adding.

Content with costunolide in every gram medicine is that index is determined to add the chloroform amount, and measurement result sees the following form:

Add the chloroform result of the test

Above result shows: add chloroform 50ml, the every gram medicine of 100ml gained content determination of costunolide is basic identical, select for use according to needs of production to add chloroform 50ml.

(2) ultrasonic time is investigated:

By method under the assay item need testing solution is detected.3 parts of need testing solution preparations, supersound process is 10 minutes, 30 minutes, 50 minutes respectively.

Content with costunolide in every gram medicine is that index is determined ultrasonic time.Measurement result sees the following form:

The ultrasonic time result of the test

Above result shows: 30 minutes, the 50 minutes every gram medicines of gained of ultrasonic time content determination of costunolide is basic identical, selects for use 30 minutes according to needs of production.

3. content assaying method is investigated:

(1) linear relationship is investigated and to be got reference substance solution (0.044mg/ml) and shake up, accurate respectively 2,4,6,8,10, the 12 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve, show that costunolide is linear between 0.088ug-0.528ug, its regression equation is:

Area＝1998.55256*Amt-0.2697686(r＝0.99958)

(2) stability test reference substance solution respectively at preparing the back 0,2,4,6,12,24 hour, is measured in accordance with the law, and the result shows that it is basicly stable in 24 hours, the results are shown in following table

(3) the accurate need testing solution of drawing of precision test, (lot number: 02070903) 10 μ l, repeat sample introduction 5 times, try to achieve relative standard deviation＜2%, the results are shown in following table

(4) the text method is pressed in the repeatability test, and (lot number: 02070903) sample is 5 parts, measures respectively, tries to achieve relative standard deviation＜2%, the results are shown in following table to get same lot number

(5) the recovery test precision take by weighing known content same lot number (lot number: sample 5g 02070601) more respectively precision take by weighing costunolide reference substance 0.35mg, press the preparation method operation of text need testing solution, measure its content, and calculate its response rate, measurement result sees the following form:

4. measurement result:

Every bag contains the Radix Aucklandiae with costunolide (C in this product ₁₅H ₂₀O ₂), meter must not be less than 0.10mg.

The present composition is the preparation of embodiment 7 described in above-mentioned discriminating, the assay experiment, but following embodiment all can realize the described effect of above-mentioned experimental example

The specific embodiment

Embodiment 1: granule

Radix Aucklandiae 70g Massa Medicata Fermentata (Jiao) 200g Rhizoma Atractylodis Macrocephalae 100g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 60g Fructus Chaenomelis 60g;

More than ten simply, Massa Medicata Fermentata is ground into 100 purpose powder; The Rhizoma Pinelliae and Rhizoma Zingiberis Recens 33g make solvent according to the percolation under fluid extract and the extractum item with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, and with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis are extracted volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Medicinal residues and Poria, Rhizoma Cyperi, Fructus Chaenomelis and Fructus Jujubae 55g decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, 1.5 parts of cane sugar powders, 2 parts in dextrin, and above-mentioned powder, ethanol an amount of, make granule, drying, add above-mentioned volatile oil, mixing is made 500g, promptly.

Embodiment 2: soft capsule

Radix Aucklandiae 35g Fructus Amomi 45g Rhizoma Atractylodis Macrocephalae 120g

Pericarpium Citri Reticulatae 120g Poria 120g Rhizoma Pinelliae (processed) 120g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 60g Herba Pogostemonis 60g Radix Glycyrrhizae 90g;

More than 12 flavors, the Rhizoma Pinelliae and Rhizoma Zingiberis Recens 36g make solvent according to the percolation under fluid extract and the extractum item with 7 0% ethanol of 6 times of amounts of medical material, flood after 24 hours, with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubae 60g such as medicinal residues and all the other Poria decoct with water secondary, and each 1.5 hours, collecting decoction, filter, filtrate and above-mentioned aqueous solution merge, and concentrate relative density 1.10 (50～55 ℃), put cold, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, and being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), drying under reduced pressure is ground into fine powder, adds vegetable oil and above-mentioned volatile oil, mixing is made 430, promptly.

Embodiment 3: effervescent

Radix Aucklandiae 50g Fructus Amomi 30g Rhizoma Atractylodis Macrocephalae 190g

Pericarpium Citri Reticulatae 200g Poria 200g Rhizoma Pinelliae (processed) 200g

Rhizoma Cyperi (vinegar system) 30g Fructus Aurantii Immaturus (stir-fry) 30g Fructus Amomi Rotundus (shelling) 50g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 60g Herba Pogostemonis 50g Radix Glycyrrhizae 70g;

More than 12 flavors, the Rhizoma Pinelliae and Rhizoma Zingiberis Recens 60g make solvent according to the percolation under fluid extract and the extractum item with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubae 100g such as medicinal residues and all the other Poria decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, and inclined and get supernatant, filter, filtrate and above-mentioned filtrate merge, reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), and drying under reduced pressure becomes dried cream, be ground into fine powder, add an amount of effervescent and adjuvant, make granule, drying, add above-mentioned volatile oil, mixing is made 460g, promptly.

Embodiment 4: capsule

Radix Aucklandiae 35g Fructus Amomi 45g Rhizoma Atractylodis Macrocephalae 120g

Pericarpium Citri Reticulatae 120g Poria 150g Rhizoma Pinelliae (processed) 160g

Rhizoma Cyperi (vinegar system) 70g Fructus Aurantii Immaturus (stir-fry) 60g Fructus Amomi Rotundus (shelling) 60g

More than 12 flavors, the Rhizoma Pinelliae and Rhizoma Zingiberis Recens 48g make solvent according to the percolation under fluid extract and the extractum item with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubae 75g such as medicinal residues and all the other Poria decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, 1.5 parts of cane sugar powders, 2 parts in dextrin, ethanol are an amount of, make granule, drying, add above-mentioned volatile oil, mixing is made 515g, promptly.

Embodiment 5: granule

Radix Aucklandiae 70g Fructus Amomi 70g Rhizoma Atractylodis Macrocephalae 100g

Pericarpium Citri Reticulatae 130g Poria 130g Rhizoma Pinelliae (processed) 130g

Rhizoma Cyperi (vinegar system) 80g Fructus Aurantii Immaturus (stir-fry) 50g Fructus Amomi Rotundus (shelling) 70g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 50g Herba Pogostemonis 50g Radix Glycyrrhizae 30g;

More than 12 flavors, the Rhizoma Pinelliae and Rhizoma Zingiberis Recens 39g make solvent according to the percolation under fluid extract and the extractum item with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubae 65g such as medicinal residues and all the other Poria decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, 1.5 parts of cane sugar powders, 2 parts in dextrin, ethanol are an amount of, make granule, drying, add above-mentioned volatile oil, mixing is made 480g, promptly.

Embodiment 6: granule

Radix Aucklandiae 500g Massa Medicata Fermentata (Jiao) 200g Rhizoma Atractylodis Macrocephalae 200g

Pericarpium Citri Reticulatae 100g Poria 100g Rhizoma Pinelliae (processed) 100g

Rhizoma Cyperi (vinegar system) 50g Fructus Aurantii Immaturus (stir-fry) 70g Fructus Amomi Rotundus (shelling) 60g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 50g Fructus Chaenomelis 80g;

More than ten simply, Massa Medicata Fermentata is ground into 100 purpose powder; The Rhizoma Pinelliae and Rhizoma Zingiberis Recens 30g make solvent according to the percolation under fluid extract and the extractum item with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, and with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis are extracted volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Medicinal residues and Poria, Rhizoma Cyperi, Fructus Chaenomelis and Fructus Jujubae 50g decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, 1.5 parts of cane sugar powders, 2 parts in dextrin, and above-mentioned powder, ethanol an amount of, make granule, drying, add above-mentioned volatile oil, mixing is made 755g, promptly.

Embodiment 7: granule

Radix Aucklandiae 70g Fructus Amomi 70g Rhizoma Atractylodis Macrocephalae 100g

Rhizoma Cyperi (vinegar system) 70g Fructus Aurantii Immaturus (stir-fry) 70g Fructus Amomi Rotundus (shelling) 70g

Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 70g Herba Pogostemonis 70g Radix Glycyrrhizae 30g;

More than 12 flavors, the Rhizoma Pinelliae and Rhizoma Zingiberis Recens 30g make solvent with 70% ethanol of 6 times of amounts of medical material, flood after 24 hours, with the speed of per minute 1～3ml, percolation slowly, the liquid of filtering is standby; The Radix Aucklandiae, Fructus Amomi, the Rhizoma Atractylodis Macrocephalae, Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus, Fructus Amomi Rotundus, Cortex Magnoliae Officinalis, Herba Pogostemonis extract volatile oil with the way of distillation, and the aqueous solution after distillation device is in addition collected; Three flavor and Fructus Jujubae 50g such as medicinal residues and all the other Poria decoct with water secondary, each 1.5 hours, collecting decoction filters, and filtrate and above-mentioned aqueous solution merge, concentrate relative density 1.10 (50～55 ℃), put coldly, add equivalent ethanol, left standstill 24 hours, incline and get supernatant, filter, filtrate and above-mentioned filtrate merge, and reclaim ethanol, being concentrated into relative density is the clear paste of 1.33～1.36 (50～55 ℃), 1 part of qinghuo reagent, 1.5 parts of cane sugar powders, 2 parts in dextrin, ethanol are an amount of, make granule, drying, add above-mentioned volatile oil, mixing is made 460g, promptly.

Differentiate

(1) get this product 15g, add ethanol-strong ammonia solution (1: 1) 5ml, moistening was placed 20 minutes, added chloroform 20ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution (12: 6: 3: 3: 1) is developing solvent, puts in the chromatography cylinder of ammonia saturated with vapor, launches, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, about 5 minutes of 105 ℃ of bakings; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical purple dot;

(2) get this product 5g, porphyrize adds water 20ml, chloroform 20ml, hydrochloric acid 3ml, and reflux 1 hour is divided and got chloroform layer, washes with water to neutrality, volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other removes hydrogen constuslactone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw control medicinal material solution 1 μ l, sample solution 2～4 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-toluene-ethyl acetate-glacial acetic acid (10: 20: 7: 0.5) be developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) get this product 15g, porphyrize adds water 50ml dissolving, adds diethyl ether and three times (25ml that at every turn adds diethyl ether, saturated nacl aqueous solution 10ml) of saturated nacl aqueous solution extraction, divides and gets ether solution, and room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1 hour filters, and the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (30～60 ℃)-acetone-ethyl acetate (84: 15: 1) is developing solvent, presaturation 15 minutes launches, and takes out, dry, uviol lamp is observed (365nm) down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

(4) get this product 5g, porphyrize, the 30ml that adds diethyl ether, reflux 30 minutes is put coldly, filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets magnolol, honokiol reference substance, adds methanol respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw the about 10 μ l of need testing solution, reference substance solution 8 μ l, put respectively on same silica gel g thin-layer plate, with toluene-methanol (27: 1) is developing solvent, launches, and takes out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

(1) according to high effective liquid chromatography for measuring,

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler (4.6 * 150mm, 5 μ m); Methanol-water (13: 7) is a mobile phase; The detection wavelength is 225nm, and theoretical cam curve is calculated by costunolide should be not less than 3000;

The accurate Rhizoma Beesiae Calthaefoliae hydrocarbon lactone reference substance 1mg that claims of the preparation of reference substance solution puts in the 25ml measuring bottle, adds the ethyl acetate dissolving, and is diluted to scale, shakes up, and filters with microporous filter membrane (0.45 μ m), promptly; (containing costunolide 0.04mg among every 1ml);

It is an amount of that granule is got under this product content uniformity in the preparation of need testing solution, porphyrize, and precision takes by weighing about 10g, puts in the tool plug Erlenmeyer flask, the accurate chloroform 100ml that adds, close plug shakes up, and claims to decide weight, and placement is spent the night, supersound process (power 250W, frequency 40KH _Z) 30 minutes, take out, put cold, close plug claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shake up, filter, precision is measured subsequent filtrate 20ml, put in the evaporating dish, volatilize, residue adds an amount of methanol, slight fever makes dissolving, is transferred in the 5ml measuring bottle, and is diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), promptly;

Accurate respectively reference substance liquid, each the 10 μ l injection chromatograph of liquid of test sample liquid drawn of algoscopy measured, promptly; Every bag contains the Radix Aucklandiae with costunolide (C in this product ₁₅H ₂₀O ₂), meter must not be less than 0.25mg.

(2) according to high effective liquid chromatography for measuring

Chromatographic condition and system suitability test are filler with the octadecylsilane chemically bonded silica; With acetonitrile-water-phosphoric acid (18: 82: 0.1) is mobile phase; The detection wavelength is 283nm, and number of theoretical plate is pressed naringin and calculated not the end in 2500.

It is an amount of that the preparation of reference substance solution is taken at 110 ℃ of naringin reference substances that are dried to constant weight, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains 80 μ g, promptly.

This product under the content uniformity item is got in the preparation of need testing solution, and porphyrize is got 5g, and accurate the title decides, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, supersound process (power 250W, frequency 40kHz) 45 minutes is put coldly, claims to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5ml, put in the 10ml measuring bottle, add methanol and be diluted to scale, shake up, filter with microporous filter membrane (0.45 μ m), get subsequent filtrate, promptly.

Algoscopy: accurate respectively reference substance solution and the need testing solution 10 μ l of drawing, inject chromatograph of liquid, measure, promptly.This product contains Pericarpium Citri Reticulatae with naringin (C for every bag ₂₇H ₃₂O ₁₄) meter, must not be less than 4.0mg.

Function with cure mainly: warming middle-JIAO for easing the stomach.Be used for anorexia, acid regurgitation, gastral cavilty is full vexed, the extremity asthenia.

Usage and consumption: boiled water is taken after mixing it with water, a 5g, 2 times on the one.

Specification: every packed 5g.

Claims

1. the detection method with pharmaceutical composition of warming middle-JIAO for easing the stomach effect is characterized in that this method comprises the steps:

The crude drug of described pharmaceutical composition consists of:

Pericarpium Citri Reticulatae 50-200g Poria 50-200g Rhizoma Pinelliae Preparata 50-200g

The Rhizoma Cyperi (processed with vinegar) 30-100g Fructus Aurantii Immaturus (parched) 30-100g Fructus Amomi Rotundus 30-100g that shells

Sichuan Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 30-100g Fructus Chaenomelis 30-100g Rhizoma Zingiberis Recens 10-60g

Fructus Jujubae 20-100g;

This detection method comprise following discrimination method and/or content assaying method one or more:

Differentiate:

(1) get the quite described composition material medicine of preparation 28-32g, add 1-2: 1-2 ethanol-strong ammonia solution 5ml, moistening was placed 15-30 minute, add chloroform 18-25ml, supersound process 15-30 minute, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with 10-15: 4-9: 2-6: 2-5: 1 toluene-ethyl acetate-methanol-isopropyl alcohol-strong ammonia solution is developing solvent, puts in the chromatography cylinder of ammonia saturated with vapor, launches, take out, dry, spray is with the 0.4-0.7% ethanol solution of ninhydrin, 105 ℃ of bakings about 4-7 minute; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical purple dot;

(2) get the quite described composition material medicine of preparation 10g,, porphyrize adds water 20ml, chloroform 20ml, hydrochloric acid 3ml, and reflux 1-2 hour, divide and get chloroform layer, wash with water to neutrality, volatilize, residue adds dehydrated alcohol 1ml makes dissolving, as need testing solution; Other removes hydrogen constuslactone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw control medicinal material solution 1 μ l, sample solution 2～4 μ l, put respectively on same silica gel g thin-layer plate, with 8-12: 18-22: 5-8: 0.4-0.7 petroleum ether (60～90 ℃)-toluene-ethyl acetate-glacial acetic acid is developing solvent, launch, take out, dry, spray is with 1% vanillin ethanol solution of sulfuric acid, and hot blast blows to clear spot; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(3) get the quite described composition material medicine of preparation 28-32 g, porphyrize, add water 50ml dissolving, add diethyl ether and saturated nacl aqueous solution extracts the 20-30ml that at every turn adds diethyl ether, saturated nacl aqueous solution 5-15ml 2-4 time, divide and get ether solution, room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1-2 hour, filter, the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with 80-90: 12-18: 1 petroleum ether (30～60 ℃)-acetone-ethyl acetate is developing solvent, presaturation 10-20 minute, launch, take out, dry, uviol lamp is observed down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

(4) get the quite described composition material medicine of preparation 10g,, porphyrize, the 30ml that adds diethyl ether, reflux 25-35 minute, put coldly, filter, filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets magnolol, honokiol reference substance, adds methanol respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw the about 10 μ l of need testing solution, reference substance solution 8 μ l, put respectively on same silica gel g thin-layer plate, with 25-30: 1 toluene-methanol is developing solvent, launches, and takes out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

According to high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler (4.6 * 150mm, 5 μ m) with octadecylsilane chemically bonded silica; 10-16: the 6-9 methanol-water is a mobile phase; The detection wavelength is 225nm, and theoretical cam curve is calculated by costunolide should be not less than 3000;

The preparation of reference substance solution: the accurate Rhizoma Beesiae Calthaefoliae hydrocarbon lactone reference substance 1mg that claims, put in the 25ml measuring bottle, add the ethyl acetate dissolving, and be diluted to scale, shake up, filter with microporous filter membrane, promptly;

The preparation of need testing solution: get the quite described composition material medicine of preparation 20g,, put in the tool plug Erlenmeyer flask, the accurate chloroform 100ml that adds, close plug shakes up, and claims to decide weight, and placement is spent the night, supersound process, power 200-400W, frequency 40KH _Z20-40 minute, take out, put cold, close plug claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shake up, filter, precision is measured subsequent filtrate 20ml, put in the evaporating dish, volatilize, residue adds an amount of methanol, slight fever makes dissolving, is transferred in the 5ml measuring bottle, and is diluted to scale, shake up, filter with microporous filter membrane, promptly; Algoscopy: accurate respectively reference substance liquid, each 10 μ l injection chromatograph of liquid of test sample liquid drawn, measure, promptly.

2. drug regimen object detecting method as claimed in claim 1 is characterized in that the crude drug in this pharmaceutical composition consists of:

Pericarpium Citri Reticulatae 110g Poria 110g Rhizoma Pinelliae Preparata 110g

Rhizoma Cyperi (processed with vinegar) 60g Fructus Aurantii Immaturus (parched) 60g Semen Myristicae (shell removed) 60g

Sichuan Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 60g Fructus Chaenomelis 60g Rhizoma Zingiberis Recens 30g

Fructus Jujubae 50g.

3. the detection method with pharmaceutical composition of warming middle-JIAO for easing the stomach effect is characterized in that this method comprises the steps:

The crude drug of described pharmaceutical composition consists of:

Rhizoma Zingiberis Recens 10-60g Fructus Jujubae 20-100g;

Differentiate:

(3) get the quite described composition material medicine of preparation 28-32g, porphyrize, add water 50ml dissolving, add diethyl ether and saturated nacl aqueous solution extracts the 20-30ml that at every turn adds diethyl ether, saturated nacl aqueous solution 5-15ml 2-4 time, divide and get ether solution, room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1-2 hour, filter, the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with 80-90: 12-18: 1 petroleum ether (30～60 ℃)-acetone-ethyl acetate is developing solvent, presaturation 10-20 minute, launch, take out, dry, uviol lamp is observed down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

Assay:

4. drug regimen object detecting method as claimed in claim 3 is characterized in that the crude drug of this pharmaceutical composition consists of:

Rhizoma Zingiberis Recens 20-40g Fructus Jujubae 40-80g.

5. as claim 3 or 4 described drug regimen object detecting methods, it is characterized in that described preparation of drug combination method is:

6. as the detection method of the arbitrary described pharmaceutical composition of claim 1-5, it is characterized in that this method comprises following discrimination method and/or content assaying method:

Differentiate:

(3) get present composition granule 15g, porphyrize adds water 50ml dissolving, add diethyl ether and three times (25ml that at every turn adds diethyl ether, saturated nacl aqueous solution 10ml) of saturated nacl aqueous solution extraction, divide and get ether solution, room temperature volatilizes, residue adds normal hexane 0.5ml dissolving, as need testing solution; Other gets Poria control medicinal material 4g, the 50ml that adds diethyl ether, and reflux 1 hour filters, and the filtrate room temperature volatilizes, and residue adds normal hexane 0.5ml dissolving, medical material solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution 2 μ l, sample solution 8 μ l, put respectively on same silica gel g thin-layer plate, with petroleum ether (30～60 ℃)-acetone-ethyl acetate (84: 15: 1) is developing solvent, presaturation 15 minutes launches, and takes out, dry, uviol lamp is observed (36 5nm) down; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

(4) get present composition granule 5g, porphyrize, the 30ml that adds diethyl ether, reflux 30 minutes is put coldly, filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets magnolol, honokiol reference substance, adds methanol respectively and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw the about 10 μ l of need testing solution, reference substance solution 8 μ l, put respectively on same silica gel g thin-layer plate, with 27: 1 toluene-methanol was developing solvent, launched, and took out, dry, spray is with 1% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

According to high effective liquid chromatography for measuring, chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica, 4.6 * 150mm, 5 μ m; 13: 7 methanol-waters are mobile phase; The detection wavelength is 225nm, and theoretical cam curve is calculated by costunolide should be not less than 3000;

The preparation of need testing solution: it is an amount of to get under the present composition granule content uniformity granule, porphyrize, and precision takes by weighing about 10g, puts in the tool plug Erlenmeyer flask, the accurate chloroform 100ml that adds, close plug shakes up, and claims to decide weight, placement is spent the night, supersound process, power 250W, frequency 40KH _Z30 minutes, take out, put cold, close plug claims to decide weight again, supplies the weight that subtracts mistake with chloroform, shake up, filter, precision is measured subsequent filtrate 20ml, put in the evaporating dish, volatilize, residue adds an amount of methanol, slight fever makes dissolving, is transferred in the 5ml measuring bottle, and is diluted to scale, shake up, filter with microporous filter membrane, promptly; Algoscopy: accurate respectively reference substance liquid, each 10 μ l injection chromatograph of liquid of test sample liquid drawn, measure, promptly.

7. the application of pharmaceutical composition in preparation treatment acetic acid type gastric ulcer medicine with warming middle-JIAO for easing the stomach effect, described pharmaceutical composition is to be made by following crude drug:

Rhizoma Zingiberis Recens 20-40g Fructus Jujubae 40-80g.