CN102091299B

CN102091299B - Method for detecting medicinal composition for treating viral hepatitis

Info

Publication number: CN102091299B
Application number: CN 201110033498
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: 2007-08-03
Filing date: 2007-08-03
Publication date: 2013-03-27
Anticipated expiration: 2027-08-03
Also published as: CN102091299A

Abstract

The invention discloses a method for detecting a medicinal composition for treating viral hepatitis. The medicinal composition is prepared from the following raw material medicaments: virgate wormwood herb, indigowoad root, male fern rhizome, Indian buead, root-tuber of aromatic turmeric, barbed skullcap herb, patchouli and the like. The method for detecting the medicinal composition is obtained by screening a great number of specific creative tests. In an identification method, the identification specificity is high by screening sample processing methods and selecting a developing solvent; and the method is economic and practical, is used for quickly acquiring results and can be applied to different thin layer plates. In a content detecting method, the quality of products can be effectively controlled by screening samples and sample supply processing methods and selecting the developing solvent. The products detected by the method have more stable medicinal effects than products detected by other methods.

Description

A kind of detection method for the treatment of the virus hepatitis pharmaceutical composition

The present invention is for dividing an application, and the original bill application number is 200710119920.5, the original bill applying date is 2007 On August 3, in, the original bill name is called A kind of pharmaceutical composition and quality control side for the treatment of virus hepatitis Method

Invention field

The present invention relates to a kind of pharmaceutical composition and preparation method and method of quality control, particularly a kind of pharmaceutical composition and preparation method and method of quality control for the treatment of virus hepatitis.

Background technology

Virus hepatitis is that the common transmittable that is caused by multiple hepatitis virus is sick, has that infectiousness is strong, the route of transmission is complicated, popular wide general, and the incidence of disease is than high.Clinically main manifestations be weak, anorexia, feel sick, vomiting, hepatomegaly and hepatic disorder, part patient can have jaundice and heating.Nettle rash, arthralgia or upper airway symptoms appear in some patient, severe patient even can develop into cirrhosis even liver cancer, the serious harm human health.Although at present the medicine of used treatment virus hepatitis has certain curative effect clinically, exist to some extent body to be developed immunity to drugs and the defective such as habituation, spinoff be obvious.

Summary of the invention

The object of the invention is to provide a kind of pharmaceutical composition; Another purpose of the present invention is to provide a kind of pharmaceutical composition for the treatment of virus hepatitis; The 3rd purpose of the present invention is to provide the preparation method of this pharmaceutical composition; The 4th purpose of the present invention is to provide the method for quality control of this pharmaceutical composition.

The present invention seeks to 1,2 to realize by following technical solution:

Technical scheme 1:

The bulk drug of pharmaceutical composition of the present invention consists of:

Poria cocos 500-1500 weight portion, oriental wormwood 500-1500 weight portion, bighead atractylodes rhizome 500-1500 weight portion, root tuber of aromatic turmeric 150-450 weight portion, Radix Angelicae Sinensis 250-750 weight portion, amber 50-150 weight portion, the root of herbaceous peony (stir-fry) 500-1500 weight portion, Sculellaria barbata 500-1500 weight portion, fructus amomi 100-300 weight portion, giant knotweed 250-750 weight portion, red sage root 500-1500 weight portion, Herba Lycopi 250-750 weight portion, radix bupleuri 150-450 weight portion, Pogostemon cablin 150-450 weight portion, eupatorium 250-750 weight portion, safflower 150-450 weight portion.

The bulk drug composition of pharmaceutical composition of the present invention is preferably:

Poria cocos 960 weight portions, oriental wormwood 960 weight portions, the bighead atractylodes rhizome 960 weight portions, root tuber of aromatic turmeric 320 weight portions, Radix Angelicae Sinensis 480 weight portions, amber 96 weight portions, the root of herbaceous peony (stir-fry) 960 weight portions, Sculellaria barbata 960 weight portions, fructus amomi 192 weight portions, giant knotweed 480 weight portions, the red sage root 960 weight portions, Herba Lycopi's 480 weight portions, radix bupleuri 320 weight portions, Pogostemon cablin 320 weight portions, eupatorium 480 weight portions, safflower 320 weight portions.

Technical scheme 2:

Poria cocos 500-1500 weight portion, oriental wormwood 500-1500 weight portion, root tuber of aromatic turmeric 150-450 weight portion, Radix Angelicae Sinensis 250-750 weight portion, amber 50-150 weight portion, the root of herbaceous peony (stir-fry) 500-1500 weight portion, Sculellaria barbata 500-1500 weight portion, fructus amomi 100-300 weight portion, giant knotweed 250-750 weight portion, red sage root 500-1500 weight portion, Herba Lycopi 250-750 weight portion, radix bupleuri 150-450 weight portion, Pogostemon cablin 150-450 weight portion, eupatorium 250-750 weight portion, safflower 150-450 weight portion, Radix Isatidis 500-1500 weight portion, thick wood-fern rhizome 250-750 weight portion, oldenlandia diffusa 500-1500 weight portion, Paris polyphylla 250-750 weight portion.

Poria cocos 960 weight portions, oriental wormwood 960 weight portions, root tuber of aromatic turmeric 320 weight portions, Radix Angelicae Sinensis 480 weight portions, amber 96 weight portions, the root of herbaceous peony (stir-fry) 960 weight portions, Sculellaria barbata 960 weight portions, fructus amomi 192 weight portions, giant knotweed 480 weight portions, the red sage root 960 weight portions, Herba Lycopi's 480 weight portions, radix bupleuri 320 weight portions, Pogostemon cablin 320 weight portions, eupatorium 480 weight portions, safflower 320 weight portions, Radix Isatidis 960 weight portions, thick wood-fern rhizome 480 weight portions, oldenlandia diffusa 960 weight portions, Paris polyphylla 480 weight portions.

Get technical solution of the present invention 1,2 pharmaceutical composition bulk drug, add conventional auxiliary material, according to common process, make the formulation of clinical acceptance, include but not limited to mixture, concentrated pill, capsule, pill, granule, tablet, soft capsule, sustained release agent, oral liquid.

Pharmaceutical composition technical scheme 1 preparation mixture method of the present invention is: Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 6-10 hour, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with the beta-schardinger dextrin-saturated water solution method inclusion of 3-5 times of weight portion, and drying gets the volatile oil beta-cyclodextrin inclusion complex, and is for subsequent use; The red sage root extracts 1-3 time with 85% alcohol reflux of 4-8 times of weight portion, and each 0.5-1.5 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The dregs of a decoction and all the other flavour of a drug boilings 1-3 time, the decocting that at every turn adds 6-10 times of weight portion boiled 1-2 hour, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, add ethanol and make and contain alcohol amount and reach 60-80%, left standstill 12-36 hour, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.10-1.15, add 80% simple syrup 900-1200 parts by volume, the volatile oil beta-cyclodextrin inclusion complex is ground into fine powder, adds in the medicinal extract; Add again Sodium Benzoate 9-10g, stir evenly, add water to the 3000-4500 parts by volume, filter, can, sterilization, and get final product.

Pharmaceutical composition technical scheme 2 preparation methods of the present invention are: above 19 flavors, and Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 6-10 hour, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with the beta-schardinger dextrin-saturated water solution method inclusion of 3-5 times of weight portion, and drying gets the volatile oil beta-cyclodextrin inclusion complex, and is for subsequent use; The red sage root extracts 1-3 time with 85% alcohol reflux of 4-8 times of weight portion, and each 0.5-1.5 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boilings such as the dregs of a decoction and all the other oriental wormwoods 1-3 time, the decocting that at every turn adds 6-10 times of weight portion boiled 1-2 hour, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, adding ethanol makes the alcohol amount of containing reach 60-80%, left standstill 12-36 hour, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, being concentrated into relative density is the thick paste II of 1.10-1.15, add the volatile oil beta-cyclodextrin inclusion complex and be ground into fine powder, add conventional auxiliary material, according to common process, make the formulation of clinical acceptance, include but not limited to mixture, concentrated pill, capsule, pill, granule, tablet, soft capsule, sustained release agent, oral liquid or freeze drying powder injection.

Pharmaceutical composition technical scheme 2 preparation mixture methods of the present invention are preferably: above 19 flavors, and Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 4 times of weight portions, and 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root extracts 2 times with 85% alcohol reflux of 6 times of weight portions, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boilings such as the dregs of a decoction and all the other oriental wormwoods 2 times add 8 times of water gagings at every turn and decocted 1.5 hours, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, add ethanol and make and contain alcohol amount and reach 70%, left standstill 24 hours, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.10-1.15, add 80% simple syrup 960 parts by volume, the volatile oil beta-cyclodextrin inclusion complex is ground into fine powder, adds in the medicinal extract; Add again Sodium Benzoate 9.6g, add water to 3200 parts by volume, filter, can, sterilization, and get final product.

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

Differentiate: A, get this drug combination preparation 1/100-3/100 day and use dosage, add water 10-30ml and make dissolving, with ethyl acetate extraction 1-3 time, 20-40ml for the first time, 10-30ml for the second time, combined ethyl acetate liquid, water bath method, residue adds ethyl acetate 0.5-1.5ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 0.5-1.5g, add water 40-60ml, decocted 0.5-1.5 hour, get supernatant, be concentrated into 10-30ml, with ethyl acetate extraction 1-3 time, for the first time 20-40ml, for the second time 10-30ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 0.5-1.5ml makes dissolving, in contrast medicinal material solution; Draw need testing solution 0.5-1.5 μ l, control medicinal material solution 4-6 μ l, put respectively on same silica gel g thin-layer plate, take 0.5-2: the 60-90 of 0.5-2 ratio ℃ petroleum ether-ethyl acetate launches as developping agent, take out, dry, put under the 365nm ultraviolet lamp and observe, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

B, get this drug combination preparation 4/1000-8/1000 day and use the dosage porphyrize, add 40-60% ethanol 5-25ml, ultrasonic processing 10-30 minute, filter, filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 0.5-1.5mg, in contrast product solution; Test according to thin-layered chromatography, draw each 5-15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 30-50: 1-10: 5-15: the methenyl choloride-ethyl acetate of 0.1-0.3 ratio-methyl alcohol-formic acid is as developping agent, launch, take out, dry, spray is with 1-10% vanillic aldehyde sulfuric acid solution, 100-110 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

C, get this drug combination preparation 1/100-5/100 day and use dosage, porphyrize, the 20-40ml that adds diethyl ether, refluxing extraction 20-40 minute, filter, filtrate volatilizes, and residue adds ethyl acetate 0.4-0.6ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 5-15ml that adds diethyl ether, and ultrasonic processing 5-15 minute, filter, filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 4-6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 5-15: the 60-90 of 0.1-1.0 ratio ℃ petroleum ether-ethyl acetate launches as developping agent, take out, dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

D, get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 1-3mg, in contrast product solution; Get this drug combination preparation 1/100-5/100 day and use dosage, porphyrize, the 20-40ml that adds diethyl ether, refluxing extraction 20-40 minute, filter, filtrate volatilizes, and residue adds ethyl acetate 0.4-0.6ml makes dissolving, as need testing solution, DrawNeed testing solution 5-15 μ l, reference substance solution 4-6 μ l puts respectively on same silica gel g thin-layer plate, and take 4-8: the cyclohexane-ethyl acetate of 1 ratio launches as developping agent, takes out, and dries; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot;

E, get this drug combination preparation 1/100-3/100 day and use dosage, porphyrize adds methyl alcohol 10-30ml, ultrasonic processing 10-30 minute filters the filtrate evaporate to dryness, residue adds 1.5-3.5mol/L sulfuric acid solution 5-15ml makes dissolving, puts in the water-bath and heats 20-40 minute, immediately cooling, extract twice with the methenyl choloride jolting, each 5-15ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 0.5-1.5ml makes dissolving, as need testing solution; Other gets the archen reference substance, adds methyl alcohol and makes the solution that every 1ml contains 0.5-1.5mg, in contrast product solution; Test according to thin-layered chromatography, draw each 4-8 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 5-25: 4-6: the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 0.5-1.5 ratio is as developping agent, launch, take out, dry, put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

Assay: shine high effective liquid chromatography for measuring: chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; 5-25: the acetonitrile-water of 70-95 ratio is mobile phase, and the detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 2600;

The preparation of reference substance solution: precision takes by weighing Paeoniflorin reference substance 4-6mg, puts in the 100ml measuring bottle, adds the methyl alcohol dissolving and is diluted to scale, shakes up, and namely gets the reference substance solution that contains Paeoniflorin 40-60 μ g among every 1ml;

The need testing solution preparation: get this drug combination preparation under the content uniformity item, porphyrize is got 6/10000-1/1000 day and is used dosage, accurately weighed, to put in the tool plug conical flask, precision adds 50-70% methyl alcohol 15-35ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 20-40 minute is taken out, and lets cool, weighed weight is supplied minimizing weight with 50-70% methyl alcohol again, shakes up, get supernatant, filter, and get final product;

Determination method: precision is drawn reference substance solution and each 5-15 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product;

The every daily dosage of this drug combination preparation contains the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter should be less than 50-70mg.

The pass of weight portion of the present invention and parts by volume is grams per milliliter.

The daily dosage of the different preparations of pharmaceutical composition of the present invention (every day taking dose or every day using dosage) is different because of preparation, but it is identical to contain suitable crude drug amount in the daily dosage of different preparations.Quality determining method of the present invention is take daily dosage as measurement unit.Amount according to crude drug converts, and the amount of the suitable crude drug of every daily dosage of being grown up is: 280-360g.

Pharmaceutical composition of the present invention confirms through experimental study: have the remarkable effect that recovers ALT, AST, the liver function leading indicators such as GGT, BiL to possess instant effect, have no side effect, use safety, obtain the advantages such as the rear difficult recurrence of effect.

The present composition is compared existing preparation and is possessed good drug effect, and the auxiliary material technique of pharmaceutical composition of the present invention is found volatile oil and beta-schardinger dextrin-inclusion under certain condition and proportioning through screening, can reach preferably curative effect; The method of quality control of Chinese medicine composition provided by the present invention, by obtaining behind a large amount of concrete creative experiment sievings, pass through the screening to sample treatment in the discrimination method, the selection of developping agent, so that differentiate that specificity is fine, and method is economic and practical, the result is quick, and can both use different thin layer plates.Pass through the screening to sample, test sample disposal route in the content assaying method, the selection of developping agent, so that content assaying method can effectivelyly carry out quality control to product, and with the product that the method is measured compare product that additive method measures drug effect show more stable.

Description of drawings

Figure of description is volatile oil beta-cyclodextrin inclusion experiment flow figure.

Embodiment

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

Experimental example 1: the medicine group to protecting liver, lowering enzymes, cholagogic, removing jaundice, engulf the pharmacodynamics test of the ability of cleaning up

Medicine group I: the pharmaceutical composition mixture of getting the embodiment of the invention 2 preparations

Medicine group II: the pharmaceutical composition mixture of getting the embodiment of the invention 13 preparations

Control group: the soothing the liver mixture of commercially available heat-clearing.

One. function for protecting liver and reducing enzyme activity

1. to the protective effect of acute liver damage

Get 84 of kunming mices, body weight 18-20g, male and female half and half are divided into 7 groups at random.First group: Normal group gavages the equivalent potable water every day; Second group: model group gavages the equivalent potable water every day; The 3rd group: medicine group I gavages medicine group I liquid 30ml (being equivalent to crude drug 9.32g)/kg body weight day; The 4th group: medicine group II gavages medicine group II liquid 30ml (being equivalent to crude drug 9.32g)/kg body weight day; The 5th group: control group gavages the soothing the liver mixture 0.3ml/10g of 0.4g/ml heat-clearing (being equivalent to contain crude drug 134g/kg) body weight day.The second～five group, after 14 days, carry out modeling with the injection of D-galactosamine 0.8g/kg body weight mouse peritoneal at successive administration.After modeling last administration in 24 hours half an hour, get blood from mouse orbit, survey SGPT, SGOT content, the results are shown in Table 1.

Table 1 medicine group of the present invention is to the protective effect of D-galactosamine induced mice acute liver damage (X ± SD)

^*Compare with model group: ^*P＜0.05; ^*P＜0.01;

Experimental result shows, with Normal group relatively, SGPT, SGOT content obviously raise (P＜0.01) in the serum of model group; Compare with model group, SGPT, the SGOT content of medicine group I of the present invention, II and control group obviously reduce (P＜0.01); And medicine group of the present invention and control group and no significant difference.

2. to the protective effect of alcoholic liver injury

Animal grouping and all the same tests of administration, in administration, the 2nd～6 group gavages 50% liquor (strong, colourless liquor distilled from sorghum, 56 °) 0.5ml/20g modeling.Continuously the modeling administration is 30 days, gets blood after last administration half an hour, surveys serum SGPT, SGOT content, the results are shown in Table 2.

Table 2 medicine group of the present invention is to the protective effect of alcoholic liver injury mouse (X ± SD)

^*Compare with model group: ^*P＜0.05; ^*P＜0.01;

Experimental result shows, with normal group relatively, SGPT, SGOT content obviously raise (P＜0.01) in the serum of model group.Compare with model group, medicine group of the present invention and control group all can reduce SGPT in the serum, SGOT content (P＜0.01); And medicine group of the present invention and control group and no significant difference.

3. to the protective effect of chronic liver injury

Get 72 of Wistar rats, body weight 140～150g, male and female half and half are divided into 6 groups at random.First group: Normal group gavages the equivalent potable water every day; Second group: model group gavages the equivalent potable water every day; The 3rd group: medicine group I gavages medicine group I liquid 30ml (being equivalent to crude drug 9.32g)/kg body weight day; The 4th group: medicine group II gavages medicine group II liquid 30ml (being equivalent to crude drug 9.32g)/kg body weight day; The 5th group: control group gavages the soothing the liver mixture 0.3ml/10g of 0.4g/ml heat-clearing (being equivalent to contain crude drug 134g/kg) body weight day.The second～five group, in administration, use 30%CC ₁₄Modeling is carried out in oil solution 0.3ml/100 hypodermic injection, twice weekly.The modeling administration was got blood and is surveyed serum SGOT, SGOT, hydroxyproline, total protein, albumin content, and calculate the A/G value after 3 months continuously, the results are shown in Table 3.

Table 3 medicine group of the present invention is to the protective effect of chronic liver injury rat (X ± SD)

^*Compare with model group; , ^*P＜0.05; ^*P＜0.01;

Experimental result shows, compares with normal group, and SGPT, SGOT content obviously raise (P＜0.01) in the serum of model group, and the A/G value obviously reduces, and hydroxyproline content obviously raises; Compare with model group, SGPT, the SGOT content of medicine group of the present invention and control group obviously reduce (P＜0.01); Hydroxyproline content obviously reduces (P＜0.01) in the obvious rising of the A/G value of medicine group of the present invention (P＜0.01) and the serum, and the A/G value notable difference (P＜0.05) of control group; And medicine group of the present invention and control group and no significant difference.

Two. on the impact of engulfing clean up ability of mouse reticuloendothelial system on inertia carbon granules in the blood flow

Animal, grouping and administration are tested with acute liver damage, more than respectively organized successive administration 20 days, after administration the 7th, 10 day, the 2nd～5 group of mouse peritoneal injection endoxan 100mg/Kg causes immune function of mice low.After 1 hour, mouse tail vein injects india ink 0.1ml/20g in the last administration, in rear 1,5 minute of injection, gets blood 20 μ l from the vena ophthalmica clump, is dissolved in 2.5ml, in the 0.1%Na2CO3 solution, shakes up, and in 680nm wavelength place colorimetric, measures the OD value.Index K value is cleaned up in calculating.The results are shown in Table 6.

Table 6 medicine group of the present invention is on the impact of macrophage phagocytic function (X ± SD)

^*Compare with model group; ^*P＜0.01

Experimental result shows, compares with normal group, and the K value of model group obviously reduces (P＜0.01).Compare with model group, medicine group of the present invention and control group K value obviously increase (P＜0.01).

Experimental example 2 auxiliary material screening experiments

Volatile oil beta-cyclodextrin inclusion process conditions preferred

Because volatile oil is easy to scatter and disappear, if therefore volatile oil is directly added, to place and with the passing of time or slightly be heated, volatile oil will lose, thereby has reduced content, affects the curative effect of medicine.For addressing this problem, adopted the method for beta-schardinger dextrin-inclusion, and adopted orthogonal experiment that the condition of inclusion has been carried out preferably.

Experiment flow is seen Figure of description.

Adopt the saturated water solution method inclusion: take by weighing a certain amount of beta-schardinger dextrin-, add an amount of distilled water, 5% beta-schardinger dextrin-aqueous solution is made in heating in 70 ℃ of water-baths, under the different temperatures (20 ℃, 40 ℃, 60 ℃) stir, rotating speed is 2500 rev/mins, add quantitative volatile oil, fully stir certain hour, get inclusion complex, stop to stir, with the inclusion complex on a small amount of distilled water flushing stirrer, airtight, to put in the refrigerator 12 hours, suction filtration gets the wet product of beta-schardinger dextrin-, through 40 ℃ of dryings of low temperature, get dry product and weigh, use the trace volatile oil extraction apparatus, measure by 2005 editions pharmacopeia (appendix X D) first method, record data calculate.

According to Preliminary experiment results, the factor of analyzing influence beta-schardinger dextrin-inclusion is set as follows test, the results are shown in Table 8-10:

Table 8 factor level table

Table 9 test findings

Table 10 analysis of variance table

F0.1(2，2)＝9，F0.05(2，2)＝19

Test findings: intuitive analysis is the result know, factor A＞C＞B, and top condition is A1B2C2; The results of analysis of variance knows, factor A impact is more remarkable, and B, C factor affecting are not remarkable.Should select A1B1C1, but from table 4 test findings, the inclusion rate of A1B2C2 is greater than A1B1C1, so be asserted A1B2C2, i.e. volatile oil: beta-schardinger dextrin-is 1: 4, and mixing time 30 minutes, inclusion temperature are 40 ℃.

The discrimination test of experimental example 3 oriental wormwoods

The preparation of blank sample is by medicine group flavour of a drug of the present invention and usage ratio, and autogamy does not contain group's medicine of oriental wormwood, the root of herbaceous peony, Radix Angelicae Sinensis, the red sage root, giant knotweed respectively, makes blank preparation by preparation technology.

The preparation of blank solution is got the blank sample of this drug combination preparation oriental wormwood and was used dosage on 14/1000th, adds water 20ml and makes dissolving, uses ethyl acetate extraction 2 times, 30ml for the first time, 20ml for the second time, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, as blank solution.

This medicine group and the matter sample of same amount got in the preparation of sample solution, makes sample solution according to the preparation method of blank solution.

Artemisia capillaris control medicinal material 1g is got in the preparation of control medicinal material solution, add water 50ml, decocted 1 hour, get supernatant, be concentrated into 20ml, with ethyl acetate extraction 2 times, for the first time 30ml, for the second time 20ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, in contrast medicinal material solution.

Following developping agent is prepared respectively in the selection of developping agent, 60-90 ℃ of petroleum ether-ethyl acetate 1: 4,60-90 ℃ of petroleum ether-ethyl acetate 1: 1,60-90 ℃ of petroleum ether-ethyl acetate 4: 1

Above three kinds of solution are put respectively on same silica gel g thin-layer plate, with 1: 4,1: Isosorbide-5-Nitrae: the 60-90 of 1 ratio ℃ petroleum ether-ethyl acetate was developping agent respectively, launch, take out, dry, put under the 365nm ultraviolet lamp and observe, take 60-90 ℃ of petroleum ether-ethyl acetate of 1: 1 ratio during as developping agent in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of obvious same color is arranged, and blank solution is showing without the fluorescence spot on corresponding position.

The discrimination test of experimental example 4 root of herbaceous peonys

The preparation of blank solution is got this pharmaceutical composition blank and preparation 4/1000-8/1000 day is used the dosage porphyrize, adds 40-60% ethanol 5-25ml, and ultrasonic processing 10-30 minute, filter, filtrate is as blank solution.

The Paeoniflorin reference substance is got in the preparation of control medicinal material solution in addition, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution.

The selection preparation of developping agent 30: 10: 5: 0.3 ratio, 40: 5: 10: 0.2 ratio, 50: 1: 15: the methenyl choloride-ethyl acetate of 0.1 ratio-methyl alcohol-formic acid was developping agent

Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, in above-mentioned developping agent, launch respectively, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and 105 ℃ of heating are in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color; Wherein, at 40: 5: 10: the most obvious in the methenyl choloride-ethyl acetate of 0.2 ratio-methyl alcohol-formic acid developping agent.

Experimental example 5 Radix Angelicae Sinensis are differentiated

The preparation of blank solution is got the blank preparation of this pharmaceutical composition and was used the dosage porphyrize on 3/100th, the 30ml that adds diethyl ether, and refluxing extraction 30 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as blank solution.

Radix Angelicae Sinensis control medicinal material 1g is got in the preparation of control medicinal material solution, the 10ml that adds diethyl ether, and ultrasonic processing 10 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution.

5: 1 ratios are prepared respectively in the selection of developping agent, 19: 1 ratios, the 60-90 of 150: 1 ratios ℃ petroleum ether-ethyl acetate is developping agent.

Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, under above-mentioned developping agent condition, launch respectively, take out, dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color; Wherein, spot is the most obvious when 60-90 ℃ of petroleum ether-ethyl acetate of 19: 1 ratios is developping agent.

The discrimination test of experimental example 6 reds sage root

The tanshinone IIA reference substance is got in the preparation of control medicinal material solution, adds ethyl acetate and makes the solution that every 1ml contains 2mg, in contrast product solution.

4: 1 ratios are prepared respectively in the selection of developping agent, 6: 1 ratios, the cyclohexane-ethyl acetate of 8: 1 ratios is developping agent.

According to the thin-layered chromatography test, draw each 10 μ l of blank solution and sample solution, reference substance solution 5 μ l put respectively on same silica gel g thin-layer plate, launch under above-mentioned developping agent condition respectively, take out, and dry; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot.Wherein, spot is the most obvious when the cyclohexane-ethyl acetate of 6: 1 ratios is developping agent.

Experimental example 7 giant knotweeds are differentiated

The giant knotweed principal ingredient is anthracene derivant, and this experiment archen after the hydrolysis in the giant knotweed is known index as inspection, has set up the TLC inspection knowledge method of giant knotweed in the preparation.

The preparation of blank solution is got the blank preparation of this pharmaceutical composition and was used the dosage porphyrize on the 14/1000th, and porphyrize adds methyl alcohol 20ml, ultrasonic processing 20 minutes filters the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml makes dissolving, puts in the water-bath and heats 30 minutes, immediately cooling, extract twice with the methenyl choloride jolting, each 10ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as blank solution.

The archen reference substance is got in the preparation of control medicinal material solution, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.

5: 6: 0.5 ratios are prepared respectively in the selection of developping agent, 15: 5: 1 ratios, the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 25: 4: 1.5 ratios.

According to the thin-layered chromatography test, draw blank solution and sample solution, each 6 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, under above-mentioned developping agent condition, launch respectively, take out, dry; Put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color; Spot is the most obvious when wherein, launching take the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 15: 5: 1 ratios as developping agent.

Experimental example 8 assays

1. instrument and medicine

The HP1050 of U.S. Hewlett-Packard type high performance liquid chromatograph, quaternary pump, DAD detecting device, chem workstation; H66025 type ultrasonic cleaning machine (Ultrasound Electronic Equipment Factory, Wuxi City); Acetonitrile is chromatographically pure, and mobile phase institute water is distilled water.

2. chromatographic condition optimization

Chromatographic column is YWGC18,10 μ m, and 4.6 * 250mm, the Dalian Chemistry and Physics Institute, the post effect is not less than 2600 in its theoretical cam curve of Paeoniflorin; Mobile phase is acetonitrile-water (15: 85); Flow velocity is 1ml/min; The detection wavelength is 230nm, and column temperature is room temperature, flow mutually in the ratio of acetonitrile and water be studied, optimized chromatographic condition.

Prepare respectively the mobile phase of following ratio: acetonitrile-water (15: 75), acetonitrile-water (15: 80), acetonitrile-water (15: 85), acetonitrile-water (15: 90), acetonitrile-water (15: 95)

By above chromatographic condition sample introduction reference substance, sample, the results are shown in following table 11 respectively:

Table 11 chromatographic condition optimization

The result as can be known, other chromatogram when mobile phase is acetonitrile-water (15: 75), acetonitrile-water (15: 80) in the sample can not be separated fully with the Paeoniflorin chromatographic peak, and with acetonitrile-water (15: 90), when acetonitrile-water (15: 95) is mobile phase, holder tail phenomenon appears in the Paeoniflorin chromatographic peak.Preferred flow is acetonitrile-water (15: 85) mutually

3. sample extraction solvent and optimization for extracting condition

The extraction solvent commonly used of Paeoniflorin is water, methyl alcohol, ethanol and rare alcohol, and it is that the sample extraction solvent can avoid water to cause the too much phenomenon generation of impurity as extracting solvent that methanol aqueous solution is adopted in this experiment.To methanol aqueous solution concentration, solubilizer multiple and ultrasonic treatment time have been done comparative study, have optimized the extraction conditions of sample.

Optimization for extracting condition sample thief powder (lot number 060529), porphyrize (crossing 50 mesh sieves), precision takes by weighing five parts of 0.25g, the accurate 60% methyl alcohol 25ml that adds of No.1～No.5 weighs close plug, according to the form below is ultrasonic processing (power 250W respectively, frequency 40KHZ), extracts completely, take out, weigh, supply the quantity of solvent that loses, get supernatant with 0.45 μ m filtering with microporous membrane, filtrate is as need testing solution, by above-mentioned chromatographic condition, sample introduction 20 μ l measure, and measurement result is added up, and the results are shown in Table 12.

Table 12 extraction time investigation (content of paeoniflorin mg/g)

The result is as can be known: no significant difference between each group.But the content with ultrasonic 10 minutes and 20 minutes is lower slightly.30 minutes, content was basically identical in 40 minutes.

According to above method, be under the condition of 30min in extraction time, investigate the concentration of extract, the results are shown in following table 13

Table 13 extract concentration is investigated (content of paeoniflorin mg/g)

The result is as can be known: no significant difference between each group, to extract with 60% methanol solution, and the Paeoniflorin yield is higher.

According to above method, be 30min in extraction time, extract is under the condition of 60% methyl alcohol, investigates the solubilizer multiple, the results are shown in Table 14.

The investigation of table 14 solubilizer multiple (content of paeoniflorin mg/g)

The result is as can be known: no significant difference between each group, it is higher to add under 100 times of amount solvent conditions the Paeoniflorin yield.

Conclusion: determine that extraction conditions is 60% methyl alcohol, 25ml, ultrasonic 30 minutes.

4. stability test

Get need testing solution (060529), respectively at after the preparation 0,2,4,6,10,16,24 hour, measure in accordance with the law, the result shows, and is basicly stable in 24 hours, the results are shown in Table 15.

Table 15 stability test

5. precision test

Accurate Paeoniflorin reference substance solution (C=0.204mg/ml) the 10 μ l that draw repeat sample introduction 5 times by aforementioned chromatographic condition, record peak area, and trying to achieve RSD is 1.13%, the results are shown in Table 16.

Table 16 Precision test result

6. linear relationship is investigated

Accurate draw the Paeoniflorin reference substance solution (2.5,5,7.5,10,12.5,15, the 20 μ l of 2.10mg → 10ml) measure by above-mentioned chromatographic condition sample introduction, measure peak area, the results are shown in Table 9.With peak area the amount of reference substance is carried out linear regression (comprising initial point), trying to achieve regression equation is Y=-39.6975+1574.5933X, and r=0.9999 shows that Paeoniflorin is linear in 0.525～4.200 μ g scope, and testing result sees Table 17.

Table 17 Paeoniflorin standard curve determination result

7. reappearance test

Get same batch sample (lot number 060529) porphyrize, precision takes by weighing 0.25g, and totally five parts, measure by the text method, try to achieve relative standard deviation.The results are shown in Table 18.

Table 18 reproducible test results

8. blank test

Do not contain group's medicine of the root of herbaceous peony in the autogamy of prescription taste of traditional Chinese medicine ratio, make blank solution by the text method.Blank solution does not show obvious chromatographic peak at the place of identical retention time with Paeoniflorin as a result, thinks noiseless.

9. recovery test

Adopt the application of sample absorption method, precision takes by weighing totally five parts of same lot number sample (lot number 060529) (content the is 6.9mg/g) 0.15g of known content, according to the form below adds respectively Paeoniflorin reference substance solution (concentration 0.234mg/ml) 5ml, precision adds 20ml60% methyl alcohol again, close plug, extract and measure by sample extraction and condition determination, with the following formula calculate recovery rate, the results are shown in Table 19.

Table 19 recovery test result

10. sample determination result

Record method by text ten batch samples are measured, the results are shown in Table 20.

Table 20 sample determination result

Lot number content of paeoniflorin (mg/g)

050524 6.20

050529 6.90

050618 5.84

060301 5.98

060306 5.17

060312 5.41

060910 5.22

060914 5.31

070312 5.30

070316 5.33

According to above data, it is that the every gram of this product contains the root of herbaceous peony by Paeoniflorin (C that content limit is fixed tentatively ₂₃H ₂₈O ₁₁) meter, should be no less than 5.0mg.

Employment and suitability test (E ﹠ ST) in the assay of experimental example 9. content assaying methods of the present invention Paeoniflorin in qizhi weitong granules

Qizhi weitong granules mainly is comprised of radix bupleuri, corydalis tuber, Fructus Aurantii, rhizoma cyperi, the root of herbaceous peony, honey-fried licorice root Six-element Chinese medicine.Paeoniflorin is measured the content of Paeoniflorin in the qizhi weitong granules for the index components of control this product quality, pilot production with the method.

(1) reappearance test

Get same batch sample (lot number 060312) porphyrize, precision takes by weighing 0.25g, and totally five parts, measure by the text method, try to achieve relative standard deviation.The results are shown in Table 21.

Table 21 reproducible test results

(2) recovery test

Adopt the application of sample absorption method, precision takes by weighing totally five parts of same lot number sample (lot number 060312) (content the is 1.5mg/g) 0.85g of known content, according to the form below adds respectively Paeoniflorin reference substance solution (concentration 0.234mg/ml) 5ml, precision adds 20ml60% methyl alcohol again, close plug, extract and measure by sample extraction and condition determination, with the following formula calculate recovery rate, the results are shown in Table 22.

Table 22 recovery test result

According to above presentation of results, content poor reproducibility, the recovery that this content assaying method is used for mensuration qizhi weitong granules Paeoniflorin are low.Although illustrate in the multiple Chinese patent drug and contain Paeoniflorin, its assay method is often different.

Following embodiment all can realize the described effect of above-mentioned experimental example

Embodiment 1

Poria cocos 960g, oriental wormwood 960g, bighead atractylodes rhizome 960g, root tuber of aromatic turmeric 320g, Radix Angelicae Sinensis 480g, amber 50-150g, the root of herbaceous peony (stir-fry) 960g, Sculellaria barbata 960g, fructus amomi 192g, giant knotweed 480g, red sage root 960g, Herba Lycopi 480g, radix bupleuri 320g, Pogostemon cablin 320g, eupatorium 480g, safflower 320g

This pharmaceutical composition adds conventional auxiliary material, makes capsule by common process.

Embodiment 2

Poria cocos 520g, oriental wormwood 1480g, bighead atractylodes rhizome 520g, root tuber of aromatic turmeric 430g, Radix Angelicae Sinensis 270g, amber 130g, the root of herbaceous peony (stir-fry) 520g, Sculellaria barbata 1480g, fructus amomi 120g, giant knotweed 730g, red sage root 520g, Herba Lycopi 730g, radix bupleuri 170g, Pogostemon cablin 430g, eupatorium 270g, safflower 430g

Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 4 times of weight portions, and 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root extracts 2 times with 85% alcohol reflux of 6 times of weight portions, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The dregs of a decoction and all the other flavour of a drug boilings 2 times add 8 times of water gagings at every turn and decocted 1.5 hours, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, add ethanol and make and contain alcohol amount and reach 70%, left standstill 24 hours, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.10-1.15, add 80% simple syrup 960 parts by volume, the volatile oil beta-cyclodextrin inclusion complex is ground into fine powder, adds in the medicinal extract; Add again Sodium Benzoate 9.6g, stir evenly, add water to 3000 parts by volume, filter, can, sterilization namely gets mixture and used dosage on the 30th.

Embodiment 3

Poria cocos 1480g, oriental wormwood 520g, bighead atractylodes rhizome 1480g, root tuber of aromatic turmeric 170g, Radix Angelicae Sinensis 730g, amber 70g, the root of herbaceous peony (stir-fry) 1480g, Sculellaria barbata 520g, fructus amomi 280g, giant knotweed 270g, red sage root 1480g, Herba Lycopi 270g, radix bupleuri 430g, Pogostemon cablin 170g, eupatorium 730g, safflower 170g

Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 4 times of weight portions, and 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root extracts 2 times with 85% alcohol reflux of 6 times of weight portions, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The dregs of a decoction and all the other flavour of a drug boilings 2 times add 8 times of water gagings at every turn and decocted 1.5 hours, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, add ethanol and make and contain alcohol amount and reach 70%, left standstill 24 hours, filter, filtrate recycling ethanol is to an amount of, merge, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.25-1.30 drying under reduced pressure under 0.08Mpa, the 65 ℃ of conditions with above-mentioned red sage root extract, add the volatile oil beta-cyclodextrin inclusion complex, be ground into fine powder; 70% ethanol softwood processed, extrusion granulator namely gets and used dosage on the 32nd.

Embodiment 4

Poria cocos 960g, oriental wormwood 960g, root tuber of aromatic turmeric 320g, Radix Angelicae Sinensis 480g, amber 96g, the root of herbaceous peony (stir-fry) 960g, Sculellaria barbata 960g, fructus amomi 192g, giant knotweed 480g, red sage root 960g, Herba Lycopi 480g, radix bupleuri 320g, Pogostemon cablin 320g, eupatorium 480g, safflower 320g, Radix Isatidis 960g, thick wood-fern rhizome 480g, oldenlandia diffusa 960g, Paris polyphylla 480g

This pharmaceutical composition adds conventional auxiliary material, makes mixture by common process.

Embodiment 5

Poria cocos 520g, oriental wormwood 1480g, root tuber of aromatic turmeric 170g, Radix Angelicae Sinensis 730g, amber 70g, the root of herbaceous peony (stir-fry) 1480g, Sculellaria barbata 520g, fructus amomi 280g, giant knotweed 270g, red sage root 1480g, Herba Lycopi 270g, radix bupleuri 430g, Pogostemon cablin 170g, eupatorium 730g, safflower 170g, Radix Isatidis 1480g, thick wood-fern rhizome 270g, oldenlandia diffusa 1480g, Paris polyphylla 270g

This pharmaceutical composition adds conventional auxiliary material, makes tablet by common process.

Embodiment 6

Poria cocos 1480g, oriental wormwood 520g, root tuber of aromatic turmeric 430g, Radix Angelicae Sinensis 270g, amber 130g, the root of herbaceous peony (stir-fry) 520g, Sculellaria barbata 1480g, fructus amomi 120g, giant knotweed 730g, red sage root 520g, Herba Lycopi 730g, radix bupleuri 170g, Pogostemon cablin 430g, eupatorium 270g, safflower 430g, Radix Isatidis 520g, thick wood-fern rhizome 730g, oldenlandia diffusa 520g, Paris polyphylla 730g

Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 4 times of weight portions, and 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root extracts 2 times with 85% alcohol reflux of 6 times of weight portions, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boilings such as the dregs of a decoction and all the other oriental wormwoods 2 times add 8 times of water gagings at every turn and decocted 1.5 hours, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, add ethanol and make and contain alcohol amount and reach 70%, left standstill 24 hours, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.10-1.15, add 80% simple syrup 1000 parts by volume, the volatile oil beta-cyclodextrin inclusion complex is ground into fine powder, adds in the medicinal extract; Add again Sodium Benzoate 9.6g, stir evenly, add water to 3600 parts by volume, filter, can, sterilization namely gets mixture.

The method of quality control of embodiment 7 preparations of the present invention

Differentiate: A, get the content of 0.014 times of daily dosage of this medicament composition granule agent of embodiment 3 preparations, add water 20ml and make dissolving, with ethyl acetate extraction 2 times, 30ml for the first time, 20ml for the second time, combined ethyl acetate liquid, water bath method, residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 1g, adds water 50ml, decocts 1 hour, gets supernatant, be concentrated into 20ml, use ethyl acetate extraction 2 times, for the first time 30ml, for the second time 20ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Draw need testing solution 1 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 1: 1 ratio as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and observe, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

B, get the content of 0.0057 times of daily dosage of this medicament composition granule agent of embodiment 3 preparation, porphyrize adds 50% ethanol 15ml, and ultrasonic processing 20 minutes filters, and filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 40: 5: 10: the methenyl choloride-ethyl acetate of 0.2 ratio-methyl alcohol-formic acid is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

C, get the content of 0.029 times of daily dosage of this medicament composition granule agent of embodiment 3 preparation, porphyrize, the 30ml that adds diethyl ether, refluxing extraction 30 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and ultrasonic processing 10 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 9.5: 0.5 ratios as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

D, get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, draw the need testing solution 10 μ l that differentiate under the C item, reference substance solution 5 μ l put respectively on same silica gel g thin-layer plate, take the cyclohexane-ethyl acetate of 6: 1 ratios as developping agent, launch, and take out, and dry; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot.

The method of quality control of embodiment 8 preparations of the present invention

Assay: shine high effective liquid chromatography for measuring: chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; The acetonitrile-water of 15: 85 ratios is mobile phase, and the detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 2600;

The preparation of reference substance solution: precision takes by weighing Paeoniflorin reference substance 5mg, puts in the 100ml measuring bottle, adds the methyl alcohol dissolving and is diluted to scale, shakes up, and namely gets the reference substance solution that contains Paeoniflorin 50 μ g among every 1ml;

The need testing solution preparation: get the content of 0.00072 times of daily dosage of this pharmaceutical composition mixture of embodiment 2 preparations under the content uniformity item, accurately weighed, put in the tool plug conical flask, precision adds 60% methyl alcohol 25ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 30 minutes is taken out, let cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again, shake up, get supernatant, filter, and get final product;

Determination method: precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product;

The every daily dosage of this pharmaceutical composition mixture contains the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter should be less than 60mg.

Embodiment 9

Differentiate: A, get the content of 0.014 times of daily dosage of this pharmaceutical composition mixture of embodiment 2 preparations, add water 20ml and make dissolving, with ethyl acetate extraction 2 times, 30ml for the first time, 20ml for the second time, combined ethyl acetate liquid, water bath method, residue adds ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 1g, adds water 50ml, decocts 1 hour, gets supernatant, be concentrated into 20ml, use ethyl acetate extraction 2 times, for the first time 30ml, for the second time 20ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Draw need testing solution 1 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 1: 1 ratio as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and observe, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

B, get the content of 0.0057 times of daily dosage of this pharmaceutical composition mixture of embodiment 2 preparation, porphyrize adds 50% ethanol 15ml, and ultrasonic processing 20 minutes filters, and filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 40: 5: 10: the methenyl choloride-ethyl acetate of 0.2 ratio-methyl alcohol-formic acid is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

C, get the content of 0.029 times of daily dosage of this pharmaceutical composition mixture of embodiment 2 preparation, porphyrize, the 30ml that adds diethyl ether, refluxing extraction 30 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and ultrasonic processing 10 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 9.5: 0.5 ratios as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

The need testing solution preparation: get 0.00072 times of daily dosage content of this pharmaceutical composition mixture of embodiment 12 preparations under the content uniformity item, accurately weighed, put in the tool plug conical flask, precision adds 60% methyl alcohol 25ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 30 minutes is taken out, let cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again, shake up, get supernatant, filter, and get final product;

The every daily dosage of this medicament composition capsule agent contains the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter should be less than 60mg.

Embodiment 10

Oriental wormwood 960g, Radix Isatidis 960g, thick wood-fern rhizome 480g, Poria cocos 960g, root tuber of aromatic turmeric 320g, Radix Angelicae Sinensis 480g, safflower 320g, amber 96g, the root of herbaceous peony (stir-fry) 960g, oldenlandia diffusa 960g, Sculellaria barbata 960g, Pogostemon cablin 320g, eupatorium 480g, fructus amomi 192g, giant knotweed 480g, red sage root 960g, Herba Lycopi 480g, radix bupleuri 320g, Paris polyphylla 480g

More than 19 flavors, Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, the aqueous solution after the distillation in addition device is collected; Volatile oil is with 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 4 times of weight portions, and 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root extracts 2 times with 85% alcohol reflux of 6 times of weight portions, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boilings such as the dregs of a decoction and all the other oriental wormwoods 2 times add 8 times of water gagings at every turn and decocted 1.5 hours, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, adding ethanol makes and contains alcohol amount and reach 70%, left standstill 24 hours, and filtered, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and the thick paste II that to be concentrated into 60～65 ℃ of relative densities be 1.10-1.15, adding 80% simple syrup 960 parts by volume, the volatile oil beta-cyclodextrin inclusion complex is ground into fine powder; Add again Sodium Benzoate 9.6g, add water to 3200 parts by volume, filter, can, sterilization namely gets mixture.

The method of quality control of embodiment 11 preparations of the present invention

The need testing solution preparation: get this medicament composition capsule agent of embodiment 1 preparation under the content uniformity item, porphyrize is got 0.00072 times daily dosage, accurately weighed, to put in the tool plug conical flask, precision adds 60% methyl alcohol 25ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 30 minutes is taken out, and lets cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again, shakes up, get supernatant, filter, and get final product;

The method of quality control of embodiment 12 preparations of the present invention

Differentiate:

A, get the daily dosage content porphyrize of 0.0057 times in this pharmaceutical composition mixture of embodiment 6 preparation, add 50% ethanol 15ml, ultrasonic processing 20 minutes filters, and filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 40: 5: 10: the methenyl choloride-ethyl acetate of 0.2 ratio-methyl alcohol-formic acid is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

B, get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, get the daily dosage content of 0.029 times in this pharmaceutical composition mixture of embodiment 6 preparations, porphyrize, the 30ml that adds diethyl ether, refluxing extraction 30 minutes filters, filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as need testing solution, draw need testing solution 10 μ l, reference substance solution 5 μ l put respectively on same silica gel g thin-layer plate, take the cyclohexane-ethyl acetate of 6: 1 ratios as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot;

C, get the daily dosage content of 0.014 times in this pharmaceutical composition mixture of embodiment 6 preparation, porphyrize adds methyl alcohol 20ml, ultrasonic place 20 minutes filters the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml makes dissolving, puts in the water-bath and heats 30 minutes, immediately cooling, extract twice with the methenyl choloride jolting, each 10ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the archen reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin-layered chromatography test, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 15: 5: 1 ratios as developping agent, launch, take out, dry, put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

Need testing solution preparation: get the daily dosage of 0.00072 times in this pharmaceutical composition mixture of embodiment 6 preparations under the content uniformity item, accurately weighed, put in the tool plug conical flask, precision adds 60% methyl alcohol 25ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 30 minutes is taken out, let cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again, shake up, get supernatant, filter, and get final product;

Embodiment 13

Oriental wormwood 960g Radix Isatidis 960g thick wood-fern rhizome 480g

Poria cocos 960g root tuber of aromatic turmeric 320g Radix Angelicae Sinensis 480g

The safflower 320g amber 96g root of herbaceous peony (stir-fry) 960g

Oldenlandia diffusa 960g Sculellaria barbata 960g Pogostemon cablin 320g

Eupatorium 480g fructus amomi 192g giant knotweed 480g

Red sage root 960g Herba Lycopi 480g radix bupleuri 320g

Paris polyphylla 480g

Method for making: above 19 flavors, Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 8 hours, and the aqueous solution after the distillation in addition device is collected, volatile oil 40 ℃ of inclusions of beta-schardinger dextrin-saturated water solution method of 40g, 40 ℃ of dryings get the volatile oil beta-cyclodextrin inclusion complex, and are for subsequent use; The red sage root is with 85% ethanol 6700ml refluxing extraction secondary, and each 1 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boiling secondaries such as the dregs of a decoction and all the other oriental wormwoods, add 8 times of water gagings decocted 1.5 hours at every turn, collecting decoction, filter, the aqueous solution after filtrate and the above-mentioned distillation merges, and being concentrated into relative density is 1.02～1.04 (60～65 ℃), adding ethanol makes and contains alcohol amount and reach 70%, left standstill 24 hours, and filtered, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, and be concentrated into the thick paste that relative density is 1.10-1.15 (60～65 ℃), add the volatile oil beta-cyclodextrin inclusion complex and be ground into fine powder, and appropriate amount of auxiliary materials, add water, can, sterilization, and get final product.

[discriminating]

(1) get this product 5g, add water 20ml and make dissolving, with ethyl acetate extraction 2 times, 30ml for the first time, 20ml for the second time, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 1g, adds water 50ml, decocts 1 hour, gets supernatant, is concentrated into 20ml, is made in the same way of control medicinal material solution with ethyl acetate; Draw need testing solution 1 μ l, control medicinal material solution 5 μ l put respectively on same silica gel g thin-layer plate, take sherwood oil (60-90 ℃)-ethyl acetate (1: 1) as developping agent, launch, and take out, and dry, and put under the ultraviolet lamp (365nm) and observe; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.

(2) get this product 2g porphyrize, add 50% ethanol 15ml, ultrasonic processing 20 minutes filters, and filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin-layered chromatography (" an appendix VI of Chinese Pharmacopoeia 2005 version B) test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take methenyl choloride-ethyl acetate-methyl alcohol-formic acid (40: 5: 10: 0.2) as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

(3) get this product 10g, porphyrize, the 30ml that adds diethyl ether, refluxing extraction 30 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and ultrasonic processing 10 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography (" an appendix VI of Chinese Pharmacopoeia 2005 version B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take sherwood oil (60-90 ℃)-ethyl acetate (9.5: 0.5) as developping agent, launch, take out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color.

(4) get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography (" an appendix VI of Chinese Pharmacopoeia 2005 version B) test, draw the need testing solution 10 μ l under the item of [discriminating] (3), reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, take cyclohexane-ethyl acetate (6: 1) as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot.

(5) get this product 5g, porphyrize adds methyl alcohol 20ml, ultrasonic processing 20 minutes filters the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml makes dissolving, puts in the water-bath and heats 30 minutes, immediately cooling, extract twice with the methenyl choloride jolting, each 10ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the archen reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin-layered chromatography (" an appendix VI of Chinese Pharmacopoeia 2005 version B) test, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the upper solution of sherwood oil (30～60 ℃)-ethyl formate-formic acid (15: 5: 1) as developping agent, launch, take out, dry, put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

Assay: according to high effective liquid chromatography for measuring

Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Acetonitrile-water (15: 85) is mobile phase, and the detection wavelength is 230nm; Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 2600;

The preparation precision of reference substance solution takes by weighing Paeoniflorin reference substance 5mg, puts in the 100ml measuring bottle, adds the methyl alcohol dissolving and is diluted to scale, shakes up, and namely gets (containing Paeoniflorin 50 μ g among every 1ml);

This product under the content uniformity item is got in the need testing solution preparation, and porphyrize is got 0.25g, and is accurately weighed, puts in the tool plug conical flask, and precision adds 60% methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 40KH _Z) 30 minutes, take out, let cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again; Shake up, get supernatant, filter, and get final product;

Determination method is accurate reference substance solution and each 10 μ l of need testing solution of drawing respectively, and the injection liquid chromatography is measured, and be get final product; The every 1ml of this product contains the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 0.6mg.

Claims

1. detection method for the treatment of the virus hepatitis pharmaceutical composition is characterized in that the content assaying method in the method is:

Chromatographic condition and system suitability: be filling agent with octadecylsilane chemically bonded silica; 5-25: the acetonitrile-water of 70-95 ratio is mobile phase, and the detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 2600;

The need testing solution preparation: get this drug combination preparation under the content uniformity item, porphyrize is got 6/10000-1/1000 day and is used dosage, accurately weighed, to put in the tool plug conical flask, precision adds 50-70% methyl alcohol 15-35ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 20-40 minute is taken out, and lets cool, weighed weight is supplied minimizing weight with 50-70% methyl alcohol again, shakes up, get supernatant, filter, and get final product; Determination method: precision is drawn reference substance solution and each 5-15 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product; The every daily dosage of this drug combination preparation contains the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter should be less than 50-70mg;

The bulk drug of described pharmaceutical composition consists of:

Poria cocos 500-1500 weight portion, oriental wormwood 500-1500 weight portion, root tuber of aromatic turmeric 150-450 weight portion, Radix Angelicae Sinensis 250-750 weight portion, amber 50-150 weight portion, stir-baked RADIX PAEONIAE ALBA 500-1500 weight portion, Sculellaria barbata 500-1500 weight portion, fructus amomi 100-300 weight portion, giant knotweed 250-750 weight portion, red sage root 500-1500 weight portion, Herba Lycopi 250-750 weight portion, radix bupleuri 150-450 weight portion, Pogostemon cablin 150-450 weight portion, eupatorium 250-750 weight portion, safflower 150-450 weight portion, Radix Isatidis 500-1500 weight portion, thick wood-fern rhizome 250-750 weight portion, oldenlandia diffusa 500-1500 weight portion, Paris polyphylla 250-750 weight portion;

Described daily dosage converts according to the amount of crude drug, and quite the amount of crude drug is: 280-360g.

2. the detection method of pharmaceutical composition as claimed in claim 1 is characterized in that the assay in the method is:

Chromatographic condition and system suitability: be filling agent with octadecylsilane chemically bonded silica; The acetonitrile-water of 15: 85 ratios is mobile phase, and the detection wavelength is 230nm, and number of theoretical plate calculates by the Paeoniflorin peak should be not less than 2600;

The need testing solution preparation: get this drug combination preparation under the content uniformity item, porphyrize is got and was used dosage on the 7/10000th, accurately weighed, to put in the tool plug conical flask, precision adds 60% methyl alcohol 25ml, close plug, weighed weight, power 250W, the ultrasonic processing of frequency 40KHZ 30 minutes is taken out, and lets cool, weighed weight is supplied minimizing weight with 60% methyl alcohol again, shakes up, get supernatant, filter, and get final product;

Determination method: precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product; The every daily dosage of this drug combination preparation contains the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter should be less than 60mg;

The bulk drug of described pharmaceutical composition consists of:

Poria cocos 500-1500 weight portion, oriental wormwood 500-1500 weight portion, root tuber of aromatic turmeric 150-450 weight portion, Radix Angelicae Sinensis 250-750 weight portion, amber 50-150 weight portion, stir-baked RADIX PAEONIAE ALBA 500-1500 weight portion, Sculellaria barbata 500-1500 weight portion, fructus amomi 100-300 weight portion, giant knotweed 250-750 weight portion, red sage root 500-1500 weight portion, Herba Lycopi 250-750 weight portion, radix bupleuri 150-450 weight portion, Pogostemon cablin 150-450 weight portion, eupatorium 250-750 weight portion, safflower 150-450 weight portion, Radix Isatidis 500-1500 weight portion, thick wood-fern rhizome 250-750 weight portion, oldenlandia diffusa 500-1500 weight portion, Paris polyphylla 250-750 weight portion.

3. the detection method of pharmaceutical composition as claimed in claim 1 or 2 is characterized in that the method also comprises one or more of following discrimination method:

Differentiate: A, get this drug combination preparation 1/100-3/100 day and use dosage, add water 10-30ml and make dissolving, with ethyl acetate extraction 1-3 time, 20-40ml for the first time, 10-30ml for the second time, combined ethyl acetate liquid, water bath method, residue adds ethyl acetate 0.5-1.5ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 0.5-1.5g, add water 40-60ml, decocted 0.5-1.5 hour, get supernatant, be concentrated into 10-30ml, with ethyl acetate extraction 1-3 time, for the first time 20-40ml, for the second time 10-30ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 0.5-1.5ml makes dissolving, in contrast medicinal material solution; Draw need testing solution 0.5-1.5 μ l, control medicinal material solution 4-6 μ l, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 0.5-2:0.5-2 ratio as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and observe, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

D, get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 1-3mg, in contrast product solution; According to thin-layered chromatography test, draw the need testing solution 5-15 μ l that differentiates under the C item, reference substance solution 4-6 μ l puts respectively on same silica gel g thin-layer plate, and take 4-8: the cyclohexane-ethyl acetate of 1 ratio launches as developping agent, takes out, and dries; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot;

E, get this drug combination preparation 1/100-3/100 day and use dosage, porphyrize adds methyl alcohol 10-30ml, ultrasonic processing 10-30 minute filters the filtrate evaporate to dryness, residue adds 1.5-3.5mol/L sulfuric acid solution 5-15ml makes dissolving, puts in the water-bath and heats 20-40 minute, immediately cooling, extract twice with the methenyl choloride jolting, each 5-15ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 0.5-1.5ml makes dissolving, as need testing solution; Other gets the archen reference substance, adds methyl alcohol and makes the solution that every 1ml contains 0.5-1.5mg, in contrast product solution; Test according to thin-layered chromatography, draw each 4-8 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 5-25: 4-6: the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 0.5-1.5 ratio is as developping agent, launch, take out, dry, put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

4. the detection of pharmaceutical composition as claimed in claim 3 is characterized in that the method comprises one or more of following discrimination method:

A, get this drug combination preparation and used dosage on 14/1000th, add water 20ml and make dissolving, with ethyl acetate extraction 2 times, 30ml for the first time, 20ml for the second time, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Artemisia capillaris control medicinal material 1g, adds water 50ml, decocts 1 hour, gets supernatant, be concentrated into 20ml, use ethyl acetate extraction 2 times, for the first time 30ml, for the second time 20ml, combined ethyl acetate liquid, water bath method, residue add ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Draw need testing solution 1 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 1: 1 ratio as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and observe, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

B, get this drug combination preparation and used the dosage porphyrize on 6/1000th, add 50% ethanol 15ml, ultrasonic processing 20 minutes filters, and filtrate is as need testing solution; Other gets the Paeoniflorin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 40: 5: 10: the methenyl choloride-ethyl acetate of 0.2 ratio-methyl alcohol-formic acid is developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

C, get this drug combination preparation and used dosage on 3/100th, porphyrize, the 30ml that adds diethyl ether, refluxing extraction 30 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 0.5ml makes dissolving, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and ultrasonic processing 10 minutes filters, and filtrate volatilizes, and residue adds ethyl acetate 1ml makes dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take 60-90 ℃ of petroleum ether-ethyl acetate of 9.5: 0.5 ratios as developping agent, launch, take out, dry, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the fluorescence spot of aobvious same color;

D, get the tanshinone IIA reference substance, add ethyl acetate and make the solution that every 1ml contains 2mg, in contrast product solution; According to thin-layered chromatography test, draw the need testing solution 10 μ l that differentiate under the C item, reference substance solution 5 μ l put respectively on same silica gel g thin-layer plate, take the cyclohexane-ethyl acetate of 6: 1 ratios as developping agent, launch, and take out, and dry; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, aobvious identical kermesinus spot;

E, get this drug combination preparation and used dosage on 14/1000th, porphyrize adds methyl alcohol 20ml, ultrasonic processing 20 minutes filters the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 10ml makes dissolving, puts in the water-bath and heats 30 minutes, immediately cooling, extract twice with the methenyl choloride jolting, each 10ml merges methenyl choloride liquid, evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the archen reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin-layered chromatography test, draw each 6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the upper solution of 30～60 ℃ of sherwood oil-ethyl formate-formic acid of 15: 5: 1 ratios as developping agent, launch, take out, dry, put in the ammonia smoked after, inspect under the daylight; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

5. the detection method of pharmaceutical composition as claimed in claim 1, the preparation method who it is characterized in that described pharmaceutical composition is: above 19 flavors, Radix Angelicae Sinensis, root tuber of aromatic turmeric, Pogostemon cablin, eupatorium, fructus amomi, Herba Lycopi extracted volatile oil 6-10 hour, and the aqueous solution after the distillation in addition device is collected; Volatile oil is with the beta-schardinger dextrin-saturated water solution method inclusion of 3-5 times of weight portion, and drying gets the volatile oil beta-cyclodextrin inclusion complex, and is for subsequent use; The red sage root extracts 1-3 time with 85% alcohol reflux of 4-8 times of weight portion, and each 0.5-1.5 hour, merge extract, filter, filtrate recycling ethanol gets red sage root extract to without the alcohol flavor, and is for subsequent use; The 12 flavor boilings such as the dregs of a decoction and all the other oriental wormwoods 1-3 time, the decocting that at every turn adds 6-10 times of weight portion boiled 1-2 hour, and collecting decoction filters; Aqueous solution after filtrate and the above-mentioned distillation merges, be concentrated into 60～65 ℃ of relative densities and be 1.02～1.04 thick paste I, adding ethanol makes the alcohol amount of containing reach 60-80%, left standstill 12-36 hour, filter, filtrate recycling ethanol is to an amount of, merge with above-mentioned red sage root extract, be concentrated into the thick paste II that relative density is 1.10-1.15, add the volatile oil beta-cyclodextrin inclusion complex and be ground into fine powder, add conventional auxiliary material, according to common process, make the formulation of clinical acceptance, include but not limited to concentrated pill, capsule, pill, granule, tablet, sustained release agent, oral liquid or freeze drying powder injection.