CN101417014B

CN101417014B - Traditional Chinese medicine composition for treating stramgutia and preparation method and quality control method thereof

Info

Publication number: CN101417014B
Application number: CN2007101762252A
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: 2007-10-23
Filing date: 2007-10-23
Publication date: 2011-12-07
Anticipated expiration: 2027-10-23
Also published as: CN101417014A

Abstract

The invention provides a traditional Chinese medicine composition for treating stranguria caused by accumulated dampness-heat and a preparation method and a quality control method; the raw materials of the medicine composition comprise Season Red and plantain; combining with the content measurement of active ingredients, the invention optimizes the extraction and separation technique; the quality control method distinguishes the Season Red and the plantain and carries out content measurement to the Season Red (calculated by quercetin) and the plantain (calculated by malol) in the formula; the quality control of products can be carried out effectively by the quality control method; and the products controlled by the method has more stable pharmacological effects.

Description

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

Technical field

The present invention relates to a kind of treatment stranguria Chinese medicine composition and method of quality control thereof, particularly relate to a kind of Chinese medicine composition for the treatment of stranguria due to the damp-heat accumulation and preparation method thereof and method of quality control.

Background technology

Gonorrhea is to infect the active chronic inflammation of the caused genitourinary system of gonorrhea diplococcus.According to investigations, in China's sexually transmitted disease (STD) sickness rate, account for the first place.Chinese medicine thinks that stranguria mainly is because the damp-heat accumulation bladder, due to the disturbance in functional activity of the urinary bladder.Disease sees that frequent micturition is short red, desires, and goes out and not smooth, and desire is ended endless, and cramp and pain in the lower abdomen drenches the puckery pain of drop etc.Though the Western medicine of common treatment stranguria can comparatively fast be cured the disease symptom of stranguria, short-term effect is relatively good, and the treatment stranguria is had certain effect, cures the symptoms, not the disease, and takes for a long time human body is had certain side effect.Therefore select efficient and comparatively safe, stay-in-grade Chinese medicine to treat gonorrhea, become the hot topic of each side's research.

Summary of the invention

The object of the invention is to provide a kind of Chinese medicine composition for the treatment of stranguria due to the damp-heat accumulation;

The object of the invention also is to provide a kind of preparation method for the treatment of the Chinese medicine composition of stranguria due to the damp-heat accumulation.

The object of the invention also is to provide a kind of method of quality control for the treatment of the Chinese medicine composition of stranguria due to the damp-heat accumulation.

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

The Chinese medicine composition of treatment stranguria of the present invention is to be made by the raw material of following weight ratio:

Herba Polygoni Capitati 34-67.5 weight portion Herba Plantaginis 88.5-135 weight portion

Herba Polygoni Capitati 67.5 weight portion Herba Plantaginiss 88.5 weight portions

Herba Polygoni Capitati 34 weight portion Herba Plantaginiss 135 weight portions

The preparation method of Chinese medicine composition of the present invention is, Herba Polygoni Capitati and Herba Plantaginis with aforementioned proportion, adding 4～6 times of water gaging circulated in countercurrent extracted 3-6 hour, collecting decoction, filter, filtrate decompression concentrates, and temperature is 60-100 ℃, and pressure is-0.08～-0.10Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 40-70%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 40-70% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08～-0.10Mpa, temperature is 50-80 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, and technology adds adjuvant and makes tablet routinely, capsule, oral liquid, soft capsule, clinical acceptable forms such as granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.

The preparation method of Chinese medicine composition of the present invention is, with the Herba Polygoni Capitati and the Herba Plantaginis of described ratio, 5 times of water gaging circulated in countercurrent were extracted collecting decoction 5 hours, filter, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, and technology adds adjuvant and makes tablet routinely, capsule, oral liquid, soft capsule, clinical acceptable forms such as granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.

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

Differentiate:

(1) get this Chinese medicinal composition preparation and be equivalent to crude drug 1-4g, add methanol 15ml, reflux 0.5-2 hour, put coldly, filter, filtrate evaporate to dryness, residue add methanol 0.5-2ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.2-1g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 1-10 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (2-6: 2-6: 0.2-1: upper solution 0.2-1) is developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 5-15 minute.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

(2) get this Chinese medicinal composition preparation and be equivalent to crude drug 6-20g, add water 20-60ml, heating for dissolving is extracted 1-3 time the 1st 10-30ml with the ethyl acetate jolting, the 2nd 5-15ml, merge ethyl acetate, wash 1-3 time, each 5-15ml volatilizes, residue adds dehydrated alcohol 0.5-2ml dissolving, as need testing solution.Get Herba Plantaginis control medicinal material 1-5g, use ethyl acetate ultrasonic 10-40 minute, filter, filtrate volatilizes, and residue adds dehydrated alcohol 0.5-2ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution, each 1-10 μ l of Herba Plantaginis control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethylcellulose, (10-40: be that developing solvent launches 1), taking-up is dried, and launches again with chloroform-methanol, take out, dry, put in the iodine vapor and develop the color, the test sample chromatograph with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

2) assay:

(1) measures according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D).

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Phosphoric acid solution (the 30-70: 30-70) be mobile phase of methanol-(0.02%-0.8%); The detection wavelength is 360nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 5000.

The preparation precision of reference substance solution takes by weighing through dry 12-48 hour Quercetin reference substance of phosphorus pentoxide an amount of, adds methanol and makes the solution that every 1ml contains 5-15 μ g, promptly.

The preparation of need testing solution is got this Chinese medicinal composition preparation and is equivalent to crude drug 0.5-2g, and accurate the title decides, and puts in the tool plug conical flask, accurate adding methanol-hydrochloric acid-water (3-10: 0.5-2: mixed solution 5-15ml 1-3), close plug shakes up, claim to decide weight, in 70 ℃ of water-baths reflux 1-3 hour, jolting in per 20 minutes once, put coldly, claim again to decide weight, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

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

This Chinese medicine composition contains Herba Polygoni Capitati Quercetin (C with each dose of being grown up ₁₅H ₁₀O ₇) meter, must not be less than 0.25mg.

(2) measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D).

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; 0.02mol/L sodium dihydrogen phosphate-methanol (5-20: 80-95) be mobile phase; The detection wavelength is 215nm.Number of theoretical plate calculates by the ursolic acid peak should be not less than 5000.Agilent 1100 type high performance liquid chromatographs; Flow velocity: 0.6-1.0mLmin ^-1Detect wavelength 215nm; Column temperature: 20 ℃.

The preparation of reference substance solution: precision takes by weighing through dry 24 hours ursolic acid reference substance of phosphorus pentoxide an amount of, adds methanol and makes the solution that every 1ml contains 5-80 μ g, promptly.

The preparation of need testing solution: get this Chinese medicinal composition preparation and be equivalent to crude drug 1.0-5.0g, the accurate title, decide, and puts in the 50ml tool plug conical flask, the accurate methanol 25ml that adds, close plug shakes up, claim to decide weight, ultrasonic 10-40 minute, put cold, claim to decide weight again, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

Algoscopy: accurate respectively reference substance solution and each 5-15 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

The method of quality control of this Chinese medicine composition is characterized in that this method of quality control comprises one or more in following discrimination method and/or the assay:

1) differentiate:

(1) get this Chinese medicinal composition preparation and be equivalent to crude drug 2g, add methanol 15ml, reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(2) get this Chinese medicinal composition preparation and be equivalent to crude drug 10g, add water 40ml, heating for dissolving is extracted 2 times with the ethyl acetate jolting, the 1st 20ml, the 2nd 10ml merges ethyl acetate, washes 2 times, each 10ml volatilizes, and residue adds dehydrated alcohol 1ml dissolving, as need testing solution.Get Herba Plantaginis control medicinal material 2g, use ultrasonic 30 minutes of ethyl acetate, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution, each 2 μ l of Herba Plantaginis control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethylcellulose, with chloroform-methanol (20: 1) is that developing solvent launches, and taking-up is dried, and launches again, take out, dry, put in the iodine vapor and develop the color, the test sample chromatograph with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

2) assay:

(1) measures according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-0.4% phosphoric acid solution (50: 50) is a mobile phase; The detection wavelength is 360nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 5000.

The preparation precision of reference substance solution takes by weighing through dry 24 hours Quercetin reference substance of phosphorus pentoxide an amount of, adds methanol and makes the solution that every 1ml contains 10 μ g, promptly.

The preparation of need testing solution is got this Chinese medicinal composition preparation and is equivalent to crude drug 1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate mixed solution 10ml that adds methanol-hydrochloric acid-water (7: 1: 2), close plug shakes up, claim to decide weight, reflux is 2 hours in 70 ℃ of water-baths, and jolting in per 20 minutes once, put coldly, claim again to decide weight, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; 0.02mol/L sodium dihydrogen phosphate-methanol (15: 85) is mobile phase; The detection wavelength is 215nm.Number of theoretical plate calculates by the ursolic acid peak should be not less than 5000.Agilent1100 type high performance liquid chromatograph; Flow velocity: 0.8mLmin ^-1Detect wavelength 215nm; Column temperature: 20 ℃.

The preparation of reference substance solution: precision takes by weighing through dry 24 hours ursolic acid reference substance of phosphorus pentoxide an amount of, adds methanol and makes the solution that every 1ml contains 60 μ g, promptly.

The preparation of need testing solution: get this Chinese medicinal composition preparation and be equivalent to crude drug 2.0g, the accurate title, decide, and puts in the 50ml tool plug conical flask, the accurate methanol 25ml that adds, close plug shakes up, claim to decide weight, ultrasonic 30 minutes, put cold, claim to decide weight again, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

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

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

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

The screening of experimental example 1. extraction and separation process

Herba Polygoni Capitati has clearing away heat-damp and promoting diuresis for the present invention's monarch drug of writing out a prescription, and detoxifcation pain relieving and blood loose and becomes silted up effects such as inducing diuresis for treating stranguria syndrome.Mainly contain flavones ingredients such as Quercetin and glycoside thereof.Still carry out the preferred of extraction process with Quercetin as the index of its effective ingredient, concrete grammar is as follows:

1. the extraction concentration technique condition is preferred

Content with Quercetin serves as to investigate index, adopts orthogonal test that amount of water, decocting time and thickening temperature parameter in the extraction concentration technique are investigated.Concrete grammar is as follows:

Take by weighing Herba Polygoni Capitati 675g, Herba Plantaginis 885g respectively, mix totally 9 parts.According to orthogonal table L ₉(3 ⁴) arrange, carry out orthogonal test, 9 samples are carried out water respectively carry, filter, concentrate, content to Quercetin in 9 extracts is measured, the result is converted into the content (mg/g) of Quercetin in the Herba Polygoni Capitati, carries out variance analysis, and the result sees Table 1 respectively, table 2, table 3.

Table 1. decocting technological factor water-glass

Table 2. decocting craft screening orthogonal experiment data table

Table 3. analysis of variance table

F _0.05(2，2)＝19.00；F _0.01(2，2)＝99.00

Show that by extreme difference R value size in the table 2 each factor effect primary and secondary is C＞A＞B; Intuitive analysis is with A ₂B ₁C ₂Be optimised process; Shown by table 3 The results of analysis of variance: the influence of C factor has the significance meaning, and the influence of A, B factor does not have the significance meaning.That is: add 5 times of water gagings, reflux, extract, 5 hours, concentrating under reduced pressure temperature are 80 ℃.

Demonstration test: press A ₂B ₁C ₂The test of the triplicate of condition shown in the scheme.The results are shown in Table 4:

Table 4. decocting process repeatability result of the test table

Conclusion: this process stabilizing is feasible.

2. the alcohol precipitation process condition is preferred

With the content of yield of extract behind the precipitate with ethanol and Quercetin serves as to investigate index, adopts orthogonal test to the relative precipitate with ethanol of the medicinal liquid time with contain the alcohol amount and the precipitate with ethanol number of times is inquired into.Concrete grammar is as follows:

Take by weighing Herba Polygoni Capitati 675g, Herba Plantaginis 885g, mix, by condition A shown in the preferred extraction preferred plan ₂B ₁C ₂Extract, the decocting liquid that obtains is equally divided into 9 parts.According to orthogonal table L ₉(3 ⁴) arrangement, carry out orthogonal test, the dry extract amount and the Herba Polygoni Capitati quercetin content of 9 parts of extracts are measured, result of the test is carried out variance analysis, the result sees Table 5 respectively, table 6, table 7, table 8.

Table 5. alcohol precipitation process factor level table

Table 6. alcohol precipitation process screening orthogonal experiment data table

Table 7. quercetin content analysis of variance table

F _0.05(2，2)＝19.00；F _0.01(2，2)＝99.00

Yield of extract analysis of variance table behind table 8. precipitate with ethanol

F _0.05(2，2)＝19.00；F _0.01(2，2)＝99.00

With the quercetin content is to investigate index, shows that by extreme difference R value size in the table 6 factor effect primary and secondary is B＞C＞A; Intuitive analysis is with A ₂B ₁C ₃Be optimised process; The results of analysis of variance shows: the B factor and the C factor affecting all have the significance meaning, the influence of A factor does not have the significance meaning, investigates index with yield of extract behind the precipitate with ethanol, by extreme difference R value size demonstration in the table 6, factor effect primary and secondary is B＞C＞A; Intuitive analysis is with A ₂B ₁C ₃Be optimised process; The results of analysis of variance shows: the B factor and the C factor affecting all have the significance meaning, the influence of A factor does not have the significance meaning, comprehensive producing feasibility and energy savings and time are considered.Preferred A ₂B ₁C ₂, also promptly: it is 1.10～1.15 that medicinal liquid is concentrated into relative density, add ethanol and make and contain alcohol amount and reach 50%, and, precipitate with ethanol 2 times, each 24 hours.

Demonstration test: press A ₂B ₁C ₂The test of the triplicate of condition shown in the preferred plan.The results are shown in Table 9

Table 9. alcohol precipitation process replica test is table as a result

Conclusion: this process stabilizing is feasible.

Experimental example 2 pharmacological testings

Medicine and reagent:

The present invention wants thing group I: Herba Polygoni Capitati 340g Herba Plantaginis 1350g

The present invention wants thing group II: Herba Polygoni Capitati 675g Herba Plantaginis 885g

Preparation method: add 5 times of water gaging countercurrent extraction 5 hours, collecting decoction filters, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts coldly, adds ethanol and make and contain the alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged, and decompression recycling ethanol, pressure are-0.08Mpa, temperature is 65 ℃, being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, adds dextrin 270g, granulates, drying is made 450g promptly.

Positive controls: TONGLINSHU PIAN, the accurate word Z20069073 of traditional Chinese medicines, Jiangxi Pozin Pharmaceutical Co., Ltd., lot number: 061204

One, diuresis

1. laboratory animal: healthy white rat, male and female are regardless of, body weight 200～300 grams, rat is provided by Peking University's medical experiment animal center.

2. method is chosen healthy white rat, is divided into ordinary water group, positive controls, the present invention at random and wants thing group I and II.Every group 6, positive controls dosage 0.4g/kg, medicine group 2.4g/kg crude drug amount of the present invention, gastric infusion, when irritating stomach 14d, every places in the glass bell jar, every rat intraperitoneal injection of saline 20mL/kg body weight water load.Write down the gain in weight of per 40 minutes filter paper with the analytical balance weighing method, observed continuously 4 hours, remove the rat faecal pellet during weighing filter paper.And the result is carried out t check.The results are shown in following table:

Table 10 pair gives influence (n=10, the X ± S) of water load rat urine amount

Compare * P＜0.05 with negative control group, △ P＜0.05 is compared with medicine group I of the present invention in * * P＜0.01.

As can be seen from the above table, medicine group I of the present invention, II have the influence of significance to water load rat urine amount, relatively have significant difference with positive controls.The influence of medicine group II of the present invention more is better than medicine group I of the present invention.

Two. antiinflammatory action

1. to the effect of granuloma induced by implantation of cotton pellets

Get 40 of Wistar kind rats, use etherization, at each Mus web portion iodine liquid disinfectant, after 95% ethanol takes off iodine, each cuts the long osculum of 10m, will be weighed as the sterilization cotton balls (on add penicillin and streptomycin mixed liquor 0.2ml) of 10 mg with the ophthalmology tweezers, puts under the fell from little otch, skin suture is divided into four groups at random after the operation immediately.The 1st group is medicine group I of the present invention, dosage 2.4g/kg crude drug amount; The 2nd group is medicine group II of the present invention, dosage 2.4g/kg crude drug amount; The 3rd group of positive matched group, dosage 0.4g/kg; The 4th group of negative matched group used the water with volume.Postoperative 24h begins administration, is gastric infusion, and successive administration 6d opened former otch on the 7th day, with cotton balls together with around connective tissue take out, reject fatty tissue and foretell the sieve ball and put people's baking oven and behind 90 ℃ of oven dry 1h, take out and weigh.With claim weight deduct the former weight of cotton balls and promptly get granulomatous weight.The results are shown in Table 11.

Table 11 Chinese medicine composition of the present invention is to the effect of granuloma induced by implantation of cotton pellets (x ± S)

Compare * P＜0.05, * * P＜0.01 with the ordinary water group; Compare △ P＜0.05 with medicine group I of the present invention

The result shows that the rat granuloma weight with medicine group II filling stomach of the present invention has highly significant difference (P＜0.01) with ordinary water group ratio, and relatively there were significant differences (P＜0.05) with medicine group I of the present invention.

2. the effect of the foot swelling that causes of on Carrageenan

Get 40 of Wistar kind rats and weigh, be divided into four groups at random, use the picric acid labelling.The 1st group is medicine group I of the present invention, dosage 2.4g/kg crude drug amount; The 2nd group is medicine group II of the present invention, dosage 2.4g/kg crude drug amount; The 3rd group of positive matched group, dosage 0.4g/kg; The 4th group of negative matched group used the water with volume.Gastric infusion 7d is stretching with the hind leg of rat at the 7th day continuously, measures foot respectively with tape and opens up girth, surveys continuously 2 times, and its meansigma methods is the girth before the medication.Respectively at rat right hind leg vola far-end inserting needle to hot-tempered juxtra-articular, after the carrageenin 0.1ml of subcutaneous injection 10g/L, in 1,2,4,6,8h measures the rat paw girth respectively.Foot swelling rate=[(the matched group foot swelling is worth an administration group foot swelling value)/matched group foot swelling value] * 100% the results are shown in Table 12.

The effect of the foot swelling that table 12 Chinese medicine composition on Carrageenan of the present invention causes (x ± S)

The result shows that the right foot swelling rate of the different time of rat after causing inflammation with medicine group II filling stomach of the present invention compares at 2h with regard to variant (P＜0.05) with the ordinary water group, illustrate that the foot swelling inhibitory action produce effects that medicine group II on Carrageenan of the present invention causes is fast, behind the 8h, medicine group II of the present invention has compared highly significant difference (P＜0.01) with the ordinary water group, with medicine group I of the present invention highly significant difference (P＜0.01) is arranged more also.

Three, external bacteriostasis

Getting sterilizes 2 * 10 in the test tube of tampon, numbering is arranged on the test tube rack, adds the aseptic broth bouillon 1.0ml of people in every Guan Zhongxian, adds medicine group I solution of the present invention (0.3g/ml) 1.0ml of sterilization then in first pipe, take out 1.0ml behind the mixing and put into second pipe, by that analogy, in the 9th pipe, take out 1.0ml and discard, make into 1: 2,1: 4,1: 8,1: 16 various concentration such as (totally 9 pipes), the 10th pipe does not add medicine and compares.Get meat soup and cultivate experimental bacteria liquid (escherichia coli, the Bacillus proteus) ring of 8h, add respectively in the above-mentioned 1-10 pipe of people, put together behind the mixing in 37 ℃ of people's the incubator and cultivate 20h.To agar plate, no bacterial growth represents that promptly this test tube scalar is subjected to the test solution minimum inhibitory concentration exactly with the interior meat soup transferred species of each pipe.Medicine group II of the present invention operates with said method.

The result: colibacillary each pipe of medicine group I inoculation of the present invention all has bacterial growth through dull and stereotyped cultivation of bouillon agar, show that this medicinal liquid 0.3g/ml is following to escherichia coli unrestraint effect, each test tube of inoculation Bacillus proteus is through the dull and stereotyped cultivation of bouillon agar, the 1-3 pipe does not have bacterial growth, so medicine group I of the present invention is 0.02g/ml to the minimum inhibitory concentration of Bacillus proteus.

Each pipe of medicine group II inoculation escherichia coli of the present invention all has bacterial growth through the dull and stereotyped cultivation of bouillon agar, and the 1st pipe does not have bacterial growth, shows that this medicinal liquid minimum inhibitory concentration is 0.075g/ml.Each test tube of inoculation Bacillus proteus is cultivated through bouillon agar is dull and stereotyped, and the 1-5 pipe does not have bacterial growth, so medicine group II of the present invention is 5mg/ml to the minimum inhibitory concentration of Bacillus proteus.

Above result, medicine group of the present invention has certain antibacterial action, and medicine group II of the present invention to the action intensity of the antibacterial action of escherichia coli, Bacillus proteus apparently higher than medicine group I of the present invention.

Experimental example 2 is differentiated screening experiment

1) thin layer of Herba Polygoni Capitati is differentiated

1. the preparation of need testing solution

Get this Chinese medicinal composition preparation and be equivalent to totally four parts of crude drug 4g, porphyrize adds methanol 15ml, and reflux 20,40,60,80 minutes is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.Compare the color developing effect of need testing solution each speckle of different extraction times with control medicinal material solution on lamellae, the results are shown in following table:

The preparation of table 13 need testing solution

As can be seen from the above table, 60 minutes prepared need testing solutions of supersound extraction and reference substance solution, it is clear that each principal spot develops the color on lamellae, met requirement of experiment.

2. the selection of developing solvent

Drawing each 5 μ l point of need testing solution and reference substance solution on the silica gel G plate, is developing solvent with the upper solution of benzene-ethyl acetate-formic acid-water, and proportioning sees the following form, and launches, and takes out, and dries, and puts in the ammonia steam smoked 10 minutes.Relatively use different proportioning developing solvents, need testing solution and the expansion effect of each speckle of control medicinal material solution on lamellae the results are shown in following table:

The selection of table 14 developing solvent

Developing solvent proportioning as can be seen from the above table is benzene-ethyl acetate-formic acid-water 4: 4: 0.5: 0.5 o'clock, on lamellae, launch effectively, and be fit to test requirements document.

3. the selection of need testing solution point sample amount

Draw need testing solution 2 μ l, 3 μ l, 5 μ l, 7 μ l, 10 μ l respectively, each 2 μ l of control medicinal material solution put on the silica gel G plate, (4: 4: 0.5: 0.5) be developing solvent, expansion was taken out with benzene-ethyl acetate-formic acid-water, dry, put in the ammonia steam and smoked 10 minutes.Compare the color developing effect of each speckle of need testing solution, the results are shown in following table:

The selection of table 15 need testing solution point sample amount

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

4. negative control test

Get the negative sample that lacks Herba Polygoni Capitati, prepare negative sample solution, launch the back and corresponding speckle on the reference substance solution correspondence position, do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method.

2) thin layer of Herba Plantaginis is differentiated

1. the selection of lamellae

Get Herba Plantaginis control medicinal material 2g, use ultrasonic 30 minutes of ethyl acetate, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution.Get control medicinal material solution 2 μ l, put respectively on silica gel G and silica gel H plate, the expansion effect of comparative control product on different lamellaes the results are shown in following table:

3) selection of lamellae in the discriminating of table 16 Herba Plantaginis

As can be seen from the above table, lamellae selects the silica gel G plate development effective, and separating degree is good.

2. the preparation of need testing solution

Get pharmaceutical preparation content of the present invention and be equivalent to crude drug 4g, grind, add water 40ml heating for dissolving, with table method processing down, get Herba Plantaginis control medicinal material 2g, with ethyl acetate ultrasonic 30 minutes, to filter, filtrate volatilizes, residue adds dehydrated alcohol 1ml makes dissolving, medical material solution compares the color developing effect of test sample extracting method principal spot on the thin layer version in contrast, the results are shown in down

Table: the preparation of table 17 need testing solution

As can be seen from the above table, press method four prepared need testing solution and control medicinal material solution by prepared need testing solution of method four and reference substance solution, it is clear that each principal spot develops the color on lamellae, meets requirement of experiment.

3. the comparison of developing solvent

Drawing each 2 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-methanol, proportioning was respectively 5: 1, and 10: 1,20: 1, launch at 40: 1, take out, dry, launch again, take out, dry, put in the iodine steam and develop the color, compare need testing solution and the expansion effect of reference substance solution on lamellae, the results are shown in following table:

The comparison of table 18 developing solvent

As can be seen from the above table, the developing solvent proportioning is chloroform-methanol 20: 1 o'clock, and need testing solution and reference substance solution are launched effective, noiseless, meet requirement of experiment.

4. the selection of test sample point sample amount

Draw need testing solution 1 μ l, 2 μ l, 3 μ l, 5 μ l, each 2 μ l of reference substance solution put respectively on same silica gel g thin-layer plate, are developing solvent with chloroform-methanol (20: 1), launch, take out, dry, launch again, take out, dry, put in the iodine steam and develop the color, compare need testing solution and the color developing effect of control medicinal material solution on lamellae, the results are shown in following table:

The selection of table 19 test sample point sample amount

As can be seen from the above table, test sample point sample amount is when 2 μ l, 3 μ l, and it is clear to develop the color on lamellae, all is fit to test requirements document, so from the practical operation angle, selecting need testing solution point sample amount is 2 μ l.

5. negative control test

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

The part thin layer of above Herba Polygoni Capitati, Herba Plantaginis is differentiated screening test, and empirical tests can effectively be controlled the quality of drug combination preparation of the present invention from the qualitative detection aspect, and the quality of drug combination preparation of the present invention is improved.

The experiment of experimental example 3 assays

1) assay of Herba Polygoni Capitati is a test sample with the embodiment of the invention 6 described capsules, adopts the high-efficient liquid phase color popularize law to measure its Quercetin (C ₁₅H ₁₀O ₇) content, improving quality determining method of the present invention, result of the test as follows:

1. the preparation of need testing solution

In to medicine of the present invention, adopted the method for being prepared as follows in the assay of Quercetin:

Get this Chinese medicinal composition preparation and be equivalent to totally four parts of crude drug 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate mixed solution 10ml that adds methanol-hydrochloric acid-water (7: 1: 2), close plug shakes up, claim to decide weight, reflux is 2 hours in 70 ℃ of water-baths, and jolting in per 20 minutes once, put coldly, claim again to decide weight, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

Compared different return times, the content of Quercetin in the need testing solution the results are shown in following table:

The preparation of table 20 need testing solution

As can be seen from the above table, reflux, extract, can be extracted the Quercetin in the medicine of the present invention fully in 120 minutes, so the preparation of need testing solution selects supersound extraction to get final product in 120 minutes.

2. the selection of mobile phase

Compared the mobile phase with the different proportionings of methanol-0.4% phosphoric acid solution, proportioning was respectively 80: 20,70: 30,60: 40,50: 50, and the separating effect at each peak in the need testing solution chromatogram the results are shown in following table:

The selection of table 21 mobile phase

As can be seen from the above table, proportion of mobile phase is the good separating effect at each peak in 50: 50 o'clock test sample chromatograms, and disturbing does not appear in main peak.

3. the methodological study of content detection

To the detection method of content that medicine of the present invention adopted, carried out related side's science of law from aspects such as linear relationship, stability, precision, repeatability, the response rate and investigated, concrete grammar and result are as follows:

Detecting instrument: with octadecylsilane chemically bonded silica is filler; Methanol-0.4% phosphoric acid solution (50: 50) is a mobile phase; The detection wavelength is 360nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 5000.Agilent 1100 type high performance liquid chromatographs; Octadecylsilane chemically bonded silica (Zobax C ₁₈, 4.6mm * 25cm * 5 μ m)

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

Mobile phase: methanol-0.4% phosphoric acid solution (50: 50); Flow velocity: 1mL min ^-1Detect wavelength 360nm; Column temperature: 20 ℃.The reference substance source: Quercetin is purchased lot number: the 0209-9202 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

(1) linear relationship is investigated and to be got reference substance solution (0.0143mg/ml) and shake up, accurate respectively 2,4,6,8,10, the 12 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve chart, show that Quercetin presents good linear relationship between 0.0286 μ g～0.1716 μ g, its regression equation is:

Area＝3446.6974×Amt+6.5677(r＝0.9999)

Table 22. linear relationship is investigated

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

Table 23. stability test

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

The test of table 24. precision

(4) the repeatability test is got with 5 parts in a collection of medicine capsule sample of the present invention by above need testing solution preparation method, measures respectively, tries to achieve relative standard deviation＜2%, the results are shown in following table:

The test of table 25. repeatability

(5) the recovery test precision take by weighing known content must be with a collection of medicine capsule sample 1.0g of the present invention, place the 50ml measuring bottle, accurate respectively Quercetin reference substance solution (0.1186mg/ml) 3ml that adds, press the preparation method operation of text need testing solution, measure its content, and calculate its response rate, measurement result sees the following form:

Table 26. application of sample recovery test

From above result of the test as can be seen, active ingredient Quercetin in the pharmaceutical preparation of the present invention is carried out content detection control, method is stable, science, can effectively guarantee drug quality and curative effect, and this also is curative effect of medication of the present invention and the more significant reason of like product.

2) assay of Herba Plantaginis

Adopt the high-efficient liquid phase color popularize law to measure content of ursolic acid in the medicine of the present invention, improving quality determining method of the present invention, result of the test as follows:

1. the preparation of need testing solution

Adopted the method for being prepared as follows during content of ursolic acid is measured in to medicine of the present invention:

Get this Chinese medicinal composition preparation and be equivalent to totally four parts of crude drug 2.0g, the accurate title, decide, and puts in the 50ml tool plug conical flask, the accurate methanol 25ml that adds, close plug shakes up, claim to decide weight, ultrasonic 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes respectively, put cold, claim to decide weight again, supply the weight that subtracts mistake with above-mentioned mixed solution, shake up, filter, get subsequent filtrate, promptly.

Compared different return times, content of ursolic acid in the need testing solution the results are shown in following table:

The preparation of table 27. need testing solution

As can be seen from the above table, supersound extraction can be extracted the ursolic acid in the medicine of the present invention fully in 30 minutes, so the preparation of need testing solution selects supersound extraction to get final product in 30 minutes.

3. the selection of mobile phase

Compared the mobile phase with the different proportionings of 0.02mol/L sodium dihydrogen phosphate-methanol solution, proportioning was respectively 10: 90,15: 85,20: 80,25: 75, and the separating effect at each peak in the need testing solution chromatogram the results are shown in following table:

The selection of table 28. mobile phase

As can be seen from the above table, disturbing does not appear in the good separating effect at each peak in test sample chromatogram during proportion of mobile phase 0.02mol/L sodium dihydrogen phosphate-methanol (15: 85), main peak.

3. the methodological study of content detection

Detecting instrument: with octadecylsilane chemically bonded silica is filler; 0.02mol/L sodium dihydrogen phosphate-methanol (15: 85) is mobile phase; The detection wavelength is 215nm.Number of theoretical plate calculates by the ursolic acid peak should be not less than 5000.Agilent 1100 type high performance liquid chromatographs; Flow velocity: 0.8mLmin ^-1Detect wavelength 215nm; Column temperature: 20 ℃.

The reference substance source: ursolic acid is purchased lot number: the 0302-9824 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

(1) linear relationship is investigated and to be got reference substance solution (0.0058mg/ml) and shake up, accurate respectively 2,4,6,8,10, the 12 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve chart, show that ursolic acid presents good linear relationship between 0.0116 μ g～0.0696 μ g, its regression equation is:

Area＝2021845.24×Amt-2067.9(r＝0.9997)

Table 29. linear relationship is investigated

(2) stability test is got reference substance solution, respectively at preparing the back 0,2,4,6,12,24 hour, gets equal volume and injects high performance liquid chromatograph, and the result shows that it is basicly stable in 24 hours, the results are shown in following table:

Table 30. stability test

The test of table 31. precision

(4) the repeatability test is got with 5 parts in a collection of pharmaceutical preparation sample of the present invention by above need testing solution preparation method, measures respectively, tries to achieve relative standard deviation＜2%, the results are shown in following table:

The test of table 32. repeatability

(5) the recovery test precision take by weighing known content must be with a collection of pharmaceutical preparation sample 2.0g of the present invention, place the 50ml measuring bottle, accurate respectively ursolic acid reference substance solution (0.0058mg/ml) 20ml that adds, press the preparation method operation of text need testing solution, measure its content, and calculate its response rate, measurement result sees the following form:

Table 33. application of sample recovery test

From above result of the test as can be seen, active ingredient ursolic acid in the pharmaceutical preparation of the present invention is carried out content detection control, method is stable, science, can effectively guarantee drug quality and curative effect, and this also is one of curative effect of medication of the present invention and the more significant reason of like product.

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

The specific embodiment

Embodiment 1.

Herba Polygoni Capitati 1350g Herba Plantaginis 1770g

Herba Polygoni Capitati and Herba Plantaginis are added 10.6L watery vomiting stream extraction 5 hours, collecting decoction, filter, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, and technology adds adjuvant and makes tablet routinely, capsule, oral liquid, drop pill, clinical acceptable forms such as granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.

Embodiment 2: granule

Herba Polygoni Capitati 680g Herba Plantaginis 2700g

Add 16.9L watery vomiting stream and extracted 5 hours, collecting decoction filters, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts coldly, adds ethanol and make and contain the alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, adds dextrin 540g, steviosin 40g, granulate, drying gets the 800g granule, promptly.

Differentiate:

(1) get this Chinese medicine composition 1g, porphyrize adds methanol 15ml, and reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(2) get this Chinese medicine composition 5g, porphyrize adds water 40ml, and heating for dissolving is extracted 2 times with the ethyl acetate jolting, the 1st 20ml, and the 2nd 10ml merges ethyl acetate, washes 2 times, and each 10ml volatilizes, and residue adds dehydrated alcohol 1ml dissolving, as need testing solution.Get Herba Plantaginis control medicinal material 2g, use ultrasonic 30 minutes of ethyl acetate, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution, each 2 μ l of Herba Plantaginis control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethylcellulose, with chloroform-methanol (20: 1) is that developing solvent launches, and taking-up is dried, and launches again, take out, dry, put in the iodine vapor and develop the color, the test sample chromatograph with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

Assay:

Herba Polygoni Capitati

Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D).

The content under this Chinese medicine composition content uniformity item is got in the preparation of need testing solution, and porphyrize is got 0.5g, the accurate title, decide, and puts in the tool plug conical flask, the accurate mixed solution 10ml that adds methanol-hydrochloric acid-water (7: 1: 2), close plug shakes up, and claims to decide weight, reflux is 2 hours in 70 ℃ of water-baths, and jolting in per 20 minutes is once put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with above-mentioned mixed solution, filter, get subsequent filtrate, promptly.

Embodiment 3: soft capsule

Herba Polygoni Capitati 680g Herba Plantaginis 2700g

Above medical material adds 16.9L watery vomiting stream and extracted collecting decoction 5 hours, filter, filtrate decompression concentrates, and temperature is 80 ℃, pressure is-0.08Mpa, and being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts cold, adding ethanol makes and contains alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continuing to add ethanol makes the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged, decompression recycling ethanol, pressure are-0.08Mpa that temperature is 65 ℃, being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, vacuum drying is crushed to 100～150 orders, presses medicated powder: PEG600: the weight ratio mix homogeneously of glycerol (6: 6: 1), be pressed into soft capsule, promptly.Soft capsule shell is with gelatin: glycerol: the preparation of water (3: 1: 2) ratio.

Differentiate:

Get 3 of this Chinese medicine compositions, get content and add kieselguhr 2g mixing, drying under reduced pressure, porphyrize adds methanol 15ml, and reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Assay:

Herba Polygoni Capitati

Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).

4 of this Chinese medicine compositions are got in the preparation of need testing solution, add kieselguhr 2g and blend mixing, drying under reduced pressure, porphyrize,, put in the tool plug conical flask the accurate mixed solution 10ml that adds methanol-hydrochloric acid-water (7: 1: 2), close plug shakes up, and claims to decide weight, reflux is 2 hours in 70 ℃ of water-baths, and jolting in per 20 minutes is once put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with above-mentioned mixed solution, filter, get subsequent filtrate, promptly.

Embodiment 4: effervescent

Herba Polygoni Capitati 680g Herba Plantaginis 2700g

Above medical material adds 16.9L watery vomiting stream and extracted collecting decoction 5 hours, filter, filtrate decompression concentrates, and temperature is 80 ℃, pressure is-0.08Mpa, and being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts cold, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, and got supernatant, supernatant is merged, decompression recycling ethanol, pressure is-0.08Mpa, and temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, vacuum drying, be crushed to 100～150 orders, get medicated powder 660g medicated powder is divided into two parts, portion adds the 66g citric acid, use alcohol granulation, another part adds sodium bicarbonate 110g, uses alcohol granulation, and be dry respectively, granulate, mixing then, tabletting gets 500, promptly.

Differentiate:

(1) get this Chinese medicine composition 1g, porphyrize adds methanol 15ml, and reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ 1 of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(2) get this Chinese medicine composition 5g, porphyrize adds water 40ml, and heating for dissolving is extracted 2 times with the ethyl acetate jolting, the 1st 2ml, and the 2nd 10ml merges ethyl acetate, washes 2 times, and each 10ml volatilizes, and residue adds dehydrated alcohol 1ml dissolving, as need testing solution.Get Herba Plantaginis control medicinal material 2g, use ultrasonic 30 minutes of ethyl acetate, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution, each 2 μ l of Herba Plantaginis control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethylcellulose, with chloroform-methanol (20: 1) is that developing solvent launches, and taking-up is dried, and launches again, take out, dry, put in the iodine vapor and develop the color, the test sample chromatograph with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

Assay:

Herba Polygoni Capitati

Herba Plantaginis

Embodiment 5: tablet

Herba Polygoni Capitati 680g Herba Plantaginis 2700g

Add 16.9L watery vomiting stream and extracted 5 hours, collecting decoction filters, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts coldly, adds ethanol and make and contain the alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, and temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, the calcium carbonate that adds 1.6 times, stir evenly 5 times of amount starch, mixing, granulate, drying, granulate, tabletting, get 1000, promptly.

Differentiate:

Get this Chinese medicine composition 5g, porphyrize adds water 40ml, and heating for dissolving is extracted 2 times with the ethyl acetate jolting, the 1st 2ml, and the 2nd 10ml merges ethyl acetate, washes 2 times, and each 10ml volatilizes, and residue adds dehydrated alcohol 1ml dissolving, as need testing solution.Get Herba Plantaginis control medicinal material 2g, use ultrasonic 30 minutes of ethyl acetate, filter, filtrate volatilizes, and residue adds dehydrated alcohol 1ml makes dissolving, in contrast medical material solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution, each 2 μ l of Herba Plantaginis control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethylcellulose, with chloroform-methanol (20: 1) is that developing solvent launches, and taking-up is dried, and launches again, take out, dry, put in the iodine vapor and develop the color, the test sample chromatograph with the corresponding position of control medicinal material chromatograph on show the speckle of same color.

Assay:

Herba Polygoni Capitati

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-0.05% phosphoric acid solution (80: 100) is a mobile phase; The detection wavelength is 360nm.Number of theoretical plate calculates by the Quercetin peak should be not less than 5000.

Herba Plantaginis

[function with cure mainly] Seedling doctor: rising sun loud, high-pitched sound flag pile crust, hollow Ju is logical to closely question: stop hollow Kai Na.

The traditional Chinese medical science: heat-clearing and toxic substances removing, inducing diuresis for treating stranguria syndrome.Be used for stranguria due to the damp-heat accumulation, card is seen: it is not smooth to urinate, odynuria.

[usage and consumption] is oral, one time 2,3 times on the one; 5 days one courses of treatment.

[specification] every heavy 0.35g

Embodiment 6: capsule

Herba Polygoni Capitati 675g Herba Plantaginis 1880g

Above medical material adds 12.8L watery vomiting stream and extracted collecting decoction 5 hours, filter, filtrate decompression concentrates, and temperature is 80 ℃, pressure is-0.08Mpa, and being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, puts cold, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged, and decompression recycling ethanol, pressure are-0.08Mpa, temperature is 65 ℃, being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, adds the 300g dextrin, mixing, granulate, drying incapsulates, and makes 1000 promptly.

Method of quality control:

Differentiate:

(1) get this Chinese medicine composition 1g, porphyrize adds methanol 15ml, and reflux 1 hour is put coldly, filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets Herba Polygoni Capitati control medicinal material 0.5g, shines medical material solution in pairs with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same that what contain 0.5% sodium hydroxide is on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, (4: 4: 0.5: upper solution 0.5) was developing solvent with benzene-ethyl acetate-formic acid-water, launch, take out, dry, put in the ammonia steam and smoked 10 minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Assay:

This Chinese medicine composition content is got in the preparation of need testing solution, and porphyrize is got 0.5g, the accurate title, decide, and puts in the tool plug conical flask, the accurate mixed solution 10ml that adds methanol-hydrochloric acid-water (7: 1: 2), close plug shakes up, and claims to decide weight, reflux is 2 hours in 70 ℃ of water-baths, and jolting in per 20 minutes is once put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with above-mentioned mixed solution, filter, get subsequent filtrate, promptly.

Embodiment 7.: oral liquid

Herba Polygoni Capitati 680g Herba Plantaginis 2700g

Add 16.9L watery vomiting stream and extracted 5 hours, collecting decoction filters, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, adds an amount of sucrose, sorbic acid, add water and adjust total amount to 1000ml, stir evenly, fill, promptly.

[function cures mainly] heat-clearing and toxic substances removing, inducing diuresis for treating stranguria syndrome.Be used for stranguria due to the damp-heat accumulation, card is seen: it is not smooth to urinate, odynuria.

[usage and dosage] is oral, one time 1,3 times on the one; 5 days one courses of treatment.

[specification] every 10ml

Claims

1. Chinese medicine composition for the treatment of stranguria due to the damp-heat accumulation is characterized in that said composition is that the raw material of following row weight portion is made:

Herba Polygoni Capitati 67.5 weight portion Herba Plantaginiss 88.5 weight portions.

2. the preparation method of Chinese medicine composition as claimed in claim 1, it is characterized in that Herba Polygoni Capitati and Herba Plantaginis with described ratio, adding 4-6 times of water gaging circulated in countercurrent extracted 3-6 hour, collecting decoction, filter, filtrate decompression concentrates, and temperature is 60-100 ℃, and pressure is-0.08～-0.10Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 40-70%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 40-70% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08～-0.10Mpa, temperature is 50-80 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, and technology adds adjuvant and makes tablet routinely, capsule, oral liquid, granule; Described adjuvant is solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant or substrate.

Chinese medicine composition as claimed in claim 2 preparation method be, with the Herba Polygoni Capitati and the Herba Plantaginis of described ratio, add 5 times of water gaging countercurrent extraction 5 hours, collecting decoction, filter, filtrate decompression concentrates, and temperature is 80 ℃, and pressure is-0.08Mpa, being concentrated into 50 ℃, to measure relative densities be 1.10～1.15 clear paste, put coldly, add ethanol and make and contain alcohol amount and reach 50%, stir evenly, left standstill 24 hours, get supernatant, with precipitation be dissolved in water to relative density be 1.10～1.15, continue to add ethanol and make the alcohol amount of containing reach 50% to stir evenly, left standstill 24 hours, get supernatant, supernatant is merged decompression recycling ethanol, pressure is-0.08Mpa, temperature is 65 ℃, and being concentrated into 50 ℃, to measure relative densities be 1.35～1.40 thick paste, and technology adds adjuvant and makes tablet routinely, capsule, oral liquid, granule; Described adjuvant is solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant or substrate.