CN101991785B

CN101991785B - Lonicerae and Forsythiae detoxication soft capsule medicine and preparation method and quality detection method thereof

Info

Publication number: CN101991785B
Application number: CN2009101684549A
Authority: CN
Inventors: 萧伟; 王振中; 毕宇安; 刘涛; 胡军华; 章晨峰; 陈凤龙
Original assignee: Jiangsu Kanion Pharmaceutical Co Ltd
Current assignee: JIANGSU KANION YANGGUANG PHARMACEUTICAL CO., LTD.; Jiangsu Kanion Pharmaceutical Co Ltd
Priority date: 2009-08-20
Filing date: 2009-08-20
Publication date: 2012-07-18
Anticipated expiration: 2029-08-20
Also published as: CN101991785A; HK1150339A1

Abstract

The invention relates to a Lonicerae and Forsythiae detoxication soft capsule medicine and a preparation method and a quality detection method thereof, belonging to the field of traditional Chinese medicines. The Lonicerae and Forsythiae detoxication soft capsule comprises the following raw materials in parts by weight: 5 parts of Lonicerae, 5 parts of Forsythiae, 3 parts of mint, 2 parts of Schizonepeta, 2.5 parts of fermented soya beans, 3 parts of burdock (fried), 3 parts of Platycodon grandiflorum, 2 parts of lophatherum gracile and 2.5 parts of liquorice. The invention provides the preparation method of the medicine and also provides the quality detection method, wherein the quality detection method comprises five identification methods and a method for carrying out content determination by adopting a high performance liquid chromatography, and the methods can effectively control the quality of the Lonicerae and Forsythiae detoxication soft capsule.

Description

A kind of Yinqiao detoxification soft gelatin pharmaceutical and preparation method thereof and quality determining method

Technical field

The present invention relates to a kind of Chinese medicine preparation and preparation method thereof and quality determining method, pharmaceutical preparation that is specifically related to a kind of Yinqiao detoxification soft capsule and preparation method thereof reaches quality determining method.

Technical background

Put down in writing some technology contents of medicines such as antitoxic bolus of honeysuckle flower and forsythia, YINQIAO JIEDU PIAN, Yinqiao detoxification capsule, YINQIAO JIEDU KELI in one one of the Pharmacopoeia of the People's Republic of China (version in 2005), as prescription, method for making, discriminating, assay, function cure mainly, content such as usage and dosage.

The Yinqiao detoxification preparation is mainly used in anemopyretic cold, fever and headache, cough xerostomia; Treatment of diseases such as have sore throat; All there is the bigger defective of dose in medicines such as existing antitoxic bolus of honeysuckle flower and forsythia, YINQIAO JIEDU PIAN, Yinqiao detoxification capsule, YINQIAO JIEDU KELI, and for example the every ball of antitoxic bolus of honeysuckle flower and forsythia weighs 3g, oral one time 1 ball; 2～3 times on the one, day taking dose is between 6g～9g; For another example, the heavy 0.52g of every of YINQIAO JIEDU PIAN, oral one time 4,2～3 times on the one, day taking dose is between 4g～6g; The heavy 0.4g of every of Yinqiao detoxification capsule, oral one time 4,2～3 times on the one, day taking dose is between 3g～5g; The taking dose of YINQIAO JIEDU KELI is bigger, and a day taking dose reaches between 15g～45g.

In addition, medicines such as antitoxic bolus of honeysuckle flower and forsythia, YINQIAO JIEDU PIAN, Yinqiao detoxification capsule, YINQIAO JIEDU KELI all exist drug dissolution bad, influence the problem of drug effect.

The present invention has carried out improving innovation to the method for preparing of prior art on the basis of existing technology, is prepared into soft capsule dosage form; Improve the medicament contg of UD, can reduce dose widely, made things convenient for the patient to take; Every heavy 0.45g of Yinqiao detoxification soft capsule provided by the invention; Oral one time 2,3 times on the one, a day taking dose is merely 2.7g; Multiple advantages such as and Yinqiao detoxification soft capsule of the present invention has rapid-actionly, and bioavailability is high, and therapeutic effect is good, and preparation is refining, attractive in appearance, health, mouthfeel are good.

Summary of the invention

The object of the invention provides a kind of Yinqiao detoxification soft gelatin pharmaceutical preparation and preparation method thereof;

Another object of the present invention provides the quality determining method of this medicine, so that control the quality of medicine of the present invention effectively.

The objective of the invention is to realize through following technical scheme:

A kind of pharmaceutical preparation of Yinqiao detoxification, this pharmaceutical preparation are soft capsule dosage form, and its medicine is to be processed by the raw material of following weight portion:

Flos Lonicerae 5, Fructus Forsythiae 5, Herba Menthae 3, Herba Schizonepetae 2, Semen Sojae Preparatum 2.5,

Fructus Arctii (stir-fry) 3, Radix Platycodonis 3, Herba Lophatheri 2, Radix Glycyrrhizae 2.5.

Described Yinqiao detoxification soft capsule is prepared by following method:

(1) extracting honeysuckle adds alcohol reflux, reclaims ethanol, and the simmer down to thick paste promptly gets the Flos Lonicerae thick paste;

(2) get Semen Sojae Preparatum and add water boil after, warm macerating filters, and promptly gets the Semen Sojae Preparatum extracting solution;

(3) get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water, decocting liquid filters, and promptly gets mixed extract II;

(4) above Semen Sojae Preparatum extracting solution, mixed extract I, mixed extract II are merged, be concentrated into extractum, centrifugal; Get extractum I, supernatant continues to be concentrated into extractum, gets extractum II; Extractum I, extractum II and Flos Lonicerae thick paste merge, and add volatile oil and edible vegetable oil and adjuvant, mixing; Sieve, be pressed into soft capsule, promptly get.

The preferred version of above-mentioned method for preparing is:

(1) extracting honeysuckle adds 70%～90% alcohol reflux 1～3 time, and each 0.5～2 hour, merge extractive liquid, reclaimed ethanol, and the simmer down to thick paste promptly gets the Flos Lonicerae thick paste;

(2) get Semen Sojae Preparatum and add water boil after, 70 ℃～90 ℃ warm macerating 1～3 time, each 1～3 hour, merge leachate, filter, promptly get the Semen Sojae Preparatum extracting solution;

(3) get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 1～3 time, each 1～3 hour, collecting decoction filtered, and promptly gets mixed extract II;

The more preferred version of above-mentioned method for preparing is:

(1) extracting honeysuckle adds 80% alcohol reflux 2 times, and each 1 hour, merge extractive liquid, reclaimed ethanol, and the simmer down to relative density is 1.28～1.30 thick paste, promptly gets the Flos Lonicerae thick paste;

(2) get Semen Sojae Preparatum and add water boil after, in 80 ℃ of warm macerating 2 times, each 2 hours, merge leachate, filter, promptly get the Semen Sojae Preparatum extracting solution;

(3) get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 2 times, each 2 hours, collecting decoction filtered, and promptly gets mixed extract II;

(4) above Semen Sojae Preparatum extracting solution, mixed extract I, mixed extract II are merged, be concentrated into relative density and be 1.15～1.20 extractum, centrifugal; Extractum I, it is 1.28～1.30 extractum that supernatant continues to be concentrated into relative density, extractum II; Extractum I, extractum II and Flos Lonicerae thick paste merge, and add volatile oil and edible vegetable oil and adjuvant, mixing; Sieve, be pressed into soft capsule, promptly get.

Yinqiao detoxification soft gelatin pharmaceutical of the present invention can adopt in the following discriminating detection method one or more to carry out quality control:

(1) get these article content 2～4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 0.6～1.0g, adds dehydrated alcohol 10～30ml, and supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 5～15 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate ratio is to be developing solvent at 15～20: 3, launches, and takes out; Dry, spray is with the anisaldehyde test solution, and it is clear to be heated to speckle colour developing at 95～115 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

(2) get these article content 2～4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as need testing solution; Get Fructus Forsythiae control medicinal material 1～3g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as control medicinal material solution; According to thin layer chromatography test, draw each 5～20 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, be to be developing solvent at 10～30: 1 with chloroform-methanol ratio; Launch, take out, dry, spray is 10～30: 1 solution with acetic anhydride-sulphuric acid ratio; It is clear to be heated to speckle colour developing at 95～115 ℃, puts coldly, puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color respectively;

(3) get these article content 2～4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as need testing solution; Get Fructus Arctii control medicinal material 0.8～1.4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 5～20 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio is 10～20: 6～10: 0.5 is developing solvent; Launch, take out, dry; Spray is with 10～30% sulphuric acid, and it is clear to be heated to the speckle colour developing at 95～115 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) get these article content 2～4g, add 10～30% hydrochloric acid 10～30ml and chloroform 20～30ml, put and reflux 0.5～1.5 hour in the water-bath; Put coldly, obtain the chloroform layer, water bath method; Residue adds dissolve with methanol and is settled to 5ml, as need testing solution; Extracting liquorice subacid reference substance adds methanol and processes the solution that every 1ml contains 0.1mg, as reference substance solution in addition; According to HPLC, use octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio is 60～80: 30 for mobile phase, and detections wavelength is 240～260nm, and number of theoretical plate should be not less than 1000 by the calculating of enoxolone peak; Draw each 5～20 μ l of above-mentioned two kinds of solution respectively and measure, should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time;

(5) get these article content 2～4g, add petroleum ether 10～30ml, supersound process 5～15 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution; Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution; According to the gas chromatography test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer, follow procedure heats up; Promptly initial with 50 ℃, keep 1min, the speed with 20 ℃/min is warming up to 100 ℃ then; And maintenance 2min; Speed with 5 ℃/min is warming up to 150 ℃ again, and keeps 5min to draw need testing solution and each 1 μ l of reference substance solution, inject gas chromatograph respectively; Should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time.

The optimal technical scheme of above-mentioned discriminating detection method is:

(1) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 0.8g, adds dehydrated alcohol 20ml, and supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate ratio be 17: 3 be developing solvent, launch, take out; Dry, spray is with the anisaldehyde test solution, and it is clear to be heated to speckle colour developing at 105 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

(2) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Get Fructus Forsythiae control medicinal material 2g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to thin layer chromatography test, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol ratio be 20: 1 be developing solvent; Launch, take out, dry, spray is 20: 1 a solution with acetic anhydride-sulphuric acid ratio; It is clear to be heated to speckle colour developing at 105 ℃, puts coldly, puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color respectively;

(3) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Get Fructus Arctii control medicinal material 1.2g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio be 15: 8: 0.5 be developing solvent; Launch, take out, dry; Spray is with 20% sulphuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) get these article content 3g, add 20% hydrochloric acid 20ml and chloroform 25ml, put in the water-bath and to reflux 1 hour, put coldly, obtain the chloroform layer, water bath method, residue add dissolve with methanol and are settled to 5ml, as need testing solution; Extracting liquorice subacid reference substance adds methanol and processes the solution that every 1ml contains 0.1mg, as reference substance solution in addition; According to HPLC, use octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio be 70: 30 for mobile phase, detections wavelength is 250nm, number of theoretical plate should be not less than 1000 by the calculating of enoxolone peak; Draw each 10 μ l of above-mentioned two kinds of solution respectively and measure, should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time;

(5) get these article content 3g, add petroleum ether 20ml, supersound process 10 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution; Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution; According to the gas chromatography test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer, follow procedure heats up; Promptly initial with 50 ℃; Keep 1min, the speed with 20 ℃/min is warming up to 100 ℃ then, and keeps 2min; Speed with 5 ℃/min is warming up to 150 ℃ again, and keeps 5min; Draw each 1 μ l of need testing solution and reference substance solution respectively, inject gas chromatograph; Should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time.

Yinqiao detoxification soft capsule specification of the present invention is every dress 0.45g, adopts HPLC to carry out assay, and detection method is:

(1) chromatographic condition and system suitability test: uses octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio is to be mobile phase at 7～11: 91, and the detection wavelength is 320～335nm;

(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, adds methanol and process the solution that every 1ml contains 20～40 μ g;

(3) preparation of need testing solution: get these article content, mixing, precision takes by weighing 1g, puts in the round-bottomed flask, adds dichloromethane 30～50ml; Water-bath refluxed 0.5～1.5 hour, filtered, and discarded dichloromethane solution, and residue adds methanol 40～60ml, supersound process 20～40 minutes; Put coldly, filter, the methanol wash medicinal residues for several times, merging filtrate and washing liquid are in the 100ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, promptly get with microporous filter membrane;

(4) algoscopy: accurate respectively reference substance solution and each 5～20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.

Above-mentioned content assaying method optimized technical scheme is:

(1) chromatographic condition and system suitability test: uses octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio be 9: 91 for mobile phase, the detection wavelength is 327nm, number of theoretical plate should be not less than 1000 by the calculating of chlorogenic acid peak;

(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds methanol and process the solution that every 1ml contains 30 μ g;

(3) preparation of need testing solution: get these article content, mixing, precision takes by weighing 1g, puts in the round-bottomed flask, adds dichloromethane 40ml; Water-bath refluxed 1 hour, filtered, and discarded dichloromethane solution, and residue adds methanol 50ml, supersound process 30 minutes; Put coldly, filter, the methanol wash medicinal residues for several times, merging filtrate and washing liquid are in the 100ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, promptly get with 0.45 μ m microporous filter membrane;

(4) algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get;

(5) measure the result: every of these article contain chlorogenic acid must not be less than 1.40mg.

The inventor has carried out experimentation to technical scheme provided by the invention, in order to the advantage of therapeutical effect and the quality determining method of the present invention of proof Chinese medicine composition of the present invention, precision, accuracy, stability etc.Following experiment is used to further specify effect of the present invention, but does not limit the present invention.

Experimentation to the present composition:

Experiment one: analgesic experiment

1. test objective

Through analgesic experiment, observe the refrigeration function of Yinqiao detoxification soft capsule, estimate its drug effect.

2. test material

2.1 experimental animal:

Rabbit, body weight 2～3kg (new zealand rabbit) is available from the quality certification is provided by the Nanjing Military Command hospital general: SCXK (Soviet Union) 2005-0029.

2.2 medicine and reagent:

(1) Yinqiao detoxification soft capsule of the present invention;

(2) YINQIAO JIEDU PIAN, source: Beijing Tongrentang Technology Development Co.ltd. Pharmaceutical Factory;

(3) Yinqiao detoxification capsule, source: Hubei Hou Detang pharmaceutcal corporation, Ltd;

(4) aspirin tablet, source: Shandong XinHua Pharmacy stock Co., Ltd;

(5) paratyphoid vaccine, the source: Shanghai Vaccine and Serum Institute, 5ml/ props up.

3. test method and result

3.1 method: select the male large ear rabbit of 38.6～39.5 ℃ of body temperature, 30, equal intravenous injection paratyphoid vaccine 1.5ml/kg; Make 0.8 ℃～1.2 ℃ of fervescence, be divided into 5 groups then at random, 6 every group; And administration simultaneously, dosage is following: Yinqiao detoxification soft capsule group is irritated clothes 0.5g crude drug/kg, and the YINQIAO JIEDU PIAN group is irritated clothes 0.5g crude drug/kg; The Yinqiao detoxification Capsules group is irritated clothes 0.5g crude drug/kg, and the aspirin group is irritated clothes 0.12mg/kg, and matched group is irritated clothes equal-volume normal saline.Every at a distance from 1 body temperature of 1h mensuration, totally 5 times after administration.Calculate the mean temperature difference of body temperature with the heating body temperature of each treated animal different time after administration, relatively with the difference mapping.

3.2 result and conclusion: experimental result is seen table 1; Experimental result shows that Yinqiao detoxification soft capsule, YINQIAO JIEDU PIAN, Yinqiao detoxification capsule all have certain refrigeration function to the fever in rabbits that the absorption Typhoid Vaccine causes; With matched group significant difference is arranged more all; Especially with the Yinqiao detoxification soft capsule to separate thermal effect best, with matched group relatively have utmost point significant difference ( ^*P＜0.01), effect obviously is superior to Yinqiao detoxification capsule and YINQIAO JIEDU PIAN.

The experiment of bringing down a fever of table 1 pair rabbit

Compare with matched group: ^*P＜0.05 ^*P＜0.01

Experiment two: analgesic experiment

1. test objective

Through analgesic experiment, observe the analgesic activity of Yinqiao detoxification soft capsule, estimate its effect, for clinical practice provides the pharmacodynamics foundation.

2. test material

2.1 experimental animal:

(1) ICR mice, body weight 20 ± 2g, male and female are all used; The quality certification is provided: SCXK (Soviet Union) 2005-0059 by the Nanjing Military Command hospital general.

(2) SD rat, body weight 200 ± 20g, male and female half and half; The quality certification is provided: SCXK (Soviet Union) 2005-0065 by the Nanjing Military Command hospital general.

2.2 medicine and reagent:

(1) Yinqiao detoxification soft capsule of the present invention;

(4) ibuprofen capsule, source: Hunan Kang Erjia Pharmaceutical group;

(5) glacial acetic acid, source: permanent Tonghua, Beijing worker's company limited.

3. test method and result

3.1 mice acetic acid twisting experiment

3.1.1 method: select 50 of ICR male mices for use, be divided into 5 groups at random, be respectively: the blank group; Ibuprofen group (0.05g/kg), and Yinqiao detoxification soft capsule group (the 0.5g crude drug/kg), the Yinqiao detoxification Capsules group (the 0.5g crude drug/kg); The YINQIAO JIEDU PIAN group (the 0.5g crude drug/kg), 10 every group.All animals are fasting 12h before experiment, and drinking-water is not limit.Press 0.2ml/10g body weight gastric infusion.The 60min mouse peritoneal is injected 0.6% acetic acid after the administration, every 0.2ml.Observe the writhing response of mice, raising with abdominal part contraction, stretching, extension hind leg or buttocks is that writhing response is positive.The number of times of mouse writhing in the 5min opening entry 20min behind the injection acetic acid.

3.1.2 result and conclusion: experimental result is seen table 2; Yinqiao detoxification pharmaceutical preparation all has the effect of obvious inhibition mouse writhing, compares with the blank group, and significant difference is all arranged; Wherein best with the analgesic effect of Yinqiao detoxification soft capsule especially, with the blank group relatively have utmost point significant difference ( ^*P＜0.01), effect obviously is superior to Yinqiao detoxification capsule and YINQIAO JIEDU PIAN.

Table 2 mice acetic acid twisting reaction result (n=10, x ± s)

Compare with the blank group: ^*P＜0.05 ^*P＜0.01

3.2 rat tail-flick method analgesic experiment

3.2.1 method: get the SD rat, male and female half and half, body weight 200 ± 20g.With 55 ± 1 ℃ of waters bath with thermostatic control is thermic pain source.After rat is fixing, hot water is put in its tail tip (5cm), the entry of record rat tails is to contracting time of dried up; Survey again once behind the 3min, select the end reaction time that contracts at 3～10s with 80 of interior rats, be divided into 8 groups at random: the blank group; The ibuprofen group (the 0.05g crude drug/kg), Yinqiao detoxification soft capsule low dose group (the 0.25g crude drug/kg), Yinqiao detoxification soft capsule high dose group (0.50g/kg); Yinqiao detoxification capsule low dose group (the 0.25g crude drug/kg), Yinqiao detoxification capsule in high dose group (the 0.50g crude drug/kg), the YINQIAO JIEDU PIAN low dose group (the 0.25g crude drug/kg); The YINQIAO JIEDU PIAN high dose group (the 0.50g crude drug/kg), 10 every group.Press 2ml/100g body weight gastric infusion, 60min measures the whipping time of rat once more after administration, relatively respectively organizes the whipping temporal differences before and after the administration.

3.2.2 result and conclusion: practice result is seen table 3, the result shows, rat the whipping reaction all occurs and prolongs phenomenon after giving the Yinqiao detoxification medicine, and with the increasing of dosage the whipping time lengthening.The whipping time after the rat administration of Yinqiao detoxification soft capsule low dose group and high dose group all is longer than the low dose group and the high dose group of Yinqiao detoxification capsule, YINQIAO JIEDU PIAN.

Table 3 Yinqiao detoxification soft capsule rat tail-flick test analgesia result (n=10, x ± s)

Compare with the blank group: ^*P＜0.01

4. conclusion: Yinqiao detoxification soft capsule of the present invention has analgesic activity preferably, and its effect is better than Yinqiao detoxification capsule and YINQIAO JIEDU PIAN.

Experiment three: Yinqiao detoxification soft capsule treatment cold, fever 120 routine clinical observations

In order to observe the clinical efficacy of Yinqiao detoxification soft capsule treatment cold, fever of the present invention, for clinical practice provides foundation.

1. data:

Observe case totally 160 examples, be divided into treatment group and matched group.Treatment is organized in 120 examples, male 74 examples, women 46 examples; Minimum 3 years old of age, maximum 62 years old, 37 years old mean age; Disease time is the shortest 1 day, and is the longest 4 days, average 1.5 days.In matched group 40 examples, male 23 examples, women 17 examples; Minimum 11 years old of age, maximum 67 years old, 41 years old mean age; Disease time is the shortest 1 day, and is the longest 4 days, average 2 days.

2. treat and observational technique

2.1 Therapeutic Method:

The treatment group: stick up the detoxifcation soft capsule, oral, be grown up each 2, every day 3 times, every heavy 0.45g, child's amount is cut down according to the circumstance.

Matched group: YINQIAO JIEDU PIAN, the source: the Beijing Tongrentang Technology Development Co.ltd. Pharmaceutical Factory, oral, be grown up each 4, every day 3 times, child's amount is cut down according to the circumstance.

2.2 diagnostic criteria:

With reference to " traditional Chinese medical science disease diagnosis criterion of therapeutical effect " relevant flu diagnosis basis; With heating fear of cold (wind), lossless headache, nasal obstruction, the red pharyngalgia of pharynx is primary symptom; And heating surpasses 38 ℃ of persons, and blood leukocytes number is normal or on the low side, and neutrophilic granulocyte reduces, lymphocyte increases relatively.

2.3 criterion of therapeutical effect: with reference to " traditional Chinese medical science disease diagnosis criterion of therapeutical effect ", additional fever time and remission scoring are judged.

Cure: temperature recovery is normal in 48 hours, clinical symptom disappearance;

Take a turn for the better: internal heat generation in 48 hours alleviates clinical symptom relief;

Invalid: heating in 48 hours does not alleviate, and clinical symptoms increases the weight of.

3. therapeutic outcome

3.1 clinical efficacy: see table 4.The cure rate of treatment group and total effective rate are all apparently higher than matched group, and through statistical procedures, the P value is all less than 0.01.

Table 4 treatment group and matched group clinical efficacy comparative example (%)

3.2 treat symptom situation after 72 hours: see table 5.Treatment group clinical symptoms is improved situation and obviously is superior to matched group, through statistical procedures, and P＜0.01.

Table 5 treatment group and treatment of control group be symptom situation comparative example (%) after 72 hours

4. conclusion:

Table 4 shows, Yinqiao detoxification soft capsule treatment cold, fever, and two of cure rate and total effective rates all obviously are superior to matched group; Table 5 shows that aspect cold symptoms alleviation improvement, the Yinqiao detoxification soft capsule also is superior to matched group, and clinical observation is to particularly evident to the heavier patient of hyperpyrexia symptom performance.Do not find any toxic and side effects and untoward reaction in the use.

Experiment five: the present invention is differentiated the research and the explanation of detection method

1. the thin layer chromatography of Herba Schizonepetae control medicinal material is differentiated:

In the thin layer chromatography of Herba Schizonepetae control medicinal material was differentiated, technical scheme provided by the invention adopted the method for ethanol ultrasonic extraction to the preparation of need testing solution and control medicinal material solution.The more existing ether jolting of this method, the simple and convenient extraction that placement is spent the night, quick, and extraction is more complete, and testing result is more accurate.

2. the thin layer chromatography of Fructus Forsythiae control medicinal material is differentiated:

In the thin layer of Fructus Forsythiae control medicinal material was differentiated, prior art was at the preparation test sample during with reference substance, the reflux, extract, method of employing or flood method for distilling; Such method length that expends time in; Complicated operating process, technical scheme provided by the invention adopts the method for ethanol ultrasonic extraction, and preparation process time is short; Easy and simple to handle, and testing result is also more accurate.

3. the thin layer chromatography of Fructus Arctii control medicinal material is differentiated:

For the Yinqiao detoxification preparation, in the thin layer of Fructus Arctii medical material was differentiated, the developing solvent of prior art was chloroform-methanol-water (40: 10: 1), and developer is the dilute sulfuric acid test solution.Because speckle diffusion with this understanding is clear inadequately after the colour developing, so adjust.The developing solvent condition is adjusted into toluene-ethyl acetate-formic acid (15: 8: 0.5), and developer is adjusted into 20% sulphuric acid.Through investigating, effect is better, and negative noiseless.

4. the chromatograph of enoxolone is differentiated:

Prior art differentiates the thin layer chromatography of enoxolone and is dehydrated alcohol extraction, extracts fully inadequately, and spot colors is shallow, unintelligible, and colour developing speckle is clearly all arranged near the enoxolone speckle in the test sample chromatograph, causes interference easily, influences result's judgement.

The present invention is revised as method for distilling and adds 20% hydrochloric acid and chloroform extracts, and can enoxolone be extracted comparatively completely, and will differentiate that revision is the liquid chromatograph discriminating; Use octadecylsilane chemically bonded silica to be filler; Acetonitrile-0.1% phosphoric acid solution ratio be 70: 30 for mobile phase, detections wavelength is 250nm, through testing; Can be good at identifying enoxolone with this understanding, and negative noiseless.

5. the discriminating of Mentholum

Because the diffusion of the discriminating speckle of Mentholum is bigger, speckle is fuzzyyer, differentiates so the present invention proposes gas chromatogram.Concrete grammar is: get these article content 3g, add petroleum ether 20ml, supersound process 10 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution.Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution.According to gas chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 E) test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer; Press following temperature programming:

Draw each 1 μ l of above-mentioned two kinds of solution respectively, inject gas chromatograph should present the chromatographic peak identical with the reference substance retention time in the test sample chromatograph.

Through experiment, with this understanding, can be good at identifying Mentholum, and negative noiseless.

Experiment five: to the research and the explanation of content assaying method of the present invention

1. chromatographic condition

Mobile phase: select second eyeball and 0.1% phosphoric acid solution for use, through different proportion research relatively, confirm that ratio is that 9: 91 o'clock separation effects are best, sample separation degree, tailing factor are all up to specification.

Wavelength:, confirm as 327nm through experimentation.

System suitability:, select to use octadecylsilane chemically bonded silica to be filler, flow velocity: 1.0ml/min through experimentation; Temperature; Room temperature; Number of theoretical plate is not less than 1000 by the chlorogenic acid peak.

2. extraction conditions is selected

Be solvent with methanol, 50% methanol, ethanol, 50% ethanol and 70% ethanol respectively, ultrasonic 30min measures, and the result sees table 6.

The different solvents that extract of table 6 are investigated table as a result

Extract solvent	Methanol	50% methanol	Ethanol	50% ethanol	70% ethanol
						Chlorogenic acid content	2.48	2.25	2.04	2.13	2.10

Judge that from the result methanol extraction effect is better.

With methanol is solvent, and ultrasonic 15min, 30min, 60min measure, and the result sees table 7.

The different extraction times investigation of table 7 result

Extract solvent	15min	30min	60min
				Chlorogenic acid content	2.11	2.48	2.46

The result shows, methanol supersound process 30min, and it is best to handle sample result.

3. the preparation of need testing solution

These article of getting content, mixing, precision takes by weighing 1g, puts in the round-bottomed flask, adds dichloromethane 40ml; Backflow 1h filters, and discards dichloromethane solution, and residue adds methanol 50ml, supersound process 30min; Put coldly, filter, with methanol wash medicinal residues several, merging filtrate and washing liquid are in the 100ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, promptly get with microporous filter membrane.

4. linear relationship is investigated

The preparation of storing solution: precision takes by weighing chlorogenic acid 10mg and puts in the 10ml measuring bottle, adds dissolve with methanol and is settled to scale, as storing solution (1mg/ml).

The preparation of reference substance solution: precision takes by weighing above-mentioned storing solution 0.2,0.4,0.6,0.8,1.0ml respectively, puts in the 10ml measuring bottle, adds methanol and is diluted to scale, shakes up, as reference substance solution.According to the high effective liquid chromatography for measuring peak area, the result sees table 8.

Table 8 chlorogenic acid linear relationship is investigated table

Sequence number	1	2	3	4	5
						Concentration (ug/ml)	20	40	60	80	100
Peak area	325752	612637	890750	1176595	1451127

Peak area (A) with the chlorogenic acid reference substance returns concentration (C), and get regression equation: with concentration is abscissa, is vertical coordinate with the peak area.

A＝14074C+46960，r＝1，

The result shows, in 20～100 μ g/ml scopes, is good linear relationship.

5. precision test

Accurate chlorogenic acid contrast liquid (30 μ g/ml) the 10 μ l that draw inject high performance liquid chromatograph, and continuous sample introduction 5 times is measured its peak area value, and experimental result is seen table 9, and RSD=0.85% shows that precision is good.

The test of table 9 precision

6. stability test

Get same need testing solution, every at a distance from 0,2,6,12,24h sample introduction twice, the record peak area, calculating shows that test sample is good at the 24h internal stability, experimental result is seen table 10, RSD=0.60% shows to have good stability.

Table 10 stability test

7. replica test

Get 5 parts of Yinqiao detoxification soft capsule contents respectively, every part of heavily about 1g, the accurate title, decide, and puts in the tool plug conical flask, adds dichloromethane 40ml, and backflow 1h filters, and discards dichloromethane solution, and residue adds methanol 50ml.Supersound process 30min is put coldly, filters, and with the methanol wash medicinal residues for several times, merging filtrate and washing liquid add methanol and are diluted to scale in the 100ml measuring bottle, shake up, with the microporous filter membrane filtration, as need testing solution.Measure by above-mentioned chromatographic condition, the result sees table 11, shows that the repeatability of experiment is good.

Table 11 replica test

8. experiment conclusion:

Comprehensive above result shows accurately chlorogenic acid contents in the working sample of detection method of the present invention.

Effect intentionally of the present invention:

1. adopt technology of the present invention that medicine is processed soft capsule dosage form; Rapid-action, bioavailability is high, and the medicine that is comprised is that the form with dispersion exists; Fully disperse with substrate; Get into the rapid disintegrate of gastrointestinal and be absorbed by the body, reach effective blood drug concentration and produce effects, thereby improved the curative effect of former dosage form.

2. adopt technology of the present invention that medicine is processed soft capsule dosage form, increased stability of drug.

3. adopt technology of the present invention that medicine is processed soft capsule dosage form, applied range does not contain sugar, is convenient to the old people and diabetics is taken.

4. Yinqiao detoxification soft capsule impurity content of the present invention is low, and refining, attractive in appearance.

5. Yinqiao detoxification soft capsule of the present invention one-shot forming, soft capsule is totally-enclosed dosage form, has reduced the chance of polluting, and guaranteed the hygiology quality of medicine, and YINQIAO JIEDU PIAN is full powder formulation, the hygiology index is difficult to guarantee.

6. Yinqiao detoxification soft capsule mouthfeel of the present invention is good, has covered the bad smell of Chinese medicine extract and the uncomfortable flavor of smelling, and is convenient to take.

7. Yinqiao detoxification soft capsule dose of the present invention is little, easy to carry, and prior art YINQIAO JIEDU PIAN or capsule etc., and dose is big, and the patient is difficult for accepting, adopt technology of the present invention to process soft capsule after, dose reduces, and is easy to product promotion.

8. technological deficiencies such as discrimination method provided by the invention has overcome the diffusion of prior art speckle, and the speckle color is shallow, speckle fuzzy, colour developing is unintelligible; The discrimination method of innovation has been proposed; These methods can identify relevant material exactly, and method is reliable and stable, and are highly sensitive; Favorable reproducibility, and negative noiseless.

9. in the content assaying method of the present invention; Adopt the method for preparing of need testing solution provided by the invention; Can extract test substance---chlorogenic acid more fully; And adopting HPLC to measure, repeatability, accuracy, precision, sensitivity are all fine, receive external influence factor less.

The specific embodiment

Embodiment 1:

Prescription: Flos Lonicerae 400g Fructus Forsythiae 400g Herba Menthae 240g

Herba Schizonepetae 160g Semen Sojae Preparatum 200g Fructus Arctii (stir-fry) 240g

Radix Platycodonis 240g Herba Lophatheri 160g Radix Glycyrrhizae 200g

Method for making: extracting honeysuckle adds 80% alcohol reflux secondary, and each 1 hour, merge extractive liquid, reclaimed ethanol, and the simmer down to relative density is 1.28～1.30 thick paste, promptly gets the Flos Lonicerae thick paste;

After getting Semen Sojae Preparatum and adding water boil, in 80 ℃ of warm macerating secondaries, each 2 hours, merge leachate, filter, promptly get the Semen Sojae Preparatum extracting solution;

Get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water secondary, each 2 hours, collecting decoction filtered, and promptly gets mixed extract II;

Semen Sojae Preparatum extracting solution, mixed extract I, mixed extract II are merged, be concentrated into relative density and be 1.15～1.20 extractum, centrifugal; Extractum I, it is 1.28～1.30 extractum that supernatant continues to be concentrated into relative density, extractum II; Extractum I, extractum II and Flos Lonicerae thick paste merge, and add volatile oil and refined edible vegetable oil and adjuvant, mixing; Sieve, be pressed into 1000 of soft capsules, promptly get.

Differentiate:

(1) get these article content 3g, add dehydrated alcohol 20ml, ultrasonic place 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 0.8g, adds dehydrated alcohol 20ml, and ultrasonic place 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate ratio be 17: 3 be developing solvent, launch, take out; Dry, spray is with the anisaldehyde test solution, and it is clear to be heated to speckle colour developing at 105 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

Assay:

Adopt HPLC to carry out assay:

(5) measure the result: it is 2.60mg that every of these article contain chlorogenic acid.The conformance with standard regulation.

Function with cure mainly: dispelling wind to relieve the exterior syndrome, heat-clearing and toxic substances removing.Be used for anemopyretic cold, card is seen fever and headache, and the cough xerostomia is had sore throat; Upper respiratory tract infection is seen above-mentioned card marquis person.

Usage and consumption: oral, one time 2,3 times on the one;

Specification: every dress 0.45g.

Embodiment 2:

Prescription: Flos Lonicerae 400g Fructus Forsythiae 400g Herba Menthae 240g

Radix Platycodonis 240g Herba Lophatheri 160g Radix Glycyrrhizae 200g

Method for making: extracting honeysuckle adds alcohol reflux, reclaims ethanol, and the simmer down to thick paste promptly gets the Flos Lonicerae thick paste;

After getting Semen Sojae Preparatum and adding water boil, warm macerating filters, and promptly gets the Semen Sojae Preparatum extracting solution;

Get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water, decocting liquid filters, and promptly gets mixed extract II;

Above Semen Sojae Preparatum extracting solution, mixed extract I, mixed extract II are merged, be concentrated into extractum, centrifugal; Get extractum I, supernatant continues to be concentrated into extractum, gets extractum II; Extractum I, extractum II and Flos Lonicerae thick paste merge, and add volatile oil and edible vegetable oil and adjuvant, mixing; Sieve, be pressed into 1000 of soft capsules, promptly get.

Differentiate:

(1) get these article content 2g, add dehydrated alcohol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 0.6g, adds dehydrated alcohol 10ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as control medicinal material solution; According to thin layer chromatography experiment, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate ratio be 15: 3 be developing solvent, launch, take out, dry, spray is with the anisaldehyde test solution, 95 ℃ be heated to speckle develop the color clear; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

(2) get these article content 2g, add dehydrated alcohol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as need testing solution; Get Fructus Forsythiae control medicinal material 1g, add dehydrated alcohol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as control medicinal material solution; According to thin layer chromatography test, draw each 5 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol ratio be 10: 1 be developing solvent; Launch, take out, dry, spray is 10: 1 a solution with acetic anhydride-sulphuric acid ratio; It is clear to be heated to speckle colour developing at 95 ℃, puts coldly, puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color respectively;

(3) get these article content 2g, add dehydrated alcohol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as need testing solution; Get Fructus Arctii control medicinal material 0.8g, add dehydrated alcohol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 1ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 5 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio be 10: 6: 0.5 be developing solvent; Launch, take out, dry; Spray is with 10% sulphuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) get these article content 2g, add 10% hydrochloric acid 10ml and chloroform 20ml, put in the water-bath and to reflux 0.5 hour, put coldly, obtain the chloroform layer, water bath method, residue add dissolve with methanol and are settled to 5ml, as need testing solution; Extracting liquorice subacid reference substance adds methanol and processes the solution that every 1ml contains 0.1mg, as reference substance solution in addition; According to HPLC, use octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio be 60: 30 for mobile phase, detections wavelength is 240nm, number of theoretical plate should be not less than 1000 by the calculating of enoxolone peak; Draw each 5 μ l of above-mentioned two kinds of solution respectively and measure, should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time;

(5) get these article content 2g, add petroleum ether 10ml, supersound process 5 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution; Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution; According to the gas chromatography test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer, follow procedure heats up; Promptly initial with 50 ℃; Keep 1min, the speed with 20 ℃/min is warming up to 100 ℃ then, and keeps 2min; Speed with 5 ℃/min is warming up to 150 ℃ again, and keeps 5min; Draw each 1 μ l of need testing solution and reference substance solution respectively, inject gas chromatograph; Should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time.

Assay:

Adopt HPLC to carry out assay:

(1) chromatographic condition and system suitability test: uses octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio is to be mobile phase at 7: 91, and the detection wavelength is 320nm;

(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, adds methanol and process the solution that every 1ml contains 30 μ g;

(3) preparation of need testing solution: get these article content, mixing, precision takes by weighing 1g, puts in the round-bottomed flask, adds dichloromethane 30ml; Water-bath refluxed 0.5 hour, filtered, and discarded dichloromethane solution, and residue adds methanol 40ml, supersound process 20 minutes; Put coldly, filter, the methanol wash medicinal residues for several times, merging filtrate and washing liquid are in the 100ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, promptly get with microporous filter membrane;

(4) algoscopy: accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.

(5) measure the result: it is 2.70mg that every of these article contain chlorogenic acid.The conformance with standard regulation.

Usage and consumption: oral, one time 2,3 times on the one;

Specification: every dress 0.45g.

Embodiment 3:

Prescription: Flos Lonicerae 400g Fructus Forsythiae 400g Herba Menthae 240g

Radix Platycodonis 240g Herba Lophatheri 160g Radix Glycyrrhizae 200g

Method for making: extracting honeysuckle adds 70%～90% alcohol reflux 1 time, and 2 hours, extracting solution reclaimed ethanol, and the simmer down to thick paste promptly gets the Flos Lonicerae thick paste;

After getting Semen Sojae Preparatum and adding water boil, 70 ℃ of warm macerating 1 time, each 3 hours, merge leachate, filter, promptly get the Semen Sojae Preparatum extracting solution;

Get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 1 time, each 3 hours, decocting liquid filtered, and promptly gets mixed extract II;

Differentiate:

(1) get these article content 4g, add dehydrated alcohol 30ml, supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 1.0g, adds dehydrated alcohol 30ml, and supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 15 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate ratio be 16: 3 be developing solvent, launch, take out; Dry, spray is with the anisaldehyde test solution, and it is clear to be heated to speckle colour developing at 115 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

(2) get these article content 4g, add dehydrated alcohol 30ml, supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as need testing solution; Get Fructus Forsythiae control medicinal material 3g, add dehydrated alcohol 30ml, supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as control medicinal material solution; According to thin layer chromatography test, draw each 20 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol ratio be 30: 1 be developing solvent; Launch, take out, dry, spray is 30: 1 a solution with acetic anhydride-sulphuric acid ratio; It is clear to be heated to speckle colour developing at 115 ℃, puts coldly, puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color respectively;

(3) get these article content 4g, add dehydrated alcohol 30ml, supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as need testing solution; Get Fructus Arctii control medicinal material 1.4g, add dehydrated alcohol 30ml, supersound process 40 minutes filters, and filtrating is concentrated into about 3ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 20 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio be 20: 10: 0.5 be developing solvent; Launch, take out, dry; Spray is with 30% sulphuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) get these article content 4g, add 30% hydrochloric acid 30ml and chloroform 30ml, put in the water-bath and to reflux 1.5 hours, put coldly, obtain the chloroform layer, water bath method, residue add dissolve with methanol and are settled to 5ml, as need testing solution; Extracting liquorice subacid reference substance adds methanol and processes the solution that every 1ml contains 0.1mg, as reference substance solution in addition; According to HPLC, use octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio be 80: 30 for mobile phase, detections wavelength is 260nm, number of theoretical plate should be not less than 1000 by the calculating of enoxolone peak; Draw each 20 μ l of above-mentioned two kinds of solution respectively and measure, should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time;

(5) get these article content 4g, add petroleum ether 30ml, supersound process 15 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution; Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution; According to the gas chromatography test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer, follow procedure heats up; Promptly initial with 50 ℃; Keep 1min, the speed with 20 ℃/min is warming up to 100 ℃ then, and keeps 2min; Speed with 5 ℃/min is warming up to 150 ℃ again, and keeps 5min; Draw each 1 μ l of need testing solution and reference substance solution respectively, inject gas chromatograph; Should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time.

Assay:

Adopt HPLC to carry out assay:

(1) chromatographic condition and system suitability test: uses octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio is to be mobile phase at 11: 91, and the detection wavelength is 335nm;

(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, adds methanol and process the solution that every 1ml contains 40 μ g;

(3) preparation of need testing solution: get these article content, mixing, precision takes by weighing 1g, puts in the round-bottomed flask, adds dichloromethane 50ml; Water-bath refluxed 1.5 hours, filtered, and discarded dichloromethane solution, and residue adds methanol 60ml, supersound process 40 minutes; Put coldly, filter, the methanol wash medicinal residues for several times, merging filtrate and washing liquid are in the 100ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, promptly get with microporous filter membrane;

(4) algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.

(5) measure the result: it is 2.65mg that every of these article contain chlorogenic acid.The conformance with standard regulation.

Usage and consumption: oral, one time 2,3 times on the one;

Specification: every dress 0.45g.

Embodiment 4:

Prescription: Flos Lonicerae 400g Fructus Forsythiae 400g Herba Menthae 240g

Radix Platycodonis 240g Herba Lophatheri 160g Radix Glycyrrhizae 200g

Method for making: extracting honeysuckle adds 90% alcohol reflux 3 times, and each 0.5 hour, merge extractive liquid, reclaimed ethanol, and the simmer down to thick paste promptly gets the Flos Lonicerae thick paste;

After getting Semen Sojae Preparatum and adding water boil, 90 ℃ of warm macerating 3 times, each 1 hour, merge leachate, filter, promptly get the Semen Sojae Preparatum extracting solution;

Get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (stir-fry), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 3 times, each 1 hour, collecting decoction filtered, and promptly gets mixed extract II;

Differentiate:

(1) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Other gets Herba Schizonepetae control medicinal material 0.8g, adds dehydrated alcohol 20ml, and supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate ratio be 18: 3 be developing solvent, launch, take out; Dry, spray is with the anisaldehyde test solution, and it is clear to be heated to speckle colour developing at 100 ℃; In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively;

(2) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Get Fructus Forsythiae control medicinal material 2g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to thin layer chromatography test, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol ratio be 20: 1 be developing solvent; Launch, take out, dry, spray is 20: 1 a solution with acetic anhydride-sulphuric acid ratio; It is clear to be heated to speckle colour developing at 100 ℃, puts coldly, puts under the 365nm ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color respectively;

(3) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Get Fructus Arctii control medicinal material 1.0g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio be 15: 8: 0.5 be developing solvent; Launch, take out, dry; Spray is with 20% sulphuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) get these article content 3g, add 20% hydrochloric acid 20ml and chloroform 25ml, put in the water-bath and to reflux 1.0 hours, put coldly, obtain the chloroform layer, water bath method, residue add dissolve with methanol and are settled to 5ml, as need testing solution; Extracting liquorice subacid reference substance adds methanol and processes the solution that every 1ml contains 0.1mg, as reference substance solution in addition; According to HPLC, use octadecylsilane chemically bonded silica to be filler, acetonitrile-0.1% phosphoric acid solution ratio be 70: 30 for mobile phase, detections wavelength is 250nm, number of theoretical plate should be not less than 1000 by the calculating of enoxolone peak; Draw each 10 μ l of above-mentioned two kinds of solution respectively and measure, should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time;

Assay:

Adopt HPLC to carry out assay:

(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown bottle, adds methanol and process the solution that every 1ml contains 20 μ g;

(5) measure the result: it is 2.68mg that every of these article contain chlorogenic acid.The conformance with standard regulation.

Usage and consumption: oral, one time 2,3 times on the one;

Specification: every dress 0.45g.

Claims

1. Yinqiao detoxification preparation of soft capsule method is characterized by:

Feedstock production by following weight portion:

Fructus Arctii (parched) 3, Radix Platycodonis 3, Herba Lophatheri 2, Radix Glycyrrhizae 2.5;

Method for preparing:

(3) get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (parched), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 1～3 time, each 1～3 hour, collecting decoction filtered, and promptly gets mixed extract II;

2. method for preparing as claimed in claim 1 is characterized in that:

(3) get Herba Menthae, Herba Schizonepetae, Fructus Forsythiae extraction volatile oil, obtain volatile oil, subsequent use, the aqueous solution after distillation device is in addition collected, and promptly gets mixed extract I; Medicinal residues after the distillation and Fructus Arctii (parched), Herba Lophatheri, Radix Glycyrrhizae, Radix Platycodonis decocte with water 2 times, each 2 hours, collecting decoction filtered, and promptly gets mixed extract II;

(4) above Semen Sojae Preparatum extracting solution, mixed extract I, mixed extract II are merged, be concentrated into relative density and be 1.15～1.20 extractum, centrifugal; It is 1.28～1.30 extractum that supernatant continues to be concentrated into relative density, and extractum I, extractum II and Flos Lonicerae thick paste merge, and add volatile oil and edible vegetable oil and adjuvant; Mixing; Sieve, be pressed into soft capsule, promptly get.

3. the discrimination method of a Yinqiao detoxification soft capsule is characterized in that adopting in the following discrimination method one or more to carry out quality testing:

(3) get these article content 2～4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as need testing solution; Get Fructus Arctii control medicinal material 0.8～1.4g, add dehydrated alcohol 10～30ml, supersound process 20～40 minutes filters, and filtrating is concentrated into about 1～3ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 5～20 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio is 10～20: 6～10: 0.5 is developing solvent; Launch, take out, dry; Spray is with 10～30% sulphuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

(5) get these article content 2～4g, add petroleum ether 10～30ml, supersound process 5～15 minutes filters, and filtrating is concentrated into about 5ml, as need testing solution; Get the Mentholum reference substance, add petroleum ether and process the solution that every 1ml contains 70ug, as reference substance solution; According to the gas chromatography test, be the capillary chromatographic column of immobile phase with 5% diphenyl-95% dimethyl-silicon alkyl copolymer, follow procedure heats up; Promptly initial with 50 ℃; Keep 1min, the speed with 20 ℃/min is warming up to 100 ℃ then, and keeps 2min; Speed with 5 ℃/min is warming up to 150 ℃ again, and keeps 5min; Draw each 1 μ l of need testing solution and reference substance solution respectively, inject gas chromatograph; Should present the chromatographic peak identical in the test sample chromatograph with the reference substance retention time.

4. the discriminating detection method of Yinqiao detoxification soft capsule as claimed in claim 3 is characterized by:

(3) get these article content 3g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Get Fructus Arctii control medicinal material 1.2g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; According to the thin layer chromatography experiment, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively on same silica gel g thin-layer plate; With toluene-ethyl acetate-formic acid ratio be 15: 8: 0.5 be developing solvent; Launch, take out, dry; Spray is with 20% sulphuric acid, and it is clear to be heated to the speckle colour developing at 95～115 ℃; In the test sample chromatograph, respectively with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color;

5. the content assaying method of a Yinqiao detoxification soft capsule is characterized in that every dress of this medicine 0.45g, adopts HPLC to carry out assay, and every of these article contain chlorogenic acid must not be less than 1.40mg, and determination step is:

6. content assaying method as claimed in claim 5 is characterized by: