CN101716260A

CN101716260A - Quality control method of capsule preparation for treating dermatosis

Info

Publication number: CN101716260A
Application number: CN200910310865A
Authority: CN
Inventors: 周强; 皮海燕; 罗阳洋; 刘艳
Original assignee: GUIYANG CHUNKE PHARMACY GROUP TECHNICAL RESEARCH Co Ltd
Current assignee: GUIYANG CHUNKE PHARMACY GROUP TECHNICAL RESEARCH Co Ltd
Priority date: 2009-12-04
Filing date: 2009-12-04
Publication date: 2010-06-02

Abstract

The invention discloses a quality control method of a capsule preparation for treating dermatosis, comprising characters, identification, examination and content measurement. The identification is used for identifying honeysuckle, panicled swallowwort root, viola japonica and dittany bark; and in the content measurement, high-efficiency chromatography phase and UV-VIS spectrophotometry are respectively used for measuring the content of paeonol and total flavonoids in the capsule preparation. Compared with the prior art, the invention has the advantages that the quality control method of the capsule preparation for treating the dermatosis is established to carry out study and screening on the characters, the identification, the examination and the content measurement of the capsule preparation, and the adopted quality control method is scientific and reasonable, has high accuracy and favorable reproducibility, can comprehensively and efficiently control the quality of the preparation and ensure the clinical effect of the preparation.

Description

Treat the method for quality control of dermopathic capsule preparations

Technical field

The present invention relates to the method for quality control of medicine, especially a kind of method of quality control for the treatment of dermopathic capsule preparations belongs to technical field of Chinese medicines.

Background technology

Dermatitis, eczema, urticaria are frequently-occurring disease, the commonly encountered diseases in the dermatosis, cause of disease complexity, many with the erythra form, be easy to ooze out, course of disease delay and tendency of recurrence is arranged is feature, because pathological changes is distributed in body surface, the patient lacks medical knowledge in addition, it is improper to nurse, bad when causing state of an illness fashion, repeatedly outbreak.Secular protracted course of disease causes the state of an illness further to increase the weight of: affected part sepage, drying, incrustation, coarse, plumpness and breach, and attend by unbearable itching, in the very long process of seeking medical advice, still not good sick, treatment is lost confidence, even misunderstood, caused fear, give the heavy thought pressure of patient's back, had a strong impact on patient's work, study, daily life.Traditional traditional Chinese medical science idea is thought, causes eczema, dermatitis, urticarial direct chief culprit, is exactly " blood poison ", and heat in blood, blood stasis, blood are dry.Yet secular clinical practice finds, patient's " blood poison " is to be difficult to remove clean, and the internal organs that reason just is the patient worthy of the name are made malicious machine just as one, carry toxin continuously in blood.So-called " sick at the blood poison, root is at dysentery ", the poison of the five internal organs is not arranged, and blood poison just can't be removed totally, causes eczema, dermatitis, urticaria in the course of time again and again, is difficult to treatment.Thus: control blood and must control dirtyly earlier, the poison that purifies the blood must be arranged dysentery earlier, and like this, the differentiation mechanism of epiderm skin cell could be recovered normally, and eczema, dermatitis, urticaria could be effected a radical cure.

The dermopathic sickness rate of China is very high, and according to interrelated data statistics, national total prevalence rate is 1.23%, promptly has 1.6 hundred million people to suffer from various degree dermatosis approximately.Through measuring and calculating, domestic theoretical market capacity reaches 32,000,000,000 yuan.Wherein antifungal agent is topmost dermatosis medication, accounts for the over half of whole market.What the treatment dermatosis was the most frequently used at present is medicine for external use, cures the symptoms, not the disease, and is easy to recurrence.Medicine for external use is selected or improper use, and is often invalid, even makes state of an illness aggravation, deterioration, and life-time service has certain drug resistance, brought painful very much to the patient.Can effectively treat the dermopathic Chinese medicines of pruritus such as eczema, urticaria, dermatitis in order to develop, the applicant has carried out a large amount of explorations and research.

The present inventor has asked for protection dermopathic medicine of a kind of treatment and preparation method thereof in number of patent application is the patent application of CN2007102030459, wherein capsule preparations is such preparation: Herba Polygoni cymosi 400g, Cortex Dictamni 200g, Flos Lonicerae 200g and Herba Violae japonicae 200g, decoct with water 3 times, add 10 times of water gagings at every turn and decocted 3 hours, filter merging filtrate, be evaporated to 60 ℃ of relative densities and be 1.25 extractum, drying under reduced pressure is pulverized, and crosses 80 mesh sieves; Radix Cynanchi Paniculati 80g pulverizes, and crosses 80 mesh sieves; With extract powder and crude drug powder mixing, adding starch is an amount of, and with 75% alcohol granulation, drying incapsulates, and every dress 0.3g makes 1000; This drug regimen mainly is made up of flavour of a drug such as Herba Polygoni cymosi, Cortex Dictamni, Flos Lonicerae, Herba Violae japonicae and Radix Cynanchi Paniculatis.Has dispeiling pathogenic wind and removing dampness, clearing away heat and cooling blood, detoxifying and relieving itching; Be used for the pruritus dermatosis such as eczema, urticaria, dermatitis due to heat in blood wind-dryness, the hot and humid ecchymosis.Because this medicine is a kind of new drug, does not also have the effective quality control method at present.The inventor studies the capsule preparations of this medicine, in the hope of exploring quality how effectively to control capsule preparations, to guarantee the clinical efficacy of capsule preparations.

Summary of the invention

Technical problem to be solved by this invention is to provide a kind of method of quality control for the treatment of dermopathic capsule preparations, comprises character, discriminating, inspection and assay.The method of quality control of being formulated can be controlled the quality of capsule preparations fully and effectively, thereby has guaranteed the clinical efficacy of capsule preparations.

For solving the problems of the technologies described above, technical scheme of the present invention: treat the method for quality control of dermopathic capsule preparations, this capsule prepares like this: Herba Polygoni cymosi 400g, Cortex Dictamni 200g, Flos Lonicerae 200g and Herba Violae japonicae 200g decoct with water 3 times, add 10 times of water gagings decocted 3 hours at every turn, filter, merging filtrate is evaporated to 60 ℃ of relative densities and is 1.25 extractum, drying under reduced pressure, pulverize, cross 80 mesh sieves, get extract powder; Get Radix Cynanchi Paniculati 80g, pulverize, cross 80 mesh sieves, get the crude drug powder; With extract powder and crude drug powder mixing, adding starch is an amount of, and with 75% alcohol granulation, drying incapsulates, and every dress 0.3g makes 1000; Method of quality control do as one likes shape, discriminating, inspection and assay are formed, wherein discriminating is the discriminating to Flos Lonicerae, Radix Cynanchi Paniculati, Herba Violae japonicae and Cortex Dictamni, and assay is respectively paeonol in the capsule preparations and content of total flavone to be measured with high performance liquid chromatography and UV-VIS spectrophotometry.

The method of quality control of the above-mentioned dermopathic capsule preparations of treatment specifically is such:

Character: this product is a hard capsule, and content is sepia and white granule and powder, and gas is special, bitter in the mouth, little puckery;

Differentiate: be contrast, be adhesive with the sodium carboxymethyl cellulose, be that developing solvent is according to thin layer chromatography discriminating Flos Lonicerae with the upper strata liquid of butyl acetate-formic acid-water with the Flos Lonicerae control medicinal material; With the Radix Cynanchi Paniculati control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with cyclohexane extraction-ethyl acetate differentiate Radix Cynanchi Paniculati according to thin layer chromatography; With the Herba Violae japonicae control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with toluene-ethyl acetate-formic acid differentiate Herba Violae japonicae according to thin layer chromatography; With the Cortex Dictamni control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with chloroform-ethyl acetate-acetone differentiate Cortex Dictamni according to thin layer chromatography;

Check: meet the every regulation under the capsule item among appendix IL of Chinese Pharmacopoeia version in 2005;

Assay:

The assay of paeonol: Radix Cynanchi Paniculati is contrast with the paeonol reference substance, is filler with the octadecylsilane chemically bonded silica, is the content of mobile phase according to paeonol in the high effective liquid chromatography for measuring capsule preparations with methanol-water (60: 40);

Content of total flavone is measured: total flavones is contrast with the control substance of Rutin, is developer with 5% sodium nitrite solution, 10% aluminum nitrate solution and sodium hydroxide test solution, measures content of total flavone in the capsule preparations according to ultraviolet visible spectrophotometry.

In the method for quality control of the dermopathic capsule preparations of aforesaid treatment, described discriminating is formed by following:

(1) Flos Lonicerae is differentiated: get capsule 's content 1g, add methanol 5ml, supersound extraction 30 minutes is filtered, and gets filtrate, as need testing solution; Extracting honeysuckle control medicinal material 0.5g makes control medicinal material solution with the need testing solution preparation method in addition; Test according to thin layer chromatography, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that the upper strata liquid of butyl acetate-formic acid-water of 7: 2.5: 2.5 is developing solvent with volume ratio, launch, take out, dry, spray was dried by the fire 5 minutes down for 105 ℃ in temperature with 5% ferric chloride-alcoholic solution, take out, inspect under the daylight; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(2) Radix Cynanchi Paniculati is differentiated: get capsule 's content 5g, add methanol 15ml, supersound extraction 30 minutes is filtered, and filtrate is concentrated into about 5ml, filters, and gets filtrate, as need testing solution; Other gets Radix Cynanchi Paniculati control medicinal material 3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that cyclohexane extraction-ethyl acetate of 3: 1 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(3) Herba Violae japonicae is differentiated: get capsule 's content 1g, add methanol 5ml, and ultrasonic 30 minutes, filter, get filtrate, as need testing solution; Other gets Herba Violae japonicae control medicinal material 0.5g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that toluene-ethyl acetate-formic acid of 5: 3: 0.5 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) Cortex Dictamni is differentiated: get capsule 's content 5g, add ethanol 20ml, water-bath refluxed 30 minutes, filtered, the filtrate water-bath volatilizes, and adds water 20ml dissolving, uses ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid, water-bath volatilizes, and adds methanol 1ml dissolving, as need testing solution; Other gets Cortex Dictamni control medicinal material 0.3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 5ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that chloroform-ethyl acetate-acetone of 10: 0.5: 0.2 is developing solvent, launch, take out, dry, spray is with the bismuth potassium iodide solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

In the method for quality control of the dermopathic capsule preparations of aforesaid treatment, the assay of paeonol is in the described capsule preparations:

Radix Cynanchi Paniculati chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-water (60: 40) is a mobile phase; Flow velocity is 1.0ml/min; The detection wavelength is 274nm; Column temperature: 35 ℃, number of theoretical plate calculates by the paeonol peak should be not less than 3000;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the paeonol reference substance, adds methanol and make the solution that contains paeonol 0.02064mg among every 1ml, promptly gets reference substance solution;

The preparation of need testing solution: get this product under the content uniformity item, mixing is got 1g, the accurate title, decide, and puts in the tool plug conical flask, adds methanol solution 25ml, weigh supersound process (power 250W, frequency 40kHz) 10 minutes, take out, put coldly, mend heavy, filter, get in subsequent filtrate 1ml to the 10ml measuring bottle, add the methanol standardize solution, filter with 0.45 μ m filter membrane, as need testing solution;

Algoscopy: get each 10 μ l of need testing solution and reference substance solution respectively, according to high performance liquid chromatography, inject hplc determination, the record chromatogram is pressed external standard method with calculated by peak area, promptly;

Content of total flavone is determined as in the described capsule preparations:

The preparation of total flavones reference substance solution: precision takes by weighing 120 ℃ of control substance of Rutin that are dried to constant weight an amount of, puts in the 25ml measuring bottle, and it is an amount of to add methanol, puts that slight fever makes dissolving in the water-bath, puts coldly, adds methanol to scale, shakes up; Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and makes the solution that contains rutin 0.20192mg among every 1ml, promptly gets reference substance solution;

The standard curve preparation: precision is measured reference substance solution 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, put respectively in the 25ml measuring bottle, respectively add water to 6.0ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, and added 10% aluminum nitrate solution 1ml, placed 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale again, shake up, placed 15 minutes, with the corresponding reagent is blank, according to ultraviolet visible spectrophotometry, measure absorbance at 510nm wavelength place, be vertical coordinate with the absorbance, concentration is abscissa, the drawing standard curve;

The preparation of need testing solution: get this product under the content uniformity item, mixing, get 0.2g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 70% ethanol 25ml that adds, weigh, ultrasonic 10min places room temperature, supply with 70% ethanol and to subtract weight loss, filter, the accurate absorption in subsequent filtrate 1ml to the 25ml volumetric flask adds water to 5ml, method under the sighting target directrix curve preparation, make need testing solution from " adding water to 6.0ml " with the method operation, measure absorbance in accordance with the law, read the weight that contains anhydrous rutin the need testing solution from standard curve, calculate, promptly.

In the method for quality control of the dermopathic capsule preparations of aforesaid treatment, every capsules contains Radix Cynanchi Paniculati in paeonol (C9H10O3), is no less than 1.02mg; Every capsules contains total flavones in anhydrous rutin (C27H30O16), is no less than 2.00mg.

In order to study the method for quality control of the dermopathic capsule preparations of treatment, the inventor has carried out a large amount of experiments with screening preferred plan, and is specific as follows:

One, the research of character

Result of the test is per sample drafted, and test agent is all described consistently with drafting the result in three batches, lists the method for quality control text in.

Two, the research of discrimination method

2.1 the thin layer chromatography of Flos Lonicerae is differentiated in the prescription

Get capsule 's content 1g, add methanol 5ml, supersound extraction 30 minutes is filtered, and gets filtrate, as need testing solution; Other gets and lacks Flos Lonicerae sample 1g, makes negative control solution with the need testing solution preparation method; Extracting honeysuckle control medicinal material 0.5g makes control medicinal material solution with the need testing solution preparation method in addition; Test according to thin layer chromatography, draw each 10 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that the upper strata liquid of butyl acetate-formic acid-water of 7: 2.5: 2.5 is developing solvent with volume ratio, launch, take out, dry, spray is with 5% ferric chloride-alcoholic solution, about 5 minutes of 105 ℃ of bakings, take out, inspect under the daylight; In the test sample chromatograph, with the corresponding position of control medicinal material solution chromatograph on, show the speckle of same color; Negative control is noiseless; Test agent checking in 3 batches, the method good reproducibility, specificity is strong, so classify it as method of quality control.

2.2 the thin layer chromatography of Radix Cynanchi Paniculati is differentiated in the prescription

Get capsule 's content 5g, add methanol 15ml, supersound extraction 30 minutes is filtered, and filtrate is concentrated into about 5ml, filters, and gets filtrate, as need testing solution; Other gets and lacks Radix Cynanchi Paniculati sample 5g, makes negative control solution with the need testing solution preparation method; Other gets Radix Cynanchi Paniculati control medicinal material 3g, makes control medicinal material solution with the need testing solution preparation method; According to thin layer chromatography test, draw above-mentioned three kinds of each 10ul of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that cyclohexane extraction-ethyl acetate of 3: 1 is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp 365nm and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material solution chromatograph on, show the speckle of same color; Negative control is noiseless; Test agent checking in 3 batches, the method good reproducibility, specificity is strong, so classify it as method of quality control.

2.3 the thin layer chromatography of Herba Violae japonicae is differentiated in the prescription

Get capsule 's content 1g, add methanol 5ml, ultrasonic 30 minutes, to filter, filtrate is as need testing solution; Other gets and lacks Herba Violae japonicae sample 1g, makes negative control solution with the need testing solution preparation method; Other gets Herba Violae japonicae control medicinal material 0.5g, makes control medicinal material solution with the need testing solution preparation method; According to thin layer chromatography test, draw above-mentioned three kinds of each 10ul of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that toluene-ethyl acetate-formic acid of 5: 3: 0.5 is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of reference substance chromatograph on show the speckle of same color; Negative control is noiseless; Test agent checking in 3 batches, the method good reproducibility, specificity is strong, so classify it as method of quality control.

2.4 the thin layer chromatography of Cortex Dictamni is differentiated in the prescription

Get capsule 's content 5g, add ethanol 20ml, water-bath refluxed 30 minutes, filtered, and the filtrate water-bath volatilizes, and added water 20ml dissolving, used ethyl acetate extraction 2 times, and each 20ml merges the water-bath of ethyl acetate liquid and volatilizes, and adds methanol 1ml dissolving, as need testing solution; Other gets and lacks Cortex Dictamni sample 5g, makes negative control solution with the need testing solution preparation method; Other gets Cortex Dictamni control medicinal material 0.3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw above-mentioned three kinds of each 5ul of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that chloroform-ethyl acetate-acetone of 10: 0.5: 0.2 is developing solvent, launches, and takes out, spray is with the bismuth potassium iodide solution of new preparation, in the test sample chromatograph, with the corresponding position of reference substance solution chromatograph on, show the speckle of same color; Negative control is noiseless; Test agent checking in 3 batches, the method good reproducibility, specificity is strong, so classify it as method of quality control.

Three, check the research of item

3.1 disintegration

By " Chinese pharmacopoeia version in 2005 one " appendix XII A inspection technique disintegration " is measured, and regulation this product should all disintegrates in 30 minutes, and three batches of testing results of this product see Table 1, and are all up to specification.

3.2 moisture inspection

By " Chinese pharmacopoeia version in 2005 one " appendix IX H aquametry first method " is measured, and it is all up to specification that measurement result sees Table 1,3 batch sample.

3.3 content uniformity inspection

According to " checking that it is all up to specification to the results are shown in Table 1,3 batch sample under " appendix IL capsule " item of Chinese pharmacopoeia version in 2005.

3.4 heavy metal inspection

According to " appendix an IX of Chinese pharmacopoeia version in 2005 E heavy metal inspection technique second method operation.

3.4.1 the preparation of standard lead solution: precision takes by weighing the plumbi nitras 0.117g that is dried to constant weight at 105 ℃, puts in the 1000ml volumetric flask, adds nitric acid 5ml, and water 50ml after the dissolving, is diluted with water to scale, shakes up, the plumbous stock solution of the standard that promptly gets.Face the time spent, precision is measured preceding stock solution 10ml, puts in the 100ml volumetric flask, and thin up shakes up to scale, promptly gets (lead that every 1ml is equivalent to 10ug).

3.4.2 the preparation of need testing solution: precision takes by weighing this product content 1g, put in the crucible, slowly blazing on the electric furnace to carbonization fully, put coldly, add sulphuric acid 1ml, make just moistening, eliminate with low-temperature heat to sulphuric acid, add nitric acid 0.5ml, evaporate to dryness is put cold, 500～600 ℃ blazingly make complete ashing in the Muffle furnace, put cold rnning hydrochloric acid 2ml, put and add water 15ml in the water-bath behind the evaporate to dryness, drip ammonia solution to phenolphthalein indicator and show neutral, add acetate buffer (pH3.5) 2ml again, after the slight fever dissolving, move in the nessler colorimetric tube, thin up becomes 25ml.

3.4.3 the preparation of positive control solution: will prepare the reagent of need testing solution, and put in another crucible behind the evaporate to dryness, and add acetate buffer (pH3.5) 2ml and water 15ml, after the slight fever dissolving, in the dislocation nessler colorimetric tube, add standard lead solution 1ml, thin up becomes 25ml again.

3.4.4 the preparation of blank liquid: will prepare the reagent of need testing solution, and put in another crucible behind the evaporate to dryness, and add acetate buffer (pH3.5) 2ml and water 15ml, after the slight fever dissolving, in the dislocation nessler colorimetric tube, thin up becomes 25ml again.

3.4.5 add each 2ml of thioacetamide test solution respectively in 3 nessler colorimetric tubes of colorimetric need testing solution, positive control solution and blank liquid, shake up, placed 2 minutes, with putting on the blank sheet of paper, from up to down have an X-rayed, color that test sample liquid shows and positive control solution compare, and color is more shallow, and blank is noiseless.The results are shown in Table 1.

The result shows that 3 batch sample content of beary metal are all less than 10ppm.

3.5 arsenic salt is checked

According to " appendix an IXF of Chinese pharmacopoeia version in 2005 arsenic salt inspection technique first method operation.

3.5.1 the preparation of standard arsenic solution: take by weighing arsenic trioxide 0.121g, put in the 1000ml measuring bottle, add 20% sodium hydroxide solution 5ml dissolving after, neutralize with an amount of dilute sulfuric acid, add dilute sulfuric acid 10ml again, be diluted with water to scale, shake up, as stock solution.Before facing usefulness, precision is measured stock solution 10ml, puts in the 1000ml measuring bottle, adds dilute sulfuric acid 10ml, is diluted with water to scale, shakes up, and promptly gets (As that every 1ml is equivalent to 1 μ g).

3.5.2 the preparation of need testing solution: precision takes by weighing this product content 1g, puts in the crucible, adds calcium hydroxide 0.5g, mixing, it is moistening to add water, and oven dry is carefully blazing on little fire, (noting content is spilt) to smog eliminates, move in the Muffle furnace 500～600 ℃ blazing to ashing, put coldly, add hydrochloric acid 5ml and water 23ml, heating for dissolving in the water-bath is as need testing solution.

3.5.3 the preparation of positive control solution: precision is measured standard arsenic solution 2ml, puts in the crucible, makes positive control solution with the 3.5.2 method.

3.5.4 the preparation of blank liquid: other gets one of crucible, adds calcium hydroxide 0.5g, makes blank liquid with the preparation method of need testing solution.

3.5.5 the preparation of arsenic speckle: need testing solution, positive control solution, blank liquid are moved to respectively in the survey arsenic bottle, add potassium iodide test solution 5ml more respectively, 5 of the inferior stannum test solutions of acid chlorization, after room temperature is placed 10 minutes, add zinc granule 2g, reaction is 45 minutes in 30 ℃ of water-baths, takes out the mercuric bromide reagent paper, observe arsenic speckle color, need testing solution arsenic speckle is shallower than standard arsenic speckle color as a result.The results are shown in Table 1.The result shows that 3 batch sample arsenic salt contents are all less than 2ppm.

Table 1 three batch sample check result tables

The sample lot number	??2007001	??2007002	??2007003
The sample lot number	??2007001	??2007002	??2007003	Disintegration time (min)	??19	??18	??19
Content uniformity	Up to specification	Up to specification	Up to specification	Disintegration time (min)	??19	??18	??19
Content uniformity	Up to specification	Up to specification	Up to specification	Moisture (%)	??1.9	??1.8	??2.0
Heavy metal (ppm)	??＜10	??＜10	??＜10	Moisture (%)	??1.9	??1.8	??2.0
Heavy metal (ppm)	??＜10	??＜10	??＜10	Arsenic salt (ppm)	??＜2	??＜2	??＜2

3.6 microbial check method validation

Experiment material: biochemical incubator, vertical pressure steam disinfecting apparatus, the real order microscope of the bitubular.Positive control bacterium and conventional Micro biological Tests chemical reagent and glass apparatus etc.

Nutrient agar, Rose Bengal Sodium agar, EMB agar, cholate lactose enriched medium, cholate lactose fermentation culture medium (more than be BeiJing, China scientific and technological development company of three sections and produce, press the preparation of CP05 version regulation and sterilize), four-methyl umbelliferone glucosiduronate (BeiJing, China's cattle cow genome technology company limited);

Escherichia coli [CMCC (B) 44102], staphylococcus aureus [CMCC (B) 26003], bacillus subtilis [CMCC (B) 63501], Candida albicans [CMCC (F) 98001], aspergillus niger [CMCC (F) 98003], above strain all derives from the institute for drug control, Guizhou Province.

The test liquid preparation: get this product capsule 's content in accordance with regulations, fully shake up the back and mix, get capsule 's content 10g, the aseptic sodium chloride-peptone buffer agent that adds pH7.0 is 1: 10 test liquid to 100ml.Checking the results are shown in Table 2 to table 5.

Table 2 antibacterial, mycete and yeast method of counting checking recovery test record

The response rate=(the average clump count of the test group-average clump count of test sample the matched group)/average clump count of bacterium liquid group * 100%.

Table 3 adopts media dilution method 0.2mL/ ware

Table 4 control bacterium inspection method checking record (escherichia coli)

Table 5 control bacterium inspection method checking record (coliform)

By demonstration test as can be known, this product can adopt conventional method to carry out mycete and yeast, escherichia coli, coliform test, with media dilution method (0.2mL/ ware) antibacterial is tested.By the checking method the three batch sample microbial limits of this product are checked that the result is all up to specification.

Conclusion: by above experiment as can be known, can adopt the every inspection of conventional method under capsule preparations is checked.

Four, the research of assay in the capsule preparations

The principal agent Radix Cynanchi Paniculati contains paeonol in paeonol this product, and modern study shows, paeonol has multiple pharmacological effect such as analgesia, hemostasis, antiinflammatory, antibiotic, blood pressure lowering; Paeonol is external to have the obvious suppression effect to multiple human pathogen, is the effective ingredient of Radix Cynanchi Paniculati, this product has adopted high effective liquid chromatography for measuring its content.

4.1 instrument and reagent:

High performance liquid chromatograph: LC-2010HT (day island proper Tianjin); SeriesIII pump (U.S.), Mode500 UV-detector (U.S.); AUW220D analytical balance (day island proper Tianjin); Methanol (chromatographically pure, Tianjin Da Mao); Water (redistillation faces and uses preceding preparation), phosphoric acid (analytical pure, Tianjin Da Mao), paeonol (middle inspection institute, lot number: 110708-200505).Three batches of (lot numbers: 2007001,2007002,2007003) of sample.

4.2 chromatographic condition:

DiamonsilC ₁₈Post (5 μ m), mobile phase: methanol-water (60: 40); Flow velocity: 1.0ml/min; Detect wavelength 274nm; Column temperature: 35 ℃.

4.3 system suitability test: each the 10 μ l of negative sample contrast solution that get paeonol reference substance solution, need testing solution and scarce Radix Cynanchi Paniculati respectively inject chromatograph of liquid, the record chromatograph.From collection of illustrative plates as can be known, under this chromatographic condition, the paeonol peak separates fully with other component peaks, and at the retention time place identical with the paeonol peak, negative control is noiseless.

4.4 the preparation of need testing solution

This test has been done following investigation to the test sample processing method:

4.4.1 extracting method is selected

Method 1: getting this product content, put in the tool plug conical flask, is to extract solvent with the methanol solution, extracts with the merceration method, has investigated different extraction time content paeonol content.The results are shown in Table 6.

Table 6 extracting method 1 is investigated the result

Extraction time (hour)	??0.5	??1	??2
Extraction time (hour)	??0.5	??1	??2	Content (mg/g)	??3.58	??3.86	??3.91

Method 2: getting this product content, is to extract solvent with methanol, is processing method with ultrasonic, has investigated different extraction time content paeonol content.The results are shown in Table 7.

Table 7 extracting method 2 is investigated the result

Extraction time (minute)	??10	??20	??30	??40
Extraction time (minute)	??10	??20	??30	??40	Content (mg/g)	??5.68	??5.69	??5.64	??5.62

Result of the test shows that this product adopts methanol supersound extraction efficient height, weak point consuming time, easy and simple to handle, and can adopt it is this product test sample processing method; According to different extraction times investigation results, determine that ultrasonic time is 10 minutes.

4.4.2 extract ratio of solvent

Get this product content; be solvent with methanol, 50% methanol and ethanol 25ml respectively; supersound extraction 10 minutes; filter; filtrate is through filtering with microporous membrane; as need testing solution; measure paeonol content; the solvent methanol extraction effect is best as a result; and 50% methanol extraction effect is taken second place, alcoholic acid extraction effect is the poorest, owing to be that the test sample of solvent preparation is that the test sample color of solvent preparation is dark than methanol with 50% methanol, considers the protection to chromatographic column; we select methanol is the sample extraction solvent, the results are shown in Table 8.

Table 8 extracts solvent and investigates the result

Extract solvent	50% methanol	Methanol	Ethanol
Extract solvent	50% methanol	Methanol	Ethanol	Content (mg/g)	??4.72	??5.53	??3.24

Result of the test shows that this product still adopts methanol comparatively suitable as extracting solvent.

4.5 linear relationship is investigated precision and is taken by weighing paeonol reference substance 25.80mg, put in the 50ml measuring bottle, add dissolve with methanol and be diluted to scale, shake up, the accurate 2ml that draws puts in the 50ml measuring bottle, add methanol and be diluted to scale, get the paeonol reference substance solution of 0.02064mg/ml, the accurate respectively paeonol reference substance solution 2 μ l that draw, 5 μ l, 10 μ l, 15 μ l, 20 μ l, injecting chromatograph, the record chromatogram, measure the peak area integrated value, measurement result sees Table 9, and is abscissa with reference substance sample size X (μ g), peak area value Y (mv.s) is a vertical coordinate, the drawing standard curve, the result shows that paeonol is good in 0.0413～0.4128 μ g scope internal linear relation.

Table 9 paeonol linear relationship is investigated

The equation that regression equation was fitted to initial point gets: y=5537711x.

Minimum point sample size (0.0413 μ g) above-mentioned two equations of substitution respectively during the line taking sexual relationship is investigated, calculating both relative deviations is 0.43%, illustrates that this product can carry out assay with one point external standard method.

4.6 precision test: the accurate paeonol reference substance solution 10 μ l (concentration is 0.02046mg/ml) that draw, repeat sample introduction 5 times, measure paeonol peak area integrated value, obtain relative standard deviation, the result shows that precision is good, the results are shown in Table 10.

Table 10 precision experimental result

4.7 stability test: get a test sample (lot number: 2007001), prepare test liquid by the preparation method of need testing solution in the method for quality control text.At room temperature sample introduction 10 μ l at set intervals measure the paeonol peak area value, ask relative standard deviation, and the result shows that paeonol basicly stable in 12 hours (RSD=0.14%) the results are shown in Table 11.

Table 11 stability test result

4.8 replica test: precision is got this product, and (lot number: 2007001) each 1g, prepares test liquid by the preparation method of need testing solution in the quality method of quality control text by totally 5 parts.Accurate each the 10 μ l of need testing solution that draw measure paeonol peak area integrated value, calculate content, ask relative standard deviation, and the result shows, repeatability good (RSD=1.53%).See Table 12.

Table 12 replica test result

4.9 middle precision: (lot number: 2007001), divide two groups on distinct device this product to be carried out assay by the different operating personnel, the result shows, middle precision good (RSD=0.58%) to get same batch sample.The results are shown in Table 13.

A group: LC-2010HT high performance liquid chromatograph; B group: SSI high performance liquid chromatograph.

Precision test result in the middle of the table 13

4.10 recovery test: adopt the test of application of sample absorption method, (lot number: 2007001) 6 parts, precision takes by weighing each about 0.5g, and the accurate paeonol reference substance that adds is an amount of to get the same batch sample of known content, be prepared into need testing solution by the need testing solution preparation method, accurate each the 10 μ l of need testing solution that draw measure the record chromatogram according to above-mentioned chromatographic condition, calculate content, the result shows that the response rate of paeonol is good, the results are shown in Table 14.

Table 14 application of sample recovery test result

4.11 sample determination: get test agent in 3 batches, measure wherein paeonol content (the results are shown in Table 15) by the method for quality control text.Be calculated as follows content:

In the formula: A _Sample-test sample paeonol peak area integrated value

A _Right-paeonol reference substance peak area integrated value

C _Right-paeonol reference substance concentration (mg/ml)

M _Sample-test sample sampling amount (g)

M _{Loading amount}-average loading amount (g)

250-test sample dilution volume (ml)

Paeonol contains the survey result in table 153 batch sample

The content limit gauge is fixed in the sample: Radix Cynanchi Paniculati is total to 80g in this product prescription, for protogenic medicinal powder is used as medicine, its paeonol content is limited to 1.3%, by pilot-scale experiment as can be known, this product paeonol rate of transform in process of production all is not less than 99%, is calculated as follows the every capsules paeonol content limit of this product:

Content limit (mg/ the grain)=medical material amount * medical material content limit * rate of transform/amount of making

That is: content limit (mg/ grain)=80g * 1.3% * 99%/1000=1.03mg/ grain

Consider the influence to medicine of production and storage process, every of regulation this product contains Radix Cynanchi Paniculati in paeonol, must not be less than 1.02mg.

Herba Polygoni cymosi, Cortex Dictamni, Flos Lonicerae and Herba Violae japonicae all contain flavone component in total flavones this product, and modern study shows, such flavone component has antiinflammatory, multiple pharmacological effect such as antibiotic, and this product adopts ultraviolet visible spectrophotometry to measure its content.

(1) instrument and reagent: TU-1901 type ultraviolet-uisible spectrophotometer (Beijing all purpose instrument equipment company); Sodium nitrite (Chuanjiang River, Chongqing), aluminum nitrate (Kingsoft, Chengdu), sodium hydroxide (Gansu Province, west, Guangdong chemical industry) are analytical pure; Control substance of Rutin (middle inspection institute, lot number: 100080-200306); Three batches of (lot numbers: 2007001,2007002,2007003) of sample.

(2) preparation of standard solution: precision takes by weighing 120 ℃ of control substance of Rutin that are dried to constant weight an amount of, puts in the 25ml measuring bottle, and it is an amount of to add methanol, puts that slight fever makes dissolving in the water-bath, puts coldly, adds methanol to scale, shakes up.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and makes the solution that contains rutin 0.20192mg among every 1ml, promptly.

(3) detect that wavelength is selected and the negative control test: take by weighing sample (lot number: 2007001) 0.2g, accurately claim surely, put in the tool plug conical flask, the accurate 70% ethanol 25ml that adds, weigh, ultrasonic 10min places room temperature, supply with 70% ethanol and to subtract weight loss, filter, the accurate absorption in the subsequent filtrate 4ml volumetric flask adds water to 6.0ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, add 10% aluminum nitrate solution 1ml, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shakes up, placed 15 minutes, as need testing solution; Precision is measured control substance of Rutin solution and each 4ml of water, puts respectively in the 25ml measuring bottle, makes reference substance solution and blank solution from " adding water to 6.0ml " with the method operation.It is an amount of to get above-mentioned three kinds of solution, in the interscan of 400～600nm wave-length coverage, reference substance solution and need testing solution have absorption maximum at the 510nm place, and the trap of negative control solution under this wavelength compared with the trap of need testing solution, less than 5%, meet relevant regulations, thus we to select 510nm be the detection wavelength of total flavones in this product.

(4) standard curve: precision is measured control substance of Rutin solution 0.5ml, 1ml, 2ml, 3ml, 4ml, 5ml puts respectively in the 25ml measuring bottle, respectively add water to 6.0ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, add 10% aluminum nitrate solution 1ml, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shake up, placing 15 minutes, is blank with the corresponding reagent, according to ultraviolet visible spectrophotometry, measure absorbance at 510nm wavelength place, obtain following equation: A=0.035552C+0.016181 (r=0.996) through recurrence.A is an absorbance in the formula, and C is control substance of Rutin solution concentration in the color solution (μ g/ml).Control substance of Rutin concentration is in 4.0384～40.3840 μ g/ml scopes, and absorbance and concentration are good linear relationship, the results are shown in Table 16.

Table 16 control substance of Rutin standard curve is investigated

Sequence number	Concentration (ug/ml)	Trap
Sequence number	Concentration (ug/ml)	Trap	??1	??4.0384	??0.109
??2	??8.0768	??0.201	??1	??4.0384	??0.109

Sequence number	Concentration (ug/ml)	Trap
Sequence number	Concentration (ug/ml)	Trap	??3	??16.1536	??0.308
??4	??24.2304	??0.411	??3	??16.1536	??0.308
??4	??24.2304	??0.411	??5	??32.3072	??0.521
??6	??40.3840	??0.689	??5	??32.3072	??0.521

(5) precision test: get " standard curve " following No. 3 reference substance solution (12.1150ug/ml) in right amount, replication is 5 times in accordance with the law, record trap value, obtain relative standard deviation, the result shows that it is good that reference substance rutin trap is measured precision, RSD=0.27% the results are shown in Table 17.

Table 17 Precision test result

(6) stability test: take by weighing sample (lot number: 2007001) 0.2g, accurate claim surely, put in the tool plug conical flask, the accurate 70% ethanol 25ml that adds, weigh, ultrasonic 10min places room temperature, supply with 70% ethanol and to subtract weight loss, filter, in the accurate absorption subsequent filtrate 4ml volumetric flask, the method under the preparation of sighting target directrix curve, from " adding water to 6.0ml ", be prepared into need testing solution; At room temperature measure trap at set intervals in accordance with the law, ask relative standard deviation; The result shows that the total flavones trap is basicly stable in 12 hours in this method working sample, and RSD=0.37% the results are shown in Table 18.

Table 18 stability test result

(7) replica test: get take by weighing sample (lot number: 2007001) 5 parts, each 0.2g, accurate claim fixed, put respectively in the tool plug conical flask, the accurate 70% ethanol 25ml that adds weighs, ultrasonic 10min, place room temperature, supply with 70% ethanol and subtract weight loss, filtration, the accurate absorption in the subsequent filtrate 4ml volumetric flask, method under the preparation of sighting target directrix curve from " adding water to 6.0ml ", is prepared into need testing solution; Replication is 5 times in accordance with the law, and record trap value is obtained relative standard deviation, and the result shows that rutin trap repeatability is good in this method working sample, and RSD=0.67% the results are shown in Table 19.

Table 19 replica test result

Sampling amount (g)	Trap	Total flavones (mg/g)	Average content (mg/g)	??RSD(％)
Sampling amount (g)	Trap	Total flavones (mg/g)	Average content (mg/g)	??RSD(％)	??0.2052??0.2012??0.2056??0.2037??0.1996	??0.351??0.348??0.355??0.353??0.346	??7.29??7.25??7.24??7.26??7.27	??7.26	??0.27

(8) recovery test: adopt the application of sample recovery test, get the same batch sample (lot number: 2007001 of the replica test of known content, the total flavones average content is 7.26mg/g) totally 5 parts, each 0.15g, the accurate title, decide, put respectively in the tool plug conical flask, accurate control substance of Rutin 5ml and the 70% ethanol 20ml of adding weighs, ultrasonic 10min, place room temperature, supply with 70% ethanol and subtract weight loss, filtration, the accurate absorption in the subsequent filtrate 4ml volumetric flask, method under the preparation of sighting target directrix curve from " adding water to 6.0ml ", is prepared into need testing solution; Measure absorbance according to above-mentioned condition, calculate content, ask relative standard deviation, RSD=1.49%, the result shows that the method has average recovery preferably, the results are shown in Table 20.

Rutin application of sample recovery test result in table 20 test sample

Sequence number	Sample volume (g)	Theoretical content (mg)	Addition (mg)	Trap	Measured value (mg)	Yield (mg)	The response rate (%)	Average recovery rate (%)	??RSD??(％)
Sequence number	Sample volume (g)	Theoretical content (mg)	Addition (mg)	Trap	Measured value (mg)	Yield (mg)	The response rate (%)	Average recovery rate (%)	??RSD??(％)	??1	??0.1524	??1.1064	??1.0096	??0.503	??2.1396	??1.1300	??102.1
??2	??0.1517	??1.1013	??1.0096	??0.501	??2.1308	??1.1212	??101.8			??1	??0.1524	??1.1064	??1.0096	??0.503	??2.1396	??1.1300	??102.1
??2	??0.1517	??1.1013	??1.0096	??0.501	??2.1308	??1.1212	??101.8			??3	??0.1493	??1.0839	??1.0096	??0.495	??2.1044	??1.0948	??101.0	??102.4	??1.49
??4	??0.1506	??1.0934	??1.0096	??0.507	??2.1571	??1.1475	??105.0			??3	??0.1493	??1.0839	??1.0096	??0.495	??2.1044	??1.0948	??101.0	??102.4	??1.49
??4	??0.1506	??1.0934	??1.0096	??0.507	??2.1571	??1.1475	??105.0			??5	??0.1514	??1.0992	??1.0096	??0.501	??2.1308	??1.1212	??102.0

(9) 3 batch samples are measured and the formulation of finished product content limit prepares need testing solution and reference substance solution down by " repeatability " mensuration item, measure absorbance, calculate wherein content of total flavone, the results are shown in Table 21.

Determination of total flavonoids result in table 213 batch sample

According to total flavones measurement result in 3 batch samples, consider the influence of factors such as production and storage, every of regulation this product contains total flavones with anhydrous rutin (C ₂₇H ₃₀O ₁₆) meter, must not be less than 2.00mg.

Five, used standard substance in the method for quality control

Paeonol reference substance: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 110708-200505 uses for assay.

Control substance of Rutin: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 100080-200306 uses for assay.

Flos Lonicerae control medicinal material: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 121060-200303 uses for differentiating.

Radix Cynanchi Paniculati control medicinal material: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 121514-200501 uses for differentiating.

Herba Violae japonicae control medicinal material: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 121429-200602 uses for differentiating.

Cortex Dictamni control medicinal material: purchase in Nat'l Pharmaceutical ﹠ Biological Products Control Institute lot number: 120978-200503 uses for differentiating.

Beneficial effect of the present invention: compared with prior art, the present invention has set up the method for quality control of the capsule preparations of treatment dermatosis, character, discriminating, inspection and assay to capsule preparations are studied and are screened, the method of quality control that is adopted is scientific and reasonable, the accuracy height, favorable reproducibility, the quality of the capsule preparations of control of quality dermatosis is fully and effectively guaranteed the clinical efficacy of said preparation.

The present invention is further illustrated below in conjunction with the specific embodiment.

The specific embodiment

Embodiment 1.The method of quality control of the capsule preparations of treatment dermatosis is as follows:

Character: this product is a hard capsule, and content is sepia and white granule and powder, and gas is special, bitter in the mouth, little puckery.

Differentiate: (1) Flos Lonicerae is differentiated: get capsule 's content 1g, add methanol 5ml, supersound extraction 30 minutes is filtered, and gets filtrate, as need testing solution; Extracting honeysuckle control medicinal material 0.5g makes control medicinal material solution with the need testing solution preparation method in addition; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 10 μ l of need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that the upper strata liquid of butyl acetate-formic acid-water of 7: 2.5: 2.5 is developing solvent with volume ratio, launch, take out, dry, spray is with 5% ferric chloride-alcoholic solution, about 5 minutes of 105 ℃ of bakings of temperature, take out, inspect under the daylight; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(2) Radix Cynanchi Paniculati is differentiated: get capsule 's content 5g, add methanol 15ml, supersound extraction 30 minutes is filtered, and filtrate is concentrated into about 5ml, filters, and gets filtrate, as need testing solution; Other gets Radix Cynanchi Paniculati control medicinal material 3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that cyclohexane extraction-ethyl acetate of 3: 1 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

(3) Herba Violae japonicae is differentiated: get capsule 's content 1g, add methanol 5ml, and ultrasonic 30 minutes, to filter, filtrate is as need testing solution; Other gets Herba Violae japonicae control medicinal material 0.5g, makes control medicinal material solution with medical material.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that toluene-ethyl acetate-formic acid of 5: 3: 0.5 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of reference substance chromatograph on show the speckle of same color.

(4) Cortex Dictamni is differentiated: get capsule 's content 5g, add ethanol 20ml, water-bath refluxed 30 minutes, filtered, the filtrate water-bath volatilizes, and adds water 20ml dissolving, uses ethyl acetate extraction 2 times, each 20ml, merge the water-bath of ethyl acetate liquid and volatilize, add methanol 1ml dissolving, as need testing solution.Other gets Cortex Dictamni control medicinal material 0.3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw need testing solution and each 5ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that chloroform-ethyl acetate-acetone of 10: 0.5: 0.2 is developing solvent, launches, and takes out, spray is with the bismuth potassium iodide solution of new preparation, in the test sample chromatograph, with the corresponding position of reference substance solution chromatograph on, show the speckle of same color.

Assay: paeonol is measured the content of paeonol in the capsule preparations 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; Methanol-water (60: 40) is a mobile phase; Flow velocity is 1.0ml/min; The detection wavelength is 274nm; Column temperature: 35 ℃.Number of theoretical plate calculates by the paeonol peak should be not less than 3000.

The preparation of reference substance solution: it is an amount of that precision takes by weighing the paeonol reference substance, adds methanol and make the solution that contains paeonol 0.02064mg among every 1ml, promptly.

The preparation of need testing solution: get this product under the content uniformity item, mixing is got 1g, the accurate title, decide, and puts in the tool plug conical flask, adds methanol solution 25ml, weigh supersound process (power 250W, frequency 40kHz) 10 minutes, take out, put coldly, mend heavy, filter, get in subsequent filtrate 1ml to the 10ml measuring bottle, add the methanol standardize solution, filter with 0.45 μ m filter membrane, as need testing solution.

Algoscopy: get each 10 μ l of need testing solution and reference substance solution respectively,, inject hplc determination, the record chromatogram according to high performance liquid chromatography.Press external standard method with calculated by peak area, promptly.

Every dermopathic capsule preparations of treatment contains Radix Cynanchi Paniculati with paeonol (C ₉H ₁₀O ₃) meter, be no less than 1.02mg.

The preparation of total flavones reference substance solution: precision takes by weighing 120 ℃ of control substance of Rutin that are dried to constant weight an amount of, puts in the 25ml measuring bottle, and it is an amount of to add methanol, puts that slight fever makes dissolving in the water-bath, puts coldly, adds methanol to scale, shakes up.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and makes the solution that contains rutin 0.20192mg among every 1ml, promptly.

The standard curve preparation: precision is measured reference substance solution 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, put respectively in the 25ml measuring bottle, respectively add water to 6.0ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, and added 10% aluminum nitrate solution 1ml, placed 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale again, shake up, placed 15 minutes, with the corresponding reagent is blank, according to ultraviolet visible spectrophotometry, measure absorbance at 510nm wavelength place, be vertical coordinate with the absorbance, concentration is abscissa, the drawing standard curve.

The preparation of need testing solution: get this product under the content uniformity item, mixing, get 0.2g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 70% ethanol 25ml that adds, weigh, ultrasonic 10min places room temperature, supply with 70% ethanol and to subtract weight loss, filter, the accurate absorption in subsequent filtrate 1ml to the 25ml volumetric flask adds water 5ml, method under the sighting target directrix curve preparation, make need testing solution from " adding water to 6.0ml " with the method operation, measure absorbance in accordance with the law, read the weight that contains anhydrous rutin the need testing solution from standard curve, calculate, promptly.

Every dermopathic capsule preparations of treatment contains total flavones with anhydrous rutin (C ₂₇H ₃₀O ₁₆) meter, be no less than 2.00mg.

Function cures mainly: dispeiling pathogenic wind and removing dampness, clearing away heat and cooling blood, detoxifying and relieving itching.Be used for the itching skin disease such as eczema, urticaria, dermatitis due to heat in blood wind-dryness, the hot and humid ecchymosis.

Usage and dosage: oral.One time 3,3 times on the one.

Specification: every dress 0.3g.

Points for attention: be not taken by pregnant women.

Storage: sealing, protection against the tide.

Embodiments of the present invention are not limited to the foregoing description, and the various variations of making under the prerequisite that does not break away from aim of the present invention all belong within protection scope of the present invention.

Claims

One kind the treatment dermopathic capsule preparations method of quality control, this capsule prepares like this: Herba Polygoni cymosi 400g, Cortex Dictamni 200g, Flos Lonicerae 200g and Herba Violae japonicae 200g, decoct with water 3 times, add 10 times of water gagings at every turn and decocted 3 hours, filter, merging filtrate, be evaporated to 60 ℃ of relative densities and be 1.25 extractum, drying under reduced pressure is pulverized, cross 80 mesh sieves, get extract powder; Get Radix Cynanchi Paniculati 80g, pulverize, cross 80 mesh sieves, get the crude drug powder; With extract powder and crude drug powder mixing, adding starch is an amount of, and with 75% alcohol granulation, drying incapsulates, and every dress 0.3g makes 1000; It is characterized in that: method of quality control do as one likes shape, discriminating, inspection and assay are formed, wherein discriminating is the discriminating to Flos Lonicerae, Radix Cynanchi Paniculati, Herba Violae japonicae and Cortex Dictamni, and assay is respectively paeonol in the capsule preparations and content of total flavone to be measured with high performance liquid chromatography and ultraviolet visible spectrophotometry.
2. the method for quality control of the dermopathic capsule preparations of treatment according to claim 1 is characterized in that:

Character: this product is a hard capsule, and content is sepia and white granule and powder, and gas is special, bitter in the mouth, little puckery;

Differentiate: be contrast, be adhesive with the sodium carboxymethyl cellulose, be that developing solvent is according to thin layer chromatography discriminating Flos Lonicerae with the upper strata liquid of butyl acetate-formic acid-water with the Flos Lonicerae control medicinal material; With the Radix Cynanchi Paniculati control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with cyclohexane extraction-ethyl acetate differentiate Radix Cynanchi Paniculati according to thin layer chromatography; With the Herba Violae japonicae control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with toluene-ethyl acetate-formic acid differentiate Herba Violae japonicae according to thin layer chromatography; With the Cortex Dictamni control medicinal material be contrast, be adhesive with the sodium carboxymethyl cellulose, to be developing solvent with chloroform-ethyl acetate-acetone differentiate Cortex Dictamni according to thin layer chromatography;

Check: meet the every regulation under the capsule item among an appendix I of Chinese Pharmacopoeia version in 2005 L;

Assay:

The assay of paeonol: Radix Cynanchi Paniculati is contrast with the paeonol reference substance, is filler with the octadecylsilane chemically bonded silica, is that 60: 40 methanol-water is the content of mobile phase according to paeonol in the high effective liquid chromatography for measuring capsule preparations with volume ratio;

Content of total flavone is measured: total flavones is contrast with the control substance of Rutin, is developer with 5% sodium nitrite solution, 10% aluminum nitrate solution and sodium hydroxide test solution, measures content of total flavone in the capsule preparations according to ultraviolet visible spectrophotometry.
3. the method for quality control of the dermopathic capsule preparations of treatment according to claim 1 and 2 is characterized in that: described discriminating is formed by following:

(1) Flos Lonicerae is differentiated: get capsule 's content 1g, add methanol 5ml, supersound extraction 30 minutes is filtered, and gets filtrate, as need testing solution; Extracting honeysuckle control medicinal material 0.5g makes control medicinal material solution with the need testing solution preparation method in addition; Test according to thin layer chromatography, draw each 10 μ l of need testing solution and control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be that the upper strata liquid of butyl acetate-formic acid-water of 7: 2.5: 2.5 is developing solvent with volume ratio, launch, take out, dry, spray was dried by the fire 5 minutes down for 105 ℃ in temperature with 5% ferric chloride-alcoholic solution, take out, inspect under the daylight; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(2) Radix Cynanchi Paniculati is differentiated: get capsule 's content 5g, add methanol 15ml, supersound extraction 30 minutes is filtered, and filtrate is concentrated into about 5ml, filters, and gets filtrate, as need testing solution; Other gets Radix Cynanchi Paniculati control medicinal material 3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that cyclohexane extraction-ethyl acetate of 3: 1 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

(3) Herba Violae japonicae is differentiated: get capsule 's content 1g, add methanol 5ml, and ultrasonic 30 minutes, filter, get filtrate, as need testing solution; Other gets Herba Violae japonicae control medicinal material 0.5g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 10ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that toluene-ethyl acetate-formic acid of 5: 3: 0.5 is developing solvent, launch, take out, dry, put under the ultra-violet lamp 365nm and inspect; With the corresponding position of reference substance chromatograph on, show the speckle of same color;

(4) Cortex Dictamni is differentiated: get capsule 's content 5g, add ethanol 20ml, water-bath refluxed 30 minutes, filtered, the filtrate water-bath volatilizes, and adds water 20ml dissolving, uses ethyl acetate extraction 2 times, each 20ml, combined ethyl acetate liquid, water-bath volatilizes, and adds methanol 1ml dissolving, as need testing solution; Other gets Cortex Dictamni control medicinal material 0.3g, makes control medicinal material solution with the need testing solution preparation method; Test according to thin layer chromatography, draw need testing solution and each 5ul of control medicinal material solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with volume ratio is that chloroform-ethyl acetate-acetone of 10: 0.5: 0.2 is developing solvent, launch, take out, dry, spray is with the bismuth potassium iodide solution of new preparation; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
4. the method for quality control of the dermopathic capsule preparations of treatment according to claim 1 and 2 is characterized in that: the assay of paeonol is in the described capsule preparations:

Radix Cynanchi Paniculati chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Volume ratio is that 60: 40 methanol-water is a mobile phase; Flow velocity is 1.0ml/min; The detection wavelength is 274nm; Column temperature: 35 ℃, number of theoretical plate calculates by the paeonol peak should be not less than 3000;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the paeonol reference substance, adds methanol and make the solution that contains paeonol 0.02064mg among every 1ml, promptly gets reference substance solution;

The preparation of need testing solution: get this product under the content uniformity item, mixing is got 1g, the accurate title, decide, and puts in the tool plug conical flask, adds methanol solution 25ml, weigh, supersound process 10 minutes is taken out, put coldly, mend heavyly, filter, get in subsequent filtrate 1ml to the 10ml measuring bottle, add the methanol standardize solution, filter with 0.45 μ m filter membrane, as need testing solution;

Algoscopy: get each 10 μ l of need testing solution and reference substance solution respectively, according to high performance liquid chromatography, inject hplc determination, the record chromatogram is pressed external standard method with calculated by peak area, promptly;

Content of total flavone is determined as in the described capsule preparations:

The preparation of total flavones reference substance solution: precision takes by weighing 120 ℃ of control substance of Rutin that are dried to constant weight an amount of, puts in the 25ml measuring bottle, and it is an amount of to add methanol, puts that slight fever makes dissolving in the water-bath, puts coldly, adds methanol to scale, shakes up; Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and makes the solution that contains rutin 0.20192mg among every 1ml, promptly gets reference substance solution;

The standard curve preparation: precision is measured reference substance solution 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, put respectively in the 25ml measuring bottle, respectively add water to 6.0ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, add 10% aluminum nitrate solution 1ml, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shaking up, placed 15 minutes, is blank with the corresponding reagent; According to ultraviolet visible spectrophotometry, measure absorbance at 510nm wavelength place, be vertical coordinate with the absorbance, concentration is abscissa, the drawing standard curve;

The preparation of need testing solution: get this product under the content uniformity item, mixing, get 0.2g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 70% ethanol 25ml that adds, weigh, ultrasonic 10min places room temperature, supply with 70% ethanol and to subtract weight loss, filter, the accurate absorption in subsequent filtrate 1ml to the 25ml volumetric flask adds water to 5ml, method under the sighting target directrix curve preparation, make need testing solution from " adding water to 6.0ml " with the method operation, measure absorbance in accordance with the law, read the weight that contains anhydrous rutin the need testing solution from standard curve, calculate, promptly.
5. according to the method for quality control of claim 1, the dermopathic capsule preparations of 2 or 4 described treatments, it is characterized in that: every capsules contains Radix Cynanchi Paniculati in paeonol (C9H10O3), is no less than 1.02mg; Every capsules contains total flavones in anhydrous rutin (C27H30O16), is no less than 2.00mg.