CN101485762A

CN101485762A - Quality control method of Chinese medicine preparation

Info

Publication number: CN101485762A
Application number: CNA2008100172977A
Authority: CN
Inventors: 韩勇; 包连勤
Original assignee: Individual
Current assignee: Individual
Priority date: 2008-01-16
Filing date: 2008-01-16
Publication date: 2009-07-22

Abstract

The invention discloses a method for controlling the quality of a golden flower acne eliminating preparation, wherein the preparation consists of Chinese traditional medicinal materials including (fried) gardenia, honeysuckle, (fried) radix scutellariae, rhubarb (stir-frying with wine), rhizoma coptidis, ballonflower, mint, bark of Chinese corktree, and liquorice. The method adopts a thin layer chromatography method to distinguish the gardenia, the rhizoma coptidis, the bark of Chinese corktree, the honeysuckle, the rhubarb, the radix scutellariae and the mint, and adopts a high-efficiency liquid phase chromatography method to perform content determination by using jasminoidin, emodin and chrysophanol as indexes. The method ensures the accuracy and the advanced property of the quality detection of the golden flower acne eliminating preparation, and controls the quality of the industrial production of the golden flower acne eliminating preparation so as to ensure the clinical curative effect of the preparation.

Description

A kind of method of quality control of Chinese medicine preparation

One, technical field

The present invention relates to the method for quality control field of Chinese medicine preparation, be specifically related to the method for quality control of little leaf deervetch herb acne removing preparation.

Two, background technology

The little leaf deervetch herb acne removing preparation crude drug is made up of Fructus Gardeniae (stir-fry), Flos Lonicerae, Radix Scutellariae (stir-fry), Radix Et Rhizoma Rhei (wine is processed), Rhizoma Coptidis, Radix Platycodonis, Herba Menthae, Cortex Phellodendri, Radix Glycyrrhizae etc., in order to control product quality effectively, we have set up the method for quality control of preparation, this method adopts according to thin layer chromatography Fructus Gardeniae, Rhizoma Coptidis, Cortex Phellodendri, Flos Lonicerae, Radix Et Rhizoma Rhei, Radix Scutellariae, Herba Menthae is differentiated, and according to high performance liquid chromatography with jasminoidin and emodin, row assay when chrysophanol is index.This method of quality control has guaranteed the accuracy and the advance of little leaf deervetch herb acne removing preparation quality inspection standard, can effectively guarantee the quality of little leaf deervetch herb acne removing preparation, and said preparation quality in industrialized great production is effectively controlled.

Three, summary of the invention

The method of quality control that the objective of the invention is to open little leaf deervetch herb acne removing preparation.

Method of quality control of the present invention comprises following discriminating and/or content assaying method.

Discrimination method of the present invention is one or more of following method:

1.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds water 10-30ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 1-3 time, each 10-30ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 1-3 time, each 10-30ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (4-6:4-6:0.5-1.5:0.5-1.5) is developing solvent, launch, take out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to the speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

2.. get little leaf deervetch herb acne removing preparation content 0.5g, porphyrize is added in the neutral alumina post (100-200 order, the 5g that have handled well, internal diameter 1.0cm, dry column-packing) on, with dehydrated alcohol 25-75ml eluting, collect eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 0.05g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Rhizoma Coptidis control medicinal material solution; Other gets Cortex Phellodendri control medicinal material 0.1g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Cortex Phellodendri control medicinal material solution; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned four kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-water (5-7:2-4:1-3:0.75-1.75:0.15-0.45) is developing solvent, put in 10-20 minute the chromatography cylinder of ammonia steam presaturation, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

3.. get little leaf deervetch herb acne removing preparation content 0.6g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds water 20-40ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 1-3 time, each 10-30ml discards ether solution, aqueous solution ethyl acetate extraction 3-5 time, each 10-30ml, merge extractive liquid,, evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the chlorogenic acid reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (4-6:0.5-1.5:0.5-1.5:0.5-1.5) 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 reference substance chromatograph on, show the fluorescence speckle of same color.

4.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 20-40ml, supersound process 5-15 minute, filter to round-bottomed flask evaporate to dryness, residue adds hydrochloric acid 1-3ml and chloroform 20-40ml, puts in the water-bath reflux 20-40 minute, puts cold, wash with water 1-3 time, each 20-40ml discards water liquid, chloroform liquid evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.3g, shines medical material solution in pairs with legal system; Get emodin, chrysophanol reference substance again, add chloroform and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-glacial acetic acid (14-16:4-6:0.15-0.45) is developing solvent, launches, and takes out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with reference substance and the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle; Put in the ammonia steam smoked after, speckle becomes redness.

5.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds that water 10-30ml is warm to make dissolving, filters while hot, and filtrate is transferred PH1-2 with dilute hydrochloric acid, water-bath 70-90 ℃ is incubated 20-40 minute, takes out, and puts cold, centrifugal (3500rpm) 5-15 minute, abandoning supernatant, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-butanone-formic acid-water (4-6:2-4:0.5-1.5:0.5-1.5) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 2%; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

6.. get little leaf deervetch herb acne removing preparation content 4g, porphyrize adds petroleum ether (60～90 ℃) and extracts 1-3 time, each 5-15ml, jolting 4-6 minute, to place 20-40 minute, merge extractive liquid, volatilizes naturally to about 1ml, as need testing solution; Other gets Herba Menthae control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the Mentholum reference substance again, add petroleum ether (60～90 ℃) and make the solution that every 1ml contains 2mg, product solution in contrast; Test according to thin layer chromatography, draw above-mentioned three kinds of each 9-11～19-21ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (18-20:0.5-1.5) is developing solvent, launch, take out, dry, spray is with the mixed solution of vanillin sulphuric acid test solution-ethanol (1:4), and it is clear to be heated to the 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.

Content assaying method of the present invention is one or more of following method:

1.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (10-20:80-90) is a mobile phase; The detection wavelength is 238 ± 2nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: it is an amount of to get little leaf deervetch herb acne removing preparation, and porphyrize is got about 0.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 20-30ml that adds, close plug claims to decide weight, supersound process 5-15 minute, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

2.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-0.1% phosphoric acid solution (70-90: 10-30) be mobile phase; The detection wavelength is 254 ± 2nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000; The preparation of reference substance solution: precision takes by weighing emodin, the chrysophanol reference substance is an amount of, adds methanol and makes the solution that contains 10ug among every 1ml respectively; The preparation of need testing solution: it is an amount of to get little leaf deervetch herb acne removing preparation, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 20-30ml that adds, claim to decide weight, reflux 0.5-1.5 hour, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, precision is measured subsequent filtrate 4-6ml, puts in the flask, fling to solvent, add 8% hydrochloric acid solution 5-15ml, supersound process 1-3 minute, add chloroform 5-15ml again, reflux 0.5-1.5 hour, put cold, put in the separatory funnel,, incorporate in the separatory funnel with a small amount of chloroform washing container, divide and get the chloroform layer, acid solution reuse chloroform extraction 2-4 time, each 5-15ml, merge chloroform liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving, be transferred in the 5-15ml measuring bottle, add methanol, shake up to scale, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

The thin layer chromatography that the invention provides Fructus Gardeniae, Rhizoma Coptidis, Cortex Phellodendri, Flos Lonicerae, Radix Et Rhizoma Rhei, Radix Scutellariae, Herba Menthae is differentiated, selected in the monarch drug Fructus Gardeniae in effective ingredient jasminoidin and the Radix Et Rhizoma Rhei active ingredient emodin, chrysophanol to set up the content assaying method of little leaf deervetch herb acne removing preparation as the assay index; Through repeated trials repeatedly, confirmation method is easy, repeatability is good, the result accurately and reliably, can be used as the little leaf deervetch herb acne removing preparation quality control index.

The used preparation of test sample adopts embodiment 1 described little leaf deervetch herb acne removing capsule in the experimental example of the present invention, and also available have other preparations that same materials is formed with the described little leaf deervetch herb acne removing capsule of embodiment.

Experimental example 1:

The content assaying method research of jasminoidin:

(1) instrument and Tianjin, reagent island LC-10ATVP type chromatograph of liquid pump main frame, Tianjin, island SPD-10ATVP type UV-detector; Electronic balance; SB2200-T type ultrasonic cleaner; Jasminoidin reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute uses for assay); Acetonitrile is a chromatographically pure, and water is redistilled water, and other reagent is analytical pure.

(2) preparation of reference substance solution: precision takes by weighing puts 24 hours jasminoidin reference substance 1.5mg of drying in the phosphorus pentoxide desiccator, puts in the 50ml volumetric flask, adds dissolve with methanol and is diluted to scale, shakes up, promptly.

(3) preparation of need testing solution: it is an amount of to get under the content uniformity item this product content, and porphyrize is got about 0.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, close plug claims to decide weight, supersound process 10 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.

(4) chromatographic condition is selected and system suitability test

1. chromatographic condition is selected

With reference to the chromatographic condition under " Fructus Gardeniae " item of 2005 editions pharmacopeia, selecting octadecyl silane for use is filler, is mobile phase with acetonitrile-water (15: 85), and the detection wavelength is 238nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500.

2. system suitability test

The number of theoretical plate of chromatographic column (n) calculates: by above selected condition, inject jasminoidin reference substance solution 10ul, record chromatogram, by formula n=5.54 (t _R/ W _H/2) ²The number of theoretical plate that calculates chromatographic column is 5820.Consider different chromatographic column conditions: differences such as column length, carrier property, filling situation, mobile phase ratio, tentative number of theoretical plate is not less than 1500 in the text.

Separating degree: this experimental condition becomes swarming to jasminoidin with other, and this has separating degree preferably: calculate with adjacent two nearest peaks; R=2 (t by formula _R2-t _R1)/W ₁+ W ₂Calculate jasminoidin separating degree 3.63, greater than 1.5 of the 2005 version Chinese Pharmacopoeia appendix VID of place regulation.

Tailing factor: T=1.04 (0.95≤T≤1.05) meets the pharmacopeia regulation.

(5) methodological study

1. the standard curve and the range of linearity: precision takes by weighing jasminoidin reference substance solution (0.1034mg/ml) 0.5,1.0,2.0,3.0,4.0,5.0ml, place the 10ml measuring bottle respectively, add methanol constant volume to scale, shake up, drawing 10ul respectively and inject chromatograph of liquid, measure its chromatographic peak area, is abscissa with jasminoidin concentration (ug), corresponding chromatographic peak area A is a vertical coordinate drawing standard curve, and learns by statistics and handle.As table 1

The table 1 jasminoidin standard curve and the range of linearity

By above-mentioned result of the test institute as can be known: jasminoidin is good linear relationship in 0.0517～0.5170ug scope.

Blank assay is in the negative sample of the scarce Fructus Gardeniae medical material of prescription ratio preparation, and the preparation method of pressing need testing solution prepares negative blank solution.

Get negative blank solution, liquid-phase chromatographic analysis is carried out in operation in accordance with the law, and the result shows that other composition is noiseless to jasminoidin mensuration.

2. stability test is got need testing solution and is measured immediately, and the need testing solution of placing different time under constant temperature is measured, and the results are shown in Table 2

Table 2 stability test result

Experimental result shows: jasminoidin is stable at 10 hours in the test sample.

3. need testing solution 10ul is got in the precision test, and continuous sample introduction 6 times is measured peak area, sees Table 3

Table 3 Precision test result

Experimental result shows: this method precision is good.

4. need testing solution 10ul is got in the repeatability test, and 5 parts, measure peak area, calculate content, the results are shown in Table 4.

Table 4 reproducible test results

Experimental result shows: this method repeatability is good.

5. recovery test is got the sample of known content (5.7502mg/ grain), porphyrize is crossed sieve No. 4, gets about 0.05g, precision takes by weighing 5 parts, put respectively in the 25ml measuring bottle, accurate respectively jasminoidin reference substance solution (0.4022mg/ml) 1.0ml that adds adds methanol to scale, below prepare need testing solution by the method under the assay item, measure in accordance with the law, calculate, the results are shown in Table 5

Table 5 average recovery result of the test

Experimental result shows: this law response rate is good.

(6) the assay data of sample and content limit determines

Through to ten batch samples, carry out the jasminoidin assay, wherein peak is the 6.4829mg/ grain, and minimum is the 4.3902mg/ grain, is 1.8% according to limiting the quantity of of jasminoidin in the Fructus Gardeniae (parched) medical material, and the theoretical content that calculates jasminoidin in this product is the 4.428mg/ grain.Consider the loss in the large-scale production process, the limit of jasminoidin content in this product is decided to be in every capsules is no less than 3.5mg, with the control inherent quality.The results are shown in Table 6

Jasminoidin assay result in ten batches of production samples of table 6

Experimental example 2:

The content assaying method research of emodin, chrysophanol:

(1) instrument and Tianjin, reagent island LC-10ATVP type chromatograph of liquid pump main frame, Tianjin, island SPD-10ATVP type UV-detector; Electronic balance; SY3200 type ultrasonic cleaner; Emodin, chrysophanol reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute uses for assay); Phosphoric acid is analytical pure, and methanol is chromatographically pure, and water is distilled water.

(2) preparation of reference substance solution: precision takes by weighing emodin, the chrysophanol reference substance is an amount of, adds methanol and makes the solution that contains 10ug among every 1ml respectively, promptly.

(3) preparation of need testing solution: depletion expense Cuo capsule is an amount of, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, claim to decide weight, reflux 1 hour is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, precision is measured subsequent filtrate 5ml, puts in the flask, fling to solvent, add 8% hydrochloric acid solution 10ml, supersound process 2 minutes, add chloroform 10ml again, reflux 1 hour is put cold, put in the separatory funnel,, incorporate in the separatory funnel with a small amount of chloroform washing container, divide and get the chloroform layer, acid solution reuse chloroform extraction 3 times, each 10ml, merge chloroform liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving, be transferred in the 10ml measuring bottle, add methanol, shake up to scale, filter, get subsequent filtrate, promptly.

(4) chromatographic condition is selected and system suitability test

1. chromatographic condition is selected

With reference to the chromatographic condition under " Radix Et Rhizoma Rhei " item of 2005 editions pharmacopeia, selecting octadecyl silane for use is filler, is mobile phase with methanol-0.1% phosphoric acid solution (80: 20), and the detection wavelength is 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000.

2. system suitability test

The number of theoretical plate of chromatographic column (n) calculates: by above selected condition, inject emodin, chrysophanol reference substance solution 10ul, record chromatogram, by formula n=5.54 (t _R/ W _H/2) ²The number of theoretical plate that calculates chromatographic column is 5023,5299.Consider different chromatographic column conditions: differences such as column length, carrier property, filling situation, mobile phase ratio, tentative number of theoretical plate is not less than 3000 in the text.

Separating degree: this experimental condition becomes swarming to emodin, chrysophanol with other, and this has separating degree preferably: calculate with adjacent two nearest peaks; R=2 (t by formula _R2-t _R1)/W ₁+ W ₂Calculate chrysophanol separating degree 2.68, greater than 1.5 of the 2005 version Chinese Pharmacopoeia appendix VID of place regulation.

(5) methodological study

1. the standard curve and the range of linearity: it is an amount of that precision takes by weighing the emodin reference substance, make the solution that every 1ml contains emodin 49.74ug with methanol, it is an amount of that precision takes by weighing the chrysophanol reference substance in addition, makes the solution that every 1ml contains chrysophanol 40.59ug with methanol, all makes the reference substance stock solution.

Precision is measured emodin stock solution 1,2,3,4,5ml places the 10ml measuring bottle respectively, add methanol constant volume to scale, shake up, respectively get 10ul and inject chromatograph of liquid, measure its chromatographic peak area, with emodin reference substance sample size C (ug/ml) is abscissa, and corresponding chromatographic peak area A is a vertical coordinate drawing standard curve, and learns by statistics and handle.Handle the chrysophanol stock solution with method, and the drawing standard curve.As table 1,2

The table 1 emodin standard curve and the range of linearity

The table 2 chrysophanol standard curve and the range of linearity

By above-mentioned result of the test institute as can be known: emodin is good linear relationship in 4.974～24.870ug/ml scope; Chrysophanol is good linear relationship in 4.059～20.295ug/ml scope.

Blank assay is in the negative sample of the scarce rhubarb medicinal material of prescription ratio preparation, and the preparation method of pressing need testing solution prepares negative blank solution.

Get negative blank solution, liquid-phase chromatographic analysis is carried out in operation in accordance with the law, and the result shows that other composition measures noiseless to emodin, chrysophanol.

2. the above-mentioned chromatographic condition of definite photograph of quantitative limit is measured than sample introduction by the noise of 10:1, as a result the lowest detectable limit of emodin, chrysophanol be 4.5 respectively, 4.0ng.

3. emodin, chrysophanol reference substance solution are got in the precision test, and continuous sample introduction 5 times is measured its peak area, and result's statistics: relative standard deviation is respectively 1.93%, 1.37%＜2.0%, sees Table 3

Table 3 Precision test result

4. stability test is got emodin, chrysophanol reference substance solution, at 0,1,2,4,6,12 hour difference sample introduction 10ul, measures its peak area, and 6 RSD are respectively 1.38%, 1.58% as a result, show that need testing solution is more stable, see Table 4

Table 4 stability test result

5. replica test is got with a need testing solution under the assay chromatographic condition, and repetitive operation is measured 5 times in accordance with the law, learns by statistics and handles, and is respectively 1.56%, 1.34% with standard deviation, sees Table 5

Table 5 replica test result

6. recovery test adopts the application of sample absorption method, get the same lot number sample of known content, accurate respectively a certain amount of emodin, the chrysophanol reference substance of adding, operate sample introduction measures in accordance with the law, with method repeated trials 5 times, the average recovery rate that records emodin, chrysophanol is respectively 98.67%, 95.47%, sees Table 6

Table 6 recovery test result

(6) the assay data of sample and content limit determines

Through to ten batch samples, carry out assay, the results are shown in Table 7.According to the sample size measurement result, characteristics in conjunction with industrialized great production, it is 1mg/g that the content of Radix Et Rhizoma Rhei in the little leaf deervetch herb acne removing capsule is fixed tentatively with the sum of emodin and chrysophanol content, be that every of this product contains Radix Et Rhizoma Rhei in emodin and chrysophanol, total amount should be less than 0.45mg, with the control inherent quality.

Emodin, chrysophanol assay result in ten batches of production samples of table 7

Following embodiment all can reach the effect of above-mentioned experimental example.

Four, the specific embodiment

Embodiment 1: get Chinese crude drug Fructus Gardeniae (stir-fry) 246g, Radix Et Rhizoma Rhei (wine is processed) 246g, Rhizoma Coptidis 61.5g and Cortex Phellodendri 61.5g according to the percolation (appendix IO of Chinese Pharmacopoeia version in 2005) under fluid extract and the extractum, make solvent with 70% ethanol, flood after 24 hours, percolation slowly, collect percolate 3600ml, the liquid of filtering is standby.Herba Menthae 246g extracts volatile oil with the way of distillation, the volatile oil beta-cyclodextrin inclusion compound, medicinal residues and Flos Lonicerae 246g, Radix Scutellariae (stir-fry) 246g, Radix Platycodonis 246g, Radix Glycyrrhizae 123g decocts with water twice, each 2 hours, collecting decoction, filter, it is 1.10-1.15 (40～45 ℃) that filtrate is concentrated into relative density, puts cold, adding ethanol makes and contains amount of alcohol and reach 70%, leave standstill, incline and get supernatant, filter, filtrate and the above-mentioned liquid of filtering merge, reclaim ethanol, being concentrated into relative density is the clear paste of 1.30-1.35 (50～55 ℃), and it is an amount of to add adjuvant silicon dioxide, mixing, drying under reduced pressure is ground into fine powder, makes granule, add above-mentioned volatile oil beta cyclodextrin inclusion complex, mixing incapsulates, and makes 1000.The method of quality control of this medicine comprises following method:

Differentiate: 1.. get this product content 1g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 2 times, each 20ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 2 times, each 20ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (5:5:1:1) is developing solvent, launch, take out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to the speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

2.. get this product content 0.5g, porphyrize is added on the neutral alumina post of having handled well (100-200 order, 5g, internal diameter 1.0cm, dry column-packing), with dehydrated alcohol 50ml eluting, collects eluent, and evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 0.05g, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate adds methanol to 5ml, as Rhizoma Coptidis control medicinal material solution; Other gets Cortex Phellodendri control medicinal material 0.1g, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate adds methanol to 5ml, as Cortex Phellodendri control medicinal material solution; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned four kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-water (6:3:2:1.5:0.3) is developing solvent, put in 15 minutes the chromatography cylinder of ammonia steam presaturation, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, with the corresponding position of reference substance chromatograph on, show identical yellow fluorescence speckle.

3.. get this product content 0.6g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 30ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 2 times, each 20ml discards ether solution, aqueous solution ethyl acetate extraction 4 times, each 20ml, merge extractive liquid,, evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the chlorogenic acid reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (5:1:1:1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical blue-fluorescence speckle.

4.. get this product content 1g, porphyrize adds methanol 30ml, supersound process 10 minutes filters to round-bottomed flask evaporate to dryness, residue adds hydrochloric acid 2ml and chloroform 30ml, puts in the water-bath reflux 30 minutes, puts cold, wash with water 2 times, each 30ml discards water liquid, chloroform liquid evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.3g, shines medical material solution in pairs with legal system; Get emodin, chrysophanol reference substance again, add chloroform and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-glacial acetic acid (15:5:0.3) is developing solvent, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with reference substance and the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle; Put in the ammonia steam smoked after, speckle becomes redness.

5.. get this product content 1g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds that water 20ml is warm to make dissolving, filters while hot, and filtrate is transferred PH1-2 with dilute hydrochloric acid, 80 ℃ of insulations of water-bath 30 minutes are taken out, and put cold, centrifugal (3500rpm) 10 minutes, abandoning supernatant, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-butanone-formic acid-water (5:3:1:1) is developing solvent, launch, take out, dry, spray ferric chloride alcoholic solution with 2%; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical dirty-green speckle.

6.. get this product content 4g, porphyrize adds petroleum ether (60～90 ℃) and extracts 2 times, each 10ml, jolting 5 minutes was placed 30 minutes, and merge extractive liquid, volatilizes naturally to about 1ml, as need testing solution; Other gets Herba Menthae control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the Mentholum reference substance again, add petroleum ether (60～90 ℃) and make the solution that every 1ml contains 2mg, product solution in contrast; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 10～20ul of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (19:1) is developing solvent, launch, take out, dry, spray is with the mixed solution of vanillin sulphuric acid test solution-ethanol (1:4), and it is clear to be heated to the 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.

Assay: 1.. measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D): chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (15: 85) is a mobile phase; The detection wavelength is 238nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: it is an amount of to get under the content uniformity item this product content, and porphyrize is got about 0.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, close plug claims to decide weight, supersound process 10 minutes, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly; Every of this product contains Fructus Gardeniae with jasminoidin (C ₁₇H ₂₄O ₁₀) meter, must not be less than 3.5mg.

2.. measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D): chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-0.1% phosphoric acid solution (80: 20) is a mobile phase; The detection wavelength is 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000; The preparation of reference substance solution: precision takes by weighing emodin, the chrysophanol reference substance is an amount of, adds methanol and makes the solution that contains 10ug among every 1ml respectively; The preparation of need testing solution: it is an amount of to get under the content uniformity item this product content, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, close plug claims to decide weight, reflux 1 hour, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5ml, put in the flask, fling to solvent, add 8% hydrochloric acid solution 10ml, supersound process 2 minutes adds chloroform 10ml again, reflux 1 hour, put coldly, put in the separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution reuse chloroform extraction 3 times, each 10ml merges chloroform liquid, and decompression and solvent recovery is to doing, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, adds methanol to scale, shake up, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly; Every of this product contains Radix Et Rhizoma Rhei with emodin (C ₁₅H ₁₀O ₅) and chrysophanol (C ₁₅H ₁₀O ₄) the total amount meter, must not be less than 0.45mg.

Embodiment 2: get Chinese crude drug Fructus Gardeniae (stir-fry) 246g, Radix Et Rhizoma Rhei (wine is processed) 246g, Rhizoma Coptidis 61.5g and Cortex Phellodendri 61.5g according to the percolation (appendix IO of Chinese Pharmacopoeia version in 2005) under fluid extract and the extractum, make solvent with 70% ethanol, flood after 24 hours, percolation slowly, collect percolate 3600ml, the liquid of filtering is standby.Herba Menthae 246g extracts volatile oil with the way of distillation, the volatile oil beta-cyclodextrin inclusion compound, medicinal residues and Flos Lonicerae 246g, Radix Scutellariae (stir-fry) 246g, Radix Platycodonis 246g, Radix Glycyrrhizae 123g decocts with water twice, each 2 hours, collecting decoction, filter, it is 1.10-1.15 (40～45 ℃) that filtrate is concentrated into relative density, puts cold, adding ethanol makes and contains amount of alcohol and reach 70%, leave standstill, incline and get supernatant, filter, filtrate and the above-mentioned liquid of filtering merge, reclaim ethanol, being concentrated into relative density is the clear paste of 1.30-1.35 (50～55 ℃), and it is an amount of to add adjuvant silicon dioxide, mixing, drying under reduced pressure is ground into fine powder, makes granule, add above-mentioned volatile oil beta cyclodextrin inclusion complex, mixing, compacting is made 1000 in flakes.The method of quality control of this medicine is with embodiment 1.

Claims

1, the method for quality control of little leaf deervetch herb acne removing preparation, wherein said preparation is made up of Fructus Gardeniae (stir-fry), Flos Lonicerae, Radix Scutellariae (stir-fry), Radix Et Rhizoma Rhei (wine is processed), Rhizoma Coptidis, Radix Platycodonis, Herba Menthae, Cortex Phellodendri, Radix Glycyrrhizae, it is characterized in that discrimination method in this method is one or more of following method:

1.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds water 10-30ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 1-3 time, each 10-30ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 1-3 time, each 10-30ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (4-6:4-6:0.5-1.5:0.5-1.5) is developing solvent, launch, take out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to the speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

2.. get little leaf deervetch herb acne removing preparation content 0.5g, porphyrize is added in the neutral alumina post (100-200 order, the 5g that have handled well, internal diameter 1.0cm, dry column-packing) on, with dehydrated alcohol 25-75ml eluting, collect eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 0.05g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Rhizoma Coptidis control medicinal material solution; Other gets Cortex Phellodendri control medicinal material 0.1g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Cortex Phellodendri control medicinal material solution; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned four kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-water (5-7: 2-4:1-3:0.75-1.75:0.15-0.45) be developing solvent, put in 10-20 minute the chromatography cylinder of ammonia steam presaturation, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

3.. get little leaf deervetch herb acne removing preparation content 0.6g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds water 20-40ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 1-3 time, each 10-30ml discards ether solution, aqueous solution ethyl acetate extraction 3-5 time, each 10-30ml, merge extractive liquid,, evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the chlorogenic acid reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (4-6:0.5-1.5:0.5-1.5:0.5-1.5) 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 reference substance chromatograph on, show the fluorescence speckle of same color;

4.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 20-40ml, supersound process 5-15 minute, filter to round-bottomed flask evaporate to dryness, residue adds hydrochloric acid 1-3ml and chloroform 20-40ml, puts in the water-bath reflux 20-40 minute, puts cold, wash with water 1-3 time, each 20-40ml discards water liquid, chloroform liquid evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.3g, shines medical material solution in pairs with legal system; Get emodin, chrysophanol reference substance again, add chloroform and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-glacial acetic acid (14-16:4-6:0.15-0.45) is developing solvent, launches, and takes out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with reference substance and the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle; Put in the ammonia steam smoked after, speckle becomes redness;

5.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds that water 10-30ml is warm to make dissolving, filters while hot, and filtrate is transferred PH1-2 with dilute hydrochloric acid, water-bath 70-90 ℃ is incubated 20-40 minute, takes out, and puts cold, centrifugal (3500rpm) 5-15 minute, abandoning supernatant, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-butanone-formic acid-water (4-6:2-4:0.5-1.5:0.5-1.5) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 2%; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

2, the capsular method of quality control of little leaf deervetch herb acne removing as claimed in claim 1 is characterized in that discrimination method in this method is one or more of following method:

1.. depletion expense Cuo capsule 's content 1g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 2 times, each 20ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 2 times, each 20ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (5:5:1:1) is developing solvent, launches, and takes out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

2.. depletion expense Cuo capsule 's content 0.5g, porphyrize is added in the neutral alumina post (100-200 order, the 5g that have handled well, internal diameter 1.0cm, dry column-packing) on, with dehydrated alcohol 50ml eluting, collect eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 0.05g, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate adds methanol to 5ml, as Rhizoma Coptidis control medicinal material solution; Other gets Cortex Phellodendri control medicinal material 0.1g, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate adds methanol to 5ml, as Cortex Phellodendri control medicinal material solution; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned four kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-water (6:3:2:1.5:0.3) is developing solvent, put in 15 minutes the chromatography cylinder of ammonia steam presaturation, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

3.. depletion expense Cuo capsule 's content 0.6g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 30ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 2 times, each 20ml discards ether solution, aqueous solution ethyl acetate extraction 4 times, each 20ml, merge extractive liquid,, evaporate to dryness, residue adds ethanol 2ml makes dissolving, as need testing solution; Other gets the chlorogenic acid reference substance, adds methanol and makes the solution that every 1ml contains 0.2mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-acetone-formic acid-water (5:1:1:1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

4.. depletion expense Cuo capsule 's content 1g, porphyrize adds methanol 30ml, supersound process 10 minutes filters to round-bottomed flask evaporate to dryness, residue adds hydrochloric acid 2ml and chloroform 30ml, puts in the water-bath reflux 30 minutes, puts cold, wash with water 2 times, each 30ml discards water liquid, chloroform liquid evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets Radix Et Rhizoma Rhei control medicinal material 0.3g, shines medical material solution in pairs with legal system; Get emodin, chrysophanol reference substance again, add chloroform and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate-glacial acetic acid (15:5:0.3), launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with reference substance and the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle; Put in the ammonia steam smoked after, speckle becomes redness;

5.. depletion expense Cuo capsule 's content 1g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds that water 20ml is warm to make dissolving, filters while hot, and filtrate is transferred PH1-2 with dilute hydrochloric acid, 80 ℃ of insulations of water-bath 30 minutes are taken out, and put cold, centrifugal (3500rpm) 10 minutes, abandoning supernatant, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-butanone-formic acid-water (5:3:1:1), launch, take out, dry, spray ferric chloride alcoholic solution with 2%; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

6.. depletion expense Cuo capsule 's content 4g, porphyrize adds petroleum ether (60～90 ℃) and extracts 2 times, each 10ml, jolting 5 minutes was placed 30 minutes, and merge extractive liquid, volatilizes naturally to about 1ml, as need testing solution; Other gets Herba Menthae control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the Mentholum reference substance again, add petroleum ether (60～90 ℃) and make the solution that every 1ml contains 2mg, product solution in contrast; Test according to thin layer chromatography, draw each 10～20ul of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (19:1) is developing solvent, launch, take out, dry, spray is with the mixed solution of vanillic acid test solution-ethanol (1:4), and it is clear to be heated to the 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.

3, the method for quality control of little leaf deervetch herb acne removing preparation, wherein said preparation is made up of Chinese crude drug Fructus Gardeniae (stir-fry), Flos Lonicerae, Radix Scutellariae (stir-fry), Radix Et Rhizoma Rhei (wine is processed), Rhizoma Coptidis, Radix Platycodonis, Herba Menthae, Cortex Phellodendri, Radix Glycyrrhizae, it is characterized in that content assaying method in this method is one or more of following method:

1.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (10-20: 80-90) be mobile phase; The detection wavelength is 238 ± 2nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: it is an amount of to get little leaf deervetch herb acne removing preparation, and porphyrize is got about 0.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 20-30ml that adds, close plug claims to decide weight, supersound process 5-15 minute, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

4, the capsular method of quality control of little leaf deervetch herb acne removing as claimed in claim 3 is characterized in that content assaying method in this method is one or more of following method:

1.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (15: 85) is a mobile phase; The detection wavelength is 238nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: depletion expense Cuo capsule is an amount of, and porphyrize is got about 0.1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, close plug claims to decide weight, and supersound process 10 minutes is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up, filter, get subsequent filtrate with methanol; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

2.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-0.1% phosphoric acid solution (80: 20) is a mobile phase; The detection wavelength is 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000; The preparation of reference substance solution: precision takes by weighing emodin, the chrysophanol reference substance is an amount of, adds methanol and makes the solution that contains 10ug among every 1ml respectively; The preparation of need testing solution: depletion expense Cuo capsule is an amount of, and porphyrize is got about 1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, claim to decide weight, reflux 1 hour is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, precision is measured subsequent filtrate 5ml, puts in the flask, fling to solvent, add 8% hydrochloric acid solution 10ml, supersound process 2 minutes, add chloroform 10ml again, reflux 1 hour is put cold, put in the separatory funnel,, incorporate in the separatory funnel with a small amount of chloroform washing container, divide and get the chloroform layer, acid solution reuse chloroform extraction 3 times, each 10ml, merge chloroform liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving, be transferred in the 10ml measuring bottle, add methanol, shake up to scale, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

5, the method for quality control of little leaf deervetch herb acne removing preparation, wherein said pharmaceutical formulation is made up of Fructus Gardeniae (stir-fry), Flos Lonicerae, Radix Scutellariae (stir-fry), Radix Et Rhizoma Rhei (wine is processed), Rhizoma Coptidis, Radix Platycodonis, Herba Menthae, Cortex Phellodendri, Radix Glycyrrhizae, it is characterized in that this method comprises following method:

Differentiate: 1.. get little leaf deervetch herb acne removing preparation content 1g, porphyrize adds methanol 25-75ml, supersound process 10-30 minute, filter the filtrate evaporate to dryness, residue adds water 10-30ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 1-3 time, each 10-30ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 1-3 time, each 10-30ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (4-6:4-6:0.5-1.5:0.5-1.5) is developing solvent, launch, take out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to the speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

2.. get little leaf deervetch herb acne removing preparation content 0.5g, porphyrize is added in the neutral alumina post (100-200 order, the 5g that have handled well, internal diameter 1.0cm, dry column-packing) on, with dehydrated alcohol 25-75ml eluting, collect eluent, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets Rhizoma Coptidis control medicinal material 0.05g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Rhizoma Coptidis control medicinal material solution; Other gets Cortex Phellodendri control medicinal material 0.1g, adds methanol 4-6ml, and supersound process 10-20 minute, filter, filtrate adds methanol to 5ml, as Cortex Phellodendri control medicinal material solution; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw above-mentioned four kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-methanol-isopropyl alcohol-water (5-7:2-4:1-3:0.75-1.75:0.15-0.45) is developing solvent, put in 10-20 minute the chromatography cylinder of ammonia steam presaturation, launch, take out, dry, place under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

6.. get little leaf deervetch herb acne removing preparation content 4g, porphyrize adds petroleum ether (60～90 ℃) and extracts 1-3 time, each 5-15ml, jolting 4-6 minute, to place 20-40 minute, merge extractive liquid, volatilizes naturally to about 1ml, as need testing solution; Other gets Herba Menthae control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the Mentholum reference substance again, add petroleum ether (60～90 ℃) and make the solution that every 1ml contains 2mg, product solution in contrast; Test according to thin layer chromatography, draw above-mentioned three kinds of each 9-11～19-21ul of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (18-20:0.5-1.5) is developing solvent, launch, take out, dry, spray is with the mixed solution of vanillin sulphuric acid test solution-ethanol (1:4), and it is clear to be heated to the 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;

Assay: 1.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (10-20: 80-90) be mobile phase; The detection wavelength is 238 ± 2nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: it is an amount of to get little leaf deervetch herb acne removing preparation, and porphyrize is got about 0.1g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 20-30ml that adds, close plug claims to decide weight, supersound process 5-15 minute, put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol, filter, get subsequent filtrate; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

6, the capsular method of quality control of little leaf deervetch herb acne removing as claimed in claim 5 is characterized in that comprising in this method following method:

Differentiate: 1.. depletion expense Cuo capsule 's content 1g, porphyrize adds methanol 50ml, supersound process 20 minutes filters the filtrate evaporate to dryness, residue adds water 20ml makes dissolving, filters, and filtrate adds diethyl ether and extracts 2 times, each 20ml discards ether solution, and aqueous solution is with water saturated n-butanol extraction 2 times, each 20ml merges n-butanol extracting liquid, evaporate to dryness, residue adds ethanol 1ml makes dissolving, as need testing solution; Other gets the jasminoidin reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw above-mentioned two kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-acetone-formic acid-water (5:5:1:1) is developing solvent, launches, and takes out, dry, spray is with sulphuric acid ethanol (5 → 10) solution, and it is clear to be heated to speckle colour developing at 110 ℃; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

6.. depletion expense Cuo capsule 's content 4g, porphyrize adds petroleum ether (60～90 ℃) and extracts 2 times, each 10ml, jolting 5 minutes was placed 30 minutes, and merge extractive liquid, volatilizes naturally to about 1ml, as need testing solution; Other gets Herba Menthae control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get the Mentholum reference substance again, add petroleum ether (60～90 ℃) and make the solution that every 1ml contains 2mg, product solution in contrast; Test according to thin layer chromatography, draw each 10～20ul of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate (19:1) is developing solvent, launch, take out, dry, spray is with the mixed solution of vanillin sulphuric acid test solution-ethanol (1:4), and it is clear to be heated to the 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;

Assay: 1.. shine high effective liquid chromatography for measuring: chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (15: 85) is a mobile phase; The detection wavelength is 238nm; Number of theoretical plate calculates by the jasminoidin peak should be not less than 1500; The preparation of reference substance solution: it is an amount of that precision takes by weighing the jasminoidin reference substance, adds methanol and make the solution that contains 30ug among every 1ml; The preparation of need testing solution: depletion expense Cuo capsule is an amount of, and porphyrize is got about 0.1g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methanol 25ml that adds, close plug claims to decide weight, and supersound process 10 minutes is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up, filter, get subsequent filtrate with methanol; Algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;