CN1857417A - Chinese medicine Shuanghuanglian ointment and its quality control method - Google Patents

Abstract

The present invention relates to Chinese medicine Shuanghuanglian ointment and its quality control method. The preparation process of Chinese medicine Shuanghuanglian ointment includes the following steps: decocting, filtering and alcohol extracting skullcap root, honeysuckle and forsythia separately to obtain water solution; mixing the water solution, regulating pH to 5.0-9.0, decompression concentrating, low temperature drying and crushing to obtain medicine powder; dissolving polyethylene glycol-400, polyethylene glycol-6000 and ethyl hydroxyl benzoate in water bath at 40-80 deg.c, adding the medicine powder gradually while stirring and lowering the temperature to room temperature to form ointment; and packing in tube. The Chinese medicine Shuanghuanglian ointment has determined curative effect.

Description

A kind of Chinese medicine Shuanghuanglian ointment and method of quality control thereof

Technical field:

The present invention relates to a kind of Chinese medicine ointment and method of quality control thereof, especially relate to a kind of Chinese medicine Shuanghuanglian ointment and method of quality control thereof that is used for the treatment of diseases such as flu due to the affection due to external wind and heat, upper respiratory tract infection, pneumonia.

Background technology:

Traditional suppository uses inconvenient, and effective ingredient absorbs slow, because of its service time longer, so the zest of being easy to generate, and production cost is also higher.Ointment has then overcome the above-mentioned shortcoming and defect of suppository, helps medicine and contacts with the affected part, only need smear during use, and the rapid stripping of effective ingredient is evenly distributed.Adopt suitable substrate, the release that helps effective ingredient absorbs, and gives full play to the curative effect of medicine more.

Summary of the invention:

Technical problem to be solved by this invention provides a kind of Chinese medicine Shuanghuanglian ointment and method of quality control thereof, to overcome the deficiency that traditional suppository exists.

The preparation method of this Chinese medicine ointment:

(1) crude drug is formed and proportioning (weight portion):

Flos Lonicerae 1000-3000 weight portion Radix Scutellariae 1000-3000 weight portion

Fructus Forsythiae 2000-6000 weight portion

Its optimum ratio is (weight portion):

Flos Lonicerae 1667 weight portion Radix Scutellariaes 1667 weight portions

Fructus Forsythiae 3334 weight portions

(2) preparation technology

The Radix Scutellariae of described weight proportion is added 4-12 times of decocting to be boiled 2-5 time, collecting decoction, filter, filtrate is concentrated into the clear paste that relative density is 1-2 (50 ~ 60 ℃), and concentrated solution adds the 1-3mol/L hydrochloric acid solution in the time of 80 ℃, regulates pH value to 1.0～2.0, be incubated and left standstill 24-48 hour after 1-3 hour, filter, precipitate adds 3-8 times of water gaging, regulates pH value to 6.0-8.5 with the 20-60% sodium hydroxide solution, add equivalent ethanol, stirring makes dissolving, filters, and filtrate is regulated pH value to 1.0-2.0 with the 1-3mol/L hydrochloric acid solution, 60 ℃ are incubated 10-50 minute, left standstill 6-24 hour, and filtered, precipitation washes with water to pH to 3.0-7.0, continue and be washed till pH value 5.0-7.0 with 50-90% ethanol, precipitate adds 3-8 times of water gaging, regulates pH value to 6.0-8.5 with the 20-40% sodium hydroxide solution, stirs and makes dissolving.Flos Lonicerae, Fructus Forsythiae adds 4-12 times of decocting and boils 1-3 time, each 1.5 hours, collecting decoction filters, and filtrate is concentrated into the clear paste that relative density is 1.0-1.4 (70～80 ℃), stir when being chilled to 20-40 ℃ and slowly add ethanol down, make to contain alcohol amount and reach 50-85%, left standstill the leaching supernatant 6-16 hour, reclaim ethanol, concentrated solution is concentrated into the clear paste that relative density is 1.0-1.3 (50～60 ℃), stirs when being chilled to 20-60 ℃ to add ethanol down again and make and contain the alcohol amount and reach 60-90%, fully stirs, left standstill 8-24 hour, the leaching supernatant reclaims ethanol to there not being the alcohol flavor, adds above-mentioned Radix Scutellariae extract aqueous solution, stir evenly, and regulate pH value to 5.0-9.0, concentrating under reduced pressure becomes thick paste, cold drying, pulverize, standby; Get 300-600 weight portion PEG400 and 100-300 part polyethylene glycol 6000,40-80 ℃ of heating in water bath dissolving, add 1-3 part ethyl hydroxybenzoate again and make dissolving, in water-bath, substrate is progressively joined in the above-mentioned fine powder, constantly stir so that mix homogeneously, stirring at room is even, and the mastic temperature drops to room temperature and congeals into the ointment shape, embedding is in the compound cream pipe of polyethylene, promptly.

Chinese medicine SHUANGHUANLIAN ointment of the present invention is yellow ointment, has dispelling wind to relieve the exterior syndrome, the effect of heat-clearing and toxic substances removing.Be used for the flu due to the affection due to external wind and heat, disease is seen heating, cough, pharyngalgia; Upper respiratory tract infection, pneumonia etc.

The method of quality control of Chinese medicine ointment of the present invention contains one or more in following discriminating and/or the content assaying method, and discriminating in the method for quality control of the present invention and content assaying method are:

Discrimination method is selected from one or more in the following method:

(1) get this product 2g, add methanol 10-30ml, supersound extraction 5-15min is put coldly, filters, and gets filtrate 1ml, as need testing solution; Other gets baicalin, the chlorogenic acid reference substance is made the solution that every 1ml contains 0.4mg with ethanol respectively, in contrast product solution.Test according to thin layer chromatography (2005 editions one appendix VI B of Chinese Pharmacopoeia), draw each 3～5 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, upper solution with butyl acetate-formic acid-water 6-8: 3-5: 2-4 is developing solvent, presaturation 30 minutes 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.

(2) get the about 2g of this product, add methanol 10-30ml, supersound extraction 5-15min is put coldly, filters, and filtrate is concentrated into about 5ml as need testing solution.Other gets Fructus Forsythiae control medicinal material 0.5g, adds methanol 10ml, puts in the water-bath reflux 10-30 minute, filters, and filtrate is medical material solution in contrast.Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2005 VI B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol 4-6: 0.5-1.5 is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 80-120 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Content assaying method is:

(1) baicalin is measured with reference to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).

With octadecyl silane is filler; Methanol-water-glacial acetic acid 40-60: 40-60: 0.5-1.5 is a mobile phase; Flow velocity: 1.0ml/min; Column temperature: room temperature; The detection wavelength is 276nm.Number of theoretical plate calculates by the baicalin peak should be not less than 1000.Precision takes by weighing baicalin reference substance 10mg, puts in the 100ml measuring bottle, and it is an amount of to add 50% methanol, puts that jolting makes its dissolving in the water-bath, is placed to room temperature, and is diluted to scale, shakes up, and is contrast device solution (containing baicalin 0.1mg among every 1ml).Get the about 2g of this product, accurate title is fixed, puts the accurate methanol 50ml that adds in the tool plug conical flask, and after weight decided in accurate title, supersound extraction 30 minutes was put and is chilled to room temperature, the weight of replenish lost.Filter, precision measures that methanol is diluted to scale in subsequent filtrate 25ml to the 50ml measuring bottle.Precision is measured 3ml and is put in the 50ml measuring bottle, adds methanol and is diluted to scale, promptly gets need testing solution.Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, and the every gram of above-mentioned traditional Chinese herb medicine preparation contains Radix Scutellariae with baicalin (C ₂₁H ₁₈O ₁₁) meter, should be no less than 50mg.

Below example further specifies the present invention by experiment:

The screening experiment of experimental example 1 ointment base

SHUANGHUANLIAN ointment is to be used for the treatment of the flu that is used for due to the affection due to external wind and heat, and disease is seen heating, cough, pharyngalgia; Disease such as upper respiratory tract infection, pneumonia.In the substrate of ointment, water-soluble base is water-soluble, and it is very fast generally to discharge medicine, and no greasy easily is coated with exhibition, to skin and mucosa nonirritant, is convenient to clean not sticking clothes.In sum, select the substrate of water-soluble base for use as SHUANGHUANLIAN ointment.In water-soluble base, modal have a polyethylene glycols, also has glycerin gelatine, glycerinum amyli, cellulose derivative, carbopol etc.Several water-soluble basees prescription screening and stability experiment have been carried out.Ointment is divided into solution-type ointment and suspension type ointment because of the dispersity difference of medicine in substrate.SHUANGHUANLIAN ointment is the suspension type ointment, because through experiment, finds that extract powder can not dissolve fully in water-soluble base, even heating also is suspended state, therefore, considers SHUANGHUANLIAN ointment is made the suspension type ointment.

Select several water-soluble basees commonly used, prescription is respectively 1. gelatin 2g, and glycerol 2g adds water to 100g; 2. methylcellulose 6g, propylene glycol 10g adds water to 100g; 3. PEG-600020g, PEG-40080g; 4. carbopol 1g, ethanol 5g, glycerol 5g, sodium hydroxide 0.4g adds water to 100g.After making ointment, put in the hermetic container.

The study on the stability index: quickening stable experiment back is to investigate index with the character, comprises pH, becomes sour, foreign odor, variable color, hardening (dehydration), granularity, layering, stretchability.

Accelerated stability experimental technique (1) centrifuging: 10g places centrifuge tube with ointment, 2500rmin ^-1, 30min; (2) heat-resistant experiment: ointment is placed 55 ℃, constant temperature 6h; (3) cold-resistant experiment: ointment is placed-5 ℃, 24h.

PH value is measured: get ointment 2g, with distilled water with 1: 10 dilution proportion after according to the mensuration of the pH value algoscopy Chinese Pharmacopoeia version appendix in 2005 VII G item under pH.

The The selection result of 4 kinds of SHUANGHUANLIAN ointment base prescriptions sees Table 1.

Table 1 SHUANGHUANLIAN ointment base prescription screening result

Prescription	Acceleration environment	pH	Foreign odor becomes sour	Color and luster	Granularity	Quality	Layering	Stretchability
									1	The cold-resistant experiment of the centrifugal property of room temperature heat-resistant experiment	5.47 4.92 5.67 5.46	Do not have	Yellow	Defective	Fineless and smooth or not	No layering does not have layering and does not have layering and do not have layering	Relatively poor
2	The cold-resistant experiment of the centrifugal property of room temperature heat-resistant experiment	5.34 4.89 5.01 5.08	Do not have	Yellow	Qualified defective qualified	It is fine and smooth that evenly inhomogeneous exquisiteness is even	No layering has drug precipitation to have drug precipitation not have layering	Good
									3	The cold-resistant experiment of the centrifugal property of room temperature heat-resistant experiment	6.91 6.86 6.83 6.01	Do not have	Yellow	Qualified	Fine and smooth uniform and smooth uniform and smooth uniform and smooth is even	No layering does not have layering and does not have layering and do not have layering	Good
4	The cold-resistant experiment of the centrifugal property of room temperature heat-resistant experiment	5.91 5.61 6.23 6.17	Do not have	Yellow	Qualified defective defective	Fine and smooth evenly inhomogeneous inhomogeneous	No layering has obvious layering to have obvious layering to have medicine to separate out	Good

Conclusion: selecting prescription 3. is ointment base.

Experimental example 2 principal agents and the test of substrate compatibility

After Polyethylene Glycol substrate and principal agent made ointment by typical concentrations, put in the hermetic container, seasoning is one month under different experimental conditions, serves as to investigate index with physical behavior and content of baicalin, whether destroyed, whether drug content changes and whether ointment is stablized if observing substrate.Experiment condition following (1) baking oven (40 ± 1 ℃) (2) room temperature (25 ± 3 ℃) (3) refrigerator (0 ± 2 ℃).Experimental result sees Table 2.

Table 2 SHUANGHUANLIAN ointment principal agent and substrate compatibility experimental result

Experiment condition	pH	Foreign odor becomes sour	Color and luster	Granularity	Quality	Layering	Stretchability	Content (mg/g)
									Prepare 0 day room temperature baking oven in January refrigerator in January January	5.97 5.92 5.87 6.05	Do not have	Yellow	Qualified	Fine and smooth uniform and smooth uniform and smooth uniform and smooth is even	No layering does not have layering and does not have layering and do not have layering	Good	96.21 96.12 95.17 95.73

Conclusion: drug content does not change, and does not have incompatibility with substrate, and the ointment of making is very stable.

The selection of experimental example 3 principal agent degree of grinding

Because SHUANGHUANLIAN ointment is the suspension type ointment, the insoluble solid medicine needs to wear into fine powder with suitable method, guarantees that granularity is up to specification.Therefore, after solid drugs pulverized, selected the sieve in different apertures to obtain varigrained fine powder, make ointment after, investigate ointment, result such as table 3 with physical behaviors such as granularity, outward appearances.

Table 3 SHUANGHUANLIAN ointment principal agent degree of grinding experimental result

Experiment condition	pH	Foreign odor becomes sour	Color and luster	Granularity	Quality	Layering	Stretchability
								Crossing 120 orders crosses 140 orders and crosses 160 orders and cross 180 orders and cross 200 orders	6.03 6.04 6.01 5.87 5.96	Do not have	Yellow yellow	Defective qualified	The fine and smooth uniform and smooth of the coarse inequality of the coarse inequality of coarse inequality is even	No layering does not have layering and does not have layering and do not have layering and do not have layering	Relatively poor general good

Conclusion: it is evenly fine and smooth that principal agent is crossed the prepared ointment of the impalpable powder that obtains behind 200 mesh sieves, and granularity is up to specification.

The selection of experimental example 4 substrate and principal agent ratio

Principal agent and substrate have suitable ratio, if drug content is too high, can influence the dispersion of principal agent in substrate, and drug content is low excessively, and the matrix content height causes dosage excessive.Adopt orthogonal design, select L for use ₉(2) ³The orthogonal table experiment arrangement, (not stratified fully is 0 with stretchability and stability, other is between 0～10) be evaluation index, the consumption of principal agent, PEG400 and polyethylene glycol 6000 (wherein the consumption of polyethylene glycol 6000 is to remove remaining ratio after the shared ratio of principal agent and PEG400) is investigated.According to experimental result, principal agent has been selected with 30.0%, 33.3% and 36.6% 3 level, and PEG400 is selected 48.0%, 50.0% and 52.0% 3 level.Experimental result sees Table 4, table 5.

Table 4 SHUANGHUANLIAN ointment prescription screening factor level table

The factor title	PEG400 consumption (%)	Principal agent consumption (%)
			Level 1 level 2 levels 3	48.0 50.0 52.0	30.0 33.3 36.6

Table 5 SHUANGHUANLIAN ointment prescription screening Orthogonal experiment results table

Factor		PEG400	The principal agent consumption	A×B	Blank column	Stretchability	Stability	Total score
									Experiment 1 experiment 2 experiments 3 experiments 4 experiments 5 experiments 6 experiments 7 experiments 8 experiments 9		1 1 1 2 2 2 3 3 3	1 2 3 1 2 3 1 2 3	1 2 3 2 3 1 3 1 2	1 2 3 2 3 1 3 1 2	8 8 8 7 9 7 6 7 7	6 7 6 9 9 9 9 9 9	14 15 14 16 18 16 15 16 16
Intuitive analysis	Average 1 average 2 averages 3 are differential	14.333 16.667 15.667 2.334	15 16.333 15.333 1.333	15.333 15.667 15.667 0.334	16 15.333 15.333 0.667
									Variance analysis	Sum of square of deviations degree of freedom F is than F marginal value significance P*	8.222 2 9.249 19.00 ＞0.05	2.889 2 3.25 19.00 ＞0.05	0.22 2 0.25 19.00 ＞0.05	0.89 2

According to last table data, by The results of analysis of variance as can be known, two factor non-interaction actions.Two factors are compared with blank column, difference that also there are no significant.The influence degree that can get two factors by intuitive analysis is followed successively by Polyethylene Glycol consumption＞principal agent consumption, determines that thus prescription is A ₂B ₂, promptly the PEG400 consumption is 50.0%, the principal agent consumption accounts for 33.3%.

Experimental example 5 confirmatory experiments

The prescription that obtains with above-mentioned orthogonal experiment experimentizes, and promptly the principal agent consumption accounts for 33.3%, and the PEG400 consumption accounts for 50.0%, and the polyethylene glycol 6000 consumption accounts for 16.7%.The ointment stretchability and the stability that make are all better.

Experimental example 6 refrigeration functions (contrast suppository)

Get normal 50 of the male rats of anus temperature and be divided into matched group, 1.2g/kg SHUANGHUANLIAN bolt positive drug group, 2.4,1.2, three administration groups of 0.6g/kg SHUANGHUANLIAN ointment at random, the anorectal administration is 5 days continuously, 4h is with subcutaneous 20% dry yeast that gives of 15ml/kg before the last administration, measure normal anus temperature 2 times with injection is preceding, getting its meansigma methods is normal body temperature.With after the administration 1,2,3,4,5h measures the anus temperature, the difference of its measured value and normal value is the heating degree.It the results are shown in Table 6.

The influence of table 6. pair rat carrageenan group swelling

Group	Dosage g/kg	The degree of heat △ ℃
								Normally	1h	2h	3h	4h	5h
		Normal group model group positive drug is subjected to reagent to be subjected to reagent to be subjected to reagent	1.2 2.4 1.2 0.6	37.6±0.3 37.6±0.5 7..5±1.3 37.5±0.5 37.4±0.4 37.5±0.4	0.23±0.15 1.22±0.43 0.89±0.20* 0.77±0.31** 0.86±0.31* 0.88±0.15*	0.12±0.19 1.41±0.54 0.99±0.27* 0.70±0.45** 0.94±0.17* 0.99±0.19*	0.02±0.29 1.04±0.53 0.63±0.26* 0.54±0.32* 0.67±0.13* 0.77±0.23*							-0.12±0.23 0.79±0.38 0.44±0.32* 0.39±0.27* 0.49±0.18* 0.69±0.13	0.02±0.44 0.77±0.61 0.60±0.29 0.33±0.22* 0.62±0.27 0.85±0.25

Annotate: compare * * P＜0.01, * P＜0.05 with matched group.

Table 6 shows that this product has significant inhibitory effect to the heating that yeast causes, promptly has tangible refrigeration function.SHUANGHUANLIAN ointment is rapid-action, refrigeration function also slightly is better than suppository.

Experimental example 7 analgesic activities (contrast suppository)

Get 50 of mices, be divided into matched group at random, positive drug group, 2.4,1.2, three administration groups of 0.6g/kg SHUANGHUANLIAN ointment, totally five groups.The anorectal administration is 5 days continuously, and every day 1 time, matched group gives substrate.Behind the last administration 45min, the abdominal cavity gives 0.9% glacial acetic acid 0.2ml/ only, and record gives behind the acetic acid mouse writhing number of times in the 15min.The results are shown in Table 7

Table 7 Dichlorodiphenyl Acetate induced mice pain influence X ± SD

Group	Dosage g/kg	Turn round the body frequency n
			Matched group SHUANGHUANLIAN bolt SHUANGHUANLIAN ointment SHUANGHUANLIAN ointment SHUANGHUANLIAN ointment	2.4 2.4 1.2 0.6	25.0±9.2 17.9±4.2* 15.5±2.2* 18.0±4.5* 18.8±6.4

Annotate: compare * * P＜0.01, * P＜0.05 with matched group.

As seen from Table 7, SHUANGHUANLIAN ointment, SHUANGHUANLIAN bolt analgesic activity under the Isodose, the ointment analgesic activity is better than suppository.

The influence (contrast suppository) of 8 pairs of ammonia induced mice coughs of experimental example

Get 50 of mices, be divided into contrast at random, 2.4g/kg SHUANGHUANLIAN bolt, 2.4,1.2, three groups of 0.6g/kg SHUANGHUANLIAN ointment (totally five groups), successive administration 5 times (1 time/day) behind the last administration 1h, utilizes spray 26% ammonia 3 seconds of ultrasound atomizer, observe and the cough latent period (promptly stopping to be sprayed to the time of cough) of record mice and mouse cough number of times in 2 minutes (with its abdominal muscle shrink, magnify mouth and hear to cough be as the criterion), the results are shown in Table 8.

The cough of table 8. pair ammonia induced mice influence X ± SD

Group	Dosage g/kg	Incubation period S	The cough frequency n
				Matched group SHUANGHUANLIAN bolt SHUANGHUANLIAN ointment SHUANGHUANLIAN ointment SHUANGHUANLIAN ointment	2.4 2.4 1.2 0.6	5.0±1.7 7.3±2.3 7.6±2.2** 6.7±1.7* 5.5±2.3	32.3±8.0 23.9±6.4* 21.8±9.2* 24.4±5.7* 27.2±8.6

As seen from Table 8, SHUANGHUANLIAN ointment significant prolongation ammonia induced mice cough latent period also can obviously reduce the mouse cough number of times.Its antitussive effect slightly is better than suppository.

SHUANGHUANLIAN ointment has heat-clearing and toxic substances removing, gently declares the function of wind heat.Be used for affection due to external wind and heat, heating, cough, pharyngalgia.Result of the test shows, that SHUANGHUANLIAN ointment has is obviously analgesic, pain relieving, antitussive effect.Anorectal gives two yellow bolts synchronously, analgesic, the antitussive of two yellow bolts, analgesic effect are also clearly, owing to SHUANGHUANLIAN ointment is changed by two yellow suppositorys, ointment is very fast by the anorectal mucosa absorption than suppository, the embodiment of pharmacological action is also than fully, by result of the test as seen, the SHUANGHUANLIAN ointment main pharmacodynamics experimental result under the Isodose all is better than two yellow bolts.

Experimental example 9 skin irritation tests

1. acute irritation experiment: get 36 of rats, be divided into matched group and administration group at random, 18 every group.Disposable anorectal gives SHUANGHUANLIAN ointment 2.4g/kg (with the clip certain hour that is lined with Cotton Gossypii), and matched group gives substrate.Respectively with administration after 24h, 48h, 7d matched group and administration group respectively put to death 6 rats, get the anorectal tissue and carry out histopathologic examination.The no abnormal reaction of rat takes place after the administration, and relatively all are normal with matched group; The result of histopathologic examination shows that SHUANGHUANLIAN ointment does not see that to rat anorectal mucosa irritant reaction takes place.

2. successive administration is to the stimulation test of rat: get 20 of rats, be divided into matched group and administration group at random, 10 every group.Anorectal gives 2.4g/kg SHUANGHUANLIAN ointment 7 days (with the clip certain hour that is lined with Cotton Gossypii) continuously, and matched group gives substrate.The no abnormal reaction of rat takes place after the administration, does not see difference with matched group; Rat body weight there is not influence yet.1h puts to death rat after the last administration, gets anorectal tissue and carries out histopathologic examination, and the result of histopathologic examination shows that SHUANGHUANLIAN ointment do not see the stimulation generation to rat anorectal mucosa.See Table 9

Table 9. anorectal continuously gives the influence of SHUANGHUANLIAN ointment to rat body weight

Group	Dosage (g/kg)	Body weight (g)
				0d	7d
		Matched group administration group	2.4			103.0±10.5 102.1±9.7	125.8±7.5 125.7±7.2

Conclusion: no matter be that disposable or continuous anorectal gives SHUANGHUANLIAN ointment and rat body and anorectal mucosa be there is no tangible irritant reaction produces.

Embodiment 1

Flos Lonicerae 1667g Radix Scutellariae 1667g Fructus Forsythiae 3334g

More than three flavors, Radix Scutellariae adds 8 times of decoctings and boils three times, 2 hours for the first time, second and third time each 1 hour, collecting decoction filters, filtrate is concentrated into the clear paste that relative density is 1.05 (50 ~ 60 ℃), and concentrated solution adds the 2mol/L hydrochloric acid solution in the time of 80 ℃, regulates pH value to 1.0～2.0, be incubated and left standstill 24 hours after 1 hour, filter, precipitate adds 6 times of water gagings, regulate pH value to 7.0～7.5 with 40% sodium hydroxide solution, add equivalent ethanol, stir and make dissolving, filter.Filtrate is regulated pH value to 2.0 with the 2mol/L hydrochloric acid solution, and 60 ℃ are incubated 30 minutes, leave standstill 12 hours, filter, and precipitation washes with water to pH to 5.0, and continuing is washed till pH value 7.0 with 70% ethanol.Precipitate adds 6 times of water gagings, regulates pH value to 7.0～7.5 with 40% sodium hydroxide solution, stirs and makes dissolving.Flos Lonicerae, Fructus Forsythiae add 8 times of decoctings and boil secondary, each 1.5 hours, collecting decoction, filter, filtrate is concentrated into the clear paste that relative density is 1.20～1.25 (70～80 ℃), stir when being chilled to 40 ℃ and slowly add ethanol down, make to contain the alcohol amount and reach 75%, left standstill 12 hours, the leaching supernatant, reclaim ethanol, concentrated solution is concentrated into the clear paste that relative density is 1.15～1.20 (50～60 ℃), stirs when being chilled to 40 ℃ to add ethanol down again and make and contain the alcohol amount and reach 85%, fully stir, left standstill 12 hours, the leaching supernatant reclaims ethanol to there not being the alcohol flavor.Add above-mentioned Radix Scutellariae extract aqueous solution, stir evenly, and regulate pH value to 7.0～7.5, concentrating under reduced pressure becomes thick paste, and cold drying is pulverized, and is standby; Get 500 gram PEG400s and 167 gram polyethylene glycol 6000s,, add 1.3 gram ethyl hydroxybenzoates again and make dissolving 65 ℃ of heating in water bath dissolvings.In water-bath substrate is progressively joined in the above-mentioned fine powder, constantly stir so that mix homogeneously, stirring at room is even, and the mastic temperature drops to room temperature and the ointment shape that congeals into, and embedding is in the compound cream pipe of polyethylene, promptly.

The discrimination method of embodiment 2 SHUANGHUANLIAN ointment of the present invention

Get this product 2g, add methanol 20ml, supersound extraction 10min is put coldly, filters, and gets filtrate 1ml, as need testing solution; Other gets baicalin, the chlorogenic acid reference substance is made the solution that every 1ml contains 0.4mg with ethanol respectively, in contrast product solution.Test according to thin layer chromatography (2005 editions one appendix VI B of Chinese Pharmacopoeia), draw each 3～5 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, with butyl acetate-formic acid-upper solution of 7: 4: 3 of water is developing solvent, presaturation 30 minutes 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.

The discrimination method of embodiment 3 SHUANGHUANLIAN ointment of the present invention

Get the about 2g of this product, add methanol 20ml, supersound extraction 10min is put coldly, filters, and filtrate is concentrated into about 5ml as need testing solution.Other gets Fructus Forsythiae control medicinal material 0.5g, adds methanol 10ml, puts in the water-bath reflux 20 minutes, filters, and filtrate is medical material solution in contrast.Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2005 VI B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-methanol is developing solvent at 5: 1, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Content of baicalin assay method in the embodiment 4 SHUANGHUANLIAN ointment of the present invention

Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).With octadecyl silane is filler; Methanol-water-glacial acetic acid is a mobile phase at 50: 50: 1; Flow velocity: 1.0ml/min; Column temperature: room temperature; The detection wavelength is 276nm.Number of theoretical plate calculates by the baicalin peak should be not less than 1000.Precision takes by weighing baicalin reference substance 10mg, puts in the 100ml measuring bottle, and it is an amount of to add 50% methanol, puts that jolting makes its dissolving in the water-bath, is placed to room temperature, and is diluted to scale, shakes up, and is contrast device solution (containing baicalin 0.1mg among every 1ml).Get the about 2g of this product, accurate title is fixed, puts the accurate methanol 50ml that adds in the tool plug conical flask, and after weight decided in accurate title, supersound extraction 30 minutes was put and is chilled to room temperature, the weight of replenish lost.Filter, precision measures that methanol is diluted to scale in subsequent filtrate 25ml to the 50ml measuring bottle.Precision is measured 3ml and is put in the 50ml measuring bottle, adds methanol and is diluted to scale, promptly gets need testing solution.Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, and the every gram of Chinese medicine ointment of the present invention contains Radix Scutellariae with baicalin (C ₂₁H ₁₈O ₁₁) meter, should be no less than 50mg.

Claims (6)

1, a kind of Chinese medicine Shuanghuanglian ointment, it is characterized in that: this ointment is made by following method:

Flos Lonicerae 1000-3000 weight portion Radix Scutellariae 1000-3000 weight portion

Fructus Forsythiae 2000-6000 weight portion

The Radix Scutellariae of described weight proportion is added 4-12 times of decocting to be boiled 2-5 time, collecting decoction, filter, filtrate is concentrated into the clear paste that relative density is 1-2 in the time of 50～60 ℃, concentrated solution adds the 1-3mol/L hydrochloric acid solution in the time of 80 ℃, regulate pH value to 1.0～2.0, be incubated and left standstill 24-48 hour after 1-3 hour, filter, precipitate adds 3-8 times of water gaging, regulates pH value to 6.0-8.5 with the 20-60% sodium hydroxide solution, adds equivalent ethanol, stirring makes dissolving, filter, filtrate is regulated pH value to 1.0-2.0 with the 1-3mol/L hydrochloric acid solution, and 60 ℃ are incubated 10-50 minute, left standstill 6-24 hour, filter, precipitation washes with water to pH to 3.0-7.0, and continuing is washed till pH value 5.0-7.0 with 50-90% ethanol, precipitate adds 3-8 times of water gaging, regulate pH value to 6.0-8.5 with the 20-40% sodium hydroxide solution, stir and make dissolving, Flos Lonicerae, Fructus Forsythiae adds 4-12 times of decocting and boils 1-3 time, each 1.5 hours, collecting decoction filters, and filtrate is concentrated into the clear paste that relative density is 1.0-1.4 in the time of 70～80 ℃, stir when being chilled to 20-40 ℃ and slowly add ethanol down, make to contain alcohol amount and reach 50-85%, left standstill the leaching supernatant 6-16 hour, reclaim ethanol, concentrated solution is concentrated into the clear paste that relative density is 1.0-1.3 in the time of 50～60 ℃, stirs when being chilled to 20-60 ℃ to add ethanol down again and make and contain the alcohol amount and reach 60-90%, fully stirs, left standstill 8-24 hour, the leaching supernatant reclaims ethanol to there not being the alcohol flavor, adds above-mentioned Radix Scutellariae extract aqueous solution, stir evenly, and regulate pH value to 5.0-9.0, concentrating under reduced pressure becomes thick paste, cold drying, pulverize, standby; Get 300-600 weight portion PEG400 and 100-300 part polyethylene glycol 6000,40-80 ℃ of heating in water bath dissolving, add 1-3 part ethyl hydroxybenzoate again and make dissolving, in water-bath, substrate is progressively joined in the above-mentioned fine powder, constantly stir so that mix homogeneously, stirring at room is even, and the mastic temperature drops to room temperature and congeals into the ointment shape, embedding is in the compound cream pipe of polyethylene, promptly.

2, a kind of Chinese medicine Shuanghuanglian ointment as claimed in claim 1, it is characterized in that: this ointment is made by following method:

Flos Lonicerae 1667 weight portion Radix Scutellariaes 1667 weight portions

Fructus Forsythiae 3334 weight portions

More than three the flavor, Radix Scutellariae adds 8 times of decoctings and boils three times, 2 hours for the first time, second, three times each 1 hour, collecting decoction filters, under 50～60 ℃ of conditions, it is 1.05 clear paste that filtrate is concentrated into relative density, concentrated solution adds the 2mol/L hydrochloric acid solution in the time of 80 ℃, regulate pH value to 1.0～2.0, is incubated and leaves standstill 24 hours after 1 hour, filter, precipitate adds 6 times of water gagings, regulates pH value to 7.0～7.5 with 40% sodium hydroxide solution, adds equivalent ethanol, stirring makes dissolving, filter, filtrate is regulated pH value to 2.0 with the 2mol/L hydrochloric acid solution, and 60 ℃ are incubated 30 minutes, left standstill 12 hours, filter, precipitation washes with water to pH to 5.0, and continuing is washed till pH value 7.0 with 70% ethanol, precipitate adds 6 times of water gagings, regulate pH value to 7.0～7.5 with 40% sodium hydroxide solution, stir and make dissolving, Flos Lonicerae, Fructus Forsythiae adds 8 times of decoctings and boils secondary, each 1.5 hours, collecting decoction filters, in the time of 70～80 ℃ filtrate to be concentrated into relative density be 1.20～1.25 clear paste, stir when being chilled to 40 ℃ and slowly add ethanol down, make to contain alcohol amount and reach 75%, left standstill the leaching supernatant 12 hours, reclaim ethanol, to be concentrated into relative density be 1.15～1.20 clear paste to concentrated solution in the time of 50～60 ℃, stirs when being chilled to 40 ℃ to add ethanol down again and make and contain the alcohol amount and reach 85%, fully stirs, left standstill 12 hours, the leaching supernatant reclaims ethanol to there not being the alcohol flavor, adds above-mentioned Radix Scutellariae extract aqueous solution, stir evenly, and regulate pH value to 7.0～7.5, concentrating under reduced pressure becomes thick paste, cold drying, pulverize, standby; Get 500 weight portion PEG400s and 167 weight portion polyethylene glycol 6000s, 65 ℃ of heating in water bath dissolvings, add 1.3 weight portion ethyl hydroxybenzoates again and make dissolving, in water-bath, substrate is progressively joined in the above-mentioned fine powder, constantly stir so that mix homogeneously, stirring at room is even, and the mastic temperature drops to room temperature and congeals into the ointment shape, embedding is in the compound cream pipe of polyethylene, promptly.

3, the method for quality control of a kind of Chinese medicine Shuanghuanglian ointment as claimed in claim 1 is characterized in that: comprise in the following discrimination method one or more in this method:

A. get this product 2g, add methanol 10-30ml, supersound extraction 5-15min is put coldly, filters, and gets filtrate 1ml, as need testing solution; Other gets baicalin, the chlorogenic acid reference substance is made the solution that every 1ml contains 0.4mg with ethanol respectively, in contrast product solution; Draw each 3～5 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 developing solvent with the upper solution of butyl acetate-formic acid-water 6-8: 3-5: 2-4, presaturation 30 minutes launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

B. get the about 2g of this product, add methanol 10-30ml, supersound extraction 5-15min is put coldly, filters, and filtrate is concentrated into about 5ml as need testing solution; Other gets Fructus Forsythiae control medicinal material 0.5g, adds methanol 10ml, puts in the water-bath reflux 10-30 minute, filters, and filtrate is medical material solution in contrast; Draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with chloroform-methanol 4-6: 0.5-1.5, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 80-120 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

4, the method for quality control of a kind of Chinese medicine Shuanghuanglian ointment as claimed in claim 3 is characterized in that: the discrimination method in this method is:

A, get this product 2g, add methanol 20ml, supersound extraction 10min is put coldly, filters, and gets filtrate 1ml, as need testing solution; Other gets baicalin, the chlorogenic acid reference substance is made the solution that every 1ml contains 0.4mg with ethanol respectively, in contrast product solution; Draw each 3～5 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 developing solvent with butyl acetate-formic acid-upper solution of 7: 4: 3 of water, presaturation 30 minutes launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

B, get the about 2g of this product, add methanol 20ml, supersound extraction 10min is put coldly, filters, and filtrate is concentrated into about 5ml as need testing solution; Other gets Fructus Forsythiae control medicinal material 0.5g, adds methanol 10ml, puts in the water-bath reflux 20 minutes, filters, and filtrate is medical material solution in contrast; Draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be at 5: 1 developing solvent with chloroform-methanol, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

5, the method for quality control of a kind of Chinese medicine Shuanghuanglian ointment as claimed in claim 1 is characterized in that: comprise following content assaying method in this method:

Content of baicalin is measured

With octadecyl silane is filler; Methanol-water-glacial acetic acid 40-60: 40-60: 0.5-1.5 is a mobile phase; Flow velocity: 1.0ml/min; Column temperature: room temperature; The detection wavelength is 276nm, and number of theoretical plate calculates by the baicalin peak should be not less than 1000; Precision takes by weighing baicalin reference substance 10mg, puts in the 100ml measuring bottle, and it is an amount of to add 50% methanol, puts that jolting makes its dissolving in the water-bath, is placed to room temperature, and is diluted to scale, shakes up, and makes the solution that contains baicalin 0.1mg among every 1ml, is contrast device solution; Get the about 2g of this product, accurate title is fixed, puts the accurate methanol 50ml that adds in the tool plug conical flask, after weight decided in accurate title, supersound extraction 30 minutes, put and be chilled to room temperature, the weight of replenish lost filters, precision measures that methanol is diluted to scale in subsequent filtrate 25ml to the 50ml measuring bottle, precision is measured 3ml and is put in the 50ml measuring bottle, adds methanol and is diluted to scale, promptly gets need testing solution; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, and the every gram of above-mentioned traditional Chinese herb medicine preparation contains Radix Scutellariae with baicalin C ₂₁H ₁₈O ₁₁Meter should be no less than 50mg.

6, the method for quality control of a kind of Chinese medicine Shuanghuanglian ointment as claimed in claim 5 is characterized in that: content assaying method is in this method:

Content of baicalin is measured

With octadecyl silane is filler; Methanol-water-glacial acetic acid is a mobile phase at 50: 50: 1; Flow velocity: 1.0ml/min; Column temperature: room temperature; The detection wavelength is 276nm, and number of theoretical plate calculates by the baicalin peak should be not less than 1000; Precision takes by weighing baicalin reference substance 10mg, puts in the 100ml measuring bottle, and it is an amount of to add 50% methanol, puts that jolting makes its dissolving in the water-bath, is placed to room temperature, and is diluted to scale, shakes up, and makes the solution that contains baicalin 0.1mg among every 1ml, is contrast device solution; Get the about 2g of this product, accurate title is fixed, puts the accurate methanol 50ml that adds in the tool plug conical flask, after weight decided in accurate title, supersound extraction 30 minutes, put and be chilled to room temperature, the weight of replenish lost filters, precision measures that methanol is diluted to scale in subsequent filtrate 25ml to the 50ml measuring bottle, precision is measured 3ml and is put in the 50ml measuring bottle, adds methanol and is diluted to scale, promptly gets need testing solution; Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid, measure, and the every gram of Chinese medicine ointment of the present invention contains Radix Scutellariae with baicalin C ₂₁H ₁₈O ₁₁Meter should be no less than 50mg.