CN102288721A - Method for detecting Chinese medicinal composition for treating chronic pharyngitis - Google Patents
Method for detecting Chinese medicinal composition for treating chronic pharyngitis Download PDFInfo
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Abstract
The invention relates to method for detecting a Chinese medicinal composition for treating chronic pharyngitis. The medicinal composition consists of raw pharmaceuticals such as figwort, aster, asparagus, cortex moutan, dwarf lilyturf root and the like. The detection method comprises methods for qualitative identification of a film chromatography and content measurement of an efficient liquid phase chromatography, and the like.
Description
The present invention is for dividing an application, and the original bill application number is 200810056634.3, and the original bill applying date is on January 23rd, 2008, and the original bill name is called: Chinese medicine composition of treatment chronic pharyngitis and preparation method thereof and method of quality control.
Technical field
The present invention relates to a kind of detection method of Chinese medicine composition, particularly a kind of detection method for the treatment of the Chinese medicine composition of chronic pharyngitis belongs to technical field of traditional Chinese medicines.
Background technology
Chronic pharyngitis is a kind of common disease, for the pharyngeal pathology of the caused diffusivity of chronic infection, mainly is the pharyngeal mucosa inflammation.Be mainly in the adult, becoming impatient property pharyngitis, long-term dust or harmful gas stimulate its main diseases because of having repeatly, tobacco and wine excessively or other bad life habits, the stimulation of nasosinusitis secretion, allergic constitution or passive protective physical fitness attenuating etc.Chronic pharyngitis also can be the topical manifestations of some systemic disease, as anaemia, diabetes, cirrhosis and chronic nephritis etc.The incidence of disease accounts for about 10% to 20% of pharyngolaryngitis in the city dweller.Because of its course of disease is long, the symptom stubbornness, the short-term difficulty is seen produce effects, so be difficult for curing.
Chronic pharyngitis does not generally need to use antibiotic therapy, because chronic pharyngitis and non-bacterial infection.Doctor trained in Western medicine is generally paid attention to local application, as: the rinse medicine, use 2% BAS, 3% salt solution and 1: 5000 furacilin solution be rinse repeatedly, perhaps use 2% jodine glycerin, 5% protargol liquid way in pharynx wall, or be contained in the oral cavity etc., the function of certain relief of symptoms is arranged with tabellae iodi gurgitis, Domiphen, peppermint etc., but general being difficult to cured, and curative effect is relatively poor.
This disease of Chinese traditional treatment focuses on effects a permanent cure, and presses the medication of medicine typing method, and curative effect is better." element is asked the negative and positive another matter " cloud: " the larynx numbness of monoyin and monoyang knot meaning." be argumentation the earliest to this disease.Ancient Chinese medicine doctor has more performance, and the etiology and pathogenesis of larynx numbness has been done discussion from different aspects, reduces phlegm heat, the fire of deficiency type, body fluid cannot flow upwards etc.Common drug has LIYANLING PIAN, clearing Ganlu pill, green plum ball, relieve sore throat tea and some buccal tablets, also has the Chinese medical spray therapy, all effect is better, so further bring into play this sick advantage of Chinese medicine, provides the better Chinese medicine composition of a kind of result of treatment to be very important.
Summary of the invention
The object of the present invention is to provide a kind of Chinese medicine composition that is used for the treatment of chronic pharyngitis;
Another object of the present invention is to provide the preparation method of this Chinese medicine composition;
The 3rd purpose of the present invention provides the method for quality control of this Chinese medicine composition.
The bright purpose of this law is achieved through the following technical solutions:
The Chinese medicine composition of treatment chronic pharyngitis of the present invention is made up of following weight portion bulk drug:
The Chinese medicine composition of treatment chronic pharyngitis of the present invention, preferred following weight portion bulk drug is formed:
The Chinese medicine composition of treatment chronic pharyngitis of the present invention, can also form by preferred following weight portion bulk drug:
Chinese medicine composition of the present invention is with yin-nourishing, moistening lung is puted forth effort, and get clearing heat and detoxicating, clearing throat, wind-dispelling heat-dissipating, relieving cough and reducing sputum, the merit that relieve sore throat is antipruritic, to the plain body deficiency of Yin, experience the heresy of wind-heat, easily the disease from dryization is very suitable, should control that pharyngitis due to the acute and chronic pharyngitis, dry throat, pharynx are itched, dry, thirsty, cough, heating, dislike wind, sweating and irritable cough, hoarsen, diseases such as aphonia.
Chinese medicine composition of the present invention, technology adding auxiliary material is made clinical acceptable forms such as tablet, capsule, oral liquid, pill, granule routinely; Described auxiliary material comprises solvent, disintegrant, flavouring, antiseptic, colorant, bonding agent, lubricant, matrix etc.
The preparation method of Chinese medicine composition tablet of the present invention is:
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boilings such as all the other radix scrophulariaes 1～3 time, each 2～4 hours, collecting decoction, left standstill 20～30 hours, and got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mix with Common Coltsfoot Flower,, be crushed to 80 orders at drying under reduced pressure below 60 ℃, add the sucrose of medicinal powder weight 8%, make particle, spray is with peppermint oil, the dolomol that adds particle weight 0.5%, mixing is pressed into 1000, sugar coating, promptly.
The method of quality control of Chinese medicine composition of the present invention comprises one or more in following qualitative checking method and/or the quantitative detecting method:
(1) qualitative detection of tussilago
Get pharmaceutical preparation content of the present invention, adding diethyl ether makes dissolving, and sonicated is filtered, and filtrate is waved to 1.5ml, as need testing solution.
The coltsfoot of withdrawing the money control medicinal material is made control medicinal material solution.
According to thin-layered chromatography test, draw above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, hot blast blow develop the color to spot clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) qualitative detection of Oroxylum indicum
Get pharmaceutical preparation content of the present invention, add the absolute ethyl alcohol sonicated, filter, the filtrate evaporate to dryness adds anhydrous alcohol solution as test sample.
Get the Oroxylum indicum control medicinal material, make control medicinal material solution.
According to thin-layered chromatography test, draw above-mentioned sample solution, control medicinal material solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with the vanillic aldehyde sulfuric acid solution, hot blast blow develop the color to spot clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(3) qualitative detection of the tuber of dwarf lilyturf
Get pharmaceutical preparation content of the present invention, add the chloroform-methanol mixed liquid dipping, sonicated is filtered, and evaporate to dryness is with the chloroform dissolving, as need testing solution.
Get the control medicinal material tuber of dwarf lilyturf, shred, make control medicinal material solution.
According to thin-layered chromatography test, draw above-mentioned solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with ethanol solution of sulfuric acid, hot blast blow develop the color to spot clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(4) detection by quantitative of normal butyl alcohol extract:
Get pharmaceutical preparation content of the present invention, the accurate title, decide, and the accurate methyl alcohol that adds claims to decide weight, put reflux in the water-bath, put coldly, supply the weight that subtracts mistake, filter with methyl alcohol, get subsequent filtrate, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water makes dissolving, extract with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight evaporate to dryness, 105 ℃ of dryings, in the dislocation exsiccator, place, weight decided in accurate rapidly title, calculates, promptly.
(5) detection by quantitative of harpagoside:
According to high effective liquid chromatography for measuring.
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, is Mobile phase B with 1% acetum, carries out gradient elution; The detection wavelength is 278nm.Number of theoretical plate calculates by the harpagoside peak should be not less than 5000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the harpagoside reference substance, adds methyl alcohol and make reference substance solution, promptly.
Chinese medicine composition content of the present invention is got in the preparation of need testing solution, gets about 1.0g, and accurate the title decides, put in the tool plug conical flask accurate 30% methyl alcohol, the close plug of adding, claim to decide weight, after the immersion, sonicated, put coldly, claim again to decide weight, supply the weight that subtracts mistake with 30% methyl alcohol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively reference substance solution and the need testing solution drawn of determination method injects liquid chromatograph, measures, promptly.
(6) detection by quantitative of Catalpol:
According to high effective liquid chromatography for measuring.
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With acetonitrile-0.1% phosphoric acid solution is moving phase; The detection wavelength is 210hm.Number of theoretical plate calculates by the Catalpol peak should be not less than 5000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the Catalpol reference substance, adds moving phase and make reference substance solution, promptly.
Chinese medicine composition phase content of the present invention is got in the preparation of need testing solution, gets about 1.0g, accurate claims surely, puts in the tool plug conical flask, and the accurate methyl alcohol that adds claims decide weight, and heating and refluxing extraction is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up filtration.Precision is measured subsequent filtrate 10ml, is concentrated near doing, and residue dissolves with moving phase, is transferred in the measuring bottle, and is diluted to scale with moving phase, shakes up, and filters, and gets subsequent filtrate, promptly.
Accurate respectively reference substance solution and the need testing solution drawn of determination method injects liquid chromatograph, measures, promptly.
The method of quality control of Chinese medicine composition of the present invention comprises one or more in following qualitative checking method and/or the quantitative detecting method:
(1) qualitative detection of tussilago
Get the pharmaceutical preparation of the present invention that is equivalent to bulk drug 10.5g, porphyrize, the 25ml that adds diethyl ether makes dissolving, and sonicated 40 minutes is filtered, and filtrate is waved to 1.5ml, as need testing solution.Other coltsfoot control medicinal material 1g that withdraws the money shines medicinal material solution in pairs with legal system.According to thin-layered chromatography test, draw each 25 μ l of above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, be developping agent with 4: 1 ratio normal hexane-ethyl acetates, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, hot blast blow develop the color to spot clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) qualitative detection of Oroxylum indicum
Get the pharmaceutical preparation of the present invention that is equivalent to bulk drug 4g, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample.Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast.According to thin-layered chromatography test, draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l are respectively and on the same silica gel g thin-layer plate, with 9: 1 ratio cyclohexane-ethyl acetate was developping agent, launched, and took out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(3) qualitative detection of the tuber of dwarf lilyturf
Get the pharmaceutical preparation of the present invention that is equivalent to bulk drug 10.5g, porphyrize adds 20 milliliters of the chloroform-methanols of 70: 30 ratios, soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of the chloroform-methanols of 70: 30 ratios, soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast.According to thin-layered chromatography test, draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, be developping agent with 5: 1 ratio normal hexane-ethyl acetate, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, hot blast blow develop the color to spot clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(4) detection by quantitative of normal butyl alcohol extract
Get the pharmaceutical preparation of the present invention that is equivalent to bulk drug 8.5g, porphyrize is got powder 2g, the accurate title, decide, and the accurate methyl alcohol 50ml that adds claims to decide weight, put in the water-bath reflux 1 hour, and put coldly, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times with water saturated normal butyl alcohol, each 25ml merges normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight, evaporate to dryness is 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculates, promptly.
Chinese medicine composition of the present invention is pressed dry product and is calculated, and contains the normal butyl alcohol extract and must not be less than 5.0%.
(5) detection by quantitative of harpagoside:
According to high effective liquid chromatography for measuring.
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, is Mobile phase B with 1% acetum, and the regulation in the according to the form below is carried out gradient elution; The detection wavelength is 278nm.Number of theoretical plate calculates by the harpagoside peak should be not less than 5000.
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0～20 | 20→50 | 80→50 |
| 20～25 | 50→20 | 50→80 |
It is an amount of that the preparation precision of reference substance solution takes by weighing the harpagoside reference substance, adds 30% methyl alcohol and make the solution that every 1ml contains 10 μ g, promptly.
The preparation of need testing solution is got Chinese medicine composition of the present invention and is equivalent to bulk drug 4.0g, and porphyrize is got about 1.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 30% methyl alcohol 50ml that adds, close plug claims to decide weight, soaks after 1 hour, at power 500W, sonicated is 30 minutes under the frequency 40kHz, puts cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with 30% methyl alcohol, filter, get subsequent filtrate, promptly.
Inaccurate reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Chinese medicine composition of the present invention is equivalent to crude drug 4.0g, contains harpagoside (C
24H
30O
11) must not be less than 0.18mg.
(6) detection by quantitative of Catalpol
According to high effective liquid chromatography for measuring.
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With 1: 99 ratio acetonitrile-0.1% phosphoric acid solution was moving phase; The detection wavelength is 210nm.Number of theoretical plate calculates by the Catalpol peak should be not less than 5000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the Catalpol reference substance, adds moving phase and make the solution that every 1ml contains 10 μ g, promptly.
The preparation of need testing solution is got Chinese medicine composition of the present invention and is equivalent to bulk drug 4.0g, and porphyrize is got about 1.0g, the accurate title, decide, put in the tool plug conical flask, the accurate methyl alcohol 25ml that adds claims to decide weight, heating and refluxing extraction 1.5 hours, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methyl alcohol, shake up, filter.Precision is measured subsequent filtrate 10ml, is concentrated near doing, and residue dissolves with moving phase, is transferred in the 10ml measuring bottle, and is diluted to scale with moving phase, shakes up, and filters, and gets subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Chinese medicine composition of the present invention is equivalent to bulk drug 4.0g, contains Catalpol (C
15H
22O
10) must not be less than 0.25mg.
Embodiment
Following examples and test example are used to further specify but are not limited to the present invention
The test of test example 1 technical study
When determining the preparation technology of Chinese medicine composition of the present invention, according to the characteristics of the contained effective ingredient of each flavour of a drug and the consideration of energy savings, determine that radix scrophulariae, the tuber of stemona (system), asparagus fern, moutan bark, the tuber of dwarf lilyturf, Oroxylum indicum, glutinous rehmannia, Radix Isatidis, Chinese olive, cicada slough water extract and gets final product, tussilago adopts full the pulverizing to be used as medicine.The kind of auxiliary material and consumption are selected, the concrete quantification of art for coating, to the ageing of the quality stability that improves medicine of the present invention, security, production and to the saving of the energy conspicuousness meaning are arranged all.
One, amount of water test
Press three parts of medicinal materials of recipe quantity configuration, every part contains radix scrophulariae 170g, the tuber of stemona (system) 130g, asparagus fern 120g, moutan bark 50g, tuber of dwarf lilyturf 70g, Oroxylum indicum 70g, glutinous rehmannia 70g, Radix Isatidis 210g, Chinese olive 140g, cicada slough 50g, and be divided into three groups and do experiment: first group of amount of water is respectively 10 times of amounts, 8 times of amounts; Second group of amount of water is 8 times of amounts, 6 times of amounts; The 3rd group of amount of water is 6 times of amounts, 4 times of amounts.With the paste volume is that index is determined amount of water.The results are shown in Table 1
Table 1: amount of water test findings
Above result shows: be that 8 times of amounts of index amount of water, 6 times of amount extractions are complete substantially with the paste volume, be defined as 8 times of amounts, 6 times of amounts according to the needs of production amount of water.
Two, the kind and the consumption that add auxiliary material
Take by weighing according to above-mentioned selection process and extract raw medicinal material, after extract concentrates, tussilago pulverizing medicinal materials to 100 order is added, mixing, drying is crushed to 80 orders, adds appropriate amount of auxiliary materials, granulate, drying, whole grain adds 0.5% dolomol again, compressing tablet, promptly.By comparing difficulty or ease, the tablet appearance of compressing tablet, select proper supplementary material and supplementary product consumption ratio, the results are shown in following table 2:
Table 2 auxiliary material is selected test findings
As can be seen from the above table, selecting to add auxiliary material during the preparation tablet is sucrose, and addition is 1: 0.08, promptly can reach the requirement of preparation tablets.Promptly add auxiliary material sucrose, its prescription consumption is 17g.
Three, art for coating
1 separation layer: place coating pan to roll plain sheet, add to mix slurry (35% peach gum: 70% syrup=1: 5) make and evenly adhere to unilateral going up (plain sheet: mix slurry=50g: 1ml), the blowing hot-air drying, for preventing that tablet is inter-adhesive or sticking on the coating pan, add talcum powder, 40～50 ℃ of hot blasts are dry down, twice of repetitive operation.
2 sub coats: make slice, thin piece continue in coating pan, to roll, add 70% syrup, after making the sheet sub-surface evenly wetting, add talcum powder, make it to adhere to the sheet sub-surface, continue to roll and blowing dry (50～55 ℃) repetitive operation, till the slice, thin piece faceted pebble disappears, heavy by plain sheet: 70% syrup: talcum powder=feed intake at 3: 1.7: 1.
3 sugarcoating layers: slice, thin piece is rolled in coating pan, add 70% syrup (plain sheet: 70% syrup=15: 1, the syrup addition is descending successively decreases), the sheet sub-surface is slowly dry, forms fine and smooth sugar crystal clothing layer, increases clothing layer fastness and sweet taste.
4 coloured sugarcoating layers: be diluted to the mill base of 2mg/ml with 70% syrup, mill base is diluted to variable concentrations with 70% syrup, concentration is ascending, adds coating pan, and is dry layer by layer.Plain sheet and pigment amount ratio are 7.5kg: 1g.
5 polishings: for making the sugar coated tablet surface-brightening attractive in appearance, have moisture-proof role concurrently, add insect wax, add, rotate coating pan insect wax is wrapped on the slice, thin piece equably, take out coating tablet by plain sheet: insect wax=3kg: 5g, dry 24 hours, promptly.
The 2 method of quality control experimental studies of test example
Medicine of the present invention has the kinds of traditional Chinese medicines bulk drug to process through extraction, grope by a large amount of tests, the thin-layer chromatography discrimination method of determining tussilago, Oroxylum indicum, the tuber of dwarf lilyturf is reappearance, good stability not only, and reliable results is noiseless, in the radix scrophulariae in harpagoside, the glutinous rehmannia foundation of catalpol content assay method then further improved the controllability of drug quality of the present invention, an assay that in checking, has added the normal butyl alcohol extract, just further perfect drug quality control method of the present invention.
The discriminating of tussilago
1) preparation of need testing solution:
Get according to 10 of the medicinal tablets of the present invention of embodiment 1 preparation for every part, remove sugar-coat, porphyrize adds ethanol, ether, chloroform respectively, and sonicated is filtered, and filtrate is waved to 1.5ml, as need testing solution.Be equipped with control medicinal material solution with legal system.Negative sample according to the scarce tussilago of embodiment 1 preparation prepares negative control product solution according to the need testing solution preparation method again.
Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 25 μ l of above-mentioned three kinds of solution, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, and it is clear that hot blast blows to the spot colour developing.Relatively more different the extraction solvent and extraction time, gained need testing solution and the color developing effect of control medicinal material solution on thin layer plate the results are shown in following table:
The preparation of table 3 need testing solution
As can be seen from the above table, during with ether ultrasonic Extraction 40min, test sample is all clear with each mottle colour developing of control medicinal material relevant position, good separating effect, and negative noiseless.Empirical tests, this method favorable reproducibility, Pass Test requirement.
2) selection of developping agent
Prepare need testing solution, control medicinal material solution and negative control product solution according to above-mentioned method for optimizing, test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), drawing each 25 μ l of above-mentioned three kinds of solution, respectively and on the same silica gel g thin-layer plate, 2: 3,3: 2,3: 1,4: 1 was developping agent with normal hexane-ethyl acetate, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, it is clear that hot blast blows to the spot colour developing, relatively the expansion effect of each spot on thin layer plate.The results are shown in following table:
The selection of table 4 developping agent
As can be seen from the above table, be at 4: 1 developping agent with normal hexane-ethyl acetate, each spot colour developing is clear, good separating effect, and negative noiseless, the Pass Test requirement.
2. the discriminating of the tuber of dwarf lilyturf
We grope having carried out the thin layer discrimination method tuber of dwarf lilyturf in the medicine of the present invention with reference to medicinal material discrimination method under 2005 an editions Chinese Pharmacopoeias items tuber of dwarf lilyturf, but sample with the tuber of dwarf lilyturf control medicinal material do not have corresponding spot, and feminine gender has interference.We are conversion developping agent on this basis again, and has carried out the check and negative investigation of three batch samples, has set up the discrimination method of the tuber of dwarf lilyturf, and the result is as follows:
Table 5 differentiates that developping agent selects test the tuber of dwarf lilyturf
So determine that the thin layer discrimination method of the tuber of dwarf lilyturf in the medicine of the present invention is as follows:
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 10.5g, porphyrize, 20 milliliters of chloroform-methanols (70: 30) soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of chloroform-methanols (70: 30), soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (5: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
3. the discriminating of Oroxylum indicum
We have carried out the thin layer discrimination method with reference to medicinal material discrimination method under 2005 editions Chinese Pharmacopoeias Oroxylum indicum items to the Oroxylum indicum in the medicine of the present invention and have groped, but sample does not have corresponding spot with the Oroxylum indicum control medicinal material, and feminine gender has interference.We are conversion developping agent on this basis again, and has carried out the check and negative investigation of three batch samples, has set up the discrimination method of the tuber of dwarf lilyturf, and the result is as follows:
Table 6 Oroxylum indicum is differentiated developping agent selection test
So determine that the thin layer discrimination method of Oroxylum indicum in the medicine of the present invention is as follows:
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 4g, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample.Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l, respectively with same silica gel g thin-layer plate on, be developping agent with cyclohexane-ethyl acetate (9: 1), the expansion, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
4, the assay of normal butyl alcohol extract
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 8.5g, porphyrize is got powder 2g, the accurate title, decide, and the accurate methyl alcohol 50ml that adds claims to decide weight, put in the water-bath reflux 1 hour, and put coldly, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times with water saturated normal butyl alcohol, each 25ml merges normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight, evaporate to dryness is 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculates, promptly.
According to above assay method, 3 batches of medicines of the present invention according to embodiment 1 preparation to be carried out extract content measure, measurement result is as follows:
Table 7 extract content measurement result
According to above measurement result, this method good stability, simple and easy to do, so being pressed dry product, calculates medicine of the present invention, contain the normal butyl alcohol extract and must not be less than 5.0%.
5, assay
(1) assay of harpagoside in the radix scrophulariae
1) chromatographic condition chromatographic column: Yi Lite C18 chromatographic column (4.6x200ram, 51xm); Moving phase: the A acetonitrile, the B1% acetum carries out gradient elution by table 2.
Table 8 moving phase ratio
| Time (min) | A(％) | B(％) |
| ?0～20 | 20→50 | 80→50 |
| ?20～25 | 50→20 | 50→80 |
Flow velocity: 1mL/min; Column temperature: 35 ℃; Detect wavelength: 278nm.Number of theoretical plate: should be not less than 5000 by the calculating of harpagoside peak.
2) it is an amount of that the harpagoside reference substance is got in the preparation of reference substance solution, adds 30% methyl alcohol and make the solution that every 1ml contains harpagoside 10 μ g, promptly.
3) it is an amount of that the medicine of the present invention for preparing according to embodiment 1 is got in the preparation of need testing solution, removes sugar-coat, and porphyrize is got 1.0g, the accurate title, decide, and puts in the tool plug conical flask, accurate 30% methyl alcohol 50ml, the close plug of adding, claim to decide weight, behind the immersion 1h, sonicated (power 5000W, frequency 40kHz) 30rain takes out, and puts cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with 30% methyl alcohol, filter, get subsequent filtrate, promptly.Respectively accurate draw reference substance solution and need testing solution each 201～1, inject high performance liquid chromatograph, measure, write down the chromatogram of 25min, promptly.
4) the negative blank sample that embodiment 1 preparation lacks radix scrophulariae is pressed in the preparation of negative control product solution, with test sample preparation method negative control solution.
5) accurate respectively each the 10 μ l of need testing solution, negative control solution and reference substance solution that draw of specificity test inject liquid chromatograph, the record chromatogram.By chromatogram as seen, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on the chromatographic peak of identical retention time is arranged, negative test is noiseless.
6) the accurate absorption of the investigation of linear relationship concentration is harpagoside reference substance solution 5,10,15,20, the 25 μ l injection high performance liquid chromatograph of 10 μ g/ml, measure its peak area integrated value, with sample size is that horizontal seat peak area integrated value is an ordinate, the drawing standard curve, its regression equation is: Y=1416.8x+12864, r=0.9998.The result shows that harpagoside has good linear relationship in 50-250 μ g scope.
Table 9 linear relationship is investigated
| Sample size (μ l) | ?5 | 10 | 15 | 20 | 25 |
| Peak area | ?20124 | 26887 | 34032 | 41099 | 48437 |
7) the accurate same need testing solution (according to the test sample of embodiment 1 preparation) of drawing of precision test repeats sample introduction 5 times, measures its peak area, RSD=1.43%.The result shows that this method precision is good.
Table 10 precision is investigated
8) accurate same need testing solution (according to the test sample of embodiment 1 preparation) the 20 μ l that draw of stability test respectively at 0,4,8,12, the 16h sample introduction, measure its peak area integrated value, and peak area mean value is 21846.6.RSD=1.57%, result show that need testing solution peak area integrated value in 16h is basicly stable.
Table 11 study on the stability
9) replica test is got 5 parts in same lot number sample (according to the test sample of embodiment 1 preparation), measures 5 times in accordance with the law, and average content is 0.2646mg/g, RSD; 1.61%, the result shows that this method repeatability better.
Table 12 repeatability is investigated
10) to take by weighing known content be the about 1.0g of 0.2637mg/g sample (according to the test sample of embodiment 1 preparation) to the recovery test precision, and totally 5 parts, accurate respectively adding harpagoside reference substance solution (1.5mg/ml) 1ml measures content in accordance with the law, the results are shown in Table 13.
Table 13 average recovery measurement result
The result shows that this method average recovery is better.
From above methodological study result as can be seen, all Pass Test requirements such as the content assaying method precision of harpagoside, reappearance, stability in the radix scrophulariae are so determine that the content to measure harpagoside in the radix scrophulariae is one of method of quality control.
(2) assay of Catalpol in the glutinous rehmannia
1 instrument and test solution
Instrument: Aglient 1100 HPLC analyzers, chem station for LC chromatographic data process software.Reagent: methyl alcohol, second is fine is chromatographically pure, and water is water for injection, other reagent be analyze pure.Reference substance: Catalpol reference substance (for assay usefulness, Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides).Sample: according to the medicine of the present invention of embodiment 1 preparation.
2 chromatographic conditions
Chromatographic column be the C18 chromatographic column (Type Waters, 4.6x250mm), moving phase is that second is fine: methyl alcohol: water (6: 3: 150), the detection wavelength is 210nm, flow velocity is 0.8mldmin.
3 detect wavelength determination
Get the Catalpol reference substance, measure its absorption curve at 200～400nm with spectrophotometer.Find that it has absorption maximum at the 210nm place, so select 210nm for detecting wavelength.
4 experimental techniques
4.1 the preparation precision of reference substance solution takes by weighing Catalpol reference substance 4.58mg, puts in the 100mL volumetric flask, adds dissolve with methanol and is diluted to scale, shakes up, and promptly gets (every 1mL reference substance solution contains Catalpol 0.0458mg).
4.2 it is an amount of that the preparation of need testing solution takes by weighing according to the medicine of the present invention of embodiment 1 preparation, removes sugar-coat, porphyrize, get 2g, accurate claim surely, put that to add methyl alcohol in the apparatus,Soxhlet's an amount of, extract 4h, take out, put cold, filter, methanol extract liquid low temperature reclaims methyl alcohol to about 10mL, and methanol extract liquid quantitatively is transferred in the 25mL volumetric flask, adds methyl alcohol and is diluted to scale, shake up, as need testing solution.
4.3 the negative blank sample that embodiment 1 preparation lacks glutinous rehmannia is pressed in the preparation of negative control solution, with test sample preparation method negative control solution.
4.4 the accurate respectively reference substance solution of drawing of assay method, need testing solution, each 20uL of negative control product solution injects liquid chromatograph, measures, promptly.Disturbing does not appear in the relevant position of test sample Catalpol in negative sample solution.
4.5 the investigation precision of linear relationship takes by weighing Catalpol reference substance solution (0.235mg/mL) 1,5,9,13,17,20mL, put in the 20mL volumetric flask, add methyl alcohol to scale, concentration be 0.01175,0.05875,0,1058,0.1528,0.1998, the Catalpol reference substance solution of 0.235mg/mL, draw above-mentioned 6 kinds of each 20uL of solution respectively, inject hplc determination, regression equation is A=94153C+198.11, related coefficient, r=0,9996.
Table 14 linear relationship is investigated
The result shows that Catalpol is good in the scope internal linear relation of 0.235-4.70 μ g.
4.6 replica test is got same batch sample (according to the medicine of the present invention of embodiment 1 preparation), parallel preparation 6 duplicate samples press sample determination method mensuration, 0.306mg/g as a result, and RSD is 1.59%.
Table 15 repeatability is investigated
4.7 accurate same Catalpol reference substance solution (0.0458mg/mL) 20uL that draws of precision test injects liquid chromatograph, METHOD FOR CONTINUOUS DETERMINATION 6 times is measured peak area value, and RSD is 0.80%.
Table 16 precision is investigated
4.8 the average recovery test is by the requirement of average recovery test, with same batch sample (according to the medicine of the present invention of embodiment 1 preparation) preparation need testing solution, precision is measured 4 parts of 5mL need testing solutions, put respectively in the 10mL measuring bottle, add Catalpol reference substance solution (0.0458mg/mL) respectively and put scale, press the sample determination method, get the test sample 5mL that does not add reference substance and put in the volumetric flask with method and measure, the total content of getting 5mL is 1.40% for background values RSD.See table 17 for details.
Table 17 recovery experimental result
From above methodological study result as can be seen, all Pass Test requirements such as the content assaying method precision of Catalpol, reappearance, stability in the glutinous rehmannia are so determine that the content to measure Catalpol in the glutinous rehmannia is one of method of quality control.
The test of test example 3 pharmacodynamic studies
1 experiment material
1.1 medicine is according to pharmaceutical preparation I of the present invention, the II of embodiment 1,2 preparations; Yanyan slice (specification: 0.25g/ sheet (sugar coated tablet), Jilin sense health pharmaceutical Co. Ltd produces; Aspirin is by Shenyang Medical College production; Ammonium chloride is pure for analyzing; According to train of thought orchid, serotonin Fluka import packing, Shanghai chemical reagent purchasing and supply station packing factory.
1.2 animal Wistar rat, ICR mouse and SD rat, ♀ ♂ all uses.
2 methods and result
Get 60 of rats, body weight 200～250g, male and female half and half, 10 every group 2.1 suppress the effect of capillary permeability.Be divided into 6 groups at random, drug component of the present invention Gei medicine I heavy dose of the present invention not organized 70g/kg, small dose group 35g/kg, medicine II small dose group 35g/kg of the present invention (the contained crude drug amount of soup), the saline control group is given physiological saline 20ml/kg, the aspirin group is given aspirin 268mg/kg, the positive drug control group is given Yanyan slice 37g/kg (the contained crude drug amount of soup), each is organized behind ig administration or salt solution 30min, at the serotonin 0.1ml of rat back center line left side hypodermic injection 1 μ g/ml, iv1% is according to the blue 4ml/kg of the train of thought immediately.Put to death animal behind the 15min, the painted skin in back is cut and shredded put into 3ml physiological saline-acetone (3: 7) mixed liquor, place 24h, get supernatant, with 721 spectrophotometers (610nm) photometry density.The results are shown in Table 18.
Table 18 suppresses the effect of capillary permeability
| Group | Dosage | Optical density value (x ± s) |
| The saline control group | 20ml/kg | 0.1233±0.0526 |
| The aspirin group | 268mg/kg | 0.0611±0.0224** |
| The positive drug control group | 37g/kg | 0.0747±0.0224* |
| Medicine group I heavy dose of the present invention | 70g/kg | 0.0570±0.0218** |
| Medicine group I low dose of the present invention | 35g/kg | 0.0628±0.0208** |
| Medicine group II low dose of the present invention | 35g/kg | 0.0651±0.0236* |
Annotate: n=10, compare * * P＜0.01, * P＜0.05 with the saline control group
Each group of medicine of the present invention all has inhibiting effect to capillary permeability due to the serotonin, the excellent positive control medicine of the inhibiting effect of medicine of the present invention, and the effect of medicine I of the present invention is better than medicine II of the present invention.
2.2 influence 60 of extracting male Wistar rats to what carrageenin caused rat paw edema, body weight 140～160g, grouping and administration are the same, 1h after the administration, in the subcutaneous injection 1% carrageenin 0.1ml/ of the right sufficient sole of the foot of rat only.Cause scorching back 1～6h and measure sufficient sole of the foot volume respectively, calculate swelling value (causing the sufficient sole of the foot difference in volume in scorching front and back).The results are shown in Table 19.
Table 19 pair carrageenin causes the influence of rat paw edema
Annotate: n=10, compare * * P＜0.01, * P＜0.05 with the saline control group
Each group of medicine of the present invention causes rat paw edema to carrageenin all inhibiting effect, and the inhibiting effect of medicine of the present invention is better than the positive control medicine, and the effect of medicine I of the present invention is better than medicine II of the present invention.
2.3 to rat granuloma hyperplasia influence 60 of extracting male Wistar rats, body weight 140-160g, grouping and administration are the same.At the descending back of narcosis median incision 1cm, at the subcutaneous diameter 9mm that buries of left shoulder, thick 1.5mm, the aseptic filter paper sheet of heavy 9.25mg.Administration every day 2 times, continuous 7 days, took out granulation tissue on the 8th day, weigh behind 60 ℃ of dry 24h, obtain average and the inhibiting rate of respectively organizing net weight.The results are shown in Table 20.
The influence of table 20 pair rat granuloma hyperplasia
| Group | Dosage | The granulation tissue net weight (mg, x ± s) | Inhibiting rate (%) |
| The saline control group | 20ml/kg | 76.9±10.6 | -- |
| The aspirin group | 268mg/kg | 41.9±9.0** | 45.5 |
| The positive drug control group | 37g/kg | 49.2±11.2* | 36.0 |
| Medicine group I heavy dose of the present invention | 70g/kg | 41.0±8.2** | 46.7 |
| Medicine group I low dose of the present invention | 35g/kg | 44.1±11.0** | 42.6 |
| Medicine group II low dose of the present invention | 35g/kg | 46.9±8.0** | 39.7 |
Annotate: n=10, compare * * P＜0.01, * P＜0.05 with the saline control group
As can be seen from the above table, each group of medicine of the present invention all has inhibiting effect to rat granuloma hyperplasia, and the inhibiting effect of medicine of the present invention is significantly higher than the positive control medicine, and the effect of medicine I of the present invention is better than medicine II of the present invention.
2.4 to influence (capillary glass-tube method) the SD rat of rat expectoration amount, ♀ ♂ half and half, body weight 200～250g is divided into 6 groups at random, and 10 every group, the positive drug control group is administration 37g/kg Yanyan slice and 1gkg ammonium chloride.Insert the normal sputum secretory volume in the 2h before the administration of glass capillary record, each group gives relative medicine with the 1mL/100g duodenum respectively then, continues the sputum secretory volume in the 2h after the record administration.Draw the effect of reducing phlegm that sputum length is estimated medicine with kapillary.The results are shown in Table 21.
The influence of table 21 pair rat expectoration amount
The result shows that pharmaceutical composition of the present invention, ammonium chloride and the Yanyan slice of 3 kinds of dosage all can significantly increase the expectoration amount of rat, and the effect of medicine I of the present invention is better than medicine II of the present invention.
2.5 to strong aqua draw cough influence the ICR mouse, ♀ ♂ half and half, body weight 20～24g is divided into 5 groups at random, 10 every group, the positive drug control group is the 37g/kg Yanyan slice.Animal is all with 0.1mL/10g body weight ig administration, one day twice, continuous 5 times.The reference literature method is observed cough latent period (spraying finished to the time that begins to occur coughing) and the interior cough number of times of 3min behind 27% the strong aqua that mouse feeds atomizing behind last administration 45min.The results are shown in Table 22.
Table 22 pair strong aqua draws the influence of coughing
| Group | Dosage (g*kg) | Cough latent period (s) | Cough number of times in the 30min |
| The saline control group | Equal-volume | 45.8±8.7 | ?75.3±6.5 |
| The positive drug control group | 37g/kg | 48.1±10.3 | ?70.0±7.4 |
| Medicine group I heavy dose of the present invention | 70g/kg | 77.8±9.8* | ?46.1±5.9* |
| Medicine group I low dose of the present invention | 35g/kg | 57.7±15.0* | ?59.7±6.2 |
| Medicine group II low dose of the present invention | 35g/kg | 53.1±7.5 | ?62.3±5.1 |
This experimental result shows, medicine I high dose of the present invention can the significant prolongation cough latent period also reduce the cough number of times, in only prolong cough latent period during dosage, to the obviously influence of cough number of times unit, the cough latent period of medicine II of the present invention and cough time number average unit marked change.But it is better that medicine of the present invention and positive control medicine relatively draw the inhibiting effect of coughing to strong aqua.
4 clinical testings of test example
Clinical data 150 routine chronic pharyngitiss all are the hospital outpatient patient, are divided into two groups at random, and 90 examples are organized in treatment, male 60 examples, women 30 examples; The oldest person 75 years old, minimum 12 years old; The course of disease 1 month～1 year.Control group 60 examples, male 45 examples, women 15 examples, maximum 72 years old age, minimum 13 years old; The course of disease 1 month～1 year.Diagnostic criteria meets " traditional Chinese medical science common disease conventional treatment ".
Methods of treatment treatment group is used the pharmaceutical preparation of the present invention according to embodiment 1 preparation, and is oral, 1 time 5,3 times on the 1st.Taking 10d continuously is 1 course of treatment, 1～3 the course of treatment observations.The Yanyan slice that control group adopts Jilin sense health pharmaceutical Co. Ltd to produce, the 0.25g/ sheet, 1d3 time, 1 time 5, taking 10d continuously is 1 course of treatment, 1～3 the course of treatment observations.
Criterion of therapeutical effect is drafted with reference to chronic throat obstruction, chronic pharyngitis standard in " traditional Chinese medical science common disease conventional treatment ".Cure: subjective symptoms disappears, and congestion of throat, oedema disappear; Produce effects: symptom obviously alleviates, congestion of throat, oedema basic controlling; Effectively: symptom makes moderate progress, and congestion of throat, oedema obviously alleviate; Invalid: doing well,improving is slight or do not have improvement, and congestion of throat water, swollen nothing obviously alleviate.
90 examples are organized in the treatment results treatment, clinical cure 23 examples, and produce effects 30 examples, effective 31 examples, invalid 6 examples, total effective rate is 93.33%.Control group 60 examples, clinical cure 6 examples, produce effects 16 examples, effective 25 examples, invalid 13 examples, total effective rate is 78.33%.Two groups relatively have significant difference (P＜0.01).
Clinical research shows that drug therapy chronic pharyngitis effect of the present invention is remarkable.
Specific embodiment
Embodiment 1
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, at drying under reduced pressure below 60 ℃, be crushed to 80 orders, add the sucrose of 25g, make particle, spray adds the 1.75g dolomol, mixing with peppermint oil, be pressed into 1000, sugar coating, promptly.
Embodiment 2
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, at drying under reduced pressure below 60 ℃, be crushed to 80 orders, add the sucrose of 25g, make particle, spray adds the 1.5g dolomol, mixing with peppermint oil, be pressed into 1000, sugar coating, promptly.
Embodiment 3
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, at drying under reduced pressure below 60 ℃, be crushed to 80 orders, add the sucrose of 15g, make particle, spray adds the 0.8g dolomol, mixing with peppermint oil, be pressed into 1000, sugar coating, promptly.
Embodiment 4
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, at drying under reduced pressure below 60 ℃, be crushed to 80 orders, add the sucrose of 15g, make particle, spray adds particle weight 0.8g dolomol, mixing with peppermint oil, be pressed into 1000, sugar coating, promptly.
Embodiment 5 is according to the method for quality control of the pharmaceutical preparation of the present invention of embodiment 1～4 preparation
Differentiate:
(1) get 10 of this product, remove sugar-coat, porphyrize, the 25ml that adds diethyl ether makes dissolving, and sonicated 40 minutes is filtered, and filtrate is waved to 1.5ml, as need testing solution.Other coltsfoot control medicinal material 1g that withdraws the money shines medicinal material solution in pairs with legal system.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 25 μ l of above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) get this product, remove sugar-coat, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample.Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l, respectively with same silica gel g thin-layer plate on, be developping agent with cyclohexane-ethyl acetate (9: 1), the expansion, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(3) sample thief is 10, removes sugar-coat, porphyrize, and 20 milliliters of chloroform-methanols (70: 30) soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of chloroform-methanols (70: 30), soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (5: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Check:
The normal butyl alcohol extract takes by weighing this product, removes sugar-coat, porphyrize, get powder 2g, the accurate title, decide, the accurate methyl alcohol 50ml that adds, claim to decide weight, put in the water-bath reflux 1 hour, put cold, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times, each 25ml with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight evaporate to dryness, 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculate, promptly.
This product is pressed dry product and is calculated, and contains the normal butyl alcohol extract and must not be less than 5.0%.
Assay:
Radix scrophulariae:
Measure according to high performance liquid chromatography (appendix VI D).
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, is Mobile phase B with 1% acetum, and the regulation in the according to the form below is carried out gradient elution; The detection wavelength is 278nm.Number of theoretical plate calculates by the harpagoside peak should be not less than 5000.
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0～20 | 20→50 | 80→50 |
| 20～25 | 50→20 | 50→80 |
It is an amount of that the preparation precision of reference substance solution takes by weighing the harpagoside reference substance, adds 30% methyl alcohol and make the solution that every 1ml contains 10 μ g, promptly.
The preparation of need testing solution is got Chinese medicine composition of the present invention and is equivalent to crude drug 4.0g, and porphyrize is got about 1.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 30% methyl alcohol 50ml that adds, close plug claims to decide weight, soaks after 1 hour, sonicated (power 500W, frequency 40kHz) 30 minutes is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with 30% methyl alcohol, filter, get subsequent filtrate, promptly.
Inaccurate reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Chinese medicine composition of the present invention is equivalent to crude drug 4.0g, contains harpagoside (C
24H
30O
11) must not be less than 0.18mg.Glutinous rehmannia:
Measure according to high performance liquid chromatography (appendix VI D).
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With acetonitrile-0.1% phosphoric acid solution (1: 99) is moving phase; The detection wavelength is 210nm.Number of theoretical plate calculates by the Catalpol peak should be not less than 5000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the Catalpol reference substance, adds moving phase and make the solution that every 1ml contains 10 μ g, promptly.
The preparation of need testing solution is got Chinese medicine composition of the present invention and is equivalent to crude drug 4.0g, and porphyrize is got about 1.0g, the accurate title, decide, put in the tool plug conical flask, the accurate methyl alcohol 25ml that adds claims to decide weight, heating and refluxing extraction 1.5 hours, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methyl alcohol, shake up, filter.Precision is measured subsequent filtrate 10ml, is concentrated near doing, and residue dissolves with moving phase, is transferred in the 10ml measuring bottle, and is diluted to scale with moving phase, shakes up, and filters, and gets subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Chinese medicine composition of the present invention is equivalent to crude drug 4.0g, contains Catalpol (C
15H
22O
10) must not be less than 0.25mg..
Embodiment 7 capsules
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boilings such as all the other radix scrophulariaes three times add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, add 6 times of water gagings for the third time and decocted 2 hours, collecting decoction left standstill 30 hours, get supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, at drying under reduced pressure below 60 ℃, be crushed to 80 orders, add the sucrose of 25g, make particle, spray adds the 1.5g dolomol with peppermint oil, mixing, 1000 capsules of packing into, promptly.
Embodiment 8 granules
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mix with Common Coltsfoot Flower, add 250g starch, 500g sucrose again, mixing, make particle, spray is made the 1000g particle, promptly with peppermint oil.
Embodiment 9 pills
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavor boiling secondaries such as all the other radix scrophulariaes add 8 times of water gagings for the first time and decocted 3 hours, add 6 times of water gagings for the second time and decoct 2 hours, collecting decoction left standstill 24 hours, got supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mix with Common Coltsfoot Flower, drying is crushed to 120 orders, mix with the Macrogol 4000 of fusion ratio according to 1: 3, add the peppermint oil mixing rapidly, drip and make dripping pill, promptly.
Embodiment 10 syrups
More than 12 flavors, except that peppermint oil, all the other radix scrophulariaes etc. ten are the boiling secondary simply, adds for the first time 8 times of water gagings and decocts 3 hours, add for the second time 6 times of water gagings and decocted 2 hours, collecting decoction left standstill 24 hours, get supernatant, be concentrated in right amount, other gets sucrose 650g and adds water boil, after the dissolving, filter, concentrate and make syrup, with above-mentioned concentrate mixing, boil, put cold, add peppermint oil, antiseptic and essence, and be diluted to 1000ml, promptly with cold boiling water.
Embodiment 11 buccal tablets
More than 12 the flavor, except that peppermint oil, all the other radix scrophulariaes etc. ten are the boiling secondary simply, add for the first time 8 times of water gagings and decocted 3 hours, add 6 times of water gagings for the second time and decocted collecting decoction 2 hours, left standstill 24 hours, and got supernatant, concentrated relative density is the thick paste of 1.35 (60-70 ℃), add sucrose 750g, vanillic aldehyde 1.5g, honey element 10g, citric acid 15g mixing, make particle, drying, whole grain sprays into volatile oil, mixing, be pressed into 1000, promptly.
Embodiment 12
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavors such as all the other radix scrophulariaes add 8 times of water gagings and decocted 4 hours, filter, and decocting liquid left standstill 24 hours, get supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, drying is crushed to 80 orders, adds the sucrose of 25g, make particle, spray adds the 1.5g dolomol with peppermint oil, mixing, be pressed into 1000, sugar coating, promptly.
Its method of quality control comprises following several:
Differentiate:
(1) get 10 of this product, remove sugar-coat, porphyrize, the 25ml that adds diethyl ether makes dissolving, and sonicated 40 minutes is filtered, and filtrate is waved to 1.5ml, as need testing solution.Other coltsfoot control medicinal material 1g that withdraws the money shines medicinal material solution in pairs with legal system.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 25 μ l of above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) get this product, remove sugar-coat, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample.Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l, respectively with same silica gel g thin-layer plate on, be developping agent with cyclohexane-ethyl acetate (9: 1), the expansion, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(3) sample thief is 10, removes sugar-coat, porphyrize, and 20 milliliters of chloroform-methanols (70: 30) soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of chloroform-methanols (70: 30), soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (5: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
Check:
The normal butyl alcohol extract takes by weighing this product, removes sugar-coat, porphyrize, get powder 2g, the accurate title, decide, the accurate methyl alcohol 50ml that adds, claim to decide weight, put in the water-bath reflux 1 hour, put cold, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, place dry evaporating dish to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times, each 25ml with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight evaporate to dryness, 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculate, promptly.
Claims (4)
1. a detection method for the treatment of the Chinese medicine composition of chronic pharyngitis is characterized in that this method comprises the steps:
This Chinese medicine composition is made up of following weight portion crude drug:
Detection method comprises one or more in following qualitative checking method and/or the quantitative detecting method:
(1) qualitative detection of tussilago
Get pharmaceutical preparation content of the present invention, adding diethyl ether makes dissolving, and sonicated is filtered, and filtrate is waved to 1.5ml, as need testing solution;
The coltsfoot of withdrawing the money control medicinal material is made control medicinal material solution;
According to thin-layered chromatography test, draw above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, hot blast blow develop the color to spot clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(2) qualitative detection of Oroxylum indicum
Get pharmaceutical preparation content of the present invention, add the absolute ethyl alcohol sonicated, filter, the filtrate evaporate to dryness adds anhydrous alcohol solution as test sample;
Get the Oroxylum indicum control medicinal material, make control medicinal material solution;
According to thin-layered chromatography test, draw above-mentioned sample solution, control medicinal material solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with the vanillic aldehyde sulfuric acid solution, hot blast blow develop the color to spot clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(3) qualitative detection of the tuber of dwarf lilyturf
Get pharmaceutical preparation content of the present invention, add the chloroform-methanol mixed liquid dipping, sonicated is filtered, and evaporate to dryness is with the chloroform dissolving, as need testing solution;
Get the control medicinal material tuber of dwarf lilyturf, shred, make control medicinal material solution;
According to thin-layered chromatography test, draw above-mentioned solution, respectively with same silica gel g thin-layer plate on, with developping agent, launch, take out, dry, spray is with ethanol solution of sulfuric acid, hot blast blow develop the color to spot clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(4) detection by quantitative of normal butyl alcohol extract:
Get pharmaceutical preparation content of the present invention, the accurate title, decide, and the accurate methyl alcohol that adds claims to decide weight, put reflux in the water-bath, put coldly, supply the weight that subtracts mistake, filter with methyl alcohol, get subsequent filtrate, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water makes dissolving, extract with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight evaporate to dryness, 105 ℃ of dryings, in the dislocation exsiccator, place, weight decided in accurate rapidly title, calculates, promptly;
(5) detection by quantitative of harpagoside:
According to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, is Mobile phase B with 1% acetum, carries out gradient elution; The detection wavelength is 278nm; Number of theoretical plate calculates by the harpagoside peak should be not less than 5000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the harpagoside reference substance, adds methyl alcohol and make reference substance solution, promptly;
Chinese medicine composition content of the present invention is got in the preparation of need testing solution, gets about 1.0g, and accurate the title decides, put in the tool plug conical flask accurate 30% methyl alcohol, the close plug of adding, claim to decide weight, after the immersion, sonicated, put coldly, claim again to decide weight, supply the weight that subtracts mistake with 30% methyl alcohol, shake up, filter, get subsequent filtrate, promptly;
Accurate respectively reference substance solution and the need testing solution drawn of determination method injects liquid chromatograph, measures, promptly;
(6) detection by quantitative of Catalpol:
According to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With acetonitrile-0.1% phosphoric acid solution is moving phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the Catalpol peak should be not less than 5000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the Catalpol reference substance, adds moving phase and make reference substance solution, promptly;
Chinese medicine composition phase content of the present invention is got in the preparation of need testing solution, gets about 1.0g, and accurate the title decides, and puts in the tool plug conical flask, the accurate methyl alcohol that adds claims to decide weight, and heating and refluxing extraction is put cold, claim again to decide weight, supply the weight that subtracts mistake, shake up, filter with methyl alcohol; Precision is measured subsequent filtrate 10ml, is concentrated near doing, and residue dissolves with moving phase, is transferred in the measuring bottle, and is diluted to scale with moving phase, shakes up, and filters, and gets subsequent filtrate, promptly; Determination method, accurate respectively reference substance solution and the need testing solution drawn injects liquid chromatograph, measures, promptly.
2. the method for claim 1 is characterized in that this Chinese medicine composition is made up of following weight portion bulk drug:
3. the method for claim 1 is characterized in that this detection method comprises one or more in following qualitative checking method and/or the quantitative detecting method:
(1) qualitative detection of tussilago
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 10.5g, porphyrize, the 25ml that adds diethyl ether makes dissolving, and sonicated 40 minutes is filtered, and filtrate is waved to 1.5ml, as need testing solution; Other coltsfoot control medicinal material 1g that withdraws the money shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, draw each 25 μ l of above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, be developping agent with 4: 1 ratio normal hexane-ethyl acetates, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, hot blast blow develop the color to spot clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(2) qualitative detection of Oroxylum indicum
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 4g, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample; Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast; According to thin-layered chromatography test, draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l are respectively and on the same silica gel g thin-layer plate, with 9: 1 ratio cyclohexane-ethyl acetate was developping agent, launched, and took out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(3) qualitative detection of the tuber of dwarf lilyturf
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 10.5g, porphyrize adds 20 milliliters of the chloroform-methanols of 70: 30 ratios, soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution; Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of the chloroform-methanols of 70: 30 ratios, soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast; According to thin-layered chromatography test, draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, be developping agent with 5: 1 ratio normal hexane-ethyl acetate, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, hot blast blow develop the color to spot clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;
(4) detection by quantitative of normal butyl alcohol extract
Get the pharmaceutical preparation of the present invention that is equivalent to raw medicinal material 8.5g, porphyrize is got powder 2g, the accurate title, decide, and the accurate methyl alcohol 50ml that adds claims to decide weight, put in the water-bath reflux 1 hour, and put coldly, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times with water saturated normal butyl alcohol, each 25ml merges normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight, evaporate to dryness is 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculates, promptly;
Chinese medicine composition of the present invention is pressed dry product and is calculated, and contains the normal butyl alcohol extract and must not be less than 5.0%;
(5) detection by quantitative of harpagoside:
According to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, is Mobile phase B with 1% acetum, and the regulation in the according to the form below is carried out gradient elution; The detection wavelength is 278nm; Number of theoretical plate calculates by the harpagoside peak should be not less than 5000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the harpagoside reference substance, adds 30% methyl alcohol and make the solution that every 1ml contains 10 μ g, promptly;
The preparation of need testing solution is got Chinese medicine composition of the present invention and is equivalent to crude drug 4.0g, and porphyrize is got about 1.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 30% methyl alcohol 50ml that adds, close plug claims to decide weight, soaks after 1 hour, at power 500W, sonicated is 30 minutes under the frequency 40kHz, puts cold, claim again to decide weight, supply the weight that subtracts mistake, shake up with 30% methyl alcohol, filter, get subsequent filtrate, promptly;
Inaccurate reference substance solution and each the 20 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly; Chinese medicine composition of the present invention is equivalent to crude drug 4.0g, contains harpagoside (C
24H
30O
11) must not be less than 0.18mg;
(6) detection by quantitative of Catalpol
According to high effective liquid chromatography for measuring; Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With 1: 99 ratio acetonitrile-0.1% phosphoric acid solution was moving phase; The detection wavelength is 210nm; Number of theoretical plate calculates by the Catalpol peak should be not less than 5000; The preparation of reference substance solution: it is an amount of that precision takes by weighing the Catalpol reference substance, adds moving phase and make the solution that every 1ml contains 10 μ g, promptly; The preparation of need testing solution: get Chinese medicine composition of the present invention and be equivalent to crude drug 4.0g, porphyrize is got about 1.0g, the accurate title, decide, put in the tool plug conical flask, the accurate methyl alcohol 25ml that adds claims to decide weight, heating and refluxing extraction 1.5 hours, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methyl alcohol, shake up, filter; Precision is measured subsequent filtrate 10ml, is concentrated near doing, and residue dissolves with moving phase, is transferred in the 10ml measuring bottle, and is diluted to scale with moving phase, shakes up, and filters, and gets subsequent filtrate, promptly; Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly; Chinese medicine composition of the present invention is equivalent to crude drug 4.0g, contains Catalpol (C
15H
22O
10) must not be less than 0.25mg.
4. a detection method for the treatment of the Chinese medicine composition of chronic pharyngitis is characterized in that this method comprises the steps:
This Chinese medicine composition is made up of following weight portion crude drug:
More than 12 the flavor, except that peppermint oil, tussilago is crushed to 100 orders, sieves; Ten flavors such as all the other radix scrophulariaes add 8 times of water gagings and decocted 4 hours, filter, and decocting liquid left standstill 24 hours, get supernatant, being condensed into relative density is the thick paste of 1.35 (60-70 ℃), mixes with Common Coltsfoot Flower, drying is crushed to 80 orders, adds the sucrose of 25g, make particle, spray adds the 1.5g dolomol with peppermint oil, mixing, be pressed into 1000, sugar coating, promptly;
Detection method comprises the steps:
Differentiate:
(1) get 10 of this product, remove sugar-coat, porphyrize, the 25ml that adds diethyl ether makes dissolving, and sonicated 40 minutes is filtered, and filtrate is waved to 1.5ml, as need testing solution.Other coltsfoot control medicinal material 1g that withdraws the money shines medicinal material solution in pairs with legal system.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 25 μ l of above-mentioned two kinds of solution, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (4: 1) is developping agent, launch, take out, dry, spray is with vanillic aldehyde concentrated sulphuric acid test solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(2) get this product, remove sugar-coat, porphyrize takes by weighing 1g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of dissolving of absolute ethyl alcohol as test sample.Other gets Oroxylum indicum control medicinal material 2g, adds 50 milliliters of absolute ethyl alcohols ultrasonic 10 minutes, filters, and the filtrate evaporate to dryness adds 1 milliliter of absolute ethyl alcohol and dissolves medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned sample solution 15 μ l, control medicinal material solution 5 μ l, respectively with same silica gel g thin-layer plate on, be developping agent with cyclohexane-ethyl acetate (9: 1), the expansion, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
(3) sample thief is 10, removes sugar-coat, porphyrize, and 20 milliliters of chloroform-methanols (70: 30) soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, as need testing solution.Other gets the control medicinal material 2g tuber of dwarf lilyturf, shreds, with 20 milliliters of chloroform-methanols (70: 30), soaked 3 hours, and ultrasonic 30 minutes, filter, evaporate to dryness is with 0.5 milliliter of dissolving of chloroform, medicinal material solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw above-mentioned solution 10 μ l respectively, respectively with same silica gel g thin-layer plate on, with normal hexane-ethyl acetate (5: 1) is developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.
The normal butyl alcohol extract detects: take by weighing this product, remove sugar-coat, porphyrize, get powder 2g, the accurate title, decide, the accurate methyl alcohol 50ml that adds, claim to decide weight, put in the water-bath reflux 1 hour, put cold, supply the weight that subtracts mistake with methyl alcohol, filter, get subsequent filtrate 25ml, put in the evaporating dish that is dried to constant weight, evaporate to dryness, residue add water 25ml, make dissolving, extract 3 times, each 25ml with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, put in the evaporating dish that is dried to constant weight evaporate to dryness, 105 ℃ of dryings 3 hours, in the dislocation exsiccator, placed 30 minutes, weight decided in accurate rapidly title, calculate, promptly.
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| CN110389181A (en) * | 2018-04-20 | 2019-10-29 | 福建省闽东力捷迅药业有限公司 | A kind of method of index component content in quick detection Zengye Oral Liquid |
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