CN103954724B - Method for detecting Jingfang granules - Google Patents
Method for detecting Jingfang granules Download PDFInfo
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Abstract
The invention relates to a method for detecting Jingfang granules. The method comprises the steps of character identifying, checking and measuring content, wherein the step of identifying comprises identification of schizonepeta, radix angelicae pubescentis, ligusticum wallichii, fructus aurantii and bupleurum longiradiatum; the step of measuring content refers to a step of measuring content of ephedrine hydrochloride. Aiming at the problem that the condition that fewer raw materials are added or corresponding raw materials are not added by illegal manufacturer or a counterfeit drug bupleurum longiradiatum is mixed cannot be monitored because active ingredients of the granules cannot be correspondingly detected by a set of perfect detection method at present, a scientific, reasonable, practical and feasible component identification and content measurement method is formulated, the clinical effects of the Jingfang granules are guaranteed, quality indexes of several medicinal materials are clear, and application of various medicinal materials in the Jingfang granules is ensured, so that the curative effect of the drug is guaranteed. Moreover, the counterfeit drug bupleurum longiradiatum in radix bupleuri can be effectively detected, and a phenomenon that health of patients is endangered by two toxic ingredients, including bupleurotoxin and acetylbupleurotoxin, contained in bupleurum longiradiatum is avoided.
Description
Technical field
The present invention relates to the detection method of the anti-particle of a kind of chaste tree.
Background technology
Anti-(electuary) particle of chaste tree is " the Sanitation Ministry medicine standard " Traditional Chinese medicine historical preparation second kind recorded, standard number is WS3-B-0328-90, prescription is schizonepeta, windproof, notopterygium root, levisticum, radix bupleuri, the root of purple-flowered peucedanum, Ligusticum wallichii, Fructus Aurantii, Poria cocos, balloonflower root, Radix Glycyrrhizae, is pure Chinese medicine and granule.It has inducing sweat and dispelling exogenous evils, the effect of loose wind clearing damp.Be used for the treatment of cold clinically, headache bodily pain, aversion to cold is lossless, nasal obstruction runny nose, cough.Be be used for the treatment of cold in the market, headache bodily pain, aversion to cold is lossless, nasal obstruction runny nose, the common drug of cough.But up to the present, a set of perfect detection method is not also had to detect accordingly its effective ingredient, illegal manufacturer so may be caused not prepare burden in strict accordance with the dosage of prescription when producing medicine, wantonly reduce the high raw material of price, the curative effect of medicine is caused obviously to decline, affect the safe and effective of medicine, the interests of grievous injury patient.On the other hand, medicinal material radix bupleuri is used in prescription, because its use amount is larger, and its limits throughput, so its adulterant bigleaf thorowax root is also had a mind to or is not intended to be mixed into use, but containing bupleurotoxin and acetylbupleurotoxin in bigleaf thorowax root, these two kinds of compositions are all noxious materials, repeatedly occur because taking the medicine and poisoning Adverse drug events that are mixed with bigleaf thorowax root, so it is necessary to control bigleaf thorowax root.The discriminating control problem of bigleaf thorowax root medicinal material in pharmacopeia also inquired in " Shandong medical industry " the 22 volume the 5th phase in 2003 " to supplementing of the Chinese Pharmacopoeia version radix bupleuri method of inspection in a 2000 " literary composition, but its method has obvious deficiency: place 24 hours, detection time is oversize, is unfavorable for that drug production process controls and drug quality supervision and check; Because of bupleurotoxin and acetylbupleurotoxin content relatively low, point sample is 2 μ l only, and point sample amount is very few, impact inspection accuracy; Because of bupleurotoxin and acetylbupleurotoxin content relatively low, inspect under uviol lamp, observe without chromogenic reagent, observing effect is bad.More importantly many, the contained complicateds of chaste tree anti-particle prescription flavour of a drug, other composition can produce to be obscured and disturbs.Other Chinese patent drug containing radix bupleuri is also the same, and because its prescription flavour of a drug are many, complicated, wherein will control bigleaf thorowax root very difficult, so finding a kind of better way to control being mixed into of bigleaf thorowax root is the task of top priority.
Summary of the invention
Object of the present invention, be to provide the detection method of the anti-particle of a kind of chaste tree, effectively can monitor illegal manufacturer throw less or do not throw corresponding raw material, monitor being mixed into of adulterant bigleaf thorowax root simultaneously, prevent the generation of taking rear poisoning Adverse drug events, make this drug safety effective, and ensure that the clinical efficacy of Yinchai Granules.
Technical scheme of the present invention is as follows:
The prescription of the anti-particle of this chaste tree: schizonepeta 75g, windproof 75g, notopterygium root 75g, levisticum 75g, radix bupleuri 75g, root of purple-flowered peucedanum 75g, Ligusticum wallichii 75g, Fructus Aurantii 75g, Poria cocos 75g, balloonflower root 75g, Radix Glycyrrhizae 25g.
The detection method of the anti-particle of this chaste tree, comprise proterties, discriminating, inspection, assay project, wherein, differentiate to comprise the discriminating of schizonepeta, assay that the discriminating of levisticum, the discriminating of Ligusticum wallichii, the discriminating of Fructus Aurantii, the discriminating of bigleaf thorowax root, assay are ephedrine hydrochloride, detection method comprises following:
The discriminating of schizonepeta is: take schizonepeta as control medicinal material, with sherwood oil-chloroform-methanol for developping agent, differentiate by thin-layered chromatography;
The discriminating of levisticum is: take levisticum as control medicinal material, with sherwood oil-chloroform-methanol for developping agent, differentiate by thin-layered chromatography;
The discriminating of Ligusticum wallichii is: take Ligusticum wallichii as control medicinal material, with sherwood oil-chloroform-methanol for developping agent, differentiate by thin-layered chromatography;
The discriminating of Fructus Aurantii is: take Fructus Aurantii as control medicinal material, places the subnatant that spends the night for developping agent, differentiate by thin-layered chromatography with chloroform-methanol-water;
The discriminating of bigleaf thorowax root is: with bupleurotoxin, acetylbupleurotoxin for reference substance, with normal hexane-ethyl acetate for developping agent, differentiate by thin-layered chromatography.
The assay of ephedrine hydrochloride: be take ephedrine hydrochloride as reference substance, with high performance liquid chromatography row assay.
Detection method comprises following:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=9 ~ 11:2 ~ 4:1.5 ~ 2.5 are developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=18 ~ 22:4 ~ 6:1.5 ~ 2.5 are developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=4 ~ 6:1.5 ~ 2.5:0.7 ~ 1.3 are developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: the mixed liquor of water=11 ~ 15:6 ~ 8:1.5 ~ 2.5, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6 ~ 10:0.5 ~ 1.5 are developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color,
(6) assay of ephedrine hydrochloride
Be filling agent with octadecylsilane chemically bonded silica, acetonitrile: 0.2% phosphoric acid=17.5:82.5 is mobile phase, column temperature 35 DEG C, determined wavelength is 283nm, flow velocity 1.0ml/min, number of theoretical plate calculates should be not less than 5000 by ephedrine hydrochloride peak, take and use the phosphorus pentoxide drying ephedrine hydrochloride reference substance 0.00503g of 24 hours in 50ml measuring bottle, add formamide dissolve and be diluted to scale, shake up, draw this reference substance solution 5ml in 10ml measuring bottle, add formamide dissolve and be diluted to scale, obtain the ephedrine hydrochloride reference substance solution of every 1ml containing 50.30 μ g, get this product 1.0g, add formamide 50ml, weighed weight, ultrasonic process 15 minutes, let cool, weighed weight again, the weight of less loss is supplied with formamide, shake up, filter with 0.45 μm of miillpore filter, get subsequent filtrate, obtain need testing solution, accurate absorption reference substance solution and each 10ul of need testing solution, injection liquid chromatography, measure peak area, calculate by external standard method, obtain.
This product every bag in ephedrine hydrochloride (C27H32O14), must not be less than 7.5mg containing Fructus Aurantii.
Detection method is as follows more specifically:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=10:3:2 is developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=20:5:2 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=5:2:1 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: water=13: the mixed liquor of 7: 2, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=8:1 is developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color,
(6) assay of ephedrine hydrochloride
Be filling agent with octadecylsilane chemically bonded silica, acetonitrile: 0.2% phosphoric acid=17.5:82.5 is mobile phase, column temperature 35 DEG C, determined wavelength is 283nm, flow velocity 1.0ml/min, number of theoretical plate calculates should be not less than 5000 by ephedrine hydrochloride peak, take and use the phosphorus pentoxide drying ephedrine hydrochloride reference substance 0.00503g of 24 hours in 50ml measuring bottle, add formamide dissolve and be diluted to scale, shake up, draw this reference substance solution 5ml in 10ml measuring bottle, add formamide dissolve and be diluted to scale, obtain the ephedrine hydrochloride reference substance solution of every 1ml containing 50.30 μ g, get this product 1.0g, add formamide 50ml, weighed weight, ultrasonic process 15 minutes, let cool, weighed weight again, the weight of less loss is supplied with formamide, shake up, filter with 0.45 μm of miillpore filter, get subsequent filtrate, obtain need testing solution, accurate absorption reference substance solution and each 10ul of need testing solution, injection liquid chromatography, measure peak area, calculate by external standard method, obtain the content of this product.
This product every bag in ephedrine hydrochloride (C27H32O14), must not be less than 7.5mg containing Fructus Aurantii.
Proterties: this product is brown particle; Gas is fragrant, sweet, the micro-hardship of taste.
Check: every regulation (" Chinese Pharmacopoeia " version in 2010 annex I C) relevant under granule item should be met.
The ratio of developping agent of the present invention and the ratio of mobile phase are all with volume basis.
Technique effect of the present invention:
The present invention is by the discriminating to the schizonepeta in the anti-particle of chaste tree, the discriminating of levisticum, the discriminating of Ligusticum wallichii, the discriminating of Fructus Aurantii, the discriminating of bigleaf thorowax root, the assay of ephedrine hydrochloride, the Main Ingredients and Appearances such as the schizonepeta in prescription, levisticum, Ligusticum wallichii, Fructus Aurantii are made to be obtained for effective monitoring, on the other hand, the adulterant bigleaf thorowax root of radix bupleuri is also effectively detected, and avoids the health of bupleurotoxin and these the two kinds of toxic substance harm patients of acetylbupleurotoxin contained in bigleaf thorowax root.The present invention can monitor the quality of the anti-particle of chaste tree better and more comprehensively; and effectively can control the anti-particle of illegal manufacturers produce chaste tree of poor quality, thus ensure that the curative effect of medicine, ensure that the safety of medicine; protect the interests of patient, this method is scientific and reasonable, practical.
The present invention is studied assay, and specific experiment data is as follows:
The investigation of content assaying method
1, instrument and reagent:
1.1HP1100 high performance liquid chromatograph, is configured to G1311A quaternary pump, G1313A automatic sampler, G1316A column oven, G1314A UV-detector, HP chem workstation.
1.2 chromatographic columns: BDS-C18 post Kromasil (250mm × 4.6mm).
1.3 ephedrine hydrochloride reference substances (research institute provides by Products in China, and lot number is 0722-201008), methyl alcohol (chromatographically pure), phosphoric acid (analyzing pure), water are self-control redistilled water.
2, chromatographic condition and system suitability: be filling agent with octadecylsilane chemically bonded silica; Acetonitrile-0.2% phosphoric acid (17.5:82.5) is mobile phase; Column temperature 35 DEG C, determined wavelength is 283nm; Flow velocity 1.0ml/min; Number of theoretical plate calculates should be not less than 5000 by ephedrine hydrochloride peak.
3, the preparation of reference substance solution: precision takes through the phosphorus pentoxide ephedrine hydrochloride reference substance 0.00503g of dry 24 hours in 50ml measuring bottle, add formamide dissolve and be diluted to scale, shake up, accurate this reference substance solution 5ml that draws, in 10ml measuring bottle, obtains the ephedrine hydrochloride reference substance solution of every 1ml containing 50.30 μ g.
4, the preparation of need testing solution: get this product 1.0g, accurately weighed, precision adds formamide 50ml, weighed weight, ultrasonic process 15 minutes, lets cool, more weighed weight, supplies the weight of less loss with formamide, shake up, filter with 0.45 μm of miillpore filter, get subsequent filtrate, to obtain final product.
5, determination method: accurate absorption reference substance solution and each 10ul of need testing solution respectively, injection liquid chromatography, measures peak area, calculates, to obtain final product by external standard method.
6, methodological study
6.1, the selection of chromatographic condition and system suitability
Be filling agent with octadecylsilane chemically bonded silica; Acetonitrile-0.2% phosphoric acid (17.5:82.5) is mobile phase; Column temperature is 35 DEG C; Flow velocity is 1.0ml/min; Selecting to measure wavelength is 283nm, detects by UV-detector.With this understanding, ephedrine hydrochloride and other components all can reach baseline separation, and experiment shows: have the peak consistent with ephedrine hydrochloride reference substance retention time in test sample, negative control is noiseless.
The investigation of 6.2 ranges of linearity:
Precision takes through the phosphorus pentoxide ephedrine hydrochloride reference substance 0.00503g of dry 24 hours in 50ml measuring bottle, adds formamide and dissolves and be diluted to scale, shake up.Accurate draw this reference substance solution 1,3,5,7,9ml in 5 10ml measuring bottles, with methanol dilution to scale, the accurate 10 μ l that draw inject high performance liquid chromatographs respectively, measure, take concentration as horizontal ordinate, peak area is that ordinate carries out recurrence process, obtains regression equation:
A＝32.9204098C-1.9912226(n＝5),γ＝1.00000
Result shows that ephedrine hydrochloride is good linear relationship between sample size 0.1006 μ g-0.9054 μ g, the results are shown in Table 1.
Table 1 ephedrine hydrochloride typical curve test findings
The investigation of 6.3 precision:
Ephedrine hydrochloride reference substance solution (concentration is 50.03 μ g/ml) is repeated sample introduction five times, measures peak area, ephedrine hydrochloride peak area mean value=1654.4886, RSD=0.24%.Result shows: precision is good, the results are shown in Table 2.
Table 2 precision investigates result
6.4 replica test
Precision takes same sample lots (050601) 5 part, and by need testing solution preparation method preparation, measure its content, mean value is 11.557mg/ bag, RSD=0.62% in accordance with the law.The results are shown in Table 3, test shows: the repeatability of assay method is good.
Table 3: sample repeatability is tested
6.5 application of sample recovery experiments
Precision takes lot number is respectively that the sample of 050601 (known content is 2.30mg/g) is about 0.50g, put in the measuring bottle of 6 50ml, add ephedrine hydrochloride reference substance solution (concentration is 1.12mg/ml) 1.0ml more respectively, by above-mentioned need testing solution preparation method and chromatographic condition, prepare application of sample reclaim need testing solution and inject high performance liquid chromatograph, measure content, with the following formulae discovery recovery, the results are shown in Table 4, test findings shows: the recovery is between 96.64% ~ 102.3%, and application of sample reclaims good.
Table 4 ephedrine hydrochloride application of sample recovery test result
6.6 stability tests: be that the sample of (050601) prepares need testing solution by need testing solution preparation method by lot number, and respectively at 0 hour, 1 hour, 2 hours, 4 hours, 8 hours, the accurate 10 μ l that draw inject high performance liquid chromatograph, measure integrating peak areas value, the results are shown in Table 5, result shows: under this experiment condition, and Determination of ephedrine hydrochloride is stable in 8 hours.
The stability test of table 5 ephedrine hydrochloride
7, the mensuration of sample size: we measure Determination of ephedrine hydrochloride in 3 batches of pilot products, the results are shown in Table 6.
Table 6 sample size measurement result
Therefore: 3 batches of anti-particle Determination of ephedrine hydrochlorides every bag of chaste tree are between 4.606mg-4.899mg, more stable.
Through above experiment, prove method reasonable of the present invention.
Embodiment:
Embodiment 1:
[prescription] schizonepeta 75g windproof 75g notopterygium root 75g levisticum 75g radix bupleuri 75g root of purple-flowered peucedanum 75g Ligusticum wallichii 75g Fructus Aurantii 75g Poria cocos 75g balloonflower root 75g Radix Glycyrrhizae 25g
More than [method for making], ten simply, and schizonepeta, windproof, notopterygium root, levisticum, the root of purple-flowered peucedanum, Ligusticum wallichii and Fructus Aurantii extract volatile oil respectively, and the another device of aqueous solution after Ligusticum wallichii, Fructus Aurantii distillation is collected; The dregs of a decoction of Ligusticum wallichii, Fructus Aurantii and Poria cocos, according to the percolation under liquid extract and extract item, be made into 25% ethanolic solution by above-mentioned aqueous solution and make solvent, carry out diacolation; The three taste boiling secondaries such as the dregs of a decoction of schizonepeta, windproof, notopterygium root, levisticum and the root of purple-flowered peucedanum and all the other radix bupleuri, balloonflower root, Radix Glycyrrhizae, each 1.5 hours, collecting decoction, filtered, and filtrate is condensed into thick paste; Merge filter liquid and thick paste, mixing, leave standstill, filter, at the temperature of 80 ~ 85 DEG C, filtrate is condensed into the clear cream that relative density is 1.30, qinghuo reagent 1 part, and with sucrose 6 parts, particle is made in mixing, dry, adds the volatile oil such as above-mentioned schizonepeta, and mixing, to obtain final product.
[proterties] this product is brown particle; Gas is fragrant, sweet, the micro-hardship of taste.
The discriminating of [discriminating] (1) schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=9:2:1.5 is developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=18:4:1.5 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=4:1.5:0.7 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: the mixed liquor of water=11:6:1.5, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6:0.5 is developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color,
Every regulation (" Chinese Pharmacopoeia " version in 2010 annex I C) that [inspection] is relevant under should meeting granule item.
[assay] is according to high effective liquid chromatography for measuring.
Be filling agent with octadecylsilane chemically bonded silica, acetonitrile: 0.2% phosphoric acid=17.5:82.5 is mobile phase, column temperature 35 DEG C, determined wavelength is 283nm, flow velocity 1.0ml/min, number of theoretical plate calculates should be not less than 5000 by ephedrine hydrochloride peak, take ephedrine hydrochloride reference substance 0.00503g in 50ml measuring bottle, add formamide dissolve and be diluted to scale, shake up, draw this reference substance solution 5ml in 10ml measuring bottle, add formamide dissolve and be diluted to scale, obtain the ephedrine hydrochloride reference substance solution of every 1ml containing 50.30 μ g, get this product 1.0g, add formamide 50ml, weighed weight, ultrasonic process 15 minutes, let cool, weighed weight again, the weight of less loss is supplied with formamide, shake up, filter with miillpore filter, get subsequent filtrate, obtain need testing solution, accurate absorption reference substance solution and each 10ul of need testing solution, injection liquid chromatography, measure peak area, calculate by external standard method, obtain the content of this product.
This product every bag in ephedrine hydrochloride (C27H32O14) 7.9mg, is no less than 7.5mg containing Fructus Aurantii.
Embodiment 2:
In the present embodiment, except discrimination method difference, all the other detection methods are all identical with embodiment 1, and concrete discrimination method is as follows:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=10:3:2 is developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=20:5:2 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=5:2:1 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: water=13: the mixed liquor of 7: 2, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=8:1 is developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color,
Embodiment 3:
In the present embodiment, except discrimination method difference, all the other detection methods are all identical with embodiment 1, and concrete discrimination method is as follows:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=11:4:2.5 is developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=22:6:2.5 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=6:2.5:1.3 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: the mixed liquor of water=15:8:2.5, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=10:1.5 is developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color.
Claims (4)
1. the detection method of the anti-particle of chaste tree, comprise proterties, discriminating, inspection, assay project, it is characterized in that: described discriminating comprises the discriminating of schizonepeta, the discriminating of levisticum, the discriminating of Ligusticum wallichii, the discriminating of Fructus Aurantii, the discriminating of bigleaf thorowax root, assay is the assay of ephedrine hydrochloride, and detection method comprises following:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=9 ~ 11:2 ~ 4:1.5 ~ 2.5 are developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=18 ~ 22:4 ~ 6:1.5 ~ 2.5 are developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=4 ~ 6:1.5 ~ 2.5:0.7 ~ 1.3 are developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: the mixed liquor of water=11 ~ 15:6 ~ 8:1.5 ~ 2.5, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=6 ~ 10:0.5 ~ 1.5 are developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color,
(6) assay of ephedrine hydrochloride
Be filling agent with octadecylsilane chemically bonded silica, acetonitrile: 0.2% phosphoric acid=17.5:82.5 is mobile phase, column temperature 35 DEG C, determined wavelength is 283nm, flow velocity 1.0ml/min, number of theoretical plate calculates should be not less than 5000 by ephedrine hydrochloride peak, take ephedrine hydrochloride reference substance 0.00503g in 50ml measuring bottle, add formamide dissolve and be diluted to scale, shake up, draw this reference substance solution 5ml in 10ml measuring bottle, add formamide dissolve and be diluted to scale, obtain the ephedrine hydrochloride reference substance solution of every 1ml containing 50.30 μ g, get this product 1.0g, add formamide 50ml, weighed weight, ultrasonic process 15 minutes, let cool, weighed weight again, the weight of less loss is supplied with formamide, shake up, filter with miillpore filter, get subsequent filtrate, obtain need testing solution, accurate absorption reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measure peak area, calculate by external standard method, obtain.
2. the detection method of the anti-particle of chaste tree according to claim 1, is characterized in that: discrimination method is as follows more specifically:
(1) discriminating of schizonepeta
Get this product 10g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, as need testing solution, separately get schizonepeta control medicinal material 1g, add diethyl ether 50ml, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds ethanol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=10:3:2 is developping agent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color,
(2) discriminating of levisticum
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get levisticum control medicinal material 1g, add 60 ~ 90 DEG C of sherwood oil 40ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate volatilizes, residue adds methyl alcohol 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=20:5:2 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(3) discriminating of Ligusticum wallichii
Get this product 10g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, as need testing solution, separately get Ligusticum wallichii control medicinal material 1g, add methyl alcohol 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds ethyl acetate 1ml makes dissolving, medicinal material solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with sherwood oil: chloroform: methyl alcohol=5:2:1 is developping agent, launch, take out, dry, putting wavelength is inspect under the ultraviolet lamp of 365nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color,
(4) discriminating of Fructus Aurantii
Get this product 10g, add acetone 30ml, ultrasonic process 20 minutes, filter, filtrate puts evaporate to dryness in water-bath, residue adds methyl alcohol 1ml makes dissolving, as need testing solution, separately get ephedrine hydrochloride reference substance, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, select chloroform: methyl alcohol: water=13: the mixed liquor of 7: 2, 10 DEG C with the subnatant of left overnight for developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color,
(5) discriminating of bigleaf thorowax root
Get this product 10g, add 60 ~ 90 DEG C of sherwood oil 30ml, soak 10 minutes, ultrasonic process 15 minutes, filter, filtrate is waved to 1ml, as need testing solution, separately get bupleurotoxin, acetylbupleurotoxin reference substance, add methyl alcohol and make the reference substance solution of every 1ml containing bupleurotoxin 1mg and acetylbupleurotoxin 1mg, product solution in contrast, test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with normal hexane: ethyl acetate=8:1 is developping agent, launch, take out, dry, spray is with 10% phosphomolybdic acid ethanol solution, spot development is heated to clear at 105 DEG C, in test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of not aobvious same color.
3. the detection method of the anti-particle of chaste tree according to claim 1, is characterized in that: the ephedrine hydrochloride reference substance in step (6) uses phosphorus pentoxide dry 24 hours.
4. the detection method of the anti-particle of chaste tree according to claim 1, is characterized in that: the miillpore filter aperture in step (6) is 0.45 μm.
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| CN109507305B (en) * | 2018-08-30 | 2021-07-16 | 河北中医学院 | Comprehensive detection and identification method for quality of schizonepeta medicinal material based on HPLC and UV-Vis technology |
| CN114903857B (en) * | 2021-02-09 | 2023-06-06 | 鲁南制药集团股份有限公司 | Jingfeng granule and preparation method thereof |
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| CN115518105A (en) * | 2021-07-02 | 2022-12-27 | 山东新时代药业有限公司 | Composition with function of treating infantile eczema and application thereof |
| CN114441680B (en) * | 2022-01-26 | 2023-11-14 | 浙江省食品药品检验研究院 | Method for distinguishing traditional Chinese medicine fructus aurantii from garden incense based on high-resolution mass spectrometry technology |
| CN114527208A (en) * | 2022-01-27 | 2022-05-24 | 鲁南厚普制药有限公司 | Method for establishing HPLC fingerprint of Jingfang preparation and standard fingerprint thereof |
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