CN104345117A - Qualitative and quantitative detection method of compound Japanese Ardisia Herb tablets - Google Patents
Qualitative and quantitative detection method of compound Japanese Ardisia Herb tablets Download PDFInfo
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- CN104345117A CN104345117A CN201310318365.4A CN201310318365A CN104345117A CN 104345117 A CN104345117 A CN 104345117A CN 201310318365 A CN201310318365 A CN 201310318365A CN 104345117 A CN104345117 A CN 104345117A
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The present invention discloses a qualitative and quantitative detection method of compound Japanese Ardisia Herb tablets, wherein microscopic identification of Japanese Ardisia Herb in the compound Japanese Ardisia Herb tablets is improved, thin-layer chromatography identification of Japanese Ardisia Herb, Dendranthema indicum and licorice root in the compound Japanese Ardisia Herb tablets is added, and a determination method of the bergenin content in the compound Japanese Ardisia Herb tablets is established. According to the qualitative and quantitative detection method, the identification is provided, wherein the microscopic identification method is adopted to identify the Japanese Ardisia Herb, and the thin-layer chromatography method is adopted to identify the Japanese Ardisia Herb, the Dendranthema indicum and the licorice root; the content determination is provided, wherein the high performance liquid chromatography method is adopted to determine the bergenin content in the Japanese Ardisia Herb; and the Japanese Ardisia Herb content in each compound Japanese Ardisia Herb tablet is not less than 1.5 mg (calculated as the bergenin (C14H16O9)). According to the present invention, with the method, the product quality controllability can be increased, and the shortcomings in the prior art can be overcome so as to well ensure the efficacy and provide good service for the majority of patients.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, be specifically related to a kind of method of quality control of composite ardisia herb piece sheet.
Background technology
Composite ardisia herb piece sheet is clearing heat and detoxicating, the medicine of preventing phlegm from forming and stopping coughing, clinically for the disease such as cough with lung heat and chronic bronchitis.Its proper mass standard is ministerial standard, standard number: WS
3-B-1793-94, only have 1 microscopical characters and 2 chemistry to differentiate in primary standard, this standard is difficult to effective control for product quality.Chinese Pharmaceutical Affairs the 24th volume the 10th phase in 2010 " HPLC method measures Determination of Bergenin in composite ardisia herb piece sheet " describes the method adopting HPLC method to measure Determination of Bergenin in composite ardisia herb piece sheet, China's medicine company the 18th volume the 22nd phase in 2009 " in rp-hplc determination composite ardisia herb piece capsule Determination of Bergenin " establishes the RP-HPLC method of Determination of Bergenin in a kind of composite ardisia herb piece capsule, Hunan Journal of Traditional Chinese Medicine the 25th volume the 6th phase in 2009 " HPLC method measures the content of Bergenin in composite ardisia herb piece sheet " describes the method that HPLC method measures the content of Bergenin in composite ardisia herb piece sheet, inventor investigates through Extraction solvent contrast, adopt the extraction ratio of methyl alcohol inferior to the extraction ratio of 80% methyl alcohol adopted in the present invention in these three sections of documents.Chinese experimental pharmacology of traditional Chinese medical formulae magazine the 16th volume the 9th phase in 2010 " HPLC measures the content of Bergenin in composite ardisia herb piece sheet " describes the method that HPLC method measures the content of Bergenin in composite ardisia herb piece sheet.Said method is all only establish Determination of Bergenin assay method, cannot carry out controlling reliably to the quality of the pharmaceutical preparations comprehensively.
Must, existing method for qualitative and quantitative detection effectively can not control the quality of composite ardisia herb piece sheet, thus will affect the production of this product and quality assurance and clinical efficacy thereof.
summary of the invention:
The object of this invention is to provide a kind of method for qualitative and quantitative detection of composite ardisia herb piece sheet, comprise the microscopical characters of ardisia japonica in composite ardisia herb piece sheet, the TLC distinguish of ardisia japonica, mother chrysanthemum, Radix Glycyrrhizae, the content assaying method of Bergenin in composite ardisia herb piece sheet; Differentiate: adopt microscopical characters method to differentiate ardisia japonica, thin-layered chromatography discriminating ardisia japonica, mother chrysanthemum, Radix Glycyrrhizae; Assay: the content adopting Bergenin in high effective liquid chromatography for measuring ardisia japonica; Every sheet contains ardisia japonica with Bergenin (C
14h
16o
9) must not count and be less than 1.5mg.The inventive method can increase the quality controllability of product, overcomes the deficiencies in the prior art, better can ensure curative effect, is better extensive patients service.
The object of the invention is to be achieved through the following technical solutions: a kind of method for qualitative and quantitative detection of composite ardisia herb piece sheet, (1) adopts microscopical characters ardisia japonica, adopts thin-layered chromatography to differentiate ardisia japonica, mother chrysanthemum, Radix Glycyrrhizae; (2) adopt the content of Bergenin in high effective liquid chromatography for measuring ardisia japonica, comprise following steps:
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 3 ~ 5 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 1 ~ 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Another extracting Radix Glycyrrhizae control medicinal material 1g, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color.
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 3000 by Bergenin peak.
The preparation of reference substance solution gets Bergenin reference substance in right amount, accurately weighed, adds 80% methyl alcohol and makes the solution of every 1ml containing 50 μ g, to obtain final product.
This product 10 is got in the preparation of need testing solution, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
The every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) meter, must not 1.5mg be less than.
For better showing essence of the present invention, by methodological study of the present invention, details are as follows:
(1) microscopical characters of ardisia japonica
Get this product, put basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).The results are shown in Figure 1.
(2) indentification by TLC of ardisia japonica
1. the preparation of need testing solution: get this product 2, removing dressing, porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.
2. the preparation of reference substance solution: get Bergenin reference substance appropriate, adds methyl alcohol and makes the solution of every 1ml containing 1mg, product solution in contrast.
3. the preparation of negative sample solution: by prescription proportioning, gets other taste medicinal materials of removing ardisia japonica, makes sample, then obtain negative sample solution by above-mentioned need testing solution preparation method by the technique under method for making item.
4. thin-layer chromatography condition and result: according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VIB) test, get above-mentioned three kinds of solution 3 ~ 5 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color, the negative control lacking ardisia japonica is noiseless, the results are shown in Figure 2.
(3) indentification by TLC of mother chrysanthemum
1. the preparation of need testing solution: get this product 3, removing dressing, porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.
2. the preparation of reference substance solution: get linarin reference substance appropriate, adds methyl alcohol and makes the solution of every 1ml containing 0.2mg, product solution in contrast.
3. the preparation of negative sample solution: by prescription proportioning, gets other taste medicinal materials of removing mother chrysanthemum, makes sample, then obtain negative sample solution by above-mentioned need testing solution preparation method by the technique under method for making item.
4. thin-layer chromatography condition and result: according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VIB) test, draw above-mentioned three kinds of solution 1 ~ 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color, the negative control lacking mother chrysanthemum is noiseless, the results are shown in Figure 3.
(4) indentification by TLC of Radix Glycyrrhizae
1. the preparation of need testing solution: get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.
2. the preparation of control medicinal material solution: extracting Radix Glycyrrhizae control medicinal material 1g, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discard ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, get normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, medicinal material solution in contrast.
3. the preparation of negative sample solution: by prescription proportioning, gets other taste medicinal materials of removing Radix Glycyrrhizae, makes sample, then obtain negative sample solution by above-mentioned need testing solution preparation method by the technique under method for making item.
4. thin-layer chromatography condition and result: according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B) test, draw each 3 ~ 5 μ l of above-mentioned three kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color, the negative control lacking Radix Glycyrrhizae is noiseless, the results are shown in Figure 4.
(3) Determination of Bergenin measures
1. instrument and reagent: Wasters 1525 high performance liquid chromatograph; CG-300 ultrasonic cleaning instrument (300W, 25kHz); Balance: AE240, CP225D; Wasters 2996(PDA); Bergenin (Nat'l Pharmaceutical & Biological Products Control Institute provides, for assay, and lot number: 111532-200202).Methyl alcohol is chromatographically pure, and it is pure that other reagent are analysis.
2. system flexibility: chromatographic column: Agient C
18(DB) (250mm × 4.6mm × 5 μm), Agilent company; Column temperature: 35 DEG C; Mobile phase: methanol-water (20: 80); Flow velocity 1.0ml/min; Determined wavelength 275nm; Chromatographic work station Empower.
3. the preparation of reference substance solution: precision takes Bergenin reference substance 24.79mg, puts in 50ml measuring bottle, adds 80% methyl alcohol and makes dissolving in right amount, adds 80% methanol dilution to scale, shakes up, and obtains reference substance stock solution (C=0.4958mg/ml).Precision measures stock solution 10ml, puts in 100ml measuring bottle, adds 80% methanol dilution to scale, shakes up, and obtains reference substance solution (C=0.04958mg/ml).Accurate absorption 10 μ l, injection liquid chromatography, measures, sees Fig. 5.
4. the preparation of need testing solution
(lot number: 20110714), removes dressing, porphyrize, for subsequent use in right amount to get this product.
5. the selection of Extraction solvent
Get above-mentioned powder 0.15g, totally 8 parts, accurately weighed, put in 25ml measuring bottle, add methyl alcohol, 80% methyl alcohol, 50% methyl alcohol, 20% methyl alcohol 20ml, ultrasonic 45 minutes, let cool, supplement corresponding solvent to scale, shake up, filter, get subsequent filtrate, to obtain final product.Accurate absorption reference substance solution 10 μ l and need testing solution 10 μ l, injection liquid chromatography, measures and calculates, the results are shown in Table 1.
table 1 Extraction solvent selection result comparison sheet
Result shows, with 4 kinds of solvent extraction test samples, it is higher that 80% methyl alcohol and 50% methyl alcohol record content results, but 50% methyl alcohol process gained need testing solution is difficult to filter, and therefore, selects 80% methyl alcohol to be Extraction solvent.
the investigation of extracting method, extraction time
Get above-mentioned powder 0.15g, totally 8 parts, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (300W, 40kHz) 15,30,45,60 minutes, lets cool, supplies the weight of less loss with 80% methyl alcohol respectively, shake up, filter, get subsequent filtrate, to obtain final product.
Get above-mentioned powder 0.15g, totally 2 parts, accurately weighed, put in tool plug conical flask, precision adds 80% methyl alcohol 25ml, close plug, weighed weight, and refluxing extraction 30 minutes, lets cool, and supplies the weight of less loss, shake up with 80% methyl alcohol, filters, gets subsequent filtrate, to obtain final product.
Determination method precision draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, and calculates, the results are shown in Table 2.
table 2 extracting method, selection of time results contrast table
Result shows, within ultrasonic 45 minutes, yield is higher, therefore selects extract for ultrasonic 45 minutes.
7. the preparation of need testing solution: get this product 10, removing dressing, accurately weighed, porphyrize, gets about 0.15g, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (300W, 40kHz) 45 minutes, lets cool, supply the weight of less loss with 80% methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.Accurate absorption 10 μ l, injection liquid chromatography, measures, sees Fig. 6.
the preparation of negative solution: all the other each flavour of a drug taking scarce ardisia japonica by prescription are appropriate, make the negative control sample of scarce ardisia japonica, with the standby negative control solution lacking ardisia japonica of legal system.Accurate absorption 10 μ l, injection liquid chromatography, measures, and the negative control lacking ardisia japonica is noiseless, the results are shown in Figure 7.
precision test: (each 10 μ l, measure, the RSD of its Bergenin integrating peak areas value is 0.68% to the same need testing solution of accurate absorption, the results are shown in Table 3, and result shows that sample introduction precision is good for lot number: 20110714), sample introduction 6 times.
table 3 Precision test result table
stability test
get same need testing solution, respectively at 0,2,4,6,12,24 hour, accurate absorption 10 μ l, injection liquid chromatography, measured, the results are shown in Table 4.Result shows: need testing solution is basicly stable in 24 hours.
table 4 stability test result table
linear relationship is investigated precision and is taken Bergenin reference substance 19.98mg, puts in 50ml measuring bottle, adds 80% methyl alcohol and make dissolving and be diluted to scale, shake up, obtain stock solution (0.3996mg/ml).Respectively accurate draw stock solution 0.5,1,2,3,5,10ml to 25,25,25,25,10, in 10ml measuring bottle, with 80% methanol dilution to scale, shake up, obtain control series product solution (0.007992,0.01598,0.03197,0.04795,0.1998,0.3996mg/ml), the each 10 μ l of the above-mentioned solution of accurate absorption, inject high performance liquid chromatograph, measure, take peak area as ordinate, sample size is horizontal ordinate drawing standard curve.Result shows: Bergenin reference substance is good in 0.07992 ~ 3.996 μ g scope internal linear relation, and its regression equation is: Y=1316272.5302X-4649.9257, r=0.9999, the results are shown in Table 5, Fig. 7.
table 5 linear relationship investigates result table
replica test gets same lot number (20071114) sample, prepares 5 parts of need testing solutions in accordance with the law, measures, the results are shown in Table 6.Result shows: this method repeatability better.
table 6 replica test result
recovery test and scope demonstration test get 3 volumetric flasks, respectively take same lot number (20110714) sample powder (containing Bergenin 7.993mg/g) the about 0.1g having measured content, accurately weighed, it is 0.3996mg/ml Bergenin reference substance solution 2ml that each precision adds concentration, prepares need testing solution in accordance with the law; Get 3 volumetric flasks, respectively take same lot number (20110714) sample powder (containing Bergenin 7.993mg/g) the about 0.15g having measured content, accurately weighed, it is 0.3996mg/ml Bergenin reference substance solution 2ml that each precision adds concentration, prepares need testing solution in accordance with the law; Get 3 volumetric flasks, respectively take same lot number (20110714) sample powder (containing Bergenin 7.993mg/g) the about 0.2g having measured content, accurately weighed, it is 0.3996mg/ml Bergenin reference substance solution 2ml that each precision adds concentration, prepares need testing solution in accordance with the law.
Measure by above-mentioned chromatographic condition, sample size is 10 μ l, and calculate the recovery, result shows, this law recovery is good, the results are shown in Table 7.
table 7 recovery test result
sample size measures: carry out assay to all the other batch sample, the results are shown in Table 8.
determination of Bergenin measurement result in table 8 composite ardisia herb piece sheet
Beneficial effect of the present invention: the present invention adopts microscopical characters ardisia japonica, thin-layered chromatography differentiates ardisia japonica, mother chrysanthemum, Radix Glycyrrhizae, adopts the Determination of Bergenin in high effective liquid chromatography for measuring ardisia japonica; By setting up the strong discrimination method of clear and definite specificity and the good content assaying method of reappearance, stability and precision, effectively can control the quality of composite ardisia herb piece sheet, composite ardisia herb piece tablet quality is made to reach stable, controlled, to product feed intake and quality control requirement stricter, overcome the deficiencies in the prior art, improve quality, the curative effect of product, meet the needs of medical treatment better.
accompanying drawing illustrates:
Fig. 1 is the microscopical characters figure of ardisia japonica in composite ardisia herb piece sheet of the present invention;
Fig. 2 is the TLC distinguish figure of ardisia japonica in composite ardisia herb piece sheet of the present invention;
Fig. 3 is the TLC distinguish figure of mother chrysanthemum in composite ardisia herb piece sheet of the present invention;
Fig. 4 is the TLC distinguish figure of Radix Glycyrrhizae in composite ardisia herb piece sheet of the present invention;
Fig. 5 is the high-efficient liquid phase chromatogram of Bergenin reference substance of the present invention;
Fig. 6 is the high-efficient liquid phase chromatogram of Determination of Bergenin in composite ardisia herb piece sheet sample of the present invention;
Fig. 7 is the high-efficient liquid phase chromatogram that composite ardisia herb piece sheet of the present invention lacks ardisia japonica negative control solution;
Fig. 8 is that composite ardisia herb piece sheet Determination of Bergenin of the present invention measures linear graph;
Embodiment
embodiment 1
Composite ardisia herb piece sheet (Film coated tablets) lot number: 20111204
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 3 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 1 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Another extracting Radix Glycyrrhizae control medicinal material 1g, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 3 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color.
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 3000 by Bergenin peak.
The preparation of reference substance solution gets Bergenin reference substance in right amount, accurately weighed, adds 80% methyl alcohol and makes the solution of every 1ml containing 50 μ g, to obtain final product.
This product 10 is got in the preparation of need testing solution, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Assay result this batch of every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) count 1.8mg.
embodiment 2
Composite ardisia herb piece sheet (Film coated tablets) lot number: 20111205
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 5 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Another extracting Radix Glycyrrhizae control medicinal material 1g, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color.
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 3000 by Bergenin peak.
The preparation of reference substance solution gets Bergenin reference substance in right amount, accurately weighed, adds 80% methyl alcohol and makes the solution of every 1ml containing 50 μ g, to obtain final product.
This product 10 is got in the preparation of need testing solution, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Assay result this batch of every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) count 1.9mg.
embodiment 3
Composite ardisia herb piece sheet (sugar coated tablet) lot number: 20111201
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 4 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Another extracting Radix Glycyrrhizae control medicinal material 1g, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 4 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color.
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 3000 by Bergenin peak.
The preparation of reference substance solution gets Bergenin reference substance in right amount, accurately weighed, adds 80% methyl alcohol and makes the solution of every 1ml containing 50 μ g, to obtain final product.
This product 10 is got in the preparation of need testing solution, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Assay result this batch of every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) count 1.8mg.
embodiment 4
Composite ardisia herb piece sheet (sugar coated tablet) lot number: 20111202
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica).
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 3 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color.
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution.Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color.
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Another extracting Radix Glycyrrhizae control medicinal material 1g, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 5 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm).In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color.
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 3000 by Bergenin peak.
The preparation of reference substance solution gets Bergenin reference substance in right amount, accurately weighed, adds 80% methyl alcohol and makes the solution of every 1ml containing 50 μ g, to obtain final product.
This product 10 is got in the preparation of need testing solution, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product.
Determination method is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
Assay result this batch of every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) count 1.9mg.
Claims (1)
1. the method for qualitative and quantitative detection of a composite ardisia herb piece sheet, it is characterized in that comprising (1) adopts microscopical characters ardisia japonica, thin-layered chromatography is adopted to differentiate ardisia japonica, mother chrysanthemum, Radix Glycyrrhizae, (2) adopt the content of Bergenin in high effective liquid chromatography for measuring ardisia japonica, described method for qualitative and quantitative detection comprises following steps:
(1) differentiate:
The microscopical characters of ardisia japonica comprises the following steps: to get this product, puts basis of microscopic observation: spiral duct diameter 7.5 ~ 25 μm, fiber bunchy, and wall is thicker, calcium oxalate prismatic crystal diameter 7.5 ~ 26 μm (ardisia japonica);
The TLC distinguish of ardisia japonica comprises the following steps: to get this product 2, removing dressing, and porphyrize, adds methyl alcohol 20ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get Bergenin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast; Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 3 ~ 5 μ l, put respectively on same silica gel g thin-layer plate, with sherwood oil (60 ~ 90 DEG C)-acetate-methanol (9: 8: 2) for developping agent, launch, take out, dry, spray with the mixed solution of 1% ferric trichloride-1% potassium ferricyanide (1: 1); In test sample chromatogram, on the position corresponding to reference substance chromatogram, the spot of aobvious same color;
The TLC distinguish of mother chrysanthemum comprises the following steps: to get this product 3, removing dressing, and porphyrize, adds methyl alcohol 15ml, ultrasonic process 30 minutes, and filter, filtrate is concentrated into about 2ml, as need testing solution; Separately get linarin reference substance appropriate, add methyl alcohol and make the solution of every 1ml containing 0.2mg, product solution in contrast; Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw above-mentioned two kinds of solution 1 ~ 2 μ l, put respectively on same polyamide film, with ethyl acetate-butanone-methenyl choloride-formic acid-water (15: 15: 6: 4: 1) for developping agent, launch, take out, dry, spray, with 2% aluminum trichloride solution, is heated to spot development at 105 DEG C clear, inspects under putting ultraviolet lamp (365nm); In test sample chromatogram, on the position corresponding to reference substance chromatogram, the fluorescence spot of aobvious same color;
The discriminating of Radix Glycyrrhizae comprises the following steps: to get this product 20, removing dressing, porphyrize, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discards ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, gets normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution; Another extracting Radix Glycyrrhizae control medicinal material 1g, add diethyl ether 40ml, ultrasonic process 15 minutes, filters, discard ether solution, the dregs of a decoction add methyl alcohol 50ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, the residue 40ml that adds water makes dissolving, extract 3 times in order to water saturated normal butyl alcohol jolting, each 20ml, merge n-butanol extracting liquid, in order to the saturated water washing of normal butyl alcohol 3 times, each 20ml, discards water liquid, get normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, medicinal material solution in contrast; Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VI B), draw each 3 ~ 5 μ l of above-mentioned two kinds of solution, put on same silica gel g thin-layer plate respectively, make into strips, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) for developping agent, launch, take out, dry, spray with 10% ethanol solution of sulfuric acid, 105 DEG C to be heated to spot development clear, inspects under putting ultraviolet lamp (365nm); In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence spot of aobvious same color;
(2) assay:
In ardisia japonica, the content assaying method of Bergenin comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D);
Chromatographic condition and system suitability: be filling agent with octadecylsilane chemically bonded silica; With methanol-water (20: 80) for mobile phase; Determined wavelength is 275nm, and number of theoretical plate calculates should be not less than 3000 by Bergenin peak;
The preparation of reference substance solution: get Bergenin reference substance appropriate, accurately weighed, add 80% methyl alcohol and make the solution of every 1ml containing 50 μ g, to obtain final product;
The preparation of need testing solution: get this product 10, removing dressing, accurately weighed, porphyrize, gets about 0.15g, accurately weighed, put in 25ml measuring bottle, add 80% methyl alcohol 20ml, ultrasonic process (power 300W, frequency 40kHz) 45 minutes, let cool, add 80% methanol dilution to scale, shake up, filter, get subsequent filtrate, to obtain final product:
Determination method: accurate absorption reference substance solution and each 10 μ l of need testing solution respectively, injection liquid chromatography, measures, to obtain final product;
The every sheet of composite ardisia herb piece sheet contains ardisia japonica with Bergenin (C
14h
16o
9) meter, must not 1.5mg be less than.
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|---|---|---|---|---|
| CN107184623A (en) * | 2017-07-06 | 2017-09-22 | 天津中医药大学 | A kind of preparation method and applications of ardisia japonica alcohol extract |
| CN111686085A (en) * | 2020-06-30 | 2020-09-22 | 贵州益佰女子大药厂有限责任公司 | Preparation method of throat-clearing preparation |
| CN115015418A (en) * | 2022-06-01 | 2022-09-06 | 湖南新汇制药股份有限公司 | Quality detection method for Japanese ardisia herb decoction |
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| CN112067739B (en) * | 2020-10-13 | 2022-04-19 | 劲牌持正堂药业有限公司 | Identification method and application of thin-layer chromatography of wild chrysanthemum flower medicinal material and formula granules |
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Application publication date: 20150211 |