CN106680414A - Detection method of compound ardisia japonica tablet - Google Patents
Detection method of compound ardisia japonica tablet Download PDFInfo
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Abstract
The invention discloses a detection method of a compound ardisia japonica tablet. Microscopic identification of ardisia japonica in the compound ardisia japonica tablet is improved, thin-layer identification of the ardisia japonica, wild chrysanthemum flowers and liquorice in the compound ardisia japonica tablet is added, and a method for measuring the content of bergenin in the compound ardisia japonica tablet is built. The ardisia japonica is identified by adopting a microscopic identification method, the ardisia japonica, the wild chrysanthemum flowers and the liquorice are identified by adopting a thin-layer chromatography; the content of the bergenin in the ardisia japonica is measured by adopting a high-performance liquid chromatography; each tablet contains not less than 1.5mg based on bergenin (C14H16O9). According to the method, the quality controllability of a product can be improved, the defects of the prior art are overcome, the curative effect can be better ensured and the majority of patients can be better served.
Description
The application is the divisional application of the patent of invention to Patent No. 201310318365.4.The applying date of original application is
On July 26th, 2013, Application No. 201310318365.4, invention and created name is a kind of qualitative, quantitative of composite ardisia herb piece piece
Detection method.
Technical field
The invention belongs to technical field of Chinese medicines, and in particular to a kind of method of quality control of composite ardisia herb piece piece.
Background technology
Composite ardisia herb piece piece is heat-clearing and toxic substances removing, the medicine of preventing phlegm from forming and stopping coughing, clinically for cough due to lung-heat and chronic tracheitiss
Deng disease.Its proper mass standard be ministry standard, standard number：WS3- B-1793-94, only has 1 microscopical identification and 2 in primary standard
Item chemistry differentiates that the standard is difficult to effective control for product quality.Chinese Pharmaceutical Affairs 2010 year the 10th phase of volume 24《HPLC methods determine multiple
Determination of Bergenin in square Herba Ardisiae Japonicae piece》Describe the side that Determination of Bergenin in composite ardisia herb piece piece is determined using HPLC methods
Method, Chinese Pharmaceutical 2009 year the 22nd phase of volume 18《Bergeninum in rp-hplc determination composite ardisia herb piece capsule
Content》Establish a kind of RP-HPLC methods of Determination of Bergenin in composite ardisia herb piece capsule, Hunan Journal of Traditional Chinese Medicine 2009 the 25th
Rolled up for the 6th phase《HPLC methods determine the content of Bergeninum in composite ardisia herb piece piece》Describe HPLC methods to determine in composite ardisia herb piece piece
The method of the content of Bergeninum, inventor investigates through Extraction solvent contrast, using the extraction ratio of methanol in this three documents
Inferior to the extraction ratio of 80% methanol adopted in the present invention.Chinese experimental pharmacology of Chinese medical formulae magazine 2010 year the 9th phase of volume 16《HPLC is surveyed
Determine the content of Bergeninum in composite ardisia herb piece piece》Describe HPLC methods and determine the content of Bergeninum in composite ardisia herb piece piece
Method.Said method is all only to establish Determination of Bergenin assay method, it is impossible to reliable control comprehensively is carried out to the quality of the pharmaceutical preparations
System.
Generally speaking, existing method for qualitative and quantitative detection can not effectively control the quality of composite ardisia herb piece piece, so as to
Production and quality assurance and its clinical efficacy of the product will be affected.
The content of the invention:
It is an object of the invention to provide a kind of method for qualitative and quantitative detection of composite ardisia herb piece piece, including composite ardisia herb piece piece
The microscopical identification of middle Herba Ardisiae Japonicae, Herba Ardisiae Japonicae, Flos Chrysanthemi Indici, the thin layer discriminating of Radix Glycyrrhizae, the content of Bergeninum in composite ardisia herb piece piece
Assay method；Differentiate：Differentiate that Herba Ardisiae Japonicae, thin layer chromatography differentiate Herba Ardisiae Japonicae, Flos Chrysanthemi Indici, Radix Glycyrrhizae using microscopical identification method；Content
Determine：Using the content of Bergeninum in high effective liquid chromatography for measuring Herba Ardisiae Japonicae；Per piece containing Herba Ardisiae Japonicae with Bergeninum
(C14H16O9) must not count less than 1.5mg.The inventive method can increase the quality controllability of product, overcome the deficiencies in the prior art,
It is preferably extensive patients service so as to preferably ensure curative effect.
The purpose of the present invention is achieved by the following technical solution：A kind of qualitative and quantitative detection side of composite ardisia herb piece piece
Method, (1) differentiates Herba Ardisiae Japonicae, Flos Chrysanthemi Indici, Radix Glycyrrhizae using microscopical identification Herba Ardisiae Japonicae using thin layer chromatography；(2) efficient liquid phase is adopted
The content of Bergeninum, comprises the steps of in chromatography determination Herba Ardisiae Japonicae：
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：Spiral duct diameter 7.5~
25 μm, fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae).
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take Bergeninum reference substance appropriate, plus methanol
Solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 3~5, puts respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid
Ethyl ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixed of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Close solution.In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color.
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take linarin reference substance appropriate, plus methanol system
Into solution of every 1ml containing 0.2mg, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 1~2, puts respectively on same polyamide film, with ethyl acetate-chloromethane of butanone-three
Alkane-formic acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, is added at 105 DEG C
Heat to spot development is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, in position corresponding with reference substance chromatograph
Put, show the fluorescence spot of same color.
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, supersound process 15
Minute, filtration discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and residue adds water 40ml
Dissolving is made, is extracted 3 times to the shaking of water saturated n-butyl alcohol, each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.Another extracting liquorice control medicinal material 1g, is made in the same way of control medicinal material solution.According to thin layer chromatography (Chinese Pharmacopoeia version one in 2010
The B of annex VI) test, each 3~5 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into strips, with second
Acetoacetic ester-formic acid-glacial acetic acid-water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid,
105 DEG C are heated to spot development clearly, put and inspected under ultra-violet lamp (365nm).In test sample chromatograph, with control medicinal material chromatograph
On corresponding position, show the fluorescence spot of same color.
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia
One D of annex VI of version in 2010) determine.
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler；With methanol-water (20:
80) it is mobile phase；Detection wavelength is 275nm.Number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000.
The preparation of reference substance solution takes Bergeninum reference substance in right amount, accurately weighed, plus 80% methanol is made every 1ml and contained
The solution of 50 μ g, obtains final product.
The preparation of need testing solution takes this product 10, removes coating, accurately weighed, finely ground, takes about 0.15g, and precision claims
It is fixed, in putting 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80%
Methanol dilution shakes up to scale, filtration, takes subsequent filtrate, obtains final product.
Algoscopy is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines,
Obtain final product.
Composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) meter, 1.5mg must not be less than.
Preferably to show the essence of the present invention, by method of the present invention investigation, details are as follows：
(1) microscopical identification of Herba Ardisiae Japonicae
This product is taken, basis of microscopic observation is put：7.5~25 μm of spiral duct diameter, fiber bunchy, wall is thicker, calcium oxalate side
7.5~26 μm of brilliant diameter (Herba Ardisiae Japonicae).As a result Fig. 1 is seen.
(2) indentification by TLC of Herba Ardisiae Japonicae
1. the preparation of need testing solution：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, supersound process 30 minutes,
Filtration, filtrate is concentrated into about 2ml, used as need testing solution.
2. the preparation of reference substance solution：Take Bergeninum reference substance in right amount, plus methanol makes solution of every 1ml containing 1mg, makees
For reference substance solution.
3. the preparation of negative sample solution：By prescription proportioning, other taste medical materials of Herba Ardisiae Japonicae are removed, by under preparation method item
Technique makes sample, then negative sample solution is obtained by above-mentioned need testing solution preparation method.
4. thin layer chromatography condition and result：According to thin layer chromatography (Chinese Pharmacopoeia one annex VIB of version in 2010) test, take
The above-mentioned μ l of three kinds of solution 3~5, put respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetate-methanol
(9: 8: 2) are developing solvent, are launched, and are taken out, and are dried, and are sprayed with the mixed solution of the potassium ferricyanide of 1% ferric chloride -1% (1: 1).For
In test product chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color, lack the negative control of Herba Ardisiae Japonicae without dry
Disturb, as a result see Fig. 2.
(3) indentification by TLC of Flos Chrysanthemi Indici
1. the preparation of need testing solution：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, supersound process 30 minutes,
Filtration, filtrate is concentrated into about 2ml, used as need testing solution.
2. the preparation of reference substance solution：Take linarin reference substance in right amount, plus methanol makes solution of every 1ml containing 0.2mg, makees
For reference substance solution.
3. the preparation of negative sample solution：By prescription proportioning, other taste medical materials of Flos Chrysanthemi Indici are removed, by under preparation method item
Technique makes sample, then negative sample solution is obtained by above-mentioned need testing solution preparation method.
4. thin layer chromatography condition and result：According to thin layer chromatography (Chinese Pharmacopoeia one annex VIB of version in 2010) test, inhale
Take the above-mentioned μ l of three kinds of solution 1~2, put respectively on same polyamide film, with ethyl acetate-butanone-chloroform-formic acid-
Water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, and at 105 DEG C speckle is heated to
Colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with reference substance chromatograph, show phase
With the speckle of color, the negative control for lacking Flos Chrysanthemi Indici is noiseless, as a result sees Fig. 3.
(4) indentification by TLC of Radix Glycyrrhizae
1. the preparation of need testing solution：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, 15 points of supersound process
Clock, filtration, discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and the residue 40ml that adds water makes
Dissolving, extracts 3 times to the shaking of water saturated n-butyl alcohol, and each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.
2. the preparation of control medicinal material solution：Extracting liquorice control medicinal material 1g, add diethyl ether 40ml, supersound process 15 minutes, filtration,
Ether solution is discarded, medicinal residues add methanol 50ml, be heated to reflux 1 hour, filtered, filtrate is evaporated, the residue 40ml that adds water makes dissolving, to
Water saturated n-butyl alcohol shaking is extracted 3 times, and each 20ml merges n-butanol extracting liquid, to the water washing 3 of n-butyl alcohol saturation
Secondary, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, used as control medicinal material solution.
3. the preparation of negative sample solution：By prescription proportioning, other taste medical materials of Radix Glycyrrhizae are removed, by the work under preparation method item
Skill makes sample, then negative sample solution is obtained by above-mentioned need testing solution preparation method.
4. thin layer chromatography condition and result：According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010) test, inhale
Each 3~5 μ l of above-mentioned three kinds of solution are taken, is put on same silica gel g thin-layer plate respectively, made into strips, with acetic ether-methanoic acid-ice
Acetic Acid-Water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid, and 105 DEG C are heated to speckle
Colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with control medicinal material chromatograph, show
The fluorescence spot of same color, the negative control for lacking Radix Glycyrrhizae is noiseless, as a result sees Fig. 4.
(3) Determination of Bergenin is determined
1. instrument and reagent：The high performance liquid chromatographs of Wasters 1525；CG-300 is cleaned by ultrasonic instrument (300W, 25kHz)；
Balance：AE240, CP225D；Wasters 2996(PDA)；(Nat'l Pharmaceutical ＆ Biological Products Control Institute provides Bergeninum, for containing
Measure fixed use, lot number：111532-200202).Methanol is chromatographically pure, and it is pure that other reagents are analysis.
2. system suitability：Chromatographic column：Agient C18(DB) (250mm × 4.6mm × 5 μm), Agilent companies；Post
Temperature：35℃；Mobile phase：Methanol-water (20: 80)；Flow velocity 1.0ml/min；Detection wavelength 275nm；Chromatographic work station Empower.
3. the preparation of reference substance solution：Precision weighs Bergeninum reference substance 24.79mg, in putting 50ml measuring bottles, plus 80%
Methanol makes in right amount dissolving, plus 80% methanol dilution shake up to scale, obtains final product reference substance stock solution (C=0.4958mg/ml).Essence
Close to measure stock solution 10ml, in putting 100ml measuring bottles, plus 80% methanol dilution shakes up to scale, obtains final product reference substance solution (C=
0.04958mg/ml).Precision draws 10 μ l, injects chromatograph of liquid, determines, and sees Fig. 5.
4. the preparation of need testing solution
Take the appropriate (lot number of this product：20110714) coating, is removed, it is finely ground, it is standby.
5. the selection of Extraction solvent
Above-mentioned powder 0.15g is taken, it is totally 8 parts, accurately weighed, in putting 25ml measuring bottles, add methanol, 80% methanol, 50% first
Alcohol, 20% methanol 20ml, ultrasound 45 minutes, let cool, and the corresponding solvent of supplement shakes up to scale, filters, and takes subsequent filtrate, obtains final product.
Precision draws the μ l of the reference substance solution 10 and μ l of need testing solution 10, injects chromatograph of liquid, determines and calculates, and the results are shown in Table 1.
The Extraction solvent selection result comparison sheet of table 1
As a result show, with 4 kinds of solvent extraction test samples, it is higher that 80% methanol and 50% methanol measure content results, but
It is that 50% methyl alcohol process gained need testing solution is difficult to filter, therefore, 80% methanol of selection is Extraction solvent.
6. the investigation of extracting method, extraction time
Above-mentioned powder 0.15g is taken, it is totally 8 parts, accurately weighed, in putting 25ml measuring bottles, 80% methanol 20ml is added, respectively ultrasound
Process (300W, 40kHz) 15,30,45,60 minutes, let cool, with 80% methanol the weight of less loss is supplied, shake up, filter, take continuous
Filtrate, obtains final product.
Above-mentioned powder 0.15g is taken, totally 2 parts, accurately weighed, in putting conical flask with cover, precision adds 80% methanol 25ml, close
Plug, weighed weight, reflux, extract, 30 minutes lets cool, and with 80% methanol the weight of less loss is supplied, and shakes up, and filters, and takes subsequent filtrate,
Obtain final product.
Algoscopy is accurate to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines, and counts
Calculate, the results are shown in Table 2.
The extracting method of table 2, selection of time results contrast table
As a result show, ultrasonic 45 minutes yield are higher, therefore select ultrasound to extract for 45 minutes.
7. the preparation of need testing solution：This product 10 is taken, coating is removed, it is accurately weighed, it is finely ground, about 0.15g is taken, put 25ml
In measuring bottle, add 80% methanol 20ml, supersound process (300W, 40kHz) 45 minutes to let cool, with 80% methanol less loss is supplied
Weight, shakes up, filtration, takes subsequent filtrate, obtains final product.Precision draws 10 μ l, injects chromatograph of liquid, determines, and sees Fig. 6.
8. the preparation of negative solution：Remaining each flavour of a drug for weighing scarce Herba Ardisiae Japonicae by prescription are appropriate, make the moon of scarce Herba Ardisiae Japonicae
Property control sample, with method prepare lack Herba Ardisiae Japonicae negative control solution.Precision draws 10 μ l, injects chromatograph of liquid, determines, and lacks
The negative control of Herba Ardisiae Japonicae is noiseless, as a result sees Fig. 7.
9. precision test：Precision draws same need testing solution (lot number：20110714), sample introduction 6 times, 10 μ l every time,
Determine, the RSD of its Bergeninum integrating peak areas value is 0.68%, the results are shown in Table 3, as a result shows that sample introduction precision is good.
The Precision test result table of table 3
10. stability test takes same need testing solution, and respectively at 0,2,4,6,12,24 hours, precision drew 10 μ l, note
Enter chromatograph of liquid, determine, the results are shown in Table 4.As a result show：Need testing solution is basicly stable in 24 hours.
The stability test result table of table 4
Linear relationship investigates precision and weighs Bergeninum reference substance 19.98mg, in putting 50ml measuring bottles, plus 80% methanol
Make to dissolve and be diluted to scale, shake up, obtain final product stock solution (0.3996mg/ml).Respectively it is accurate draw stock solution 0.5,1,2,3,
5th, 10ml to 25,25,25,25,10, in 10ml measuring bottles, with 80% methanol dilution to scale, shake up, obtain final product control series product molten
Liquid (0.007992,0.01598,0.03197,0.04795,0.1998,0.3996mg/ml), precision draws each 10 μ of above-mentioned solution
L, injects high performance liquid chromatograph, determines, and with peak area as vertical coordinate, sample size is that abscissa draws standard curve.As a result table
It is bright：Bergeninum reference substance linear relationship in 0.07992~3.996 μ g ranges is good, and its regression equation is：Y=
1316272.5302X-4649.9257, r=0.9999 the results are shown in Table 5, Fig. 7.
The linear relationship of table 5 investigates result table
Replica test takes same lot number (20071114) sample, and 5 parts of need testing solutions are prepared in accordance with the law, determines, as a result
It is shown in Table 6.As a result show：This method repeatability is preferably.
The replica test result of table 6
Recovery test and scope checking test take 3 volumetric flasks, respectively weigh the same lot number for having determined content
(20110714) sample powder (7.993mg/g containing Bergeninum) about 0.1g, accurately weighed, and each accurate addition concentration is
0.3996mg/ml Bergeninum reference substance solution 2ml, prepares need testing solution in accordance with the law；3 volumetric flasks are taken, is respectively weighed and is determined
Same lot number (20110714) sample powder (7.993mg/g containing Bergeninum) the about 0.15g of content, accurately weighed, each precision
Addition concentration is 0.3996mg/ml Bergeninum reference substance solution 2ml, and need testing solution is prepared in accordance with the law；3 volumetric flasks are taken, respectively
Same lot number (20110714) sample powder (7.993mg/g containing Bergeninum) the about 0.2g for determining content is weighed, precision claims
Fixed, each accurate addition concentration is 0.3996mg/ml Bergeninum reference substance solution 2ml, and need testing solution is prepared in accordance with the law.
Determine by above-mentioned chromatographic condition, sample size is 10 μ l, calculates the response rate, is as a result shown, this law response rate is good, knot
Fruit is shown in Table 7.
The recovery test result of table 7
Sample size is determined：Assay is carried out to remaining batch sample, 8 are the results are shown in Table.
Determination of Bergenin measurement result in the composite ardisia herb piece piece of table 8
Beneficial effects of the present invention：The present invention differentiates Herba Ardisiae Japonicae, Herba Dendranthematis indici using microscopical identification Herba Ardisiae Japonicae, thin layer chromatography
Flower, Radix Glycyrrhizae, using the Determination of Bergenin in high effective liquid chromatography for measuring Herba Ardisiae Japonicae；It is strong by setting up clear and definite specificity
The good content assaying method of discrimination method and repeatability, stability and precision, is capable of effective control composite ardisia herb piece piece
Quality, reaches composite ardisia herb piece tablet quality stable, controllable, to product feed intake and quality control requirement is stricter, overcome
The deficiencies in the prior art, improve quality, the curative effect of product, preferably meet the needs of medical treatment.
Description of the drawings:
Fig. 1 is the microscopical identification figure of Herba Ardisiae Japonicae in composite ardisia herb piece piece of the invention；
Fig. 2 is the thin layer discriminating figure of Herba Ardisiae Japonicae in composite ardisia herb piece piece of the invention；
Fig. 3 is the thin layer discriminating figure of Flos Chrysanthemi Indici in composite ardisia herb piece piece of the invention；
Fig. 4 is the thin layer discriminating figure of Radix Glycyrrhizae in composite ardisia herb piece piece of the invention；
Fig. 5 is the high-efficient liquid phase chromatogram of the Bergeninum reference substance of the present invention；
Fig. 6 is the high-efficient liquid phase chromatogram of Determination of Bergenin in composite ardisia herb piece piece sample of the invention；
Fig. 7 is the high-efficient liquid phase chromatogram that the composite ardisia herb piece piece of the present invention lacks Herba Ardisiae Japonicae negative control solution；
Fig. 8 is that the composite ardisia herb piece piece Determination of Bergenin of the present invention determines linear graph；
Specific embodiment
Embodiment 1
Composite ardisia herb piece piece (Film coated tablets) lot number：20111204
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：Spiral duct diameter 7.5~
25 μm, fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae).
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take Bergeninum reference substance appropriate, plus methanol
Solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 3, puts respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid second
Ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixing of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Solution.In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color.
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take linarin reference substance appropriate, plus methanol system
Into solution of every 1ml containing 0.2mg, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 1, puts respectively on same polyamide film, with ethyl acetate-butanone-chloroform-first
Acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, and at 105 DEG C speckle is heated to
Point colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with reference substance chromatograph, show
The fluorescence spot of same color.
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, supersound process 15
Minute, filtration discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and residue adds water 40ml
Dissolving is made, is extracted 3 times to the shaking of water saturated n-butyl alcohol, each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.Another extracting liquorice control medicinal material 1g, is made in the same way of control medicinal material solution.According to thin layer chromatography (Chinese Pharmacopoeia version one in 2010
The B of annex VI) test, each 3 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into strips, with acetic acid
Ethyl ester-formic acid-glacial acetic acid-water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid, 105
DEG C it is heated to spot development clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, with control medicinal material chromatograph phase
On the position answered, show the fluorescence spot of same color.
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia
One D of annex VI of version in 2010) determine.
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler；With methanol-water (20:
80) it is mobile phase；Detection wavelength is 275nm.Number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000.
The preparation of reference substance solution takes Bergeninum reference substance in right amount, accurately weighed, plus 80% methanol is made every 1ml and contained
The solution of 50 μ g, obtains final product.
The preparation of need testing solution takes this product 10, removes coating, accurately weighed, finely ground, takes about 0.15g, and precision claims
It is fixed, in putting 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80%
Methanol dilution shakes up to scale, filtration, takes subsequent filtrate, obtains final product.
Algoscopy is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines,
Obtain final product.
Assay result this batch composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) be calculated as
1.8mg。
Embodiment 2
Composite ardisia herb piece piece (Film coated tablets) lot number：20111205
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：Spiral duct diameter 7.5~
25 μm, fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae).
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take Bergeninum reference substance appropriate, plus methanol
Solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 5, puts respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid second
Ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixing of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Solution.In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color.
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take linarin reference substance appropriate, plus methanol system
Into solution of every 1ml containing 0.2mg, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 2, puts respectively on same polyamide film, with ethyl acetate-butanone-chloroform-first
Acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, and at 105 DEG C speckle is heated to
Point colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with reference substance chromatograph, show
The fluorescence spot of same color.
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, supersound process 15
Minute, filtration discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and residue adds water 40ml
Dissolving is made, is extracted 3 times to the shaking of water saturated n-butyl alcohol, each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.Another extracting liquorice control medicinal material 1g, is made in the same way of control medicinal material solution.According to thin layer chromatography (Chinese Pharmacopoeia version one in 2010
The B of annex VI) test, each 5 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into strips, with acetic acid
Ethyl ester-formic acid-glacial acetic acid-water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid, 105
DEG C it is heated to spot development clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, with control medicinal material chromatograph phase
On the position answered, show the fluorescence spot of same color.
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia
One D of annex VI of version in 2010) determine.
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler；With methanol-water (20:
80) it is mobile phase；Detection wavelength is 275nm.Number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000.
The preparation of reference substance solution takes Bergeninum reference substance in right amount, accurately weighed, plus 80% methanol is made every 1ml and contained
The solution of 50 μ g, obtains final product.
The preparation of need testing solution takes this product 10, removes coating, accurately weighed, finely ground, takes about 0.15g, and precision claims
It is fixed, in putting 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80%
Methanol dilution shakes up to scale, filtration, takes subsequent filtrate, obtains final product.
Algoscopy is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines,
Obtain final product.
Assay result this batch composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) be calculated as
1.9mg。
Embodiment 3
Composite ardisia herb piece piece (coated tablet) lot number：20111201
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：Spiral duct diameter 7.5~
25 μm, fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae).
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take Bergeninum reference substance appropriate, plus methanol
Solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 4, puts respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid second
Ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixing of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Solution.In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color.
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take linarin reference substance appropriate, plus methanol system
Into solution of every 1ml containing 0.2mg, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 2, puts respectively on same polyamide film, with ethyl acetate-butanone-chloroform-first
Acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, and at 105 DEG C speckle is heated to
Point colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with reference substance chromatograph, show
The fluorescence spot of same color.
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, supersound process 15
Minute, filtration discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and residue adds water 40ml
Dissolving is made, is extracted 3 times to the shaking of water saturated n-butyl alcohol, each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.Another extracting liquorice control medicinal material 1g, is made in the same way of control medicinal material solution.According to thin layer chromatography (Chinese Pharmacopoeia version one in 2010
The B of annex VI) test, each 4 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into strips, with acetic acid
Ethyl ester-formic acid-glacial acetic acid-water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid, 105
DEG C it is heated to spot development clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, with control medicinal material chromatograph phase
On the position answered, show the fluorescence spot of same color.
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia
One D of annex VI of version in 2010) determine.
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler；With methanol-water (20:
80) it is mobile phase；Detection wavelength is 275nm.Number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000.
The preparation of reference substance solution takes Bergeninum reference substance in right amount, accurately weighed, plus 80% methanol is made every 1ml and contained
The solution of 50 μ g, obtains final product.
The preparation of need testing solution takes this product 10, removes coating, accurately weighed, finely ground, takes about 0.15g, and precision claims
It is fixed, in putting 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80%
Methanol dilution shakes up to scale, filtration, takes subsequent filtrate, obtains final product.
Algoscopy is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines,
Obtain final product.
Assay result this batch composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) be calculated as
1.8mg。
Embodiment 4
Composite ardisia herb piece piece (coated tablet) lot number：20111202
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：Spiral duct diameter 7.5~
25 μm, fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae).
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take Bergeninum reference substance appropriate, plus methanol
Solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 3, puts respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid second
Ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixing of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Solution.In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color.
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, ultrasound
Process 30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution.Separately take linarin reference substance appropriate, plus methanol system
Into solution of every 1ml containing 0.2mg, as reference substance solution.According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010)
Test, draws the μ l of above two solution 2, puts respectively on same polyamide film, with ethyl acetate-butanone-chloroform-first
Acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, and at 105 DEG C speckle is heated to
Point colour developing is clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, on position corresponding with reference substance chromatograph, show
The fluorescence spot of same color.
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, supersound process 15
Minute, filtration discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and residue adds water 40ml
Dissolving is made, is extracted 3 times to the shaking of water saturated n-butyl alcohol, each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid.Another extracting liquorice control medicinal material 1g, is made in the same way of control medicinal material solution.According to thin layer chromatography (Chinese Pharmacopoeia version one in 2010
The B of annex VI) test, each 5 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into strips, with acetic acid
Ethyl ester-formic acid-glacial acetic acid-water (15: 1: 1: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 10% ethanol solution of sulfuric acid, 105
DEG C it is heated to spot development clear, puts and inspected under ultra-violet lamp (365nm).In test sample chromatograph, with control medicinal material chromatograph phase
On the position answered, show the fluorescence spot of same color.
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia
One D of annex VI of version in 2010) determine.
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler；With methanol-water (20:
80) it is mobile phase；Detection wavelength is 275nm.Number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000.
The preparation of reference substance solution takes Bergeninum reference substance in right amount, accurately weighed, plus 80% methanol is made every 1ml and contained
The solution of 50 μ g, obtains final product.
The preparation of need testing solution takes this product 10, removes coating, accurately weighed, finely ground, takes about 0.15g, and precision claims
It is fixed, in putting 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80%
Methanol dilution shakes up to scale, filtration, takes subsequent filtrate, obtains final product.
Algoscopy is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, injects chromatograph of liquid, determines,
Obtain final product.
Assay result this batch composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) be calculated as
1.9mg。
Claims (1)
1. a kind of method for qualitative and quantitative detection of composite ardisia herb piece piece, it is characterised in that including (1) using microscopical identification Herba Ardisiae Japonicae,
Herba Ardisiae Japonicae, Flos Chrysanthemi Indici, Radix Glycyrrhizae are differentiated using thin layer chromatography, (2) are using Rhizoma Seu Herba Bergeniae in high effective liquid chromatography for measuring Herba Ardisiae Japonicae
The content of element, the method for qualitative and quantitative detection is comprised the steps of：
(1) differentiate：
The microscopical identification of Herba Ardisiae Japonicae comprises the following steps：This product is taken, basis of microscopic observation is put：7.5~25 μm of spiral duct diameter,
Fiber bunchy, wall is thicker, 7.5~26 μm of prism of calcium oxalate diameter (Herba Ardisiae Japonicae)；
The thin layer of Herba Ardisiae Japonicae differentiates to comprise the following steps：This product 2 is taken, coating is removed, it is finely ground, plus methanol 20ml, supersound process
30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution；Separately take Bergeninum reference substance appropriate, plus methanol is made
The solution containing 1mg per 1ml, as reference substance solution；According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010) examination
Test, draw the μ l of above two solution 3~5, put respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-acetic acid second
Ester-methanol (9: 8: 2) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with the mixing of the potassium ferricyanide of 1% ferric chloride -1% (1: 1)
Solution；In test sample chromatograph, on position corresponding with reference substance chromatograph, show the speckle of same color；
The thin layer of Flos Chrysanthemi Indici differentiates to comprise the following steps：This product 3 is taken, coating is removed, it is finely ground, plus methanol 15ml, supersound process
30 minutes, filtration, filtrate is concentrated into about 2ml, used as need testing solution；Separately take linarin reference substance appropriate, plus methanol is made often
Solution of the 1ml containing 0.2mg, as reference substance solution；According to thin layer chromatography (Chinese Pharmacopoeia one B of annex VI of version in 2010) examination
Test, draw the μ l of above two solution 1~2, put respectively on same polyamide film, with ethyl acetate-butanone-chloroform-
Formic acid-water (15: 15: 6: 4: 1) is developing solvent, is launched, and is taken out, and is dried, and is sprayed with 2% aluminum trichloride solution, is heated at 105 DEG C
Spot development is clear, puts and inspected under ultra-violet lamp (365nm)；In test sample chromatograph, on position corresponding with reference substance chromatograph,
The fluorescence spot of aobvious same color；
The discriminating of Radix Glycyrrhizae comprises the following steps：This product 20 is taken, coating is removed, finely ground, add diethyl ether 40ml, 15 points of supersound process
Clock, filtration, discards ether solution, and medicinal residues add methanol 50ml, are heated to reflux 1 hour, filtration, and filtrate is evaporated, and the residue 40ml that adds water makes
Dissolving, extracts 3 times to the shaking of water saturated n-butyl alcohol, and each 20ml merges n-butanol extracting liquid, to n-butyl alcohol saturation
Water washing 3 times, each 20ml discards aqueous, takes n-butyl alcohol liquid and is evaporated, and residue adds methanol 5ml to make dissolving, molten as test sample
Liquid；Another extracting liquorice control medicinal material 1g, add diethyl ether 40ml, supersound process 15 minutes, filtration, discards ether solution, and medicinal residues add methanol
50ml, is heated to reflux 1 hour, filtration, and filtrate is evaporated, and the residue 40ml that adds water makes dissolving, carries to the shaking of water saturated n-butyl alcohol
Take 3 times, each 20ml, merge n-butanol extracting liquid, to the water washing 3 times of n-butyl alcohol saturation, each 20ml discards aqueous,
Take n-butyl alcohol liquid to be evaporated, residue adds methanol 5ml to make dissolving, used as control medicinal material solution；According to thin layer chromatography (Chinese Pharmacopoeia 2010
Year one B of annex VI of version) test, each 3~5 μ l of above two solution are drawn, put on same silica gel g thin-layer plate respectively, make into
Strip, with acetic ether-methanoic acid-glacial acetic acid-water (15: 1: 1: 2) as developing solvent, launches, and takes out, and dries, and sprays with 10% sulphuric acid
Ethanol solution, 105 DEG C are heated to spot development clearly, put and inspected under ultra-violet lamp (365nm)；In test sample chromatograph, with it is right
According on the corresponding position of medical material chromatograph, show the fluorescence spot of same color；
(2) assay：
The content assaying method of Bergeninum comprises the following steps in Herba Ardisiae Japonicae：According to high performance liquid chromatography (Chinese Pharmacopoeia 2010
Year one D of annex VI of version) determine；
Chromatographic condition and system suitability：It is filler with octadecylsilane chemically bonded silica；With methanol-water (20: 80)
For mobile phase；Detection wavelength is 275nm, and number of theoretical plate is calculated by Bergeninum peak and should be not less than 3000；
The preparation of reference substance solution：Take Bergeninum reference substance appropriate, it is accurately weighed, plus 80% methanol makes every 1ml containing 50 μ g
Solution, obtain final product；
The preparation of need testing solution：This product 10 is taken, coating is removed, it is accurately weighed, it is finely ground, about 0.15g is taken, it is accurately weighed, put
In 25ml measuring bottles, plus 80% methanol 20ml, supersound process (power 300W, frequency 40kHz) 45 minutes, let cool, plus 80% methanol
Scale is diluted to, is shaken up, filtered, take subsequent filtrate, obtained final product：
Algoscopy：It is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, chromatograph of liquid is injected, determine, obtain final product；
Composite ardisia herb piece piece is per piece containing Herba Ardisiae Japonicae with Bergeninum (C14H16O9) meter, 1.5mg must not be less than.
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| CN112067739A (en) * | 2020-10-13 | 2020-12-11 | 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 |
| 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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| CN111686085B (en) * | 2020-06-30 | 2022-09-06 | 贵州益佰女子大药厂有限责任公司 | Preparation method of throat clearing preparation |
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| CN112067739A (en) * | 2020-10-13 | 2020-12-11 | 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 |
| 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 |
| 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 |
| CN115015418B (en) * | 2022-06-01 | 2023-12-15 | 湖南新汇制药股份有限公司 | Quality detection method of Japanese ardisia herb decoction |
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