CN107037168A - A kind of quality determining method of the brave fruit Chinese herbal granules of the rock of Sugarless type two - Google Patents
A kind of quality determining method of the brave fruit Chinese herbal granules of the rock of Sugarless type two Download PDFInfo
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- G—PHYSICS
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- 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 relates to a kind of quality determining method of the brave fruit Chinese herbal granules of rock of Sugarless type two.Mainly include character, inspection, differentiate and assay；Qualitive test is differentiated using thin-layered chromatography to fleshyleaf lysionotus herb, purple bergenia herb, giant knotweed in preparation；Check and required so that pharmacopeia is relevant, granule is checked；And assay mainly carries out assay to monarch drug in a prescription fleshyleaf lysionotus herb principle active component nevadensin in preparation, so as to set up the quality standard of said preparation.Traditional party is clinically used with decoction, there is very inconvenience in transport and storing process, and taking dose is big, is unfavorable for batch production, great limitation has been carried out to using and developing with.It is of the invention that traditional decoction is mainly changed system into granule, formulate corresponding quality of the pharmaceutical preparations control standard.The method of the present invention can effectively control the quality standard of granule, for exploitation safely, effectively, stably, it is controllable, adapt to clinical and market demands, granular preparation convenient to take and lay the foundation.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, it is more particularly to a kind of with cough-relieving, heat-clearing, resolving sputum, relieving asthma the effect of
The method of quality control of granula medicine.
Background technology
This product is the prescription of the rich day training of old docter of TCM, and prescription is made up of fleshyleaf lysionotus herb, purple bergenia herb, giant knotweed, the taste Chinese medicine of denseflower bulbophyllum herb four；
Lung resolving sputum is supported with fleshyleaf lysionotus herb, purple bergenia herb, giant knotweed heat-clearing expelling phlegm and arresting coughing is relievingd asthma；Denseflower bulbophyllum herb nourishing the stomach to improve the production of body fluid, moistening lung for removing phlegm.Complete square compatibility 5
It is rigorous, determined curative effect；The party tradition is clinically used with soup with decoction, in transport and storing process exist very it is many not
Just, and taking dose is big, it is unfavorable for batch production, great limitation has been carried out to using and developing with.Wei Full are enough to vast trouble
The need for person, clinic is more easily served；Because our day dosing is larger, therefore is prepared into granule, and formulates corresponding system
Agent quality control standard, drafts quality standard draft, and the base of science is established for the granule hospital preparation and industrialized production
Plinth.
The content of the invention
It is an object of the present invention to provide Sugarless type cough-relieving, resolving sputum, the method for quality control of Pingchuan Granule.The present invention
For with cough-relieving, anti-inflammatory, the medicine relievingd asthma, the quality of the product after sugar free granule is changed system into particular for conventional formulation
Control method, drafts the TLC Identification of fleshyleaf lysionotus herb in granule, purple bergenia herb, giant knotweed；Monarch drug in a prescription fleshyleaf lysionotus herb is mainly effective
Composition nevadensin HPLC content assaying methods, so as to formulate the quality standard of said preparation, for exploitation safely, effectively, stably, can
Control, adapt to clinical and take convenient Sugarless type cough-relieving, resolving sputum, Pingchuan Granule preparation and lay the foundation.
A kind of quality determining method of the brave fruit Chinese herbal granules of the rock of Sugarless type two
【Prescription】Fleshyleaf lysionotus herb 30g purple bergenia herb 30g giant knotweed 30g denseflower bulbophyllum herbs 30g
【Preparation method】It is well mixed in 4 taste medicinal materials in the prescription ratio side of weighing, plus 8 times of amount decoctings are boiled, decoction 3 times, 1 hour every time,
Merging filtrate after three decoctions, is concentrated into the concentrate that relative density is 1.125；Ethanol is added into concentrate again makes ethanol body
Fraction is 70% progress alcohol precipitation removal of impurities, and alcohol precipitation 24 hours removes residue and waved in water-bath to without alcohol taste, obtains thick paste；By thick paste
With dextrin, soluble starch, sweetener presses 10：17:3:After 0.3 ratio uniform mixing, then appropriate 80% ethanol is added to be made
Grain, 70 DEG C of dryings are packed in the environment of relative air humidity is 69%, produced.
Method of quality control of the present invention includes：Character, inspection, discriminating and the part or complete in assay project
Portion；Wherein Qualitive test is differentiated using thin-layered chromatography to fleshyleaf lysionotus herb, purple bergenia herb, giant knotweed in preparation；Inspection has with pharmacopeia
Close and require, granule is checked；And assay is mainly to monarch drug in a prescription fleshyleaf lysionotus herb principle active component lysiontus pauciflorus in preparation
Element carries out assay.
The Sugarless type cough-relieving, resolving sputum, the method for quality control of Pingchuan Granule, it is characterised in that the character mirror of granule
Not Wei yellowish-brown to the particle of yellowish-brown uniform color, gas is fragrant, the micro- sweet tea of taste.
The Sugarless type cough-relieving, resolving sputum, the method for quality control of Pingchuan Granule, it is characterised in that the check item of granule
It is main to include checking granularity, moisture, melting, heavy metal, arsenic etc..
The discrimination method of fleshyleaf lysionotus herb is using nevadensin reference substance as control, with toluene-ethyl acetate-formic acid（2~10：2~
8：1~5）For solvent, deploy, take out, dry, spray with appropriate 2% ferric trichloride ethanol solution, be positioned in 105 DEG C of baking ovens,
It is heated to taking out after colour developing is clear.
The discrimination method of purple bergenia herb is using Bergenin reference substance as control, with chloroform-acetic ether-methanoic acid（2~
10：2~8：1~5）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under the TLC Identification inspected.
The discrimination method of giant knotweed is using rheum emodin reference substance as control, with benzene-acetate-methanol（2~10：2~8：1~5）
For solvent, expansion is dried, puts the thin-layer identification method inspected under ultraviolet lamp (254nm).
Described discrimination method includes the part or all of of following items：
（1）Take this product appropriate, it is finely ground, plus methanol, ultrasonically treated, filtration, filtrate is evaporated, and residue adds methanol to make dissolving, is used as confession
Test sample solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography《Middle traditional Chinese medicines
Allusion quotation》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, puts respectively in same silica gel
GF254On lamellae, with toluene-ethyl acetate-formic acid（2~10：2~8：1~5）For solvent, deploy, take out, dry, spray is with suitable
2% ferric trichloride ethanol solution is measured, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and negative control chromatogram is corresponding herein
Immaculate on position.
（2）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, take filtrate as need testing solution.Take
Bergenin reference substance and appropriate negative sample, with method obtained solution.According to《Chinese Pharmacopoeia》Under 2015 editions four general rules 0502
Experiment carries out point sample, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively in same silica G F254On lamellae, with chloroform-second
Acetoacetic ester-formic acid（2~10：2~8：1~5）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under inspect.Test sample color
In spectrum, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram is on this relevant position
Immaculate.
（3）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, be evaporated filtrate, residue obtained addition sulphur
Acid solution, is heated to reflux half an hour, cooling, then is extracted twice with chloroform, merges solution, and drying, residue adds three again
Chloromethanes dissolves it, to make need testing solution.Weigh rheum emodin reference substance a little, addition methanol solution is prepared as every solution and contained
The solution of rheum emodin, is used as the reference substance solution of giant knotweed.It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati.According to thin-layered chromatography《In
State's pharmacopeia》2015 editions four general rules 0502 are tested, and are drawn above-mentioned each 5 μ L of four kinds of solution, are put respectively in same silica G F254Thin layer
On plate, using benzene-acetate-methanol as solvent, expansion is dried, puts and inspected under ultraviolet lamp (254nm).Test sample chromatogram
In, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram.
Specific discrimination method includes the part or all of of following items：
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography
《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, put respectively in
Same silica G F254On lamellae, with toluene-ethyl acetate-formic acid（5：4：1）For solvent, deploy, take out, dry, spray with
Appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color, and negative control chromatogram phase herein on position corresponding with reference substance chromatogram and control medicinal material chromatogram
Answer immaculate on position.
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as confession
Test sample solution.Bergenin reference substance and appropriate negative sample are taken, with method obtained solution.According to《Chinese Pharmacopoeia》2015 editions four lead to
Then experiment carries out point sample under 0502, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively in same silica G F254On lamellae, with
Chloroform-acetic ether-methanoic acid（5：4：2）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under inspect.For examination
In product chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram is in this corresponding positions
Put immaculate.
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, gained
Residue adds 2.5 mo1/L sulfuric acid solution 10ml, is heated to reflux half an hour, cools down, then is extracted twice altogether with 5ml chloroforms
10ml, merges solution, dries, residue adds chloroform again dissolves it, to make need testing solution.Weigh rheum emodin control
Product are a little, add the solution that methanol solution is prepared as every 1ml solution 48ug containing rheum emodin, are used as the reference substance solution of giant knotweed.Separately
Prepare the negative sample for lacking Rhizoma Polygoni Cuspidati.According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, in absorption
Each 5 μ L of four kinds of solution are stated, are put respectively in same silica G F254On lamellae, with benzene-acetate-methanol（7：2：1）For expansion
Agent, expansion, dries, puts and inspected under ultraviolet lamp (254nm).In test sample chromatogram, with reference substance chromatogram and comparison medicine wood color
Compose the spot for showing same color on corresponding position.
The assay of nevadensin contained by this product fleshyleaf lysionotus herb is as control, with mobile phase with nevadensin reference substance：
Methanol-water（50~80:50~20）；Column temperature：20~35 ℃；Detection wavelength：280~350 nm；Flow velocity：1.0 ml.min-1；Sample introduction
Amount：20ul high effective liquid chromatography for measuring.
Described fleshyleaf lysionotus herb content shines Chinese Pharmacopoeia high effective liquid chromatography for measuring, including following：
Chromatographic column：DIKMA-C18 posts；Mobile phase：Methanol-water（68:32）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：
1 ml.min-1；Sample size：20ul；And nevadensin theoretical cam curve must not be less than 3000, under these conditions, nevadensin
It is not interfere with each other with remaining composition chromatographic peak.
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol is dissolved as every 1m1 containing 50 μ g stones
The solution of bracketplant element, it is standby.
The preparation of need testing solution：Take Sugarless type cough-relieving, resolving sputum, Pingchuan Granule a little, grind, accurate measuring 1g is put
Enter in vial conical flask, precision adds 50 mL 75% methanol, weighs, the min of ultrasonic dissolution 20.Supply less loss weight, mistake
Filter, it is standby.
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined.
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
Described quality prosecutor method includes：
【Character】Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste；
【Check】Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules；
【Differentiate】
（1）Take this product appropriate, it is finely ground, plus methanol, ultrasonically treated, filtration, filtrate is evaporated, and residue adds methanol to make dissolving, is used as confession
Test sample solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography《Middle traditional Chinese medicines
Allusion quotation》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, puts respectively in same silica gel
GF254On lamellae, with toluene-ethyl acetate-formic acid（2~10：2~8：1~5）For solvent, deploy, take out, dry, spray is with suitable
2% ferric trichloride ethanol solution is measured, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and negative control chromatogram is corresponding herein
Immaculate on position.
（2）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, take filtrate as need testing solution.Take
Bergenin reference substance and appropriate negative sample, with method obtained solution.According to《Chinese Pharmacopoeia》Under 2015 editions four general rules 0502
Experiment carries out point sample, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively in same silica G F254On lamellae, with chloroform-second
Acetoacetic ester-formic acid（2~10：2~8：1~5）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under inspect.Test sample color
In spectrum, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram is on this relevant position
Immaculate.
（3）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, be evaporated filtrate, residue obtained addition sulphur
Acid solution, is heated to reflux half an hour, cooling, then is extracted twice with chloroform, merges solution, and drying, residue adds three again
Chloromethanes dissolves it, to make need testing solution.Weigh rheum emodin reference substance a little, addition methanol solution is prepared as every solution and contained
The solution of rheum emodin, is used as the reference substance solution of giant knotweed.It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati.According to thin-layered chromatography《In
State's pharmacopeia》2015 editions four general rules 0502 are tested, and are drawn above-mentioned each 5 μ L of four kinds of solution, are put respectively in same silica G F254Thin layer
On plate, using benzene-acetate-methanol as solvent, expansion is dried, puts and inspected under ultraviolet lamp (254nm).Test sample chromatogram
In, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram.
【Assay】Nevadensin assay presses Chinese Pharmacopoeia high effective liquid chromatography for measuring；
Chromatographic condition：With mobile phase：Methanol-water（50~80:50~20）；Column temperature：20~35 ℃；Detection wavelength：280~350
nm；Flow velocity：1.0 ml.min-1；Sample size：Assay is carried out under conditions of 20ul.
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol dissolving, standby.
The preparation of need testing solution：Take this product appropriate, grind, precision measures 1g, is put into vial conical flask, add first
Alcohol, weighs, ultrasonically treated.Less loss weight is supplied, is filtered, it is standby.
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined.
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
For the method for quality control science for ensureing the present invention, reasonable, effective, mirror of the applicant to each medicine in method
Do not studied with content assaying method, it is specific as follows：
【Character】This product is particle of the yellowish-brown to yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste
【Check】Reference《Chinese Pharmacopoeia》2015 editions lower relevant requirements of four annex granules, to test agent in three batches
Granularity, moisture, melting, heavy metal, arsenic etc. are checked.As a result：
【Differentiate】
（1）Fleshyleaf lysionotus herb differentiates research
With reference to the composition of pharmacopeia and fleshyleaf lysionotus herb chemical composition, this method is right using test sample, reference substance, control medicinal material and feminine gender
According to methanol ultrasonic extraction resulting solution, respectively with petroleum ether-ethyl acetate-acetic acid of methanol -36%（9：8：3：1）Acetic acid second
Ester-n-butanol-methyl alcohol-formic acid（6：6：1：1）Chloroform-acetic ether-methanoic acid（5：4：2）；Chloroform-ethyl acetate-
Formic acid（5：5：2）Solvent is made, is deployed, is dried, put ultraviolet lamp（254nm）Under inspect.As a result, toluene-ethyl acetate-formic acid
Development system is preferable to the separating effect of nevadensin, spot is more visible, and negative control is noiseless, boundless edge effect, therefore choosing
Toluene-ethyl acetate-formic acid is selected as the developing solvent system of fleshyleaf lysionotus herb in Sugarless type cough-relieving, resolving sputum, Pingchuan Granule, wherein
Toluene-ethyl acetate-formic acid（5：4：1）It is the optimal solvent ratio condition of fleshyleaf lysionotus herb indentification by TLC.
（2）The discriminating research of purple bergenia herb
With reference to Chinese Pharmacopoeia standard and purple bergenia herb component, this law is used test sample, reference substance and negative control, carried with methanol ultrasound
Resulting solution is taken, respectively from petroleum ether-ethyl acetate-acetic acid of methanol -36%（9：8：3：1）；Ethyl acetate-n-butanol-first
Alcohol-formic acid（6：6：1：1）；Chloroform-acetic ether-methanoic acid（5：4：2）；Chloroform-acetic ether-methanoic acid（5：5：2）
For solvent, expansion takes out silica gel thin-layer plate after expansion completely, dries, be positioned over ultraviolet lamp（254nm）Under inspect.As a result,
Different developing solvent systems influence larger, oil to the thin-layer chromatography of Bergenin in Sugarless type cough-relieving, resolving sputum, Pingchuan Granule
Ether-the acetic acid of acetate-methanol -36%（9：8：3：1）Although Rf values are moderate in developing solvent, inferior separating effect, hangover are tight
Weight；Ethyl acetate-n-butanol-methyl alcohol-formic acid（6：6：1：1）Solvent is not clear enough to the separating effect of Bergenin, and Rf
Value is bigger than normal, and negative control has interference；Chloroform-acetic ether-methanoic acid solvent（5：4：2）Separation of the system to Bergenin
Effect is not good, but Rf values are between 0.2 ~ 0.8 and boundless edge effect, therefore temporarily selects chloroform-acetic ether-methanoic acid as sugar-free
The developing solvent system of purple bergenia herb in type cough-relieving, resolving sputum, Pingchuan Granule.
（3）The discriminating research of giant knotweed
With reference to the main chemical compositions of Chinese Pharmacopoeia standard and giant knotweed, this law is using test sample, reference substance, right
Medicinal material and negative control, with methanol ultrasonic extraction, resulting solution filtration is evaporated, 2.5 mo1/L sulfuric acid of residue obtained addition
Solution 10ml, is heated to reflux half an hour, cooling, then extracts common 10ml twice with 5ml chloroforms, merges solution, drying, residue
Adding chloroform 1mL again dissolves it, produces.Solvent is selected respectively：Petroleum ether (30-60 DEG C)-formic acid-Ethyl formate
(15：1：5), petroleum ether (30-60 DEG C)-formic acid-Ethyl formate (13：1：5), petroleum ether (30-60 DEG C)-formic acid-Ethyl formate
(10：1：5), petroleum ether (30-60 DEG C)-formic acid-Ethyl formate (10：1：4)；Benzene-methanol（2：1）, benzene-methanol（3：1）, benzene-
Methanol（4：1）, benzene-methanol（5：1）；Benzene-acetate-methanol（7：1：0.2）, benzene-acetate-methanol（5：1：0.2）,
Benzene-acetate-methanol（7：2：1）, benzene-acetate-methanol（8：1.5：0.5）Expansion, takes out silica gel thin after expansion completely
Laminate, is dried, and is positioned over ultraviolet lamp（254nm）Under inspect.As a result, different developing solvent systems are to the brave fruit sugar-free particle of two rocks
The separating effect of the active ingredient emodin of middle giant knotweed influences smaller, petroleum ether（30-60℃）- Ethyl formate-formic acid expansion is molten
Although in agent system Rf values it is moderate between 2-8, good separating effect, negative control is noiseless, there is slight edge effect,
And reappearance is not sufficiently stable；Benzene-methanol solvent system is not good to the separating effect of giant knotweed, has conditions of streaking, and have slight
Edge effect；The separating effect of benzene-acetate-methanol solvent system rheum emodin is preferable, spot is more visible, and negative control
It is noiseless, therefore selection benzene-acetate-methanol is used as the developing solvent system of giant knotweed in Sugarless type cough-relieving, resolving sputum, Pingchuan Granule
System.Wherein benzene-acetate-methanol（7：2：1）It is the optimal solvent ratio condition of giant knotweed indentification by TLC.
【Assay】
Sugarless type cough-relieving, resolving sputum, Pingchuan Granule prescription traditional Chinese medicinal materials assortment have 4 kinds.According to Chinese medicine traditional monarch, minister, help, make reason
By it is assay Con trolling index to select monarch drug in a prescription first, and secondly selection ministerial drug, selects adjuvant again.According to production and quality
Actual conditions are examined, by monarch drug in a prescription fleshyleaf lysionotus herb in the side of analysis and the known chemical composition of ministerial drug purple bergenia herb and its pharmacological action, then
With reference to the relevant theory of Chinese medicine, intend selecting the effective main component nevadensin of monarch drug in a prescription fleshyleaf lysionotus herb, carrying out assay to preparation grinds
Study carefully；It is applied in production and quality of the pharmaceutical preparations control, effective control for product quality.
1 chromatographic condition
With reference to《Chinese Pharmacopoeia》The method containing survey of the lower nevadensin of version one lysiontus pauciflorus in 2015 is tested.Chromatographic column：
DIKMA-C18 posts；Mobile phase：Methanol-water（68:32）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：1 ml.min-1；
Sample size：20ul；Under these conditions, nevadensin and remaining composition chromatographic peak are not interfere with each other.
（1）The preparation of reference substance solution
Precise nevadensin reference substance is a small amount of, plus methanol is dissolved as the solution that every 1m1 contains 50 μ g nevadensins, standby.
（2）The preparation of need testing solution
Take the brave fruit sugar-free particle of two rocks a little, grind, accurate measuring 1g is put into vial conical flask, precision adds 50 mL's
75% methanol, weighs, the min of ultrasonic dissolution 20.Less loss weight is supplied, is filtered, it is standby.
（3）The preparation of negative controls solution
In prescription ratio, other flavour of a drug of lysiontus pauciflorus are removed, sugar free granule is made, negative control solution presses test sample preparation side
It is prepared by method.Feminine gender is test result indicates that going out at peak position without other interference in nevadensin.
2 methodological studies
（1）Linear relationship is investigated
Standard items mother liquor is accurately pipetted with pipette（50 μ g/ml) each 1ml, 2ml, 3ml, 4ml, 5ml, plus methanol is dissolved to
10ml, filtering, to select chromatographic condition, surveys the peak area at its nevadensin peak successively.Made with peak area and nevadensin concentration
Linear regression, obtains standard curve and regression equation.
The linear relationship of table 1 investigates result
Measurement result is shown in Table 1, and the equation of linear regression of calculating is Y=45.52X+14.833 (R2=0.9992), experimental result table
Bright, nevadensin concentration is good with nevadensin peak area linear relationship in the range of 5 ~ 50ug/ml.
（2）Precision test
Precision draws need testing solution, continuous sample introduction 6 times, by high performance liquid chromatography（Four general rules of Chinese 2015 pharmacopeia version
0512 determines）, measure peak area and calculate RSD values.
The Precision Experiment result (n=6) of table 2
As shown in Table 2, the peak area of nevadensin in need testing solution is determined, average value is 2345.7, calculating RSD values is
0.278, result of the test shows that the equipment precision is good.
（3）Stability test
This product 1g that lot number is 20160701 is taken, it is accurately weighed, prepared by method, accurate injection 20ul sample solutions, Yi Biao is set
Its peak area is determined in the fixed time, RSD values are calculated.
The stability test result of table 3（n=6）
As shown in Table 3, the peak area average value of stability test nevadensin is that 1464.4, RSD values are 0.349, specify that confession
Test sample solution is stable within 24h.
（4）Average recovery is tested
Take same lot number（20160701）6 parts of particle, every part of 1g, precision adds nevadensin reference substance standard liquid in right amount, presses
Prepared by " under 1.2.3.3 ", draw in 20ul injection high performance liquid chromatographs, determine.Calculate the gained rate of recovery and RSD values.
The average recovery result of the test (n=6) of table 4
Found out by the above results, mean sample recovery rate is 101.16%, in 95% ~ 105% normal range (NR).RSD values are
1.87%, RSD are less than 2.0%.Show that this method can be used for the measure of nevadensin in Sugarless type cough-relieving, resolving sputum, Pingchuan Granule.
（5）Repeated experiment
Totally 6 parts of same lot number granule is taken, test liquid is prepared as stated above, precision draws the solution 20 μ L, according to chromatostrip
Part sample introduction, calculates nevadensin content（ug/ml）With RSD values.
The repeated experiment result of table 5（n=6）
Table 2-6 understands that the average value for calculating nevadensin content in need testing solution is 31.624, and the method is reproducible
RSD is 0.325%.
（6）Test sample（Nevadensin）Determine and content limit is tentatively customized
Three batches of self-control samples are taken respectively（Lot number：20160701,20160702,20160703）, test sample is prepared as stated above
Solution, measures the μ L sample introductions of need testing solution 20, measures peak area, calculates the content and RSD values of nevadensin, every batch determines 2
It is secondary to average.
The nevadensin assay result of table 6
Determine three batches self-control samples in contents, every batch determine twice, as a result as shown in table 2-7, Sugarless type cough-relieving, resolving sputum, put down
The content for breathing heavily nevadensin in granule is up to 32.17 ug/g and is not less than 29.62ug/g.Average content is 31.17ug/
G, it is 3.34% to calculate RSD values.Multiplied according to the average value of nevadensin content in three batches of Sugarless type cough-relievings, resolving sputum, Pingchuan Granules
24.9ug must not be less than with the content of nevadensin in 80%, i.e. Sugarless type cough-relieving, resolving sputum, Pingchuan Granule.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
The various changes carried out on the premise of invention spirit and scope to the material component in these embodiments and consumption or change
Belong to protection scope of the present invention.Raw materials used and reagent of the invention is commercially available.
Embodiment 1
Sugarless type cough-relieving, resolving sputum, the quality control of Pingchuan Granule
Method of quality control of the present invention is mainly included the following steps that：
【Character】Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste.
【Check】Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules.
【Differentiate】
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography
《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, put respectively in
Same silica G F254On lamellae, with toluene-ethyl acetate-formic acid（5：4：1）For solvent, deploy, take out, dry, spray with
Appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color, and negative control chromatogram phase herein on position corresponding with reference substance chromatogram and control medicinal material chromatogram
Answer immaculate on position.
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as confession
Test sample solution.Bergenin reference substance and appropriate negative sample are taken, with method obtained solution.According to《Chinese Pharmacopoeia》2015 editions four lead to
Then experiment carries out point sample under 0502, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively in same silica G F254On lamellae, with
Chloroform-acetic ether-methanoic acid（5：4：2）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under inspect.For examination
In product chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram is in this corresponding positions
Put immaculate.
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, gained
Residue adds 2.5 mo1/L sulfuric acid solution 10ml, is heated to reflux half an hour, cools down, then is extracted twice altogether with 5ml chloroforms
10ml, merges solution, dries, residue adds chloroform again dissolves it, to make need testing solution.Weigh rheum emodin control
Product are a little, add methanol solution and are prepared as solution of every solution containing rheum emodin, are used as the reference substance solution of giant knotweed.Another prepare lacks tiger
The negative sample of cane medicinal material.According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and above-mentioned four kinds of absorption is molten
Each 5 μ L of liquid, put in same silica G F respectively254On lamellae, with benzene-acetate-methanol（7：2：1）For solvent, deploy,
Dry, put and inspected under ultraviolet lamp (254nm).In test sample chromatogram, corresponding with reference substance chromatogram and control medicinal material chromatogram
Show the spot of same color on position.
【Assay】According to《Chinese Pharmacopoeia》High performance liquid chromatography is lower to be determined.
Chromatographic condition：With mobile phase：Methanol-water（68:32）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：1.0
ml.min-1；Sample size：Assay is carried out under conditions of 20ul.
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol dissolving, standby.
The preparation of need testing solution：Take this product a little, grind, precision measures 1g, is put into vial conical flask, add first
Alcohol, weighs, ultrasonically treated.Less loss weight is supplied, is filtered, it is standby.
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined.
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
【Cure mainly function】With cough-relieving, heat-clearing, resolving sputum and relieving asthma function.Cure mainly wind-heat affecting lung, accumulation phlegm-heat in the interior it is cough caused,
Asthma.
【Specification】Per packed gram of 15g
【Usage and dosage】Warm boiled water, one after each meal, one time 15 grams, 3 times a day.
【Storage】Sealing, puts at shady and cool dry.
Embodiment 2
Sugarless type cough-relieving, resolving sputum, the quality control of Pingchuan Granule
Method of quality control of the present invention is mainly included the following steps that：
【Character】Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste.
【Check】Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules.
【Differentiate】
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography
《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, put respectively in
Same silica G F254On lamellae, with toluene-ethyl acetate-formic acid（6：4：1）For solvent, deploy, take out, dry, spray with
Appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color, and negative control chromatogram phase herein on position corresponding with reference substance chromatogram and control medicinal material chromatogram
Answer immaculate on position.
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as confession
Test sample solution.Bergenin reference substance and appropriate negative sample are taken, with method obtained solution.According to《Chinese Pharmacopoeia》2015 editions four lead to
Then experiment carries out point sample under 0502, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively on same silica GF254 lamellae, with
Petroleum ether-ethyl acetate-the acetic acid of methanol -36%（9：8：3：1）For solvent, deploy, take out, put ultraviolet lamp（254nm）Lower inspection
Depending on.In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram.
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, gained
Residue adds 2.5 mo1/L sulfuric acid solution 10ml, is heated to reflux half an hour, cools down, then is extracted twice altogether with 5ml chloroforms
10ml, merges solution, dries, residue adds chloroform again dissolves it, to make need testing solution.Weigh rheum emodin control
Product are a little, add methanol solution and are prepared as solution of every solution containing rheum emodin, are used as the reference substance solution of giant knotweed.Another prepare lacks tiger
The negative sample of cane medicinal material.According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and above-mentioned four kinds of absorption is molten
Each 5 μ L of liquid, put in same silica G F respectively254On lamellae, with benzene-acetate-methanol（5：1：0.2）For solvent, exhibition
Open, dry, put and inspected under ultraviolet lamp (254nm).In test sample chromatogram, corresponding to reference substance chromatogram and control medicinal material chromatogram
Position on show same color spot.
【Assay】According to《Chinese Pharmacopoeia》High performance liquid chromatography is lower to be determined.
Chromatographic condition：With mobile phase：Methanol-water（50：50）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：1.0
ml.min-1；Sample size：Assay is carried out under conditions of 20ul.
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol dissolving, standby.
The preparation of need testing solution：Take this product a little, grind, precision measures 1g, is put into vial conical flask, add first
Alcohol, weighs, ultrasonically treated.Less loss weight is supplied, is filtered, it is standby.
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined.
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
【Cure mainly function】With cough-relieving, heat-clearing, resolving sputum and relieving asthma function.Cure mainly wind-heat affecting lung, accumulation phlegm-heat in the interior it is cough caused,
Asthma.
【Specification】Per packed gram of 15g
【Usage and dosage】Warm boiled water, one after each meal, one time 15 grams, 3 times a day.
【Storage】Sealing, puts at shady and cool dry.
Embodiment 3
Sugarless type cough-relieving, resolving sputum, the quality control of Pingchuan Granule, method of quality control of the present invention mainly include following
Step：
【Character】Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste.
【Check】Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules.
【Differentiate】
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution.Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution.According to thin-layered chromatography
《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 10 μ L of four kinds of solution, put respectively in
Same silica G F254On lamellae, with toluene-ethyl acetate-formic acid（8：4：1）For solvent, deploy, take out, dry, spray with
Appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to taking out after colour developing is clear.In test sample chromatogram,
Show the spot of same color, and negative control chromatogram phase herein on position corresponding with reference substance chromatogram and control medicinal material chromatogram
Answer immaculate on position.
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as confession
Test sample solution.Bergenin reference substance and appropriate negative sample are taken, with method obtained solution.According to《Chinese Pharmacopoeia》2015 editions four lead to
Then experiment carries out point sample under 0502, draws above-mentioned each 5 μ L of three kinds of solution, puts respectively on same silica GF254 lamellae, with
Development system：Chloroform-acetic ether-methanoic acid（5：5：2）For solvent, deploy, take out, put ultraviolet lamp（254nm）Under
Inspect.In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram.
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, gained
Residue adds 2.5 mo1/L sulfuric acid solution 10ml, is heated to reflux half an hour, cools down, then is extracted twice altogether with 5ml chloroforms
10ml, merges solution, dries, residue adds chloroform again dissolves it, to make need testing solution.Weigh rheum emodin control
Product are a little, add methanol solution and are prepared as solution of every solution containing rheum emodin, are used as the reference substance solution of giant knotweed.Another prepare lacks tiger
The negative sample of cane medicinal material.According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and above-mentioned four kinds of absorption is molten
Each 5 μ L of liquid, put in same silica G F respectively254On lamellae, with benzene-acetate-methanol（8：1.5：0.5）For solvent, exhibition
Open, dry, put and inspected under ultraviolet lamp (254nm).In test sample chromatogram, corresponding to reference substance chromatogram and control medicinal material chromatogram
Position on show same color spot.
【Assay】According to《Chinese Pharmacopoeia》High performance liquid chromatography is lower to be determined.
Chromatographic condition：With mobile phase：Methanol-water（30：70）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：1.0
ml.min-1；Sample size：Assay is carried out under conditions of 20ul.
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol dissolving, standby.
The preparation of need testing solution：Take this product a little, grind, precision measures 1g, is put into vial conical flask, add first
Alcohol, weighs, ultrasonically treated.Less loss weight is supplied, is filtered, it is standby.
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined.
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
【Cure mainly function】With cough-relieving, heat-clearing, resolving sputum and relieving asthma function.Cure mainly wind-heat affecting lung, accumulation phlegm-heat in the interior it is cough caused,
Asthma.
【Specification】Per packed gram of 15g
【Usage and dosage】Warm boiled water, one after each meal, one time 15 grams, 3 times a day.
【Storage】Sealing, puts at shady and cool dry.
Using the method for quality control of above example 1, it will be easier to control the quality of the granule, newly-built quality control
Method sensitivity height processed, favorable reproducibility, separating degree are big, practical, can preferably meet containing for granule index composition
It is fixed to measure.
Finally it should be noted that above-described embodiment is only used for the inventive concept of the present invention, rather than to rights protection of the present invention
Restriction, to those skilled in the art according to description of the invention, can to the present invention carry out equivalent substitution, combination, improve or
Modification, the change done without departing from theon the basis of the spirit of the present invention all should fall into protection scope of the present invention.
Claims (8)
1. a kind of quality determining method of the brave fruit Chinese herbal granules of the rock of Sugarless type two, its constitute by fleshyleaf lysionotus herb, purple bergenia herb, giant knotweed,
Denseflower bulbophyllum herb is prepared from, wherein fleshyleaf lysionotus herb 30g purple bergenia herbs 30g giant knotweeds 30g denseflower bulbophyllum herb 30g, and the character of granule differentiates as Huang
Brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste.
2. the quality determining method described in claim 1, it is characterised in that the check item of granule mainly includes to granularity, water
Point, melting, heavy metal, arsenic etc. checked；The thin-layer identification method of wherein fleshyleaf lysionotus herb be using nevadensin as control, with
Toluene-ethyl acetate-formic acid（3~10：2~8：1~5）The thin-layer identification method deployed for solvent；The thin layer discriminating side of purple bergenia herb
Method is using Bergenin as control, with chloroform-acetic ether-methanoic acid (3 ~ 10：2~8：1 ~ 5) be solvent, deploy, take out
Thin-layer identification method；The thin-layer identification method of giant knotweed is using rheum emodin as control, with benzene-acetate-methanol（3~15：1~
5：1~5）For solvent, expansion is dried, puts the thin-layer identification method inspected under ultraviolet lamp (254nm).
3. the quality determining method described in claim 1, it is characterised in that discrimination method includes the part or all of of following items：
Take this product appropriate, it is finely ground, plus methanol, ultrasonically treated, filtration, filtrate is evaporated, and residue adds methanol to make dissolving, is used as test sample
Solution；Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution；
According to thin-layered chromatography《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned four kinds of solution
Each 10 μ L, put in same silica G F respectively254On lamellae, with toluene-ethyl acetate-formic acid（2~10：2~8：1~5）For expansion
Agent, deploys, and takes out, dries, and sprays with appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to colour developing clear
Taken out after clear；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and
Negative control chromatogram immaculate on this relevant position；
Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, take filtrate as need testing solution；
Bergenin reference substance and appropriate negative sample are taken, with method obtained solution；
According to《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 5 μ L of three kinds of solution, respectively
Point is in same silica G F254On lamellae, with chloroform-acetic ether-methanoic acid（2~10：2~8：1~5）For solvent, deploy,
Take out, put ultraviolet lamp（254nm）Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram exists
Immaculate on this relevant position；
Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, be evaporated filtrate, residue obtained addition sulfuric acid solution,
Half an hour is heated to reflux, is cooled down, then is extracted twice with chloroform, merges solution, dries, residue adds chloroform again to be made
It dissolves, to make need testing solution；Weigh rheum emodin reference substance a little, add methanol solution and be prepared as every solution containing rheum emodin
Solution, is used as the reference substance solution of giant knotweed；It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati；
According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and draw above-mentioned each 5 μ L of four kinds of solution, respectively point
In same silica G F254On lamellae, using benzene-acetate-methanol as solvent, expansion is dried, puts ultraviolet lamp (254nm)
Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram.
4. the quality determining method described in claim 1, it is characterised in that specific discrimination method includes the part of following items
Or all：
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution；Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution；
According to thin-layered chromatography《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned four kinds of solution
Each 10 μ L, put in same silica G F respectively254On lamellae, with toluene-ethyl acetate-formic acid（5：4：1）For solvent, deploy,
Take out, dry, spray with appropriate 2% ferric trichloride ethanol solution, be positioned in 105 DEG C of baking ovens, be heated to taking after colour developing is clear
Go out；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and
Negative control chromatogram immaculate on this relevant position；
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as test sample
Solution；Bergenin reference substance and appropriate negative sample are taken, with method obtained solution；
According to《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 5 μ L of three kinds of solution, respectively
Point is in same silica G F254On lamellae, with chloroform-acetic ether-methanoic acid（5：4：2）For solvent, deploy, take out, put
Ultraviolet lamp（254nm）Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram exists
Immaculate on this relevant position；
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, it is residue obtained
2.5 mo1/L sulfuric acid solution 10ml are added, half an hour is heated to reflux, cooled down, then common 10ml twice is extracted with 5ml chloroforms,
Merge solution, dry, residue adds chloroform again dissolves it, to make need testing solution；
Weigh rheum emodin reference substance a little, add methanol solution and be prepared as solution of every solution containing rheum emodin, be used as pair of giant knotweed
According to product solution；It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati；
According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and draw above-mentioned each 5 μ L of four kinds of solution, respectively point
In same silica G F254On lamellae, with benzene-acetate-methanol（7：2：1）For solvent, expansion is dried, puts ultraviolet lamp
Inspected under (254nm)；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram.
5. the quality determining method described in claim 1, it is characterised in that the content assaying method of fleshyleaf lysionotus herb is with reference《In
State's pharmacopeia》Efficient liquid-phase chromatography method is determined, using Bergenin as reference substance, with mobile phase：Methanol-water（50~80:50~20）；
Column temperature：20~35 ℃；Detection wavelength：280~350 nm；Flow velocity：1.0 ml.min-1；Sample size：Contained under conditions of 20ul
It is fixed to measure.
6. the quality determining method described in claim 1, it is characterised in that the content assaying method of fleshyleaf lysionotus herb includes following：
Chromatographic column：DIKMA-C18 posts；Mobile phase：Methanol-water（68:32）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：
1 ml.min-1；Sample size：20ul；And nevadensin theoretical cam curve must not be less than 3000, under these conditions, nevadensin
It is not interfere with each other with remaining composition chromatographic peak；
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol is dissolved as every 1m1 containing 50 μ g lysiontus pauciflorus
The solution of element, it is standby；
The preparation of need testing solution：Take this product a little, grind, accurate measuring 1g is put into vial conical flask, precision adds 50
ML 75% methanol, weighs, the min of ultrasonic dissolution 20；
Less loss weight is supplied, is filtered, it is standby；
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined；
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
7. the quality determining method described in claim 1, it is characterised in that the content assaying method of fleshyleaf lysionotus herb includes：
Character：Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste；
Check：Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules；
Differentiate：
（1）Take this product appropriate, it is finely ground, plus methanol, ultrasonically treated, filtration, filtrate is evaporated, and residue adds methanol to make dissolving, is used as confession
Test sample solution；
Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution；
According to thin-layered chromatography《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned four kinds of solution
Each 10 μ L, put in same silica G F respectively254On lamellae, with toluene-ethyl acetate-formic acid（2~10：2~8：1~5）For expansion
Agent, deploys, and takes out, dries, and sprays with appropriate 2% ferric trichloride ethanol solution, is positioned in 105 DEG C of baking ovens, is heated to colour developing clear
Taken out after clear；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and
Negative control chromatogram immaculate on this relevant position；
（2）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, take filtrate as need testing solution；
Bergenin reference substance and appropriate negative sample are taken, with method obtained solution；
According to《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 5 μ L of three kinds of solution, respectively
Point is in same silica G F254On lamellae, with chloroform-acetic ether-methanoic acid（2~10：2~8：1~5）For solvent, deploy,
Take out, put ultraviolet lamp（254nm）Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram exists
Immaculate on this relevant position；
（3）Take this product appropriate, it is finely ground, plus methanol, it is ultrasonically treated, let cool, filter, be evaporated filtrate, residue obtained addition sulfuric acid is molten
Liquid, is heated to reflux half an hour, cooling, then is extracted twice with chloroform, merges solution, and drying, residue adds three chloromethanes again
Alkane dissolves it, to make need testing solution；
Weigh rheum emodin reference substance a little, add methanol solution and be prepared as solution of every solution containing rheum emodin, be used as pair of giant knotweed
According to product solution；It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati；
According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and draw above-mentioned each 5 μ L of four kinds of solution, respectively point
In same silica G F254On lamellae, using benzene-acetate-methanol as solvent, expansion is dried, puts ultraviolet lamp (254nm)
Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram；
Assay：Nevadensin assay presses Chinese Pharmacopoeia high effective liquid chromatography for measuring；
Chromatographic condition：With mobile phase：Methanol-water（50~80:50~20）；Column temperature：20~35 ℃；Detection wavelength：280~350
nm；Flow velocity：1.0 ml.min-1；Sample size：Assay is carried out under conditions of 20ul；
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol dissolving, standby；
The preparation of need testing solution：Take this product a little, grind, precision measures 1g, is put into vial conical flask, add methanol,
Weigh, it is ultrasonically treated；
Less loss weight is supplied, is filtered, it is standby；
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined；
This product fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
8. the quality determining method described in claim 1, it is characterised in that the content assaying method of fleshyleaf lysionotus herb includes：
Character：Yellowish-brown is to the particle of yellowish-brown uniform color, and gas is fragrant, the micro- sweet tea of taste；
Check：Granule should be met《Chinese Pharmacopoeia》Relevant every regulation under 2015 editions four annex granules；
Differentiate：
（1）This product 1g is taken, finely ground, plus methanol 20ml, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds methanol to make dissolving,
It is used as need testing solution；
Fleshyleaf lysionotus herb reference substance, control medicinal material and negative sample separately are taken, with method obtained solution；
According to thin-layered chromatography《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned four kinds of solution
Each 10 μ L, put in same silica G F respectively254On lamellae, with toluene-ethyl acetate-formic acid（5：4：1）For solvent, deploy,
Take out, dry, spray with appropriate 2% ferric trichloride ethanol solution, be positioned in 105 DEG C of baking ovens, be heated to taking after colour developing is clear
Go out；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram, and
Negative control chromatogram immaculate on this relevant position；
（2）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 40min, let cool, filter, take filtrate as test sample
Solution；
Bergenin reference substance and appropriate negative sample are taken, with method obtained solution；
According to《Chinese Pharmacopoeia》Experiment carries out point sample under 2015 editions four general rules 0502, draws above-mentioned each 5 μ L of three kinds of solution, respectively
Point is in same silica G F254On lamellae, with chloroform-acetic ether-methanoic acid（5：4：2）For solvent, deploy, take out, put
Ultraviolet lamp（254nm）Under inspect；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram, and negative control chromatogram exists
Immaculate on this relevant position；
（3）This product appropriate 1g is taken, it is finely ground, plus methanol 20ml, ultrasonically treated 15min, let cool, filter, be evaporated filtrate, it is residue obtained
2.5 mo1/L sulfuric acid solution 10ml are added, half an hour is heated to reflux, cooled down, then common 10ml twice is extracted with 5ml chloroforms,
Merge solution, dry, residue adds chloroform again dissolves it, to make need testing solution；
Weigh rheum emodin reference substance a little, add methanol solution and be prepared as solution of every solution containing rheum emodin, be used as pair of giant knotweed
According to product solution；It is another to prepare the negative sample for lacking Rhizoma Polygoni Cuspidati；
According to thin-layered chromatography《Chinese Pharmacopoeia》2015 editions four general rules 0502 are tested, and draw above-mentioned each 5 μ L of four kinds of solution, respectively point
In same silica G F254On lamellae, with benzene-acetate-methanol（7：2：1）For solvent, expansion is dried, puts ultraviolet lamp
Inspected under (254nm)；
In test sample chromatogram, the spot of same color is shown on position corresponding with reference substance chromatogram and control medicinal material chromatogram；
Assay：Nevadensin assay presses Chinese Pharmacopoeia high effective liquid chromatography for measuring；
Chromatographic condition：With mobile phase：Methanol-water（68:32）；Column temperature：30 ℃；Detection wavelength：334 nm；Flow velocity：1.0
ml.min-1；Sample size：Assay is carried out under conditions of 20ul；
The preparation of reference substance solution：Precise nevadensin reference substance is a small amount of, plus methanol is dissolved as every 1m1 lysiontus pauciflorus containing 50ug
The solution of element, it is standby；
The preparation of need testing solution：Take this product a little, grind, accurate measuring 1g is put into vial conical flask, precision is added
The ml of 75% methanol 50, weighs, the min of ultrasonic dissolution 20；
Less loss weight is supplied, is filtered, it is standby；
Determination method：It is accurate respectively to draw reference substance solution and each 20ul of need testing solution, liquid chromatograph is injected, is determined, this product
Fleshyleaf lysionotus herb is in terms of nevadensin, and content must not be less than 24.9ug/g.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113791165A (en) * | 2021-09-13 | 2021-12-14 | 湖南新汇制药股份有限公司 | Quality detection method for fried fructus viticis standard decoction |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113791165A (en) * | 2021-09-13 | 2021-12-14 | 湖南新汇制药股份有限公司 | Quality detection method for fried fructus viticis standard decoction |
| CN113791165B (en) * | 2021-09-13 | 2023-09-26 | 湖南新汇制药股份有限公司 | Quality detection method for stir-fried fructus viticis standard decoction |
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