CN107315061A - A kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding - Google Patents
A kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding Download PDFInfo
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Abstract
The invention discloses a kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding, the method for quality control includes the discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese medicine preparation；Discriminating be to the madder control medicinal material in alizarin root of Dahurian angelica Chinese medicine preparation, rubimaillin, purpurin, pseudo-ginseng control medicinal material, notoginsenoside, ginsenoside, root of Dahurain angelica control medicinal material, Imperatorin, Isomperatorin, radix cyathulae control medicinal material, cyasterone indentification by TLC.The present invention is a kind of active component by controlling alizarin root of Dahurian angelica class Chinese medicine preparation and then controls the analysis method of patent medicine quality, the quality standard of the alizarin root of Dahurian angelica improve, it is perfect, on the basis of ministry standard, improve the quality control of medicine, ensure the stability and security of its quality, product technology level is improved, using the quality standard of the alizarin root of Dahurian angelica Chinese medicine preparation of modern analysis means General Promotion, makes the production technology of the alizarin root of Dahurian angelica more strict reasonable.
Description
Technical field
The invention belongs to quality control technologies for traditional Chinese medicine field, it is related to a kind of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding
Method of quality control.
Background technology
In existing technology, alizarin root of Dahurian angelica class Chinese medicine preparation is mainly by radix cyathulae, and pseudo-ginseng, madder, the root of Dahurain angelica is made.Preparation method:
Radix cyathulae, the root of Dahurain angelica are ground into coarse powder, are mixed and stirred with the ethanol of equivalent 70%, vexed profit, according to the percolation under liquid extract and extract
（One annex of China's coastal port）, slowly diacolation, liquid of filtering filtration, is returned after measuring 70% ethanol as solvent soaking 24h using 10 times
Ethanol is received, and it is about 1.10 to be concentrated into relative density（35℃）, spray drying.Madder（System）, pseudo-ginseng be ground into fine powder, it is and above-mentioned
Cream powder and starch merge, and mix, and add 80% ethanol, are mixed with proper auxiliary materials, and corresponding system is made according to conventional formulation method
Agent.Major function: promoting blood circulation and hemostasis, removing blood stasis for promoting tissue regeneration, swelling and pain relieving, for caused by energy stagnation and blood stasis flooding, time lengthening,
It is dripping more than, underbelly pain;The common above-mentioned disease person of commotio retinae uterine hemorrhage amount.
Alizarin root of Dahurian angelica class Chinese medicine preparation proper mass standard only has a content discriminating, the Indexes Comparison list controlled final product quality
One.Preferably to control this quality, the compositions of our Chinese medicines using in preparation carries out assay as Con trolling index, and to containing
Quantity measuring method carries out the investigation of the methodologies such as linear relationship, stability, repeatability, the rate of recovery, as a result shows that this method is feasible,
Favorable reproducibility, specificity is strong, and controllability is good, can preferably control the inherent quality of this product, it is ensured that preparation process it is stable and
Quality it is controllable.
The content of the invention
It is an object of the invention to provide the method for quality control of alizarin root of Dahurian angelica class Chinese medicine preparation, to solve existing alizarin root of Dahurian angelica Chinese medicine preparation
Control method exist index components are single, characteristic differentiation not comprehensively, the problems such as assay specificity is poor, so as to more
The quality of scientific evaluation madder root of Dahurian angelica Chinese medicine preparation, it is ensured that the stability and security of its quality, improves product technology level, realizes profit
With the quality standard of the alizarin root of Dahurian angelica class Chinese medicine preparation of modern analysis means General Promotion, move towards the gateway of a country for the alizarin root of Dahurian angelica and more preferable guarantee is provided.
To achieve the above object, present invention employs following technical scheme:
A kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding, the method for quality control is included in the alizarin root of Dahurian angelica
The discrimination method and content assaying method of medicine preparation；Discriminating is to the madder control medicinal material in alizarin root of Dahurian angelica Chinese medicine preparation, rubia leiocaulis
Element, purpurin, pseudo-ginseng control medicinal material, notoginsenoside, ginsenoside, root of Dahurain angelica control medicinal material, Imperatorin, different Imperatoria ostruthium
Element, radix cyathulae control medicinal material, the indentification by TLC of cyasterone；The discrimination method is：
Differentiate：
（1）The method of madder is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes in the alizarin root of Dahurian angelica equivalent to 15.25~20.55g of raw medicinal herbs
Medicine preparation, removes and is coated, finely ground, adds 20~40ml of ether, places 1 hour, constantly shaking is extracted, filtration, and filtrate is flung to, residual
Slag adds ethyl acetate 1ml to make dissolving, standby as madder need testing solution；Madder control medicinal material 0.8~1.5g of powder separately is taken, together
Control drug solution is made in madder need testing solution preparation method；According to thin-layered chromatography experiment, each 5 μ of above two solution is drawn
L, puts on same silica gel g thin-layer plate respectively, the cyclohexane-acetone-two using volume ratio as 8~15: 4~10: 1~3: 4~10
Ethamine-water be solvent expansion, take out, dry, put and inspected under ultraviolet lamp (365nm), in test sample chromatogram, with comparison medicine
Wood color is composed on corresponding position, shows the spot of same color；
It is reference substance to take rubimaillin, adds the solution that 1ml 0.1mg containing rubimaillin are made in methanol, molten as reference substance
Liquid；According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of the madder need testing solution are drawn, is put respectively in same silica G
On lamellae, using volume ratio as 20~25: 3~6: 0.1~0.8 petroleum ether（60~90 DEG C）- ethyl acetate-ice vinegar
Acid is solvent expansion, takes out, dries, and spray is with dinitrophenylhydrazine ethanol test solution, in test sample chromatogram, with control medicinal material chromatogram
On corresponding position, show the spot of same color；
It is reference substance to take purpurin, adds methanol and every μ g of 1ml hydroxyls rubican 40 are made, be used as reference substance solution；According to
Thin-layered chromatography is tested, and is drawn reference substance solution and each 5 μ l of the madder need testing solution, is put respectively in same silica G thin layer
On plate, by 15~20: 5~12: 1~2 ethyl acetate of volume ratio-acetone-formic acid is solvent, is deployed, take out, dry,
Put and inspected under ultraviolet lamp (365nm), in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Spot；
（2）The method of pseudo-ginseng is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes the alizarin root of Dahurian angelica Chinese medicine equivalent to 13.6~18.6g of raw medicinal herbs
Preparation, removes and is coated, finely ground, adds 15~30ml of methanol solution of 5% formic acid, is heated to reflux 30 minutes, lets cool, and filters, filtrate
Fling to, residue adds methanol 2ml to make dissolving, it is standby as pseudo-ginseng need testing solution；It is another take pseudo-ginseng control medicinal material powder 0.8~
1.5g, control drug solution is made with pseudo-ginseng need testing solution preparation method；According to thin-layered chromatography experiment, above two is drawn molten
Each 5 μ l of liquid, put on same silica gel g thin-layer plate respectively, by 8~12: 6~9: 1~3 benzene of volume ratio-acetic ether-methanoic acid is
Solvent, deploy, take out, dry, put and inspected under ultraviolet lamp (365nm), in test sample chromatogram, with control medicinal material chromatogram phase
On the position answered, show the spot of same color；
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Solution of every 1ml containing 1mg is made in reference substance, plus methanol, molten as reference substance
Liquid；According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of pseudo-ginseng need testing solution are drawn, is put respectively in same silica G thin layer
On plate, by 5~12: 3~7: 0.5~1.5 chloroform-methanols of volume ratio-upper solution of glacial acetic acid is solvent, expansion takes
Go out, dry, spray with ethanol solution of sulfuric acid, be heated to spot development at 105 DEG C clear；In test sample chromatogram, with reference substance color
Compose on corresponding position, show the spot of same color；
Take ginsenoside Rb1's reference substance and ginsenoside Re's reference substance, plus methanol that mixed solutions of every 1ml containing 0.5mg is made, make
For reference substance solution；According to thin-layered chromatography experiment, reference substance solution and each 3 μ l of pseudo-ginseng need testing solution are drawn.Put respectively in same
On one silica gel g thin-layer plate, by 10~18: 35~45: 17~25: 8~12 chloroforms of volume ratio-acetate-methanol-water
10 DEG C of lower floor's solution arranged below are solvent, are deployed, and take out, dry, and spray with sulfuric acid solution, spot is heated at 105 DEG C
Colour developing is clear；In test sample chromatogram, on position corresponding with reference substance chromatogram, show identical colored speckles；
（3）The method of the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes the alizarin root of Dahurian angelica Chinese medicine equivalent to 14.6~19.8g of raw medicinal herbs
Preparation, removes and is coated, finely ground, plus petroleum ether（60~90 DEG C）10~40ml, shaking is extracted 30 minutes, and filtration, filter residue is standby, filter
Liquid flings to petroleum ether to 2ml, standby as root of Dahurain angelica need testing solution；Separately take the root of Dahurain angelica 0.4~0.9g of control medicinal material, plus petroleum ether
（60~90 DEG C）5~10ml, shaking is extracted 30 minutes, is stood, and supernatant is used as control medicinal material solution.According to thin-layered chromatography examination
Test, draw each 5 μ l of above two solution, put respectively on same silica gel g thin-layer plate, using volume ratio as 12~20: 2~5: 0.1
~0.8 petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid is solvent, is deployed below 25 DEG C, takes out, dries, put purple
Inspected under outer lamp (365nm)；In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color；
Take Imperatorin reference substance and Isomperatorin reference substance, adding ethyl acetate, that the mixing that every 1ml contains 1mg is made is molten
Liquid, is used as reference substance solution；According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of root of Dahurain angelica need testing solution are drawn, respectively point
In on same silica gel g thin-layer plate, using volume ratio as 5~9: 2~5: 0~1 petroleum ether（30~60 DEG C）- ether-formic acid is expansion
Agent, deploys below 25 DEG C, takes out, dries, put and inspected under ultraviolet lamp；In test sample chromatogram, with control medicinal material chromatogram phase
On the position answered, show the spot of same color；
（4）The method of radix cyathulae is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes in the alizarin root of Dahurian angelica equivalent to 12.7~18.9g of raw medicinal herbs
Medicine preparation, plus ethanol 30~50ml, ultrasonically treated 30 minutes, filtration, filtrate was evaporated, and residue adds diethyl ether 15~25ml, and shaking is carried
Take, volatilize, residue adds ethyl acetate 1ml to make dissolving, be used as radix cyathulae need testing solution；It is another take radix cyathulae control medicinal material 1~
4g, plus 20~40ml of ethanol, control drug solution is made with radix cyathulae need testing solution preparation method；According to TLC test,
The μ l of radix cyathulae need testing solution 6~8, the μ l of control medicinal material solution 3~5 are drawn, is put respectively on same silica G lamellae, with body
Product is deployed than being 10~25: 8~18: 1~3 toluene: ethyl acetate: formic acid is solvent, takes out, dries, and sprays with volume ratio
For 1: 1% ferric trichloride: it is clear that the mixed solution of 1% potassium ferricyanide is heated to spot development；In test sample chromatogram, with compareing
On the corresponding position of medicinal material chromatogram, show the spot of same color；
Take cyasterone reference substance, plus methanol that solution of every 1ml containing 0.5mg is made, be used as reference substance solution；According to thin-layered chromatography
Experiment, draws the μ l of need testing solution 5~10 and μ l of reference substance solution 5, puts respectively on same silica gel g thin-layer plate, with volume ratio
It is solvent for 13~18: 3~5: 1~2 chloroforms-acetone-formic acid, deploys, take out, dry, sprays with 10% ethanol solution of sulfuric acid,
Spot development is heated at 105 DEG C clear, puts and inspected under ultraviolet lamp；In test sample chromatogram, corresponding to control medicinal material chromatogram
Position on, show same color spot；
The content assaying method of the alizarin root of Dahurian angelica Chinese medicine preparation, it is main using root of Dahurain angelica institute in high effective liquid chromatography for measuring Chinese medicine composition
Amount containing Imperatorin；
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: from the chromatographic column that octadecylsilane chemically bonded silica is filler；
Mobile phase: using volume ratio as（30∶70）~（65∶35）Component A and component B constitute mixed solvent be used as flowing
Phase；The component A is the acetonitrile and methanol solution that volume ratio is 5: 1；Component B is 0.01~0.05mol/L biphosphates
Potassium solution or sodium dihydrogen phosphate or ammonium dihydrogen phosphate（3ml triethylamine is added per 1000ml）, adjusted with phosphoric acid solution
PH values are saved to 3.0~4.5；
Detection wavelength: 275~315nm；
Column temperature: 25~45 DEG C；
Flow velocity: 0.8~1.2ml/min of flow velocity；
The preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contains 20 μ g
Solution, shake up, produce.
The preparation of need testing solution: this product equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken,
It is accurately weighed, put in conical flask with cover, the methanol 25ml of precision addition 50%, weighed weight, ultrasonic extraction 30~40 minutes,
Let cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filters, discards primary filtrate, remaining filtrate is by aperture
0.45 μm of miillpore filter filtration, produces need testing solution.
Determination method: it is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, is determined,
Produce.
The present invention is a kind of active component by controlling alizarin root of Dahurian angelica class Chinese medicine preparation and then controls the analysis side of patent medicine quality
Method, to the quality standard of the alizarin root of Dahurian angelica improve, perfect, on the basis of ministry standard, improves the quality control of medicine, it is ensured that
The stability and security of its quality, improve product technology level, utilize the alizarin root of Dahurian angelica Chinese medicine preparation of modern analysis means General Promotion
Quality standard, make the production technology of the alizarin root of Dahurian angelica more it is strict rationally.
Present invention understands that the index components of certainly existing alizarin root of Dahurian angelica Chinese medicine preparation control method presence are single, characteristic differentiation is not comprehensive, contain
The problems such as specificity is poor is determined in measurement, so as to the quality of the alizarin root of Dahurian angelica Chinese medicine preparation of more scientific evaluation, it is ensured that the stabilization of its quality
Property and security, improve product technology level, realize the quality using the alizarin root of Dahurian angelica class Chinese medicine preparation of modern analysis means General Promotions
Standard, moves towards the gateway of a country for the alizarin root of Dahurian angelica and provides more preferable guarantee.
The present invention has done following research to the content of alizarin root of Dahurian angelica Chinese medicine preparation Imperatorin:
First, experimental drug
1. the prescription of experimental drug: radix cyathulae 600g pseudo-ginseng 85g madders（System）85g roots of Dahurain angelica 85g
2. the preparation method of experimental drug: four tastes above, radix cyathulae, the root of Dahurain angelica be ground into coarse powder, is mixed and stirred with the ethanol of equivalent 70%, vexed profit, photograph
Liquid extract and the percolation under extract（One annex of China's coastal port, is soaked using 10 times of 70% ethanol of amount as solvent
Slowly diacolation, liquid of filtering filtration after 24h is steeped, ethanol is reclaimed, and it is about 1.10 to be concentrated into relative density（35℃）, spray drying.It is alizarin
Grass（System）, pseudo-ginseng be ground into fine powder, merge with above-mentioned cream powder and starch, mix, add 80% alcohol granulation, dry, stiffened resin acid
Magnesium is mixed, and is arranged, is pressed into 1000, film coating is produced.
2nd, research process:
1st, instrument, reagent, sample: Waters515 types high performance liquid chromatograph, Waters2487UV-Vis detectors, WDL-95
Chromatographic work station, UV-1800 type ultraviolet-uisible spectrophotometers, 800 type settling vessels（Shanghai Surgical Operation Equipment Factory）, QX type ultrasonic waves
Shaker.Reagent: acetonitrile, methanol, potassium dihydrogen phosphate, triethylamine are chromatographically pure, and water is double distilled water.Imperatorin reference substance
（Chinese pharmaceutical biological product inspection institute）.Alizarin root of Dahurian angelica piece（Gansu Lanyao Pharmaceuticals Co., Ltd.）.
2nd, the selection of wavelength is determined：Imperatorin reference substance solution is taken, with ultraviolet-uisible spectrophotometer, in wave-length coverage
Measure is scanned in 200~400nm wave-length coverages.As a result show: wavelength has preferable effect in 275nm~315nm scopes
Really, its Imperatorin maximum absorption wavelength is 300nm.
3rd, chromatographic condition: chromatographic column：μ-Bondapak2 C18(4.6nm × 250nm, 5 μm, Chinese Academy of Sciences's Dalian Chemical Physics
Chromatogram research analysis center of country of research institute)；Mobile phase: the acetonitrile: methanol that volume ratio is 45: 55（5∶1）— 0.02mol/
L potassium dihydrogen phosphate mixed solvents are used as mobile phase（3ml triethylamine is added per 1000ml）, pH is adjusted with phosphoric acid solution
It is worth to 4.0.Flow velocity: 1.0ml/min.Column temperature: 30 DEG C of sample sizes: 5 μ L.
4th, the preparation of need testing solution:
（1）The determination of extraction time:
Alizarin root of Dahurian angelica Chinese medicine preparation is taken, it is finely ground, about 1.5g is taken, it is four parts, accurately weighed, put in conical flask with cover, precision plus 50% methanol
25ml, weighed weight is ultrasonically treated respectively（Power 250W, frequency 40kHz）20th, 30,40,50 minutes, let cool, then it is weighed heavy
Amount, the weight of less loss is supplied with 50% methanol, is shaken up, and is filtered.Subsequent filtrate (0.45 μm) filtration of miillpore filter, is determined.As a result
It is shown in Table 1.
The comparison of the extraction time of table 1
It can be seen from the result of table 1, extract 40 minutes and the content of 50 minutes Imperatorins is approached, therefore the ultrasonic conduct in 40 minutes of choosing
Extraction time.
（2）This product equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken, it is accurately weighed, put tool plug
In conical flask, precision adds 50% methanol 50ml, and weighed weight, ultrasonic extraction 40 minutes is let cool, then weighed weight, with 50%
Methanol supplies the weight of less loss, filtration, discards primary filtrate, and remaining filtrate is that 0.45 μm of miillpore filter is filtered by aperture, i.e.,
Obtain need testing solution.
5th, the preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contained
There is the solution of 20 μ g Imperatorins, shake up, produce.
6th, the preparation of negative need testing solution: by prescription（Remove the root of Dahurain angelica）, pharmaceutical preparation is prepared into according to preparation technology.Press
According to the preparation method of need testing solution, negative need testing solution is similarly made.Analyzed according to content determination, blank sample chromatogram
Figure does not interfere with peak in Imperatorin peak retention time.As a result show, it is negative noiseless.
7th, the investigation of linear relationship
Precision draws Imperatorin reference substance solution（20 μg /ml）8th, 10,12,16,20,24 μ l, are entered by above-mentioned chromatographic condition
Sample, determines peak area, the results are shown in Table 2.
The Imperatorin linear relationship measurement result of table 2
Using respective sample size as abscissa, peak area is absorbed as ordinate using reference substance, standard curve is drawn, obtains standard curve
Equation Y=5937.0X+98.61（r=0.9998）, Fig. 1 is made to sample size (μ g) to absorb peak area value, a straight line is obtained.More than
As a result show, Imperatorin absorbs peak area value with sample size in good linear pass in the range of 0.16~0.48 μ g
System.
8th, precision test precision draws Imperatorin reference substance solution 20ul, continuous sample introduction 5 times, with reference substance peak face
Product is inspection target, and as a result the RSD of Imperatorin peak area is 1.10%, shows that the precision of this method is good.
9th, stability test precision draws above-mentioned need testing solution 20ul, and respectively in 0h, 4h, 8h, 12h, 20h and 24h enter
Sample, using test sample calculated by peak area RSD as 1.12%.Measurement result shows that Imperatorin is basicly stable in 24h.
10th, the alizarin root of Dahurian angelica piece fine powder that replica test precision weighs same lot number in right amount, does 5 parts, by need testing solution altogether
Prepare and carry out time-and-motion study with the method determined under item.
The replica test result of table 3
Content average value is measured as shown in Table 3 for 0.0762mg/ pieces, RSD is 1.92%, illustrate that this method determines repetition
Property is good.
11st, recovery test precision weighs the sample about 8g of known content, 6 parts, accurately weighed, is respectively placed in tool plug cone
In shape bottle, precision weighs addition Imperatorin reference substance respectively, according to the content assaying method of sample, the results are shown in Table 4.
The recovery test result of table 4
As a result show, this method determines Imperatorin and shows that average recovery is good.
12nd, the measure of sample size takes each 3 parts of 3 batches of samples, as stated above every gram of raw medicinal herbs Imperatoria ostruthium in determination sample
The content of element, measurement result is shown in Table 5.
The sample size determination experiment of table 5
Average content value by three batches of sample Imperatorins of results showed that of table 5 is 0.076mg, and RSD is 4.2%.Knot
Actual state and international food Drug Administration standard are produced in symphysis, therefore require that this Chinese medicine preparation equivalent to 1g raw medicinal herbs contains
The root of Dahurain angelica is with Imperatorin（C18H14O4Meter）Meter, must not be less than 0.067mg.
Brief description of the drawings
Fig. 1 is Imperatorin linear relationship measurement result figure of the present invention.
Embodiment
The present invention is further illustrated below by embodiment.
A kind of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding of the present invention it is main by radix cyathulae, pseudo-ginseng, madder,
The root of Dahurain angelica is constituted, and being made various according to the conventional method of pharmacy is applied to the acceptable capsule of clinical treatment, tablet, particle
Agent, pill and pill etc..By promoting blood circulation and hemostasis, removing blood stasis for promoting tissue regeneration, swelling and pain relieving, for caused by energy stagnation and blood stasis uterine hemorrhage mistake
Many, time lengthening is dripping more than, underbelly pain；The common above-mentioned disease person of commotio retinae uterine hemorrhage amount.The system provided
The method of quality control of agent is stable, reliable, better than ministry standard, more effectively ensure that the quality of alizarin root of Dahurian angelica class Chinese medicine preparation.Its
Method of quality control includes the discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese medicine preparation.
Embodiment 1, is a kind of quality control of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding of capsule for formulation
Method processed, the discrimination method is：
Character: capsule shells are removed, alizarin root of Dahurian angelica Chinese medicine preparation content is the powder of brown；Gas fragrance, pungent, slight bitter.
Differentiate:
（1）The method of madder is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: 20 this product are taken, capsule shells are removed, ether 20ml is added,
Place 1 hour, constantly shaking is extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution.
Madder control medicinal material powder 0.8g separately is taken, control drug solution is made with need testing solution preparation method.According to thin-layered chromatography examination
Test, draw each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively on same silica gel g thin-layer plate, with hexamethylene
Alkane-acetone-diethylamine-water（8∶4∶1∶5）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp (365nm), supply
In test product chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
It is reference substance to take rubimaillin, adds the solution that 1ml 0.1mg containing rubimaillin are made in methanol, is used as control
Product solution.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica G thin layer
On plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（20∶3∶0.2）For solvent, deploy, take out, dry in the air
Dry, spray in test sample chromatogram, on position corresponding with control medicinal material chromatogram, shows identical face with dinitrophenylhydrazine ethanol test solution
The spot of color.
It is reference substance to take purpurin, adds methanol and every 1ml hydroxyls rubican 40 μ g are made, molten as reference substance
Liquid.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid（15∶7∶1）For solvent, deploy, take out, dry, put ultraviolet lamp（365nm）
Under inspect, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color spot.
（2）The method of pseudo-ginseng is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes 17 this product, removes capsule shells, adds 5% first
The methanol solution 20ml of acid, is heated to reflux 30 minutes, lets cool, and filters, and filtrate is flung to, and residue adds methanol 2ml to make dissolving, is used as confession
Test sample solution.Pseudo-ginseng control medicinal material powder 0.8g separately is taken, control drug solution is made with need testing solution preparation method.According to thin layer
Chromatography experiment, draws each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively in same silica gel g thin-layer plate
On, with benzene-acetic ether-methanoic acid（8∶7∶1）For solvent, deploy, take out, dry, put ultraviolet lamp（365nm）Under inspect,
In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Solution of every 1ml containing 1mg is made in reference substance, plus methanol, is used as reference substance
Solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with chloroform-methanol-water-glacial acetic acid (7: 2: 4: 1）Upper solution be solvent, deploy, take out, dry, spray with sulfuric acid second
Alcoholic solution（1→10）, it is heated to spot development at 105 DEG C clear.In test sample chromatogram, in position corresponding with reference substance chromatogram
On, show the spot of same color.
Take ginsenoside Rb1's reference substance and ginsenoside Re's reference substance, plus that mixing of every 1ml containing 0.5mg is made is molten for methanol
Liquid, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 3 μ l of need testing solution are drawn.Put respectively in
On same silica gel g thin-layer plate, with chloroform-acetate-methanol-water（10∶35∶17∶8）10 DEG C of lower floors arranged below are molten
Liquid is solvent, is deployed, and takes out, dries, and is sprayed with sulfuric acid solution（1→10）, it is heated to spot development at 105 DEG C clear.For examination
In product chromatogram, on position corresponding with reference substance chromatogram, show identical colored speckles.
（3）The method of the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes 18 this product, removes capsule shells, plus petroleum ether
（60~90 DEG C）20ml, shaking is extracted 30 minutes, and filtration, filter residue is standby, and filtrate flings to petroleum ether to about 2ml, is used as test sample
Solution.Separately take root of Dahurain angelica control medicinal material 0.4g, plus petroleum ether（60~90 DEG C）5ml, shaking is extracted 30 minutes, is stood, and supernatant is made
For control medicinal material solution.According to thin-layered chromatography experiment, each 5 μ l of above two solution are drawn（Need testing solution gives over to standby）, point
Other point is on same silica gel g thin-layer plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（12∶2∶0.3）For solvent,
Deploy below 25 DEG C, take out, dry, put uviol lamp（365nm）Under inspect.In test sample chromatogram, with control medicinal material chromatogram
On corresponding position, show the spot of same color.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added and the mixing that every 1ml contains 1mg is made
Solution, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of three kinds of solution of need testing solution are drawn,
Put respectively on same silica gel g thin-layer plate, with petroleum ether（30~60 DEG C）- ether（5∶2）For solvent, opened up below 25 DEG C
Open, take out, dry, put ultraviolet lamp（365nm）Under inspect.In test sample chromatogram, in position corresponding with control medicinal material chromatogram
On, show the spot of same color.
（4）The method of radix cyathulae is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes 14 this product, removes capsule shells, plus ethanol
30ml, ultrasonically treated 30 minutes, filtration, filtrate is evaporated, and residue adds diethyl ether 15ml, and shaking is extracted, volatilized, residue adds ethyl acetate
1ml makes dissolving, is used as need testing solution；Separately take radix cyathulae control medicinal material 2g, plus ethanol 20ml, need testing solution preparation method system
Shine drug solution in pairs.According to TLC test, the μ l of need testing solution 6, the μ l of control medicinal material solution 3 are drawn, is put respectively in same
On silica G lamellae, with toluene: ethyl acetate: formic acid（10∶9∶1）For solvent, deploy, take out, dry, spray with 1% 3
Iron chloride: 1% potassium ferricyanide（1∶1）Mixed solution to be heated to spot development clear.In test sample chromatogram, with comparison medicine
Wood color is composed on corresponding position, shows the spot of same color.
Take cyasterone reference substance, plus methanol that solution of every 1ml containing 0.5mg is made, be used as reference substance solution.According to thin layer color
Spectrometry is tested, and is drawn the μ l of the need testing solution 5 and μ l of reference substance solution 5, is put respectively on same silica gel g thin-layer plate, with chloroform-the third
Ketone-formic acid (13: 3: 1) is solvent, is deployed, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear, puts ultraviolet lamp（365nm）Under inspect.In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show
The spot of same color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: from the chromatographic column that octadecylsilane chemically bonded silica is filler；
Mobile phase: the acetonitrile: methanol that volume ratio is 45: 55（5∶1）- 0.02mol/l potassium dihydrogen phosphate mixed solvents
It is used as mobile phase（3ml triethylamine is added per 1000ml）, pH values are adjusted with phosphoric acid solution to 4.0；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contained
20 μ g solution, shakes up, produces.
The preparation of need testing solution: this product equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken,
It is accurately weighed, put in conical flask with cover, the methanol 25ml of precision addition 50%, weighed weight, ultrasonic extraction 30~40 minutes,
Let cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filters, discards primary filtrate, remaining filtrate is by aperture
0.45 μm of miillpore filter filtration, produces need testing solution.
Determination method: it is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, is determined,
Produce.
This Chinese medicine preparation equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin（C18H14O4Meter）Meter, must not be less than
0.067mg。
Embodiment 2, is a kind of quality control of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding of tablet for formulation
Method, the discrimination method is：
Character: this product is Film coated tablets, is removed brown after being coated：Gas fragrance, pungent, slight bitter.
Differentiate:
（1）The method of madder is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: 25 this product are taken, removes and is coated, it is finely ground, and add ether
40ml, is placed 1 hour, constantly shaking is extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make dissolving, is used as test sample
Solution.Madder control medicinal material powder 1.5g separately is taken, control drug solution is made with need testing solution preparation method.According to thin-layer chromatography
Method is tested, and draws each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively on same silica gel g thin-layer plate, with
Cyclohexane-acetone-diethylamine-water（15∶10∶3∶10）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp, supply
In test product chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
It is reference substance to take rubimaillin, adds the solution that 1ml 0.1mg containing rubimaillin are made in methanol, is used as control
Product solution.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica G thin layer
On plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（25∶5∶0.8）For solvent, deploy, take out, dry,
Spray is with dinitrophenylhydrazine ethanol test solution, in test sample chromatogram, on position corresponding with control medicinal material chromatogram, shows same color
Spot.
It is reference substance to take purpurin, adds methanol and every 1ml hydroxyls rubican 40 μ g are made, molten as reference substance
Liquid.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid（20∶11∶2）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp,
In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
（2）The method of pseudo-ginseng is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: 22 this product are taken, removes and is coated, it is finely ground, and add 5%
The methanol solution 30ml of formic acid, is heated to reflux 30 minutes, lets cool, and filtration, filtrate is flung to, and residue adds methanol 2ml to make dissolving, as
Need testing solution.Pseudo-ginseng control medicinal material powder 1.5g separately is taken, control drug solution is made with need testing solution preparation method.According to thin
Layer chromatography is tested, and draws each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively in same silica G thin layer
On plate, with benzene-acetic ether-methanoic acid（12∶9∶2.5）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp, supply
In test product chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Solution of every 1ml containing 1mg is made in reference substance, plus methanol, is used as reference substance
Solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with chloroform-methanol-water-glacial acetic acid（12∶7∶5∶1）Upper solution be solvent, deploy, take out, dry, spray with sulfuric acid
Ethanol solution（1→10）, it is heated to spot development at 105 DEG C clear.In test sample chromatogram, in position corresponding with reference substance chromatogram
Put, show the spot of same color.
Take ginsenoside Rb1's reference substance and ginsenoside Re's reference substance, plus that mixing of every 1ml containing 0.5mg is made is molten for methanol
Liquid, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 3 μ l of need testing solution are drawn.Put respectively in same
On one silica gel g thin-layer plate, with chloroform-acetate-methanol-water（18∶45∶25∶12）10 DEG C of lower floors arranged below are molten
Liquid is solvent, is deployed, and takes out, dries, and sprays with sulfuric acid solution, is heated to spot development at 105 DEG C clear.Test sample chromatogram
In, on position corresponding with reference substance chromatogram, show identical colored speckles.
（3）The method of the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: 22 this product are taken, removes and is coated, it is finely ground, plus oil
Ether（60~90 DEG C）40ml, shaking is extracted 30 minutes, and filtration, filter residue is standby, and filtrate flings to petroleum ether to about 2ml, as trying
Product solution.Separately take root of Dahurain angelica control medicinal material 0.9g, plus petroleum ether（60~90 DEG C）10ml, shaking is extracted 30 minutes, is stood, supernatant
It is used as control medicinal material solution.According to thin-layered chromatography experiment, each 5 μ l of above two solution are drawn（Need testing solution gives over to standby）,
Put respectively on same silica gel g thin-layer plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（20∶5∶0.8）For expansion
Agent, deploys below 25 DEG C, takes out, dries, put and inspected under uviol lamp.In test sample chromatogram, corresponding to control medicinal material chromatogram
Position on, show same color spot.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added and the mixing that every 1ml contains 1mg is made
Solution, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of three kinds of solution of need testing solution are drawn,
Put respectively on same silica gel g thin-layer plate, with petroleum ether（30~60 DEG C）- ether-formic acid（9∶5∶1）For solvent, at 25 DEG C
Deploy below, take out, dry, put and inspected under ultraviolet lamp.In test sample chromatogram, in position corresponding with control medicinal material chromatogram
On, show the spot of same color.
（4）The method of radix cyathulae is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes this product equivalent to 20, plus ethanol
50ml, ultrasonically treated 30 minutes, filtration, filtrate is evaporated, and residue adds diethyl ether 25ml, and shaking is extracted, volatilized, residue adds ethyl acetate
1ml makes dissolving, is used as need testing solution；Separately take radix cyathulae control medicinal material 4g, plus ethanol 40ml, need testing solution preparation method
Control drug solution is made.According to TLC test, the μ l of need testing solution 8, the μ l of control medicinal material solution 5 are drawn, is put respectively in same
On one silica G lamellae, with toluene: ethyl acetate: formic acid（25 ∶18 ∶3）For solvent, deploy, take out, dry, spray with
1% ferric trichloride: 1% potassium ferricyanide（1 ∶ 1）Mixed solution to be heated to spot development clear.In test sample chromatogram, with
On the corresponding position of control medicinal material chromatogram, show the spot of same color.
Take cyasterone reference substance, plus methanol that solution of every 1ml containing 0.5mg is made, be used as reference substance solution.According to thin layer color
Spectrometry is tested, and draws the μ l of the need testing solution 10 and μ l of reference substance solution 5, is put respectively on same silica gel g thin-layer plate, with chloroform-
Acetone-formic acid (18: 5: 2) is solvent, is deployed, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Point colour developing is clear, puts and is inspected under ultraviolet lamp.In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show identical
The spot of color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: from the chromatographic column that octadecylsilane chemically bonded silica is filler；
Mobile phase: the acetonitrile: methanol that volume ratio is 45: 55（5∶1）- 0.02mol/l potassium dihydrogen phosphate mixed solvents
It is used as mobile phase（3ml triethylamine is added per 1000ml）, pH values are adjusted with phosphoric acid solution to 4.0；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation of need testing solution: this product equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken,
It is accurately weighed, put in conical flask with cover, the methanol 25ml of precision addition 50%, weighed weight, ultrasonic extraction 30~40 minutes,
Let cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filters, discards primary filtrate, remaining filtrate is by aperture
0.45 μm of miillpore filter filtration, produces need testing solution.
The preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contained
20 μ g solution, shakes up, produces.
Determination method: it is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, is determined,
Produce.
This Chinese medicine preparation equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin（C18H14O4Meter）Meter, must not be less than
0.067mg。
Embodiment 3, is a kind of quality control of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding of pill for formulation
Method, the discrimination method is：
Character: alizarin root of Dahurian angelica pill is brown：Gas fragrance, pungent, slight bitter.
Differentiate:
（1）The method of madder is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: this product of 23 balls is taken, it is finely ground, and ether 30ml is added, is placed
1 hour, constantly shaking was extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make dissolving, is used as need testing solution.Separately take
Madder control medicinal material powder 1.2g, control drug solution is made with need testing solution preparation method.According to thin-layered chromatography experiment, inhale
Take each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively on same silica gel g thin-layer plate, with hexamethylene-the third
Ketone-diethylamine-water（12∶5∶2∶5）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp, in test sample chromatogram,
On position corresponding with control medicinal material chromatogram, show the spot of same color.
It is reference substance to take rubimaillin, adds the solution that 1ml 0.1mg containing rubimaillin are made in methanol, is used as control
Product solution.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica G thin layer
On plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（22∶4∶0.5）For solvent, deploy, take out, dry in the air
Dry, spray in test sample chromatogram, on position corresponding with control medicinal material chromatogram, shows identical face with dinitrophenylhydrazine ethanol test solution
The spot of color.
It is reference substance to take purpurin, adds methanol and every 1ml hydroxyls rubican 40 μ g are made, molten as reference substance
Liquid.According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid（17∶9∶1）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp, supply
In test product chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color.
（2）The method of pseudo-ginseng is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: this product of 20 balls is taken, it is finely ground, and add 5% formic acid
Methanol solution 25ml, is heated to reflux 30 minutes, lets cool, and filtration, filtrate is flung to, and residue adds methanol 2ml to make dissolving, is used as test sample
Solution.Pseudo-ginseng control medicinal material powder 1.1g separately is taken, control drug solution is made with need testing solution preparation method.According to thin-layer chromatography
Method is tested, and draws each 5 μ l of above two solution（Need testing solution gives over to standby）, put respectively on same silica gel g thin-layer plate, with
Benzene-acetic ether-methanoic acid（10∶7∶2）For solvent, deploy, take out, dry, put and inspected under ultraviolet lamp, test sample chromatogram
In, on position corresponding with control medicinal material chromatogram, show the spot of same color.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Solution of every 1ml containing 1mg is made in reference substance, plus methanol, is used as reference substance
Solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, with chloroform-methanol-glacial acetic acid（9∶5∶1）Upper solution be solvent, deploy, take out, dry, spray it is molten with sulfuric acid ethanol
Liquid, is heated to spot development clear at 105 DEG C.In test sample chromatogram, on position corresponding with reference substance chromatogram, show identical face
The spot of color.
Take ginsenoside Rb1's reference substance and ginsenoside Re's reference substance, plus that mixing of every 1ml containing 0.5mg is made is molten for methanol
Liquid, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 3 μ l of need testing solution are drawn.Put respectively in same
On one silica gel g thin-layer plate, with chloroform-acetate-methanol-water（14∶40∶22∶9）10 DEG C of lower floor's solution arranged below
For solvent, deploy, take out, dry, spray with sulfuric acid solution, be heated to spot development at 105 DEG C clear.In test sample chromatogram,
On position corresponding with reference substance chromatogram, show identical colored speckles.
（3）The method of the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: 21 ball this product are taken, it is finely ground, plus petroleum ether（60~90
℃）25ml, shaking is extracted 30 minutes, and filtration, filter residue is standby, and filtrate flings to petroleum ether to about 2ml, is used as need testing solution.Separately
Take root of Dahurain angelica control medicinal material 0.6g, plus petroleum ether（60~90 DEG C）8ml, shaking is extracted 30 minutes, is stood, and supernatant is used as comparison medicine
Material solution.According to thin-layered chromatography experiment, each 5 μ l of above two solution are drawn（Need testing solution gives over to standby）, put respectively in same
On one silica gel g thin-layer plate, with petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid（15∶4∶0.5）For solvent, at 25 DEG C
Deploy below, take out, dry, put and inspected under uviol lamp.In test sample chromatogram, on position corresponding with control medicinal material chromatogram,
The spot of aobvious same color.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added and the mixing that every 1ml contains 1mg is made
Solution, is used as reference substance solution.According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of three kinds of solution of need testing solution are drawn,
Put respectively on same silica gel g thin-layer plate, with petroleum ether（30~60 DEG C）- ether（7∶4）For solvent, opened up below 25 DEG C
Open, take out, dry, put and inspected under ultraviolet lamp.In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show phase
With the spot of color.
（4）The method of radix cyathulae is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes at 18 ball this product, plus ethanol 40ml, ultrasound
Reason 30 minutes, filtration, filtrate is evaporated, and residue adds diethyl ether 20ml, and shaking is extracted, volatilized, residue adds ethyl acetate 1ml to make dissolving,
It is used as need testing solution；Radix cyathulae control medicinal material 2.5g, plus ethanol 30ml separately are taken, comparison medicine is made in need testing solution preparation method
Product solution.According to TLC test, the μ l of need testing solution 7, the μ l of control medicinal material solution 4 are drawn, is put respectively thin in same silica G
On laminate, with toluene: ethyl acetate: formic acid（18 ∶13 ∶2）For solvent, deploy, take out, dry, spray with 1% ferric trichloride
: 1% potassium ferricyanide（1 ∶ 1）Mixed solution to be heated to spot development clear.In test sample chromatogram, with comparison medicine wood color
Compose on corresponding position, show the spot of same color.
Take cyasterone reference substance, plus methanol that solution of every 1ml containing 0.5mg is made, be used as reference substance solution.According to thin layer color
Spectrometry is tested, and is drawn the μ l of the need testing solution 8 and μ l of reference substance solution 5, is put respectively on same silica gel g thin-layer plate, with chloroform-the third
Ketone-formic acid (15: 4: 1) is solvent, is deployed, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, spot is heated at 105 DEG C
Colour developing is clear, puts and is inspected under ultraviolet lamp.In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show identical face
The spot of color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: from the chromatographic column that octadecylsilane chemically bonded silica is filler；
Mobile phase: the acetonitrile: methanol that volume ratio is 45: 55（5∶1）- 0.02mol/l potassium dihydrogen phosphate mixed solvents
It is used as mobile phase（3ml triethylamine is added per 1000ml）, pH values are adjusted with phosphoric acid solution to 4.0；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation of need testing solution: this product equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken,
It is accurately weighed, put in conical flask with cover, precision adds 50% methanol 25ml, weighed weight, ultrasonic extraction 40 minutes is let cool, then
Weighed weight, the weight of less loss is supplied with 50% methanol, filtration, discards primary filtrate, and remaining filtrate is 0.45 μm micro- by aperture
Hole filter membrane filtration, produces need testing solution.
The preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contained
20 μ g solution, shakes up, produces.
Determination method: it is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, is determined,
Produce.
This Chinese medicine preparation equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin（C18H14O4Meter）Meter, must not be less than
0.067mg。
Claims (2)
1. a kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding, it is characterised in that the quality control
Method includes the discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese medicine preparation；Discriminating is to the madder control in alizarin root of Dahurian angelica Chinese medicine preparation
Medicinal material, rubimaillin, purpurin, pseudo-ginseng control medicinal material, notoginsenoside, ginsenoside, root of Dahurain angelica control medicinal material, Imperatoria ostruthium
Element, Isomperatorin, radix cyathulae control medicinal material, the indentification by TLC of cyasterone；The discrimination method is：
Character: the alizarin root of Dahurian angelica Chinese medicine preparation of the treatment uterus bleeding is brown, gas fragrance, pungent, slight bitter；
Differentiate：
（1）The method of madder is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes in the alizarin root of Dahurian angelica equivalent to 15.25~20.55g of raw medicinal herbs
Medicine preparation, removes and is coated, finely ground, adds 20~40ml of ether, places 1 hour, constantly shaking is extracted, filtration, and filtrate is flung to, residual
Slag adds ethyl acetate 1ml to make dissolving, standby as madder need testing solution；Madder control medicinal material 0.8~1.5g of powder separately is taken, together
Control drug solution is made in madder need testing solution preparation method；According to thin-layered chromatography experiment, each 5 μ of above two solution is drawn
L, puts on same silica gel g thin-layer plate respectively, the cyclohexane-acetone-two using volume ratio as 8~15: 4~10: 1~3: 4~10
Ethamine-water be solvent expansion, take out, dry, put and inspected under ultraviolet lamp (365nm), in test sample chromatogram, with comparison medicine
Wood color is composed on corresponding position, shows the spot of same color；
It is reference substance to take rubimaillin, adds the solution that 1ml 0.1mg containing rubimaillin are made in methanol, molten as reference substance
Liquid；According to thin-layered chromatography experiment, reference substance solution and each 5 μ l of the madder need testing solution are drawn, is put respectively in same silica G
On lamellae, using volume ratio as 20~25: 3~6: 0.1~0.8 petroleum ether（60~90 DEG C）- ethyl acetate-ice vinegar
Acid is solvent expansion, takes out, dries, and spray is with dinitrophenylhydrazine ethanol test solution, in test sample chromatogram, with control medicinal material chromatogram
On corresponding position, show the spot of same color；
It is reference substance to take purpurin, adds methanol and every μ g of 1ml hydroxyls rubican 40 are made, be used as reference substance solution；According to
Thin-layered chromatography is tested, and is drawn reference substance solution and each 5 μ l of the madder need testing solution, is put respectively in same silica G thin layer
On plate, by 15~20: 5~12: 1~2 ethyl acetate-acetones of volume ratio-formic acid is solvent, is deployed, take out, dry, put
Inspected under ultraviolet lamp (365nm), in test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color
Spot；
（2）The method of pseudo-ginseng is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes the alizarin root of Dahurian angelica Chinese medicine equivalent to 13.6~18.6g of raw medicinal herbs
Preparation, removes and is coated, finely ground, adds 15~30ml of methanol solution of 5% formic acid, is heated to reflux 30 minutes, lets cool, and filters, filtrate
Fling to, residue adds methanol 2ml to make dissolving, it is standby as pseudo-ginseng need testing solution；It is another take pseudo-ginseng control medicinal material powder 0.8~
1.5g, control drug solution is made with pseudo-ginseng need testing solution preparation method；According to thin-layered chromatography experiment, above two is drawn molten
Each 5 μ l of liquid, put on same silica gel g thin-layer plate respectively, by 8~12: 6~9: 1~3 benzene of volume ratio-acetic ether-methanoic acid is
Solvent, deploy, take out, dry, put and inspected under ultraviolet lamp (365nm), in test sample chromatogram, with control medicinal material chromatogram phase
On the position answered, show the spot of same color；
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Solution of every 1ml containing 1mg is made in reference substance, plus methanol, is used as reference substance solution；
According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of pseudo-ginseng need testing solution are drawn, is put respectively in same silica gel g thin-layer plate
On, by 5~12: 3~7: 0.5~1.5 chloroform-methanols of volume ratio-upper solution of glacial acetic acid is solvent, deploy, take out,
Dry, spray with ethanol solution of sulfuric acid, be heated to spot development at 105 DEG C clear；In test sample chromatogram, with reference substance chromatogram pair
On the position answered, show the spot of same color；
Take ginsenoside Rb1's reference substance and ginsenoside Re's reference substance, plus methanol that mixed solutions of every 1ml containing 0.5mg is made, make
For reference substance solution；According to thin-layered chromatography experiment, reference substance solution and each 3 μ l of pseudo-ginseng need testing solution are drawn;
Put respectively on same silica gel g thin-layer plate, by 10~18: 35~45: 17~25: 8~12 chloroforms of volume ratio-second
10 DEG C of acetoacetic ester-methanol-water lower floor's solution arranged below is solvent, is deployed, and takes out, dries, and is sprayed with sulfuric acid solution,
105 DEG C to be heated to spot development clear；In test sample chromatogram, on position corresponding with reference substance chromatogram, show identical color
Spot；
（3）The method of the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes the alizarin root of Dahurian angelica Chinese medicine equivalent to 14.6~19.8g of raw medicinal herbs
Preparation, removes and is coated, finely ground, plus petroleum ether（60~90 DEG C）10~40ml, shaking is extracted 30 minutes, and filtration, filter residue is standby, filter
Liquid flings to petroleum ether to 2ml, standby as root of Dahurain angelica need testing solution；Separately take the root of Dahurain angelica 0.4~0.9g of control medicinal material, plus petroleum ether
（60~90 DEG C）5~10ml, shaking is extracted 30 minutes, is stood, and supernatant is tried as control medicinal material solution according to thin-layered chromatography
Test, draw each 5 μ l of above two solution, put respectively on same silica gel g thin-layer plate, using volume ratio as 12~20: 2~5: 0.1
~0.8 petroleum ether（60~90 DEG C）- ethyl acetate-glacial acetic acid is solvent, is deployed below 25 DEG C, takes out, dries, put purple
Inspected under outer lamp (365nm)；In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the spot of same color；
Take Imperatorin reference substance and Isomperatorin reference substance, adding ethyl acetate, that the mixing that every 1ml contains 1mg is made is molten
Liquid, is used as reference substance solution；According to thin-layered chromatography experiment, reference substance solution and each 4 μ l of root of Dahurain angelica need testing solution are drawn, respectively point
In on same silica gel g thin-layer plate, using volume ratio as 5~9: 2~5: 0~1 petroleum ether（30~60 DEG C）- ether-formic acid is expansion
Agent, deploys below 25 DEG C, takes out, dries, put and inspected under ultraviolet lamp；In test sample chromatogram, with control medicinal material chromatogram phase
On the position answered, show the spot of same color；
（4）The method of radix cyathulae is identified in alizarin root of Dahurian angelica Chinese medicine preparation prescription is: takes in the alizarin root of Dahurian angelica equivalent to 12.7~18.9g of raw medicinal herbs
Medicine preparation, plus ethanol 30~50ml, ultrasonically treated 30 minutes, filtration, filtrate was evaporated, and residue adds diethyl ether 15~25ml, and shaking is carried
Take, volatilize, residue adds ethyl acetate 1ml to make dissolving, be used as radix cyathulae need testing solution；It is another take radix cyathulae control medicinal material 1~
4g, plus 20~40ml of ethanol, control drug solution is made with radix cyathulae need testing solution preparation method；According to TLC test,
The μ l of radix cyathulae need testing solution 6~8, the μ l of control medicinal material solution 3~5 are drawn, is put respectively on same silica G lamellae, with body
Product is deployed than being 10~25: 8~18: 1~3 toluene: ethyl acetate: formic acid is solvent, takes out, dries, and sprays with volume ratio
For 1: 1% ferric trichloride: it is clear that the mixed solution of 1% potassium ferricyanide is heated to spot development；In test sample chromatogram, with compareing
On the corresponding position of medicinal material chromatogram, show the spot of same color；
Take cyasterone reference substance, plus methanol that solution of every 1ml containing 0.5mg is made, be used as reference substance solution；According to thin-layered chromatography
Experiment, draws the μ l of need testing solution 5~10 and μ l of reference substance solution 5, puts respectively on same silica gel g thin-layer plate, with volume ratio
It is solvent for 13~18: 3~5: 1~2 chloroforms-acetone-formic acid, deploys, take out, dry, sprays with 10% ethanol solution of sulfuric acid,
Spot development is heated at 105 DEG C clear, puts and inspected under ultraviolet lamp；In test sample chromatogram, corresponding to control medicinal material chromatogram
Position on, show same color spot；
In the content assaying method of the alizarin root of Dahurian angelica Chinese medicine preparation, the main alizarin root of Dahurian angelica Chinese medicine composition using high effective liquid chromatography for measuring
The amount of Imperatorin contained by the root of Dahurain angelica；
Chromatographic condition and system suitability:
Flow stationary phase: from the chromatographic column that octadecylsilane chemically bonded silica is filler；
Mobile phase: using volume ratio as（30∶70）~（65∶35）Component A and component B constitute mixed solvent be used as stream
Dynamic phase；The component A is the acetonitrile and methanol solution that volume ratio is 5: 1, and component B is 0.01~0.05mol/L phosphoric acid
Dihydro potassium solution or sodium dihydrogen phosphate or ammonium dihydrogen phosphate（3ml triethylamine is added per 1000ml), it is molten with phosphoric acid
Liquid adjusts pH values to 3.0~4.5；
Detection wavelength: 275~315nm；
Column temperature: 25~45 DEG C；
Flow velocity: 0.8~1.2ml/min of flow velocity；
The preparation of need testing solution: the alizarin root of Dahurian angelica Chinese medicine preparation equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, take
1.8g, it is accurately weighed, put in conical flask with cover, precision adds 50% methanol 50ml, weighed weight, ultrasonic extraction 30~40
Minute, let cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filters, discards primary filtrate, remaining filtrate passes through hole
Footpath is 0.45 μm of miillpore filter filtration, produces need testing solution；
The preparation: precision of reference substance solution weighs Imperatorin reference substance, accurately weighed, plus methanol is made every 1ml and contains 20 μ g
Solution, shake up, produce；
Determination method: it is accurate respectively to draw reference substance solution and each 10 μ l of need testing solution, liquid chromatograph is injected, determines, produces.
2. a kind of method of quality control of alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding according to claim 1, its
It is characterised by：The chromatographic condition mixed with component A in system suitability and component B using volume ratio as 45: 55 after as
Mobile phase and the triethylamine that 3ml is added per 1000ml, pH values are adjusted to 4.0 with phosphoric acid solution；The component A is volume
Than the acetonitrile and methanol solution for 5: 1, the component B is 0.02mol/l potassium dihydrogen phosphates；The Detection wavelength
300nm ；The column temperature is 30 DEG C, and the flow velocity is 1.0ml/min.
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