CN106198789A - Cowherb red tablet quality control method - Google Patents
Cowherb red tablet quality control method Download PDFInfo
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- CN106198789A CN106198789A CN201610504410.9A CN201610504410A CN106198789A CN 106198789 A CN106198789 A CN 106198789A CN 201610504410 A CN201610504410 A CN 201610504410A CN 106198789 A CN106198789 A CN 106198789A
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- cowherb
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- flos carthami
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- hydroxysafflor yellow
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
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Abstract
The invention discloses a kind of cowherb red tablet quality control method, carry out Flos Carthami discriminating including utilizing thin layer chromatography and utilize high performance liquid chromatography to carry out S-A Hydroxysafflor yellow A assay.The cowherb red tablet quality control method of the present invention, it is provided that differentiate the method for Flos Carthami in medicine, and use main flavour of a drug Flos Carthami in high-efficient liquid phase technique the other side to carry out assay with S-A Hydroxysafflor yellow A for index.This method of quality control can better control over the quality of medicine, has stronger specificity and good repeatability, and real embodiment drug safety is effective, quality controllable.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, specifically, relate to a kind of cowherb red tablet quality control method.
Background technology
Cardiovascular and cerebrovascular disease is serious harm people's health, life-threatening, a kind of commonly encountered diseases of impact work, frequently-occurring disease,
Sickness rate is high.Traditional Chinese medical science cerebrovascular is called " apoplexy ".The cause of disease of apoplexy is the deficiency of vital energy, expectorant Sheng turbid, fiery, blood stasis etc..Major part patient
Belong to apoplex involving the channels and collaterals scope, occur meridians disease, minority serious symptom person can cause damage of kidney-YIN, extreme hyperactivity of the liver-yang, hold under the arm expectorant hold under the arm fire, horizontal alter through
Network, the disturbance mind, formed into dirty enter internal organs close disease or de-disease." the big opinion of Plain Questions YIN YANG classification of natural phenomena " " promoting circulation of QI-blood, makes its tune reach,
And it is flat to make peace ", " hard masses should be resolved gradually ", the theory of " disease with retention of pathogen in the body should be treated by elimination therapy ", with blood circulation promoting and blood stasis dispelling as principle for the treatment of.
Rhizoma Curcumae is warm in nature, bitter in the mouth, pungent, function broken knot eliminating stagnation, and promoting the circulation of blood pain relieving is the highest strong.Finding through pharmacological research, Rhizoma Curcumae can
Increase blood flow.Flos Carthami is feverfew, enters the heart, Liver Channel, has promoting blood circulation to restore menstrual flow, the effect of stasis-dispelling and pain-killing.Cowherb uses cowherb in red side
The power of its high strong removing blood stasis of art, " Bencao Tujing " " Rhizoma Curcumae, ancient prescription loses user, and modern doctor controls all gas of accumulation, for ministerial drug." therefore with
Rhizoma Curcumae is monarch drug.Flos Carthami " Amplification on Materia Medica addendum ": " Flos Carthami, broken stagnant blood nourish blood ", for ministerial drug.The monarch and his subjects make a concerted effort, and removing blood stasis with potent drugs is controlled blood and nourished blood
To uncharm, make vim and vigour adjust and reach.
For ensureing stability and the curative effect of preparation, red of cowherb having been formulated comprehensive quality standard, Flos Carthami is in red of cowherb
Main flavour of a drug, this law uses Flos Carthami in high-efficient liquid phase technique the other side to carry out assay with S-A Hydroxysafflor yellow A for index, with thin
Layer chromatography is respectively to Flos Carthami qualitative identification.Preparation has been carried out long-term stable experiment, and result shows preparation in 24 months
Quality, without significantly change, provides strong evidence for formulating the effect duration of preparation.
Summary of the invention
In view of this, the technical problem to be solved is to provide the method for quality control of red of a kind of cowherb, the party
Method can differentiate the Flos Carthami in red of cowherb fast and accurately, and by the middle hydroxyl Flos Carthami of red of high effective liquid chromatography for measuring cowherb
The content of safflor yellow A.
The invention discloses a kind of cowherb red tablet quality control method, carry out Flos Carthami discriminating and profit including utilizing thin layer chromatography
S-A Hydroxysafflor yellow A assay is carried out by high performance liquid chromatography.
Further, described Flos Carthami discrimination method is: take cowherb red 10, removes coating, finely ground one-tenth powder；Take described powder
End 0.5g, adds 80% acetone soln 5ml, shakes 15min, stand after close plug；Aspirate supernatant, as need testing solution；
Take Flos Carthami control medicinal material 0.5g, be made in the same way of control medicinal material solution；
Take blank sample, be made in the same way of placebo solution；
Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively and in same with sodium carboxymethyl cellulose be
On the silica gel H lamellae of adhesive, it is that developing solvent launches with acetic ether-methanoic acid-water-methanol (7:2:3:0.4), takes out, dry in the air
Dry；
Observe silica gel H lamellae, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, show in the sunlight
The speckle of same color.
Further, described S-A Hydroxysafflor yellow A content assaying method is:
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With volume ratio for 26:
The methanol of 2:72-second eyeball-0.7% phosphoric acid solution is flowing phase；Detection wavelength is 403nm, and number of theoretical plate presses hydroxyl Flos Carthami yellow
Element A peak calculates >=1500；
The preparation of reference substance solution: precision weighs S-A Hydroxysafflor yellow A reference substance, adds 25% methanol and makes every 1ml and contain
The solution of 0.13mg, to obtain final product；
The preparation of need testing solution: take cowherb red 20, removes coating, finely ground one-tenth powder, takes described powder 2g, accurate title
Fixed, put in tool plug conical flask, accurate addition 25% methanol 25ml, close plug, weighed weight, supersound process, let cool, more weighed heavy
Amount, adds 25% methanol and supplies the weight of less loss, shake up, and filters, takes subsequent filtrate, to obtain final product；
Assay method: precision draws reference substance solution and each 20 μ l of need testing solution respectively, injects chromatograph of liquid, surveys
Fixed, calculates and get final product, this product every contains Flos Carthami in terms of S-A Hydroxysafflor yellow A, no less than 0.045mg.
Further, in described S-A Hydroxysafflor yellow A content assaying method, the ultrasonic place of need testing solution preparation process
Reason power is 300W, and supersonic frequency is 50kHz, ultrasonic time 40 minutes.
Compared with prior art, the present invention can obtain and include techniques below effect:
1) cowherb of the present invention red tablet quality control method, it is provided that differentiate the method for Flos Carthami in medicine, and use high-efficient liquid
In phase method the other side, main flavour of a drug Flos Carthami carries out assay with S-A Hydroxysafflor yellow A for index.
2) method of quality control of the present invention can better control over the quality of medicine, has stronger specificity and good
Repeatability, real embodiment drug safety is effective, quality controllable.
Certainly, the arbitrary product implementing the present invention must be not necessarily required to reach all the above technique effect simultaneously.
Accompanying drawing explanation
Accompanying drawing described herein is used for providing a further understanding of the present invention, constitutes the part of the present invention, this
Bright schematic description and description is used for explaining the present invention, is not intended that inappropriate limitation of the present invention.In the accompanying drawings:
Fig. 1 show Flos Carthami in the embodiment of the present invention 1 and differentiates thin-layer chromatogram；
Fig. 2 show S-A Hydroxysafflor yellow A assay reference substance high-efficient liquid phase chromatogram in the embodiment of the present invention 2；
Fig. 3 show S-A Hydroxysafflor yellow A assay test sample high-efficient liquid phase chromatogram in the embodiment of the present invention 2；
Fig. 4 show S-A Hydroxysafflor yellow A assay negative control high performance liquid chromatography in the embodiment of the present invention 2
Figure；
Fig. 5 show S-A Hydroxysafflor yellow A content linearity curve in the embodiment of the present invention 2.
Detailed description of the invention
Describe embodiments of the present invention in detail below in conjunction with drawings and Examples, thereby how the present invention is applied
Technological means solves technical problem and reaches the process that realizes of technology effect and can fully understand and implement according to this.
Embodiment 1, the discriminating of Flos Carthami
Take cowherb red 10, remove coating, finely ground, take powder 0.5g, add 80% acetone soln 5ml, shake after close plug
15min, stands；Aspirate supernatant, as need testing solution.
Take Flos Carthami control medicinal material 0.5g, be made in the same way of control medicinal material solution.
Take blank sample, be made in the same way of placebo solution.
Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively and in same with sodium carboxymethyl cellulose be
On the silica gel H lamellae of adhesive, the acetic ether-methanoic acid-water-methanol solution with volume ratio as 7:2:3:0.4 is as developing solvent
Launching, taking-up is dried, and inspects in the sunlight.
The Flos Carthami of the red tablet quality control method of result cowherb as of the present invention in Fig. 1 differentiates shown in thin-layer chromatogram, in figure 1,3,5,7
For Flos Carthami control medicinal material, 2,4,6 is test sample, and 8 is blank.In figure in test sample chromatograph, with Flos Carthami control medicinal material color
Compose on corresponding position, the fluorescence speckle of aobvious same color.In blank control sample chromatograph, with Flos Carthami control medicinal material chromatograph phase
Answer on position, have no corresponding speckle.Show in red of cowherb containing Flos Carthami.
Embodiment 2, the assay of S-A Hydroxysafflor yellow A
(1) method of assay
Flos Carthami is main flavour of a drug in red of cowherb, and in this law employing high-efficient liquid phase technique the other side, Flos Carthami is with S-A Hydroxysafflor yellow A
Assay is carried out for index.
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With volume ratio for 26:
The methanol of 2:72-second eyeball-0.7% phosphoric acid solution is flowing phase；Detection wavelength is 403nm, and number of theoretical plate presses hydroxyl Flos Carthami yellow
Element A peak calculates and should be not less than 1500.
The preparation of reference substance solution: precision weighs S-A Hydroxysafflor yellow A reference substance, adds 25% methanol and makes every 1ml and contain
The solution of 0.13mg, to obtain final product.
The preparation of need testing solution: take cowherb red 20, removes coating, finely ground, takes 2g, accurately weighed, puts tool plug taper
In Ping, accurate addition 25% methanol 25ml, close plug, weighed weight, supersound process (power 300W, frequency 50kHz) 40 minutes, put
Cold, more weighed weight, supply the weight of less loss with 25% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
The preparation of negative control solution: take the negative sample without Flos Carthami, according to the preparation method of need testing solution, same to legal system
?.
Assay method: precision measures need testing solution and each 20 μ l of reference substance solution are injected separately into chromatograph of liquid, record
Chromatogram, goes out the content of S-A Hydroxysafflor yellow A in test sample by external standard method with calculated by peak area, to obtain final product.
(2) methodological study of assay
The determination of 2.1 detection wavelength
With methanol-second eyeball-0.7% phosphoric acid solution=26:2:72 as solvent, to hydroxyl Flos Carthami in the range of 350-450nm
Safflor yellow A standard substance are scanned, and S-A Hydroxysafflor yellow A standard substance, in this solvent, have absorption maximum at 403nm.
2.2 system suitability
Under above-mentioned chromatographic condition, take reference substance solution, need testing solution and negative control solution, each sampling volume respectively
20 μ l, record chromatogram, and measurement result is as in Figure 2-4.In sample, S-A Hydroxysafflor yellow A theoretical cam curve calculates: n=
3797 (＞ 1500)；Main peak is 1.3 with the separating degree of impurity peaks；Retention time is 13.83min；Negative control solution is in comparison
Without absorbing at product absworption peak；Show that adjuvant and various catabolite are less to main peak interference, meet wanting of system suitability
Ask.
2.3 linear relationship tests
Accurately weighed 2.0mg S-A Hydroxysafflor yellow A reference substance, in 50ml volumetric flask, adds 25% methanol to scale,
Shake up, respectively precision measure 0.5,1,2,3,4,5,6ml in 10ml measuring bottle, add 25% methanol dilution to scale, shake up.Accurate
Measure each 20 μ l of above-mentioned solution, be injected separately into chromatograph of liquid according to above-mentioned chromatographic condition, measure peak area.Right with peak area (Y)
Sample introduction concentration (X, μ g/ml) carries out linear regression, and regression equation is y=0.2804x+0.0255, coefficient R2=0.9997,
Result shows, S-A Hydroxysafflor yellow A concentration is in the range of 2.12～25.43 μ g/ml, and peak area is good with concentration linear relationship
Good, it is shown in Table 1 and Fig. 5.
Table 1 S-A Hydroxysafflor yellow A linear relationship result of the test
2.4 minimum detectable activity
Precision measures the sample 20 μ l that concentration in standard solution is 2 μ g/ml, injects in chromatograph of liquid, records chromatograph
Figure, the response value of results peaks area, in the range of linear, illustrate that minimum detectable activity should be less than 2 μ g/ml, meets the requirements.
2.5 precision test
Precision measures the sample 20 μ l that concentration in standard solution is 8 μ g/ml, injects in chromatograph of liquid, records chromatograph
Figure, METHOD FOR CONTINUOUS DETERMINATION 6 times.The results are shown in Table 2, meansigma methods is 30.018 ten thousand, RSD=0.94%, and instrument precision is good.
Table 2 Precision test result
2.6 stability test
Take cowherb red 10, be prepared into need testing solution by method below assay item；Accurate absorption need testing solution 20 μ
L, sample introduction measures once at regular intervals, measures 8 hours altogether, the results are shown in Table 3.RSD is 1.8%, and result shows, test sample is molten
Liquid is basicly stable in 8 hours.
Table 3 stability test result
The cowherb red tablet quality control method of the present invention, it is provided that differentiate the method for Flos Carthami in medicine, and use efficient liquid phase
In method the other side, main flavour of a drug Flos Carthami carries out assay with S-A Hydroxysafflor yellow A for index.The method of quality control of the present invention
Can better control over the quality of medicine, have stronger specificity and good repeatability, real embodiment drug safety is effective, matter
Measure controlled.
As employed some vocabulary in the middle of description and claim to censure special component or method.Art technology
Personnel are it is to be appreciated that same composition may be called with different nouns in different regions.This specification and claims are not
In the way of the difference of title is used as distinguishing composition." comprising " as mentioned by the middle of description and claim in the whole text is
One open language, therefore " comprise but be not limited to " should be construed to." substantially " refer in receivable range of error, this area
Technical staff can solve described technical problem in the range of certain error, basically reaches described technique effect.Description is follow-up
It is described as implementing the better embodiment of the present invention, for the purpose of right described description is the rule so that the present invention to be described, not
In order to limit the scope of the present invention.Protection scope of the present invention is when being as the criterion depending on the defined person of claims.
Also, it should be noted term " includes ", " comprising " or its any other variant are intended to nonexcludability
Comprise, so that include that the commodity of a series of key element or system not only include those key elements, but also include the most clearly
Other key elements listed, or also include the key element intrinsic for this commodity or system.In the feelings not having more restriction
Under condition, statement " including ... " key element limited, it is not excluded that in the commodity including described key element or system also
There is other identical element.
Described above illustrate and describes some preferred embodiments of the present invention, but as previously mentioned, it should be understood that the present invention
Be not limited to form disclosed herein, be not to be taken as the eliminating to other embodiments, and can be used for other combinations various,
Amendment and environment, and can be in invention contemplated scope described herein, by above-mentioned teaching or the technology of association area or knowledge
It is modified.And the change that those skilled in the art are carried out and change are without departing from the spirit and scope of the present invention, the most all should be at this
In the protection domain of bright claims.
Claims (4)
1. cowherb red tablet quality control method, it is characterised in that include utilizing thin layer chromatography carry out Flos Carthami discriminating and utilize efficiently
Liquid chromatography carries out S-A Hydroxysafflor yellow A assay.
2. cowherb red tablet quality control method as claimed in claim 1, it is characterised in that described Flos Carthami discrimination method is: take cowherb
Red 10, remove coating, finely ground one-tenth powder；Take described powder 0.5g, add 80% acetone soln 5ml, shake after close plug
15min, stands；Aspirate supernatant, as need testing solution；
Take Flos Carthami control medicinal material 0.5g, be made in the same way of control medicinal material solution；
Take blank sample, be made in the same way of placebo solution；
Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same with sodium carboxymethyl cellulose for binding
On the silica gel H lamellae of agent, the acetic ether-methanoic acid-water-methanol solution with volume ratio as 7:2:3:0.4 launches for developing solvent,
Take out, dry；
Observe silica gel H lamellae, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, show identical in the sunlight
The speckle of color.
3. cowherb red tablet quality control method as claimed in claim 1, it is characterised in that described S-A Hydroxysafflor yellow A content
Assay method is:
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica as filler；With volume ratio as 26:2:72
Methanol-second eyeball-0.7% phosphoric acid solution for flowing phase；Detection wavelength is 403nm, and number of theoretical plate presses S-A Hydroxysafflor yellow A peak
Calculate >=1500；
The preparation of reference substance solution: precision weighs S-A Hydroxysafflor yellow A reference substance, adds 25% methanol and makes every 1ml and contain
The solution of 0.13mg, to obtain final product；
The preparation of need testing solution: take cowherb red 20, removes coating, finely ground one-tenth powder, takes described powder 2g, accurately weighed,
Put in tool plug conical flask, accurate addition 25% methanol 25ml, close plug, weighed weight, supersound process, let cool, more weighed weight, add
25% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product；
Assay method: precision draws reference substance solution and each 20 μ l of need testing solution respectively, injects chromatograph of liquid, measures, meter
Calculate and get final product, this product every containing Flos Carthami in terms of S-A Hydroxysafflor yellow A, no less than 0.045mg.
4. cowherb red tablet quality control method as claimed in claim 3, it is characterised in that described S-A Hydroxysafflor yellow A content
In assay method, the sonification power of need testing solution preparation process is 300W, and supersonic frequency is 50kHz, ultrasonic time 40
Minute.
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Application publication date: 20161207 |