CN108956835A - A kind of fingerprint atlas detection method of the antipyretic oral drugs of clearing - Google Patents
A kind of fingerprint atlas detection method of the antipyretic oral drugs of clearing Download PDFInfo
- Publication number
- CN108956835A CN108956835A CN201710358947.3A CN201710358947A CN108956835A CN 108956835 A CN108956835 A CN 108956835A CN 201710358947 A CN201710358947 A CN 201710358947A CN 108956835 A CN108956835 A CN 108956835A
- Authority
- CN
- China
- Prior art keywords
- solution
- reference substance
- mobile phase
- clearing
- fingerprint
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a kind of fingerprint atlas detection methods of the antipyretic oral drugs of clearing, wherein the measuring method includes the preparation of reference substance solution, the step of the preparation of test solution, measurement.Wherein, the efficient liquid phase detects chromatographic condition are as follows: for chromatographic column using octadecylsilane chemically bonded silica as filler, mobile phase A is acetonitrile, Mobile phase B is 0.2% phosphoric acid, carries out gradient elution, and the flow velocity of mobile phase is 1.2ml/min, 35 DEG C of column temperature, Detection wavelength: 206nm.
Description
Technical field
The present invention relates to drug detection fields, more particularly, to a kind of high performance liquid chromatography detection finger-print of Chinese patent drug
Method
Background technique
The antipyretic oral solution of children's clearing is sub- precious medicine company Sichuan pharmaceutical Co. Ltd production for treating children's acute pharynx
The new Chinese medicine of scorching lung stomach excess heat symptom-complex, by radix bupleuri, scutelloside, viola mandshurica, calculus bovis factitius, sowthistle, cordate houttuynia, reed root, red
8 taste Chinese medicine of red bean composition, has the effect of clearing heat and detoxicating, relieving sore-throat of reducing swelling, and belongs to lung stomach for children's acute pharyngitis (acute throat obstruction)
Excessive heat syndrome describes its main component and composition in patent CN1296088C.Clinical practice proves, the antipyretic oral solution of children's clearing
For children's acute pharyngitis (lung stomach excess heat symptom-complex) pharyngalgia, stool is dry, pharyngitis symptom and sign is effective, and safety is preferable,
It is worth clinical application.
The quality inspection standard of the kind is not recorded in " Chinese Pharmacopoeia ".Shortcomings in existing standard are as follows: stress
In Qualitive test and quantitative detection to the antipyretic oral solution main component of children's clearing, lacks and the quality of drug entirety is controlled.
Through Literature Consult, no researcher carried out global quality control research to the kind.
Finger-print is the understanding based on centering medicinal substances group's mass action, institute in certain Chinese medicine or Chinese patent drug of foundation
Chromatography or spectrum atlas shared, with certain characteristic class or several classes of ingredients, have contain much information, characteristic is strong, whole
The features such as property and ambiguity.At this stage, the middle the effective elements of the medicine overwhelming majority does not have in specific situation, and finger-print can be compared with
The relativeness for comprehensively reflecting chemical component contained by drug, embodies the complexity and correlation of traditional Chinese medicine ingredients, with traditional Chinese medicine
Traditional theory be adapted, can really to Chinese medicine inherent quality carry out Efficient Characterization, overall merit and comprehensively control.For effective
The quality of control Chinese medicine or Chinese patent drug is of great significance.
The antipyretic oral solution of children's clearing belongs to children special-purpose drug, and quality control is particularly important.In order to control this comprehensively
The product quality of product, it is necessary to establish the quality testing side of a kind of new, quick, accurate and suitable industrialized production demand
Method meets the global quality control requirement to the product.
Summary of the invention
The present invention is various for the antipyretic oral solution flavour of a drug of children's clearing, complicated component, the qualitative and quantitative analysis of individual components
It is difficult to reflect the comprehensive information of drug comprehensively, establishes the fingerprint atlas detection method of said preparation, more preferably, more effectively to control
The total quality of the antipyretic oral solution of children's clearing.
The present invention provides a kind of fingerprint atlas detection methods of antipyretic oral preparation of children's clearing, which is characterized in that packet
Include following steps:
(1) preparation of reference substance solution
Scutelloside reference substance is weighed, adds methanol that reference substance solution is made, weighs aesculetin reference substance, adds 50% methanol system
At reference substance solution；
(2) preparation of test solution
The antipyretic oral solution of children's clearing is measured, adds the dilution of 70% ethyl alcohol to be settled to scale, shakes up, filter, take subsequent filtrate, i.e.,
For test solution；
(3) measuring method:
The reference substance solution and the test solution are drawn, high performance liquid chromatograph is injected, obtains chromatogram,
Wherein, the chromatographic condition of the high performance liquid chromatograph is as follows:
For chromatographic column using octadecylsilane chemically bonded silica as filler, mobile phase A is acetonitrile, and Mobile phase B is phosphoric acid solution,
Gradient elution is carried out, Gradient program is as follows:
Flow velocity is 0.8~1.2ml/min；Detection wavelength is 192~230nm；Column temperature is 25~40 DEG C.
In an embodiment of measuring method of the present invention, the Mobile phase B be 0.2% phosphoric acid solution.
In one embodiment of measuring method of the invention, scutelloside is dense in reference substance solution described in step (1)
0.2mg/ml is spent, the concentration of aesculetin reference substance solution is 0.08mg/ml.
In an embodiment of measuring method of the present invention, a certain concentration is added in pharmaceutical composition described in step (2)
Ethyl alcohol so that in every 10ml dilute solution contained original liquid be 0.5ml~2.0ml.
In an embodiment of measuring method of the present invention, the chromatographic column is Phenomenex Gemini-C18 column.
In an embodiment of measuring method of the present invention, in the chromatographic condition, preferably 35 DEG C of column temperature.
In an embodiment of measuring method of the present invention, in the chromatographic condition, the flow velocity of mobile phase is preferred
1.2ml/min。
In an embodiment of measuring method of the present invention, in the chromatographic condition, Detection wavelength is preferably 206nm.
In the second aspect of the present invention, a kind of fingerprint of antipyretic oral preparation of children's clearing is provided, is wrapped
Include following steps:
(1) preparation of reference substance solution
Scutelloside reference substance is weighed, adds methanol that reference substance solution is made, weighs aesculetin reference substance, adds 50% methanol system
At reference substance solution；
(2) preparation of test solution
The antipyretic oral solution of children's clearing is measured, adds the dilution of 70% ethyl alcohol to be settled to scale, shakes up, filter, take subsequent filtrate, i.e.,
For test solution；
(3) measuring method:
The reference substance solution and the test solution are drawn, high performance liquid chromatograph is injected, obtains chromatogram,
Wherein, the chromatographic condition of the high performance liquid chromatograph is as follows:
For chromatographic column using octadecylsilane chemically bonded silica as filler, mobile phase A is acetonitrile, and Mobile phase B is phosphoric acid solution,
Gradient elution is carried out, Gradient program is as follows:
Flow velocity is 0.8~1.2ml/min；Detection wavelength is 192~230nm；Column temperature is 25~40 DEG C；
(4) it generates reference fingerprint: selecting the finger-print of the multiple batches of antipyretic oral solution of children's clearing, be with scutelloside
Reference peak obtains the map at 10 shared peaks, generates reference fingerprint with median calculating method；Wherein, finger-print is opposite
Retention time are as follows:
In an embodiment of fingerprint of the invention, Mobile phase B is in the step (3)
0.2% phosphoric acid solution.
In an embodiment of fingerprint of the invention, reference substance described in the step (1) is molten
The concentration 0.2mg/ml of scutelloside in liquid, the concentration of aesculetin reference substance solution are 0.08mg/ml.
In an embodiment of fingerprint of the invention, pharmaceutical composition described in the step (2)
70% ethyl alcohol is added in object, so that contained original liquid is 0.5ml~2.0ml in every 10ml dilute solution.
In an embodiment of fingerprint of the invention, chromatographic column is Phenomenex Gemini-
C18 column, preferably 35 DEG C of column temperature, the preferred 1.2ml/min of the flow velocity of mobile phase, Detection wavelength is preferably 206nm.
The selected testing conditions of the present invention are compared and verify repeatedly by experiment, and accuracy is high, may be implemented half
Quantitative determination.The present invention takes HPLC finger-print to detect the kind for the first time, by mobile phase and gradient elution
The methods of condition optimizes, so that the main peak separation of map is good, while the peak shape for compareing peak also obviously improves, can be comprehensive
The total quality of this product is controlled, while meeting the requirement of industrialized production.
Beneficial effects of the present invention
(1) establish the antipyretic oral solution finger-print of children's clearing with method provided by the present invention, in conjunction with scutelloside and
Two main components of aesculetin are pointed out, and the product quality can be effectively characterized, and are conducive to control drug quality comprehensively.
(2) finger-print focuses on each tandem and correlation for constituting fingerprint characteristic peak, and it is special to focus on whole looks
Sign, not only avoided and determined the one-sidedness of the antipyretic oral solution quality of children's clearing because measuring individual chemical ingredient, but also reduce for
Requisite quality and a possibility that artificially handle.
(3) the method for the present invention has the advantages that method is easy, stablizes, precision is high, favorable reproducibility.
Detailed description of the invention
1 acetonitrile of attached drawing is mobile phase A, and water is that children's clearing of Mobile phase B progress efficient liquid phase chromatographic analysis foundation is antipyretic
Oral solution finger-print.
2 acetonitrile of attached drawing is mobile phase A, and 0.2% phosphoric acid solution is the youngster that Mobile phase B carries out efficient liquid phase chromatographic analysis foundation
The virgin antipyretic oral solution finger-print of clearing.
3 acetonitrile of attached drawing is mobile phase A, and 0.05% trifluoroacetic acid solution is that Mobile phase B progress efficient liquid phase chromatographic analysis is built
The antipyretic oral solution finger-print of vertical children's clearing.
4 methanol of attached drawing is mobile phase A, and 0.2% phosphoric acid solution is the youngster that Mobile phase B carries out efficient liquid phase chromatographic analysis foundation
The virgin antipyretic oral solution finger-print of clearing.
The antipyretic oral solution finger-print-control map of 5 children's clearing of attached drawing, wherein No. 4 peaks are aesculetin, 8 peaks (S)
For scutelloside.
Specific embodiment
1 finger-print of embodiment is established
1.1 instrument
LC-20AT Shimadzu high performance liquid chromatograph, 1200 high performance liquid chromatograph of Agilent, Agilent 1260 are efficient
Liquid chromatograph, chromatographic column (Agilent Extend C18,4.6 × 250mm, 5 μm, S/N:USHR007237), chromatographic column
(Phenomenex Gemini-C18,4.6 × 250mm, 5 μm, S/N:540529-7), chromatographic column (Phenomenex Gemini-
C18,4.6 × 250mm, 5 μm, S/N:540542-28), chromatographic column (Phenomenex Gemini-C18,4.6 × 250mm, 5 μ
M, S/N:540608-4), chromatographic column (Kromasil 100-5C18,4.6 × 250mm, 5 μm, S/N:E59208).
1.2 reagent
(AR, Sinopharm Chemical Reagent Co., Ltd., lot number: 20130318), (AR, Beijing essence seek chemical industry to phosphoric acid to formic acid
Co., Ltd, lot number: 20101107), acetic acid (AR, Tianjin good fortune morning chemical reagent factory, lot number: 20130702), and ethyl alcohol
(AR, Sinopharm Chemical Reagent Co., Ltd., lot number: 20130801) pure water (Hangzhou Wahaha Group Co., Ltd, lot number
20130520), acetonitrile (HPLC, SIGMA-ALDRICH, lot number: S92130), methanol (HPLC, Honeywell, lot number
20130729)。
Reference substance: aesculetin reference substance (National Institute for Food and Drugs Control, lot number: 110741-200506), radix scutellariae
Glycosides reference substance (National Institute for Food and Drugs Control, lot number: 110715-200815).
1.3 test sample
Children's clearing solution that the lot number of sub- treasured medicine company Sichuan pharmaceutical Co. Ltd production is 130201,130202,130203
In hot oral solution and the north Ya Bao the lot number of big (Beijing) pharmaceutical Co. Ltd production be 18002B, 28008,28001B, 28019B,
The antipyretic oral solution of children's clearing of 28024B, 38003B, 08004B, 98015B, 08005B.
1.4 test method
High performance liquid chromatography (four general rules 0512 of Chinese Pharmacopoeia version in 2015) measurement.
The preparation of reference solution: precision weighs scutelloside reference substance 10mg, sets in 50ml measuring bottle, and methanol is added to dissolve and determine
Hold to scale, shake up, filters, take subsequent filtrate to get reference solution；Precision weighs aesculetin reference substance 10mg, sets 50ml
In measuring bottle, add 50% methanol to dissolve and be settled to scale, shake up, precision measures 10ml, sets in 25ml measuring bottle, adds 50% methanol fixed
Hold to scale, shake up, filter, take subsequent filtrate to get.
The preparation of test solution: precision measures the antipyretic oral solution 1ml of children's clearing, sets in 10ml measuring bottle, adds 70% second
Alcohol dilution is settled to scale, shakes up, and filters, takes subsequent filtrate, as test solution.
Chromatographic condition: using octadecylsilane chemically bonded silica as filler (chromatographic column: Phenomenex Gemini-C18,
250mm × 4.6mm, 5 μm)；Using acetonitrile as mobile phase A, 0.2% phosphoric acid solution be Mobile phase B, by 2 gradient elution program of table into
Row gradient elution:
2 eluent gradient elution requirement of table
Flow velocity is 1.2ml/min；Detection wavelength is 206nm；Column temperature is 35 DEG C.Theoretical cam curve is answered by the calculating of scutelloside peak
Not less than 10000.
Measuring method: it is accurate respectively to draw reference solution and each 10 μ l of test solution, inject high performance liquid chromatograph, note
Record 80 minutes in chromatogram to get.
2 optimum chromatogram condition of embodiment is investigated
Instrument and reagent are the same as embodiment 1.
The selection of 2.1 Detection wavelengths
The preparation of reference substance solution, the preparation of test solution are the same as embodiment 1.
192nm, 206nm, 230nm, 254nm, 278nm, 353nm, 450nm is respectively set to the antipyretic oral solution of children's clearing
Finger-print analyzed, wherein chromatographic peak separation is preferable under 192nm~230nm wavelength.With the map under other 6 wavelength
It compares, the finger-print information content of 206nm is most, can more fully embody the chemical component of this product, and baseline drift
Less, therefore select 206nm as optimal detection wavelength.
The preparation of 2.2 test solutions is investigated
Preparation, chromatographic condition and the measuring method of reference solution are the same as embodiment 1.
It takes this product stoste, water to dilute 10 times of solution, 70% ethyl alcohol dilution, 10 times of solution, 70% ethyl alcohol and dilutes 25 times of solution,
Test solution is respectively prepared.
Chromatographic peak main in the obtained chromatogram of the above different dilution process is integrated, compares its separating degree and divides
From effect, the results show that the separating degree and map whole separation effect of sample are better than at others after 10 times of 70% ethyl alcohol dilution
Reason method, it is thus determined that the antipyretic oral solution preparation method of test article of children's clearing is as follows: this product 1ml is taken, is placed in 10ml measuring bottle,
Add 70% ethyl alcohol dilution be settled to scale, shake up to get.
The selection of 2.3 chromatographic columns
The filler and type of feed of chromatographic column may have an impact drug chemistry ingredient separating effect.Inventor couple
The C18 chromatographic column (5 μm, 250 × 4.6mm) of tri- different brands of Agilent, Kromasil and Phenomenex is investigated.
Other chromatographic conditions are the same as embodiment 1.The separating effect and peak type of Phenomenex Gemini-C18 column are superior to remaining two kinds,
Therefore preferentially select Phenomenex Gemini-C18 chromatographic column.
The selection of 2.4 column temperatures
In chromatogram finger print measuring, column temperature often will affect separating effect, and inventor is respectively in 25 DEG C, 30 DEG C, 35
DEG C, under the conditions of 40 DEG C of four column temperatures, the finger-print of the antipyretic oral solution test solution of children's clearing is analyzed.As a result table
It is bright, it can get test article fingerprint under the conditions of four column temperatures, wherein the separating effect of each chromatographic peak compares at 35 DEG C
It is good, therefore 35 DEG C of optimal selection are detection column temperature.
2.5 flow velocitys are investigated
In chromatogram finger print measuring, flow velocity will affect separating effect to a certain extent, this test exists respectively
Under tri- flow conditions of 0.8ml/min, 1.0ml/min, 1.2ml/min, to the antipyretic oral solution test solution of children's clearing
Finger-print is analyzed.The result shows that can get test article fingerprint under three flow conditions, wherein in 1.2ml/
The separating effect of each chromatographic peak is relatively good when min, therefore selects 1.2ml/min for most suitable flow velocity.
2.6 Chromatographic information acquisition times determine
To investigate Chromatographic information acquisition time, this product is detected, chromatographic condition is as follows: chromatographic column: Phenomenex
Gemini-C18 chromatographic column (250mm × 4.6mm, 5 μm)；Detection wavelength: 206nm；Column temperature: 35 DEG C；Flow velocity: 1.2ml/min；Into
Sample amount: 10 μ l；Mobile phase: using acetonitrile as mobile phase A, 0.2% phosphoric acid solution is Mobile phase B, carries out gradient elution by table 3.It adopts
Collect the Chromatographic information in 120min.
3 eluent gradient elution requirement of table
Because without absorption peak after 80min, therefore acquiring the Chromatographic information in 80min.
The verifying of 3 fingerprint spectrum method of embodiment
According to preliminary result, different flow visualizings is investigated, screening chromatographic peak information is most abundant, and separating degree is best
Chromatographic condition.
The preparation of test solution: taking this product 1ml, is placed in 10ml measuring bottle, adds the dilution of 70% ethyl alcohol to be settled to scale, shakes
Even, filtration takes subsequent filtrate as test solution.
Chromatographic condition: using Phenomenex Gemini-C18 chromatographic column, investigates acetonitrile-aqueous solution, -0.2% phosphorus of acetonitrile
Acid solution, -0.05% trifluoroacetic acid solution of acetonitrile, methanol 4 systems of -0.2% phosphoric acid solution are tested
(1) system 1: acetonitrile is mobile phase A, and water is Mobile phase B, referring to attached drawing 1
4 eluent gradient elution requirement of table
(2) system 2: acetonitrile is mobile phase A, and 0.2% phosphoric acid solution is Mobile phase B, referring to attached drawing 2
5 eluent gradient elution requirement of table
(3) system 3: acetonitrile is mobile phase A, and 0.05% trifluoroacetic acid solution is Mobile phase B, referring to attached drawing 3
6 eluent gradient elution requirement of table
(4) system 4: methanol is mobile phase A, and 0.2% phosphoric acid solution is Mobile phase B, referring to attached drawing 4
7 eluent gradient elution requirement of table
- 0.2% phosphoric acid solution system appearance information content of acetonitrile is larger it can be seen from the map of system above, and baseline is flat
Surely, it can preferably reflect the total quality information of the antipyretic oral solution of children's clearing；And its excess-three system baseline drift it is serious or
Appearance is on the low side, can not comprehensive representation this product quality information, therefore screened, determine that selection -0.2% phosphoric acid solution system of acetonitrile is made
For the mobile phase condition of this product finger-print.
The verifying of 4 fingerprint spectrum method of embodiment
4.1 blank test
70% ethyl alcohol is taken, by 1 chromatographic condition direct injected of embodiment, 80 minutes chromatograms of record, the results showed that system
There is no residuals and interference.
4.2 system suitability
This product is taken, is operated according to 1 method of embodiment, is prepared into test solution, direct injected 6 times, with No. 8 peak scutellosides
To calculate the relative retention time and relative peak area of each main chromatographic peak referring to peak.10 shared peaks is opposite as the result is shown
Retention time is stablized, and RSD is respectively less than 2%, and relative peak area is relative constant, and RSD is respectively less than 5%；Show that the system of method is applicable in
Property is good.It the results are shown in Table 8,9.
8 precision test relative retention time of table
9 precision test relative peak area of table
4.3 repetitive test
Testing crew A takes same batch this product, operates according to 1 method of embodiment, prepares 6 parts of test solutions, detects,
It is to calculate the relative retention time and relative peak area of each main chromatographic peak referring to peak with No. 8 peak scutellosides.10 as the result is shown
The relative retention time at shared peak is stablized, and RSD is respectively less than 2%, and relative peak area is relative constant, and RSD is respectively less than 5%；Show this
Method repeatability is good.It the results are shown in Table 10,11.
10 repetitive test relative retention time of table
11 repetitive test relative peak area of table
4.4 stability test
This product is taken, is operated according to 1 method of embodiment, is prepared 0,3,6,9,12,18,24,30,36 hour respectively at test sample
Sample detection is to calculate the relative retention time and relative peak area of each main chromatographic peak referring to peak with No. 8 peak scutellosides.Knot
Fruit shows that the relative retention time at 10 shared peaks is stablized, and RSD is respectively less than 2%, and relative peak area is relative constant, and RSD is respectively less than
5%；Show that the test solution of this product is stablized in 36 hours.It the results are shown in Table 12,13.
12 stability test relative retention time of table
13 stability test relative peak area of table
4.5 serviceability test
4.5.1 different instrument durabilities
This product is taken, operates, is detected using the instrument of different model, with Agilent 1200 according to 1 method of embodiment
Instrument map is reference, to calculate the similarity of map between different instruments referring to map, the results showed that measure between different instruments
As a result roughly the same, fingerprint similarity is all larger than 0.99, illustrates the model of high performance liquid chromatograph to this finger-print side
Method influences little.It the results are shown in Table 14.
14 instrument serviceability test result of table
4.5.2 different batches chromatographic column durability
This product is taken, operates, is detected using same model different batches chromatographic column, with wherein according to 1 method of embodiment
One batch-wise chromatography column (sequence number 540529-7) map is reference, calculates similarity, the results showed that different batches chromatography intercolumniation
Measurement result is roughly the same, and fingerprint similarity is all larger than 0.99, and it is little to illustrate that the batch of chromatographic column influences this method.Knot
Fruit is shown in Table 15.
15 chromatographic column serviceability test result of table
5 reference fingerprint of embodiment is established
Children's clearing that lot number to sub- precious medicine company Sichuan pharmaceutical Co. Ltd production is 130201,130202,130203
In antipyretic oral solution and the north Ya Bao the lot number of big (Beijing) pharmaceutical Co. Ltd production be 18002B, 28008,28001B,
The antipyretic oral solution of children's clearing of 28019B, 28024B, 38003B, 08004B, 98015B, 08005B amount to 12 batch samples,
It is detected respectively by the finger print measuring method of above-mentioned foundation, using Chinese Pharmacopoeia Commission's " chromatographic fingerprints of Chinese materia medica
Similarity evaluation system " (2012.0 version) fitting generation reference fingerprint, referring to attached drawing 5.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (10)
1. a kind of fingerprint atlas detection method of the antipyretic oral preparation of children's clearing, which comprises the following steps:
(1) preparation of reference substance solution
Scutelloside reference substance is weighed, adds methanol that reference substance solution is made, weighs aesculetin reference substance, 50% methanol is added to be made pair
According to product solution；
(2) preparation of test solution
The antipyretic oral solution of children's clearing is measured, adds the dilution of 70% ethyl alcohol to be settled to scale, shakes up, filter, take subsequent filtrate, as supply
Test sample solution；
(3) measuring method:
The reference substance solution and the test solution are drawn, high performance liquid chromatograph is injected, obtains chromatogram,
Wherein, the chromatographic condition of the high performance liquid chromatograph is as follows:
For chromatographic column using octadecylsilane chemically bonded silica as filler, mobile phase A is acetonitrile, and Mobile phase B is phosphoric acid solution, is carried out
Gradient elution, Gradient program are as follows:
Flow velocity is 0.8~1.2ml/min；Detection wavelength is 192~230nm；Column temperature is 25~40 DEG C.
2. measuring method according to claim 1, which is characterized in that the phosphorus that Mobile phase B is 0.2% in the step (3)
Acid solution.
3. measuring method according to claim 1, wherein the concentration of scutelloside in reference substance solution described in step (1)
0.2mg/ml, the concentration of aesculetin reference substance solution are 0.08mg/ml.
4. measuring method according to claim 1, which is characterized in that pharmaceutical composition described in step (2) is added 70%
Ethyl alcohol, so that contained original liquid is 0.5ml~2.0ml in every 10ml dilute solution.
5. measuring method according to claim 1, which is characterized in that chromatographic column is Phenomenex Gemini-C18 column,
Preferably 35 DEG C of column temperature, the preferred 1.2ml/min of the flow velocity of mobile phase, Detection wavelength is preferably 206nm.
6. a kind of fingerprint of the antipyretic oral preparation of children's clearing, comprising the following steps:
(1) preparation of reference substance solution
Scutelloside reference substance is weighed, adds methanol that reference substance solution is made, weighs aesculetin reference substance, 50% methanol is added to be made pair
According to product solution；
(2) preparation of test solution
The antipyretic oral solution of children's clearing is measured, adds the dilution of 70% ethyl alcohol to be settled to scale, shakes up, filter, take subsequent filtrate, as supply
Test sample solution；
(3) measuring method:
The reference substance solution and the test solution are drawn, high performance liquid chromatograph is injected, obtains chromatogram,
Wherein, the chromatographic condition of the high performance liquid chromatograph is as follows:
For chromatographic column using octadecylsilane chemically bonded silica as filler, mobile phase A is acetonitrile, and Mobile phase B is phosphoric acid solution, is carried out
Gradient elution, Gradient program are as follows:
Flow velocity is 0.8~1.2ml/min；Detection wavelength is 192~230nm；Column temperature is 25~40 DEG C；
(4) it generates reference fingerprint: the finger-print of the multiple batches of antipyretic oral solution of children's clearing is selected, using scutelloside as reference
Peak obtains the map at 10 shared peaks, generates reference fingerprint with median calculating method；Wherein, finger-print is opposite retains
Time are as follows:
7. fingerprint according to claim 6, which is characterized in that Mobile phase B is in the step (3)
0.2% phosphoric acid solution.
8. fingerprint according to claim 6, wherein radix scutellariae in reference substance solution described in step (1)
The concentration 0.2mg/ml of glycosides, the concentration of aesculetin reference substance solution are 0.08mg/ml.
9. fingerprint according to claim 6, which is characterized in that pharmaceutical composition described in step (2)
70% ethyl alcohol is added, so that contained original liquid is 0.5ml~2.0ml in every 10ml dilute solution.
10. fingerprint according to claim 6, which is characterized in that chromatographic column Phenomenex
Gemini-C18 column, preferably 35 DEG C of column temperature, the preferred 1.2ml/min of the flow velocity of mobile phase, Detection wavelength is preferably 206nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710358947.3A CN108956835B (en) | 2017-05-19 | 2017-05-19 | Fingerprint detection method of oral medicine for clearing heat from throat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710358947.3A CN108956835B (en) | 2017-05-19 | 2017-05-19 | Fingerprint detection method of oral medicine for clearing heat from throat |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108956835A true CN108956835A (en) | 2018-12-07 |
| CN108956835B CN108956835B (en) | 2022-02-25 |
Family
ID=64462748
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710358947.3A Active CN108956835B (en) | 2017-05-19 | 2017-05-19 | Fingerprint detection method of oral medicine for clearing heat from throat |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108956835B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113437A (en) * | 2021-12-06 | 2022-03-01 | 苏州市药品检验检测研究中心 | HPLC (high Performance liquid chromatography) fingerprint of heat-clearing and detoxifying oral liquid and application of HPLC fingerprint in quality control of heat-clearing and detoxifying oral liquid |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1689627A (en) * | 2004-04-30 | 2005-11-02 | 北京中医药大学药厂 | Medicine for treating acute pharyngitis of children and preparation method thereof |
| WO2006024545A1 (en) * | 2004-09-03 | 2006-03-09 | Stichting Voor De Technische Wetenschappen | Fused bicyclic natural compounds and their use as inhibitors of parp and parp-mediated inflammatory processes |
| CN101904948A (en) * | 2010-07-27 | 2010-12-08 | 广东众生药业股份有限公司 | Traditional Chinese medicine preparation of new Zhongsheng pill and preparation method thereof |
| CN105510488A (en) * | 2016-01-25 | 2016-04-20 | 亚宝药业集团股份有限公司 | Fingerprint spectrum of pain-relieving patch and quality detection method thereof |
| CN105911161A (en) * | 2016-04-12 | 2016-08-31 | 吉林修正药业新药开发有限公司 | Anti-inflammatory tablet HPLC fingerprint construction method |
-
2017
- 2017-05-19 CN CN201710358947.3A patent/CN108956835B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1689627A (en) * | 2004-04-30 | 2005-11-02 | 北京中医药大学药厂 | Medicine for treating acute pharyngitis of children and preparation method thereof |
| WO2006024545A1 (en) * | 2004-09-03 | 2006-03-09 | Stichting Voor De Technische Wetenschappen | Fused bicyclic natural compounds and their use as inhibitors of parp and parp-mediated inflammatory processes |
| CN101904948A (en) * | 2010-07-27 | 2010-12-08 | 广东众生药业股份有限公司 | Traditional Chinese medicine preparation of new Zhongsheng pill and preparation method thereof |
| CN105510488A (en) * | 2016-01-25 | 2016-04-20 | 亚宝药业集团股份有限公司 | Fingerprint spectrum of pain-relieving patch and quality detection method thereof |
| CN105911161A (en) * | 2016-04-12 | 2016-08-31 | 吉林修正药业新药开发有限公司 | Anti-inflammatory tablet HPLC fingerprint construction method |
Non-Patent Citations (3)
| Title |
|---|
| 滕会会 等: "基于中药指纹图谱结合化学模式识别的清开灵软/硬胶囊的成分分析", 《环球中医药》 * |
| 白冰等: "消炎片的指纹图谱分析及多指标成分含量测定 ", 《中国现代应用药学》 * |
| 陈红林 等: "高效液相色谱技术在中药配方颗粒质量控制中应用现状", 《中国中医药信息杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113437A (en) * | 2021-12-06 | 2022-03-01 | 苏州市药品检验检测研究中心 | HPLC (high Performance liquid chromatography) fingerprint of heat-clearing and detoxifying oral liquid and application of HPLC fingerprint in quality control of heat-clearing and detoxifying oral liquid |
| CN114113437B (en) * | 2021-12-06 | 2024-04-12 | 苏州市药品检验检测研究中心 | HPLC fingerprint spectrum of oral liquid for clearing away heat and toxic materials and application thereof in quality control of oral liquid for clearing away heat and toxic materials |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108956835B (en) | 2022-02-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109633001B (en) | Fingerprint detection method of wine glossy privet fruit | |
| CN110031570B (en) | Fingerprint spectrum detection method of anti-cold granules | |
| CN110501435B (en) | Fingerprint quality detection method for moringa oleifera leaf, lotus leaf and ginseng solid beverage | |
| CN111487344B (en) | Method for detecting fingerprint spectrum of motherwort particles | |
| CN108872410A (en) | A kind of method for building up and its finger-print of lung-nourishing semifluid extract finger-print | |
| CN107576739B (en) | HPLC fingerprint detection method of Longmu Zhuanggu granules | |
| CN110031564B (en) | Quality detection method of natural plant anticoccidial feed additive based on HPLC fingerprint | |
| CN109709222B (en) | Component detection method of Ganmaoling and compound Ganmaoling | |
| CN108956835A (en) | A kind of fingerprint atlas detection method of the antipyretic oral drugs of clearing | |
| CN111487351B (en) | Method for detecting fingerprint of blood-activating pain-relieving capsule | |
| CN110441443B (en) | UPLC characteristic spectrum construction method and identification method of pyrrosia peduncularis, pyrrosia lingua, pyrrosia cottonii and pyrrosia huabeiensis | |
| CN114563496A (en) | Quantitative fingerprint analysis method for components in ginger, ginger and pinellia tuber percolate | |
| CN109061024B (en) | Construction method and application of standard fingerprint spectrum of infant spleen-supporting granules | |
| CN109374771B (en) | Fingerprint spectrum detection method of snow pear syrup | |
| CN114034798A (en) | Red water dendrobium stem flower fingerprint construction and content determination method | |
| CN114034797A (en) | Method for measuring content of flower components of dendrobium nobile lindl | |
| Chen Wei et al. | Determination of AF and AFG in red ginseng by high performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD). | |
| CN113092640A (en) | Method for detecting benzyl alcohol and benzaldehyde in heparin sodium injection | |
| CN111505156A (en) | Fingerprint spectrogram quality determination method for herba Cirsii formulation granules | |
| CN100416271C (en) | Effective liquid phase chromatography for measuring alkali content of dita leaves | |
| CN112415115B (en) | Detection method of blood-replenishing and milk-producing preparation | |
| CN113219095B (en) | Construction method of fingerprint of Fukean tablet and fingerprint thereof | |
| CN110118841A (en) | A kind of construction method of the HPLC characteristic spectrum of oral liquid for clearing liver and gallbladder | |
| CN109725100B (en) | Method for detecting fingerprint spectrum of hawthorn granules | |
| CN113391002B (en) | Establishing method and application of Xintong granule UPLC fingerprint spectrum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |