CN109030641A - A kind of construction method of yangweishu granules finger-print - Google Patents
A kind of construction method of yangweishu granules finger-print Download PDFInfo
- Publication number
- CN109030641A CN109030641A CN201810596195.9A CN201810596195A CN109030641A CN 109030641 A CN109030641 A CN 109030641A CN 201810596195 A CN201810596195 A CN 201810596195A CN 109030641 A CN109030641 A CN 109030641A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- solution
- yangweishu
- surplus
- peaks
- 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of construction methods of yangweishu granules finger-print, take yangweishu granules by extracting as test solution, corresponding chromatogram is obtained by chromatographic condition measurement at least 10 batches of qualified yangweishu granules of optimization, similarity evaluation as defined in Chinese Pharmacopoeia Commission is imported to get yangweishu granules finger-print is arrived.The present invention has the characteristics that method is easy, stability is good, precision is high, favorable reproducibility, can reflect the material information of Chinese materia medica preparation comprehensively, provides a kind of new analysis method for the quality testing of yangweishu granules and quality control.
Description
Technical field
The present invention relates to a kind of construction methods of yangweishu granules finger-print, can be used for different batches yangweishu granules sample
The quality evaluation of product belongs to Pharmaceutical Analysis field.
Background technique
Traditional Chinese medicine fingerprint refers to certain or certain place of production Chinese medicine or Chinese patent drug after processing appropriate, using certain
Analysis method, what is obtained can indicate the chromatogram or spectrogram of this Chinese medicine or Chinese patent drug feature.Wherein with chromatography side
Based on method.
Yangweishu granules are to obtain country according to 105 hospital Jin Naishi of liberation army professor and Anhui Chinese Medicine College professor Su Chunlang
The new product of the scientific achievement exploitation of traditional Chinese medicine major scientific and technological achievement prize, tonifying-Yin and nourishing-stomach.For chronic gastritis, gastral cavilty is scorching hot, hidden
It is hidden to have a pain, it is the core kind of Hefei China Resources mind Smilacina japonica industry.Since the listing of approval in 1985, by clinical application in 30 years, show
Distinctive clinical application advantage is shown, many honor are also therefore obtained.
Yangweishu granules are by Chinese yam, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae (stir-fry), fructus crataegi cuneatae (stir-fry), dried orange peel, rhizoma zingiberis, radix scrophulariae, Radix Glehniae, Huang
The compound Chinese medicinal preparation of smart (steaming), Semen Cuscutae and dark plum composition.It is contained at complex chemical composition in the composing prescription preparation, it executes at present
Quality standard are as follows: ministry standard Traditional Chinese medicine historical preparation the 18th and pharmacopeia industry send out (2000) No. 028.Due to testing at that time
The limitation of condition, standard formulation is simple, is not well positioned to meet the requirement of quality evaluation.Both at home and abroad still not about Yang Weishu
The report of grain finger-print research, it is uniform, stable and patient medication safe and effective for what is ensured drug quality, establish a kind of energy
The method of enough quality for comprehensively, rapidly detecting yangweishu granules, has the detection of its total quality and global quality control
It is significant.Based on this, the present invention establishes a kind of detection method of yangweishu granules finger-print.
Summary of the invention
The object of the present invention is to provide the high yangweishu granules finger-prints of a kind of easy to operate, favorable reproducibility, precision
Construction method, the construction method of the finger-print can be used in yangweishu granules quality testing and quality control.
The bulk pharmaceutical chemicals group of the yangweishu granules become Chinese yam, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae (stir-fry), fructus crataegi cuneatae (stir-fry), dried orange peel, rhizoma zingiberis,
Radix scrophulariae, Radix Glehniae, rhizoma polygonati (steaming), Semen Cuscutae and dark plum.
The construction method of yangweishu granules finger-print of the present invention, includes the following steps:
Step 1: the preparation of test solution
It weighs 5.0-10.0g yangweishu granules to be added in 25-100mL methanol aqueous solution, is ultrasonically treated 10- at 10-60 DEG C
60min, cooled and filtered, gained filtrate are test solution；
Step 2: the preparation of reference substance solution
Using methanol as solvent, weighs aurantiamarin, harpagoside, chlorogenic acid and be configured to reference substance solution respectively；
Step 3: chromatography detection
Test solution and each 5-20 μ L of reference substance solution are drawn respectively, and injection high performance liquid chromatograph detection obtains respectively
To the chromatogram of test solution and control solution；
Chromatography detection parameters are as follows:
Using octadecylsilane chemically bonded silica as filler；Using acetonitrile as mobile phase A, with the phosphoric acid of volume fraction 0.1%
Aqueous solution is that Mobile phase B carries out gradient elution；Detection wavelength is 275-285nm；25-40 DEG C of column temperature；Flow velocity is 0.5-1.5mL/
min。
Gradient elution program is provided that
0min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
0-10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10-35min, volume shared by mobile phase A are gradually incremented to 28% by 10%, and surplus is Mobile phase B；
35min, volume shared by mobile phase A are 28%, and surplus is Mobile phase B；
35-45min, volume shared by mobile phase A are gradually incremented to 40% by 28%, and surplus is Mobile phase B；
45min, volume shared by mobile phase A are 40%, and surplus is Mobile phase B；
45-46min, volume shared by mobile phase A are gradually incremented to 80% by 40%, and surplus is Mobile phase B；
46min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
46-55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55-60min, volume shared by mobile phase A are gradually decremented to 10% by 80%, and surplus is Mobile phase B；
60min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
60-65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B.
Step 4: utilize Chinese Pharmacopoeia Commission's similarity evaluation, to test solution and
The chromatogram of reference substance solution carries out data importing, Supplements and Data Matching respectively, and using median method, time width is
0.3min is to get finger-print.
Preferably, in step 1, the methanol that 25-50mL volumetric concentration 40-80% is added in 5.0-8.0g yangweishu granules is weighed
In aqueous solution, 30-50min, cooled and filtered are ultrasonically treated at 10-60 DEG C, gained filtrate is test solution.
Further preferably, in step 1, the methanol-water that 25mL volumetric concentration 50-70% is added in 7.5g yangweishu granules is weighed
In solution, 45min, cooled and filtered are ultrasonically treated at 10-60 DEG C, gained filtrate is test solution.
Preferably, in step 2, in reference substance solution, the concentration of aurantiamarin solution is 0.06mg/mL, harpagoside it is dense
Degree is 0.05mg/mL, and the concentration of chlorogenic acid is 0.05mg/mL.
Preferably, in step 3, the sample volume of test solution and reference substance solution is 5-15 μ L, further preferably 10 μ
L；Detection wavelength is 280nm；30 DEG C of column temperature；Flow velocity is 1.0mL/min.
Preferably, in step 3,250mm × 4.6mm, 5 μm of Agilent Eclipse XDB- are used when chromatography detects
C18 chromatographic column.
In the finger-print for the yangweishu granules that the present invention obtains, there are 13 shared peaks, compared through reference substance map, determines
Wherein No. 2 peaks are chlorogenic acid, No. 8 peaks are aurantiamarin, No. 11 peaks are harpagoside；It is referring to peak, each peak number with No. 8 peak aurantiamarins
Relative retention time be respectively as follows: the peak 0.346,3 of No. 1 peak 0.307,2 peak 0.397,5 of peak 0.374,4 peak 0.522,6
Number peak 0.931,8 of peak 0.610,7 peak 1.031,10 of peak 1.000,9 peak 1.259,12 of peak 1.246,11 peak
1.309 and No. 13 peaks 1.380.
The application for the yangweishu granules finger-print that the present invention constructs is to detect to support using the finger-print that the present invention constructs
The quality of stomach relaxation grain preparation, specifically comprises the following steps:
Step 1: the preparation of product to be tested solution
Precision weighs 5.0-10.0g yangweishu granules to be measured, and 0~100% methanol solution of 25-100mL volumetric concentration is added,
10-60min, cooled and filtered are ultrasonically treated at 10-60 DEG C, gained filtrate is product to be tested solution；
Step 2: the preparation of reference substance solution
Using methanol as solvent, the concentration for being configured to aurantiamarin solution respectively is 0.06mg/mL, and the concentration of harpagoside is
0.05mg/mL, the concentration of chlorogenic acid are the reference substance solution of 0.05mg/mL.
Step 3: chromatography detection
Product to be tested solution and each 5-20 μ L of reference substance solution are drawn respectively, and injection high performance liquid chromatograph detection obtains respectively
To the chromatogram of product to be tested solution and reference substance solution；
Step 4: utilizing Chinese Pharmacopoeia Commission's similarity evaluation, gained product to be tested is referred to
Line map calculates fingerprint similarity compared with the standard finger-print of building pair, if similarity >=0.9 and standard occur and referring to
13 characteristic peaks in line map, peak corresponding with reference peak are the peak S, and ± 5% principle is not more than according to retention time deviation,
Retention time is provided are as follows:
No. 1 peak: 0.307 ± 0.015；No. 2 peaks: 0.346 ± 0.017；No. 3 peaks: 0.374 ± 0.019；No. 4 peaks: 0.397
±0.020；No. 5 peaks: 0.522 ± 0.026；No. 6 peaks: 0.610 ± 0.030；No. 7 peaks: 0.931 ± 0.047；No. 8 peaks: 1.000；
No. 9 peaks: 1.031 ± 0.052；No. 10 peaks: 1.246 ± 0.062；No. 11 peaks: 1.259 ± 0.063；No. 12 peaks: 1.309 ±
0.065；No. 13 peaks: 1.380 ± 0.069.Wherein No. 2 peaks are chlorogenic acid, No. 8 peaks are aurantiamarin, No. 11 peaks are harpagoside.
Within the above range, then the yangweishu granules to be measured are up-to-standard for the relative retention time at each shared peak, it is on the contrary then
It is unqualified.
Yangweishu granules of the present invention are that 10 batches of qualified samples are examined through current standard.
The present invention provides a kind of buildings of the high yangweishu granules finger-print of easy to operate, favorable reproducibility, precision
Method can be used to control the quality of preparation by the building of finger-print, and stability and repeatability are good, can be used for Yang Weishu
Detection in grain processing technique monitoring, quality evaluation is conducive to the detection of its total quality and global quality control, to ensure to support
The quality of stomach relaxation grain complies with standard.
The preparation method of test sample of the present invention is easy, and chromatographic condition is easy to accomplish.
The method for the yangweishu granules finger-print that the present invention constructs, has that stability is high, precision is high, reproducible etc.
Advantage, the finger-print of foundation can be used in the production process monitoring and quality control of yangweishu granules, so as to ensure
The stability and homogeneity of yangweishu granules quality, to help to improve the validity and safety of the clinical drug medication.
Detailed description of the invention
Fig. 1 is the liquid chromatogram of aurantiamarin reference substance solution in experimental example 3 of the present invention；
Fig. 2 is the liquid chromatogram of harpagoside reference substance solution in experimental example 3 of the present invention；
Fig. 3 is the liquid chromatogram of 3 Content of Chlorogenic Acid reference substance solution of experimental example of the present invention；
Fig. 4 is 3 yangweishu granules test solution of experimental example (lot number: liquid chromatogram 1705202) of the present invention；
Fig. 5 is that 10 batches of yangweishu granules test solutions importing fingerprint similarities calculating are soft in experimental example 3 of the present invention
Derived superposition chromatogram after part；
Fig. 6 is the finger-print for the yangweishu granules established in experimental example 3 of the present invention.
Specific embodiment
In order to make the content of the present invention more clearly understood, it below according to specific embodiments of the present invention and combines
Attached drawing, the present invention is described in further detail.
Instrument of the embodiment of the present invention and reagent:
Instrument: FA2104 a ten thousandth electronic balance (manufacture of Shanghai balance)；AU-220 a ten thousandth electronic balance (Japan
Shimadzu Corporation)；Ultrasound Instrument (Tianjin Ao Tesaiensi precision instrument company)；LC-20AT high performance liquid chromatograph, SPD-20A inspection
It surveys device (Japanese Shimadzu Corporation)；Waters e2695 high performance liquid chromatograph, 2998PDA detector (water generation company of the U.S.).
Reagent: methanol is to analyze pure (Shanghai Su Yi chemical reagent Co., Ltd)；Acetonitrile is chromatographically pure (Sweden's Ou Sen Bark
Environmental Chemistry company)；Phosphoric acid is to analyze pure (Shanghai Su Yi chemical reagent Co., Ltd)；Ultrapure water is water purification machine self-control；Nourishing the stomach
Relaxation grain lot number is respectively 1705202,1704213,1704208,1703220,1703216,1703207,1701219,
1701210、1701206、1701201。
Embodiment 1: the preparation of test solution
1, the investigation of Extraction solvent and sample volume
Yangweishu granules sample (1705202) about 7.5g is taken, 4 parts, is set in 150mL stuffed conical flask respectively, it is accurate respectively
Methanol, 75% methanol, 50% methanol, water 25mL is added, ultrasonic extraction 30min is put to room temperature, crosses 0.45 μm of filter membrane, take continuous
Filtrate to obtain the final product.The above test sample distinguishes 20 μ L of sample introduction, 10 μ L.Compare the peak area difference of different solvents unit mass.
The results show that 20 μ L peak shape of sample introduction is poor, 10 μ L peak shapes are preferable, and theoretical pedal number is higher, therefore select 10 μ L's
Sample volume；The peak area for the main chromatographic peak unit mass of test sample that 50% methanol solution extracts is larger, extracts more complete；With first
Alcohol extracting will lead to sample package, and recovery rate is lower, therefore selects 50% methanol as Extraction solvent.
2, the investigation of extraction time
Yangweishu granules sample (1705202) about 7.5g is taken, 3 parts, is set in 150mL plug conical flask respectively, precision is added
50% methanol 25mL, difference ultrasonic extraction 30min, 45min, 60min.It puts to room temperature, crosses 0.45 μm of filter membrane, take subsequent filtrate i.e.
?.Compare the peak area difference of different extraction time unit masses.
The results show that ultrasonic extraction 45min extraction is more complete, the peak area of 60min unit mass is not further added by, therefore is selected
Selecting extraction time is 45min.
Embodiment 2: chromatographic condition optimization
1, the investigation of Detection wavelength
" HPLC method measures containing for yangweishu granules Content of Chlorogenic Acid, vitexin rhamnoside and aurantiamarin simultaneously to reference literature
Amount ", the maximum absorption wavelength of chlorogenic acid, vitexin rhamnoside and aurantiamarin are respectively 327nm, 335nm, 283nm, are used
PDA detector measures yangweishu granules sample (1705202) in 220nm-400nm wave-length coverage, respectively extract 220nm,
Chromatogram at 254nm, 280nm, 300nm, 330nm knows color at 220nm and 280nm by the comparison of the above different wave length
Spectral peak information is most abundant, and 280nm baseline preferably and have several main chromatographic peaks (RT=39.745,31.891,
39.954) there is absorption maximum near 280nm, therefore select 280nm as the Detection wavelength of yangweishu granules finger-print.
2, gradient is investigated
Because including the compound of various polarity in yangweishu granules, therefore using octadecylsilane chemically bonded silica as filler；
- 0.1% phosphate aqueous solution of acetonitrile is mobile phase；Column temperature: 30 DEG C, Detection wavelength 220nm, 10 μ L primary condition of sample volume is to gradient
Elution program optimizes, preferably optimal gradient out are as follows:
Embodiment 3: yangweishu granules fingerprint map construction
Qualified 10 batches, yangweishu granules sample of current standard of learning from else's experience detection, lot number be respectively 1705202 (S1),
1704213(S2)、1704208(S3)、1703220(S4)、1703216(S5)、1703207(S6)、1701219(S7)、
1701210 (S8), 1701206 (S9), 1701201 (S10) carry out yangweishu granules fingerprint image according to the testing conditions after optimization
Spectrum detection, specifically comprises the following steps:
(1) preparation of test solution: taking yangweishu granules powder, and precision weighs 7.5g, and the volumetric concentration of 25mL is added
50% methanol solution is ultrasonically treated 45min, lets cool, and filters, takes subsequent filtrate up to test solution；
(2) preparation of reference substance solution: precision weighs aurantiamarin reference substance, adds methanol that the reference substance of 0.06mg/mL is made
Solution is as reference solution.The reference substance for separately taking harpagoside, chlorogenic acid to add methanol that 0.05mg/mL is made respectively in right amount is molten
Liquid；
(3) chromatographic condition: using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase A, with volume fraction
Phosphate aqueous solution for 0.1% is that Mobile phase B carries out gradient elution according to the procedure below:
0min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
0-10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10-35min, volume shared by mobile phase A are gradually incremented to 28% by 10%, and surplus is Mobile phase B；
35min, volume shared by mobile phase A are 28%, and surplus is Mobile phase B；
35-45min, volume shared by mobile phase A are gradually incremented to 40% by 28%, and surplus is Mobile phase B；
45min, volume shared by mobile phase A are 40%, and surplus is Mobile phase B；
45-46min, volume shared by mobile phase A are gradually incremented to 80% by 40%, and surplus is Mobile phase B；
46min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
46-55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55-60min, volume shared by mobile phase A are gradually decremented to 10% by 80%, and surplus is Mobile phase B；
60min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
60-65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B.
Detection wavelength is 280nm, 30 DEG C of column temperature, flow velocity 1.0mL/min；
(4) accurate respectively to draw test sample and each 10 μ L of reference substance solution, high performance liquid chromatograph is injected, is measured, respectively
Obtain the chromatogram of test solution and control solution；
(5) " chromatographic fingerprints of Chinese materia medica is similar in the fingerprint similarity evaluation software using pharmacopoeia commission's establishment
Spend evaluation system ", generate compare feature map.Liquid chromatogram, the lot number of reference substance solution are the confession of 1705202 yangweishu granules
The liquid chromatogram of test sample solution, 10 batches of yangweishu granules test solutions export after importing fingerprint similarity software for calculation
Superposition chromatogram and foundation yangweishu granules reference fingerprint difference it is as shown in Figs. 1-5.
(6) in the finger-print for detecting resulting yangweishu granules, share peak are as follows: No. 1 peak, No. 2 peak chlorogenic acids, No. 3 peaks,
No. 4 peaks, No. 5 peaks, No. 6 peaks, No. 7 peaks, No. 8 peak aurantiamarins, No. 9 peaks, No. 10 peaks, No. 11 peak harpagosides, No. 12 peaks and No. 13
Peak；It is referring to peak with No. 8 peaks, relative retention time 1 calculates the relative retention time at other each peaks, as a result see the table below.
1 10 batches of yangweishu granules of table share peak relative retention time
It is obtained by table 1: with No. 8 peak aurantiamarins for referring to peak, the average relative retention time at each peak number is respectively as follows: No. 1 peak
0.307, No. 2 peaks 0.346,3 peak 0.397,5 of peak 0.374,4 peak 0.610,7 of peak 0.522,6 peak of peak 0.931,8
1.000, No. 9 peaks 1.031,10 1.246, No. 11 peaks in peak 1.259, No. 12 peaks 1.309 and No. 13 peak 1.380.Relative standard is inclined
Difference is smaller.It is not more than ± 5% principle according to retention time deviation, retention time is provided are as follows:
No. 1 peak: 0.307 ± 0.015；No. 2 peaks: 0.346 ± 0.017；
No. 3 peaks: 0.374 ± 0.019；No. 4 peaks: 0.397 ± 0.020；
No. 5 peaks: 0.522 ± 0.026；No. 6 peaks: 0.610 ± 0.030；
No. 7 peaks: 0.931 ± 0.047；No. 8 peaks: 1.000；
No. 9 peaks: 1.031 ± 0.052；No. 10 peaks: 1.246 ± 0.062；
No. 11 peaks: 1.259 ± 0.063；No. 12 peaks: 1.309 ± 0.065；
No. 13 peaks: 1.380 ± 0.069；
And wherein No. 2 peaks are chlorogenic acid, No. 8 peaks are aurantiamarin, No. 11 peaks are harpagoside.
Embodiment 4: yangweishu granules fingerprint spectrum method verifying
1, precision test
It takes with a collection of yangweishu granules sample (1705202), prepares test solution by sample solution preparation method, even
Continuous sample introduction 6 times, the relative retention time and relative peak area at each shared peak and internal standard peak are investigated, as a result as shown in Tables 1 and 2.
1 precision test of table-relative retention time
2 precision tests of table-relative peak area
The result shows that relative retention time RSD < 1%, relative peak area RSD < 3%, this method precision are preferable.
2, repetitive test
(1705202) 6 parts of sample of same lot number are taken, prepare test solution by sample solution preparation method respectively,
Chromatogram is measured, the results are shown in Table 3 and 4.
3 repetitive tests of table-relative retention time
4 repetitive tests of table-relative peak area
The result shows that relative retention time RSD < 2% of each shared chromatographic peak, relative peak area RSD < 3%, this method weight
Renaturation is preferable.
3, stability test
Take same test solution, respectively 0,2,4,8,12h, detected for 24 hours, when recording each shared chromatographic peak and retaining
Between and peak area, the results are shown in Table 5 and 6.
5 stability tests of table-relative retention time
6 stability tests of table-relative peak area
The result shows that relative retention time RSD < 1% of interior each shared chromatographic peak, relative peak area RSD < 3% supply for 24 hours
Test sample solution is at least stablized interior for 24 hours.
Embodiment 5: the similarity evaluation of yangweishu granules finger-print
Take 10 batches, yangweishu granules sample, lot number be respectively 1705202 (S1), 1704213 (S2), 1704208 (S3),
1703220(S4)、1703216(S5)、1703207(S6)、1701219(S7)、1701210(S8)、1701206(S9)、
1701201 (S10) carry out the detection of finger-print according to the method provided in embodiment 3, and 10 batches of sample finger-prints are imported
Pharmacopoeia commission provides similarity evaluation, is referring to spectrum, using median with 1705202 spectrograms
Method, time width 0.3min, is matched, and similarity is calculated.Similarity result is shown in Table 7.
7 fingerprint similarity result of table
Find out from the similarity result of table 7,10 batches of yangweishu granules samples are given birth to referring to spectrum and according to median method
At control map between similarity be all larger than 0.9, show that the similarity of the yangweishu granules of different batches is preferable.It can rest
Stomach relaxation grain finger-print is included in its quality evaluation, its similarity is set to not less than 0.9.
Embodiment 6:
The difference of the method and embodiment 3 of the finger-print of the present embodiment building yangweishu granules is only that: test sample is molten
In the preparation step of liquid, the yangweishu granules of 8.0g are weighed, remaining experiment condition and experimental implementation are same as Example 3.
Embodiment 7:
The difference of the method and embodiment 3 of the finger-print of the present embodiment building yangweishu granules is only that: test sample is molten
In the preparation step of liquid, for the methanol solution for being 70% using volume fraction as Extraction solvent, remaining experiment condition and experimental implementation are equal
It is same as Example 3.
Embodiment 8:
The difference of the method and embodiment 3 of the finger-print of the present embodiment building yangweishu granules is only that: test sample is molten
In the preparation step of liquid, extraction ultrasonic time is 40min, remaining experiment condition and experimental implementation are same as Example 3.
Embodiment 9: the quality testing of yangweishu granules
Sample lot number: 1706201,1706202
Test solution preparation, reference substance solution preparation and chromatographic condition: with embodiment 3.
The test solution and control solution of accurate draws equal amounts inject high performance liquid chromatograph respectively, by above-mentioned chromatography
Condition records 65min to get the liquid chromatogram of each test solution and control solution.Chinese medicine is provided using pharmacopoeia commission
Chromatographic fingerprinting similarity evaluation system evaluates map, calculates the similarity with reference fingerprint.With reference substance
Aurantiamarin retention time is 1 relative retention time for calculating other characteristic peaks, and calculated result is as shown in table 8.
8 lot number of table is the relative retention time and fingerprint similarity of the characteristic peak of 1706201,1706202 samples
Table 8 is it is found that lot number is the relative retention time of the characteristic peak of 1706201,1706202 samples in defined reservation
In time range, similarity > 0.9 illustrates the satisfactory quality of the two batches yangweishu granules.
Claims (8)
1. a kind of construction method of yangweishu granules finger-print, it is characterised in that include the following steps:
Step 1: the preparation of test solution
It weighs 5.0-10.0g yangweishu granules to be added in 25-100mL methanol aqueous solution, is ultrasonically treated 10- at 10-60 DEG C
60min, cooled and filtered, gained filtrate are test solution；
Step 2: the preparation of reference substance solution
Using methanol as solvent, weighs aurantiamarin, harpagoside, chlorogenic acid and be configured to reference substance solution respectively；
Step 3: chromatography detection
Test solution and each 5-20 μ L of reference substance solution are drawn respectively, and injection high performance liquid chromatograph detection respectively obtains confession
The chromatogram of test sample solution and reference substance solution；
Step 4: Chinese Pharmacopoeia Commission's similarity evaluation is utilized, to test solution and control
The chromatogram of product solution carries out data importing, Supplements and Data Matching respectively, and using median method, time width is
0.3min is to get finger-print.
2. construction method according to claim 1, it is characterised in that:
In step 1, weighs 5.0-8.0g yangweishu granules and is added in the methanol aqueous solution of 25-50mL volumetric concentration 40-80%,
30-50min, cooled and filtered are ultrasonically treated at 10-60 DEG C, gained filtrate is test solution.
3. construction method according to claim 1, it is characterised in that:
In step 2, in reference substance solution, the concentration of aurantiamarin solution is 0.06mg/mL, and the concentration of harpagoside is 0.05mg/
ML, the concentration of chlorogenic acid are 0.05mg/mL.
4. construction method according to claim 1, it is characterised in that:
In step 3,250mm × 4.6mm, 5 μm of Agilent Eclipse XDB-C18 chromatographic column are used when chromatography detects.
5. construction method according to claim 1, it is characterised in that:
In step 3, chromatography detection parameters are as follows:
Using octadecylsilane chemically bonded silica as filler；Using acetonitrile as mobile phase A, it is water-soluble with the phosphoric acid of volume fraction 0.1%
Liquid is that Mobile phase B carries out gradient elution；Detection wavelength is 275-285nm；25-40 DEG C of column temperature；Flow velocity is 0.5-1.5mL/min.
6. construction method according to claim 5, it is characterised in that:
Gradient elution program is provided that
0min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
0-10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
10-35min, volume shared by mobile phase A are gradually incremented to 28% by 10%, and surplus is Mobile phase B；
35min, volume shared by mobile phase A are 28%, and surplus is Mobile phase B；
35-45min, volume shared by mobile phase A are gradually incremented to 40% by 28%, and surplus is Mobile phase B；
45min, volume shared by mobile phase A are 40%, and surplus is Mobile phase B；
45-46min, volume shared by mobile phase A are gradually incremented to 80% by 40%, and surplus is Mobile phase B；
46min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
46-55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55min, volume shared by mobile phase A are 80%, and surplus is Mobile phase B；
55-60min, volume shared by mobile phase A are gradually decremented to 10% by 80%, and surplus is Mobile phase B；
60min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
60-65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B；
65min, volume shared by mobile phase A are 10%, and surplus is Mobile phase B.
7. a kind of application for the yangweishu granules finger-print that claim 1 constructs, it is characterised in that: utilize the fingerprint image of building
The quality of spectrum detection yangweishu granules preparation.
8. application according to claim 7, it is characterised in that include the following steps:
Step 1: the preparation of product to be tested solution
Precision weighs 5.0-10.0g yangweishu granules to be measured, and 0~100% methanol solution of 25-100mL volumetric concentration, 10- is added
10-60min, cooled and filtered are ultrasonically treated at 60 DEG C, gained filtrate is product to be tested solution；
Step 2: the preparation of reference substance solution
Using methanol as solvent, the concentration for being configured to aurantiamarin solution respectively is 0.06mg/mL, and the concentration of harpagoside is
0.05mg/mL, the concentration of chlorogenic acid are the reference substance solution of 0.05mg/mL；
Step 3: chromatography detection
Draw product to be tested solution and each 5-20 μ L of reference substance solution respectively, injection high performance liquid chromatograph detection, respectively obtain to
The chromatogram of survey product solution and reference substance solution；
Step 4: Chinese Pharmacopoeia Commission's similarity evaluation is utilized, by gained product to be tested fingerprint image
Spectrum is with the standard finger-print of building compared with pair, calculating fingerprint similarity, if similarity >=0.9 and there is standard fingerprint figure
13 characteristic peaks in spectrum, peak corresponding with reference peak are the peak S, and ± 5% principle is not more than according to retention time deviation, will be protected
Stay time rule are as follows:
No. 1 peak: 0.307 ± 0.015；No. 2 peaks: 0.346 ± 0.017；No. 3 peaks: 0.374 ± 0.019；No. 4 peaks: 0.397 ±
0.020；No. 5 peaks: 0.522 ± 0.026；No. 6 peaks: 0.610 ± 0.030；No. 7 peaks: 0.931 ± 0.047；No. 8 peaks: 1.000；9
Number peak: 1.031 ± 0.052；No. 10 peaks: 1.246 ± 0.062；No. 11 peaks: 1.259 ± 0.063；No. 12 peaks: 1.309 ±
0.065；No. 13 peaks: 1.380 ± 0.069；
Within the above range, then the yangweishu granules to be measured are up-to-standard for the relative retention time at each shared peak.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810596195.9A CN109030641B (en) | 2018-06-11 | 2018-06-11 | Method for constructing fingerprint of Yangweishu granules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810596195.9A CN109030641B (en) | 2018-06-11 | 2018-06-11 | Method for constructing fingerprint of Yangweishu granules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109030641A true CN109030641A (en) | 2018-12-18 |
| CN109030641B CN109030641B (en) | 2021-04-06 |
Family
ID=64612565
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810596195.9A Active CN109030641B (en) | 2018-06-11 | 2018-06-11 | Method for constructing fingerprint of Yangweishu granules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109030641B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376092A (en) * | 2019-08-28 | 2019-10-25 | 西南医科大学 | A kind of compound haw thorn Content of Chlorogenic Acid and content of hesperidin measurement and integrated evaluating method |
| CN114306466A (en) * | 2021-12-30 | 2022-04-12 | 合肥华润神鹿药业有限公司 | Traditional Chinese medicine for losing weight and reducing fat and detection method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703748A (en) * | 2009-11-06 | 2010-05-12 | 合肥神鹿双鹤药业有限责任公司 | Method for detecting quality of Yangweishu granules |
| WO2016201299A1 (en) * | 2015-06-12 | 2016-12-15 | Indiana University Research & Technology Corporation | Predicting suicidality using a combined genomic and clinical risk assessment |
| CN106770785A (en) * | 2017-01-11 | 2017-05-31 | 合肥华润神鹿药业有限公司 | A kind of method of the finger-print for setting up madder rattan medicinal material |
-
2018
- 2018-06-11 CN CN201810596195.9A patent/CN109030641B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703748A (en) * | 2009-11-06 | 2010-05-12 | 合肥神鹿双鹤药业有限责任公司 | Method for detecting quality of Yangweishu granules |
| WO2016201299A1 (en) * | 2015-06-12 | 2016-12-15 | Indiana University Research & Technology Corporation | Predicting suicidality using a combined genomic and clinical risk assessment |
| CN106770785A (en) * | 2017-01-11 | 2017-05-31 | 合肥华润神鹿药业有限公司 | A kind of method of the finger-print for setting up madder rattan medicinal material |
Non-Patent Citations (11)
| Title |
|---|
| HU, FU-LIAN: "A multicenter study of Chinese patent medicine wenweishu/yangweishu in the treatment of Helicobacter pylori positive patients with chronic gastritis and peptic ulcer", 《ZHONGHUA YI XUE ZA ZHI》 * |
| MARÍA GALVEZ-LLOMPART 等: "Modeling Natural Anti-Inflammatory Compounds by Molecular Topology", 《INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES》 * |
| 刘小花 等: "咳立止滴丸的指纹图谱研究", 《第三届中国中西部地区色谱学术交流会论文集》 * |
| 屈巧玲 等: "急支糖浆指纹图谱鉴别方法研究", 《中国中药杂志》 * |
| 彭文莉 等: "高效液相色谱法测定养胃舒颗粒中橙皮苷的含量", 《安徽医药》 * |
| 李宪刚 等: "HPLC-DAD法同时测定清肺宁嗽丸中4种成分的含量", 《中国医药科学》 * |
| 江萌 等: "HPLC法同时测定养胃舒颗粒中绿原酸、牡荆素鼠李糖苷及橙皮苷的含量", 《中国药师》 * |
| 程健 等: "怀牛膝"性善下行"对通塞脉微丸中绿原酸、异甘草苷、哈巴俄苷和甘草素体内药代动力学影响研究", 《中国中药杂志》 * |
| 胡怀明 等: "麻黄宣肺止咳糖浆指纹图谱研究", 《中国药师》 * |
| 薛鑫 等: "HPLC法同时测定银翘散煎液中七种有效成分含量", 《2013年"好医生杯"中药制剂创新与发展论坛论文集》 * |
| 陈永刚 等: "五参二连颗粒高效液相色谱指纹图谱研究及多成分定量分析", 《中国医药工业杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376092A (en) * | 2019-08-28 | 2019-10-25 | 西南医科大学 | A kind of compound haw thorn Content of Chlorogenic Acid and content of hesperidin measurement and integrated evaluating method |
| CN114306466A (en) * | 2021-12-30 | 2022-04-12 | 合肥华润神鹿药业有限公司 | Traditional Chinese medicine for losing weight and reducing fat and detection method |
| CN114306466B (en) * | 2021-12-30 | 2023-02-21 | 合肥华润神鹿药业有限公司 | Traditional Chinese medicine for losing weight and reducing fat and detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109030641B (en) | 2021-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107356691A (en) | Build the detection method of bent finger-print | |
| CN102778516B (en) | Rheum officinale and eupolyphaga capsule finger-print spectrum identification method | |
| CN107328872A (en) | The construction method and its standard finger-print of radix scrophulariae HPLC-FPS | |
| CN110187041A (en) | A kind of method for building up and its finger-print of tanreqin injection finger-print | |
| CN110441441B (en) | UPLC characteristic spectrum construction method and identification method of rhizoma cibotii and rhizoma cibotii scalding medicinal materials | |
| CN109030641A (en) | A kind of construction method of yangweishu granules finger-print | |
| CN106802327B (en) | A method of it establishing youngster and rushes down the finger-print for stopping pharmaceutical preparation | |
| CN106290599A (en) | A kind of content assaying method of Chinese medicine composition | |
| CN104155383B (en) | The detection method of blue or green Pu granule | |
| CN109507312A (en) | A kind of identification method of Cortex Phellodendri and its application | |
| CN113759035A (en) | Method for constructing fingerprint of Xiaochengqi decoction | |
| CN104007198B (en) | A kind of glossy ganoderma emperor's preparation HPLC standard finger-print and construction method thereof and application | |
| CN108760928A (en) | A kind of construction method of temperature stomach relaxation grain finger-print | |
| CN106290645B (en) | A kind of construction method and its standard finger-print of Lhasa rhubarb finger-print | |
| CN106918673B (en) | A kind of method for building up of the finger-print of Chinese medicine composition | |
| CN102133333A (en) | Quality control method for shenmai injection mass spectrum finger prints | |
| CN107643343A (en) | A kind of HPLC finger print measuring methods of yunü decoction standard soup | |
| CN106770785B (en) | A method of establishing the finger-print of madder rattan medicinal material | |
| CN110274970A (en) | The method for building up for melting poor finger-print and its application in Yixiesheng capsule Quality Control | |
| CN109633035A (en) | The HPLC finger-print and its construction method of a kind of hawthorn broken wall medicine materical crude slice and application | |
| CN114660204A (en) | Method for establishing fingerprint of Sanbai decoction | |
| CN103257191A (en) | Method for assaying kidney tonifying and life lengthening capsule fingerprint | |
| CN109298091B (en) | Standard characteristic spectrum construction method and quality detection method of solanum torvum asthma relieving ointment | |
| CN112557553A (en) | Fingerprint spectrum construction method and detection method of angelica sinensis Liuhuang decoction composition | |
| CN114113356A (en) | Fingerprint spectrum detection method of Xiaoyao pills |
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 |