CN1808114A - Quality control method for injection containing breviscapini and dipyridamole - Google Patents
Quality control method for injection containing breviscapini and dipyridamole Download PDFInfo
- Publication number
- CN1808114A CN1808114A CN 200610200039 CN200610200039A CN1808114A CN 1808114 A CN1808114 A CN 1808114A CN 200610200039 CN200610200039 CN 200610200039 CN 200610200039 A CN200610200039 A CN 200610200039A CN 1808114 A CN1808114 A CN 1808114A
- Authority
- CN
- China
- Prior art keywords
- dipyridamole
- finger
- solution
- scutellarin
- 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
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a quality control method for an injection. The method comprises: fingerprint pattern testing of the injection which takes breviscapine and dipyridamole as main ingredients; identification testing method of all or part ingredients in scutellarin and dipyridamole; content measuring method of all or part ingredients in scutellarin and dipyridamole. Compared with prior technology, the invention has more perfect quality control and has better precision and stability.
Description
Technical field:
The present invention is a kind of method of quality control of injection containing breviscapini and dipyridamole, belongs to the technical field that the quality in the drug production process is controlled.
Background technology:
Cardiovascular and cerebrovascular disease such as coronary heart disease, cerebral thrombus, hypertension, cerebral infarction etc. all are one of the most common and diseases that harm is maximum in the world today, also are common disease, the frequently-occurring disease of harm China people ' s health, have become human mortality's one of the main reasons; It was reported that the incidence of disease in recent years has and increases trend year by year, and in, young patient constantly increases.In order to reach the purpose of control, the applicant had once submitted a number of patent application and had been " 200410022790X ", name is called the application of " a kind of Chinese medicine preparation for the treatment of cardiovascular and cerebrovascular disease and preparation method thereof ", relate to a kind of treatment cardiovascular and cerebrovascular disease traditional Chinese medicine of making by Breviscapinun and Dipyridamole prescription, but pharmaceutical preparation must be on the basis that guarantees the constant product quality controllable safety, the more new development that could not lack, in order better to control the quality of said preparation, guarantee the security of medication, better instruct and produce, make technology controlling and process rationally strict more, make consumer's energy full appreciation product quality, need this parenteral solution method for quality of research control.
Summary of the invention:
The objective of the invention is to: a kind of method of quality control of injection containing breviscapini and dipyridamole is provided, and at by Breviscapinun: Dipyridamole=1: the medicine that the 0.3-3 prescription is made carries out quality control.This method provides means, technical method of the index that detects, detection or the like to relevant production, testing agency; So that better control the quality of said preparation, guarantee the security of medication, can better instruct production, make controlling of production process rationally strict more, make consumer's energy full appreciation product quality.
The present invention constitutes like this:
The method of quality control of injection containing breviscapini and dipyridamole is characterized in that: this method comprises following all or part of content:
(1) finger-print of injection containing breviscapini and dipyridamole test comprises the finger-print based on Breviscapinun, Dipyridamole composition characteristics;
(2) the differential test method of all or part of composition in scutellarin, the Dipyridamole;
(3) content test method of all or part of composition in scutellarin, the Dipyridamole.
The method of quality control of described injection containing breviscapini and dipyridamole is characterized in that this method comprises following finger-print:
Adopt the finger-print of liquid phase chromatography test based on Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: ejection preparation is not an amount of to get lamp to be measured, adds water or methyl alcohol or dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: get a kind of in an amount of scutellarin, the Dipyridamole, water or methyl alcohol or dissolve with ethanol are settled to suitable concn, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: moving phase is that acetonitrile or methyl alcohol-pH value are that 5.0～2.0 0.01mol/L～2mol/L biphosphate sodium water solution or pH value are that 5.0～2.0 0.01mol/L～2mol/L potassium dihydrogen phosphate aqueous solution or pH value are 5.0～2.0 0.01mol/L～2mol/L sodium hydrogen phosphate aqueous solution or water or 0.1%～5% glacial acetic acid solution or 0.1%～5% formic acid solution or 0.02%～5% phosphoric acid solution, gradient elution, flow velocity is 0.5～2.0ml/min, the detection wavelength is one or several in 190～410nm scope, and column temperature is in 20～60 ℃ of scopes;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 1～5;
(5) with the described method in (1)～(3) as in the injection to be measured based on the means of testing of the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with the finger-print of injection to be measured and the contrast of above-mentioned standard finger-print, in should meeting the following requirements partly or entirely:
I. calculate the similarity of the finger-print and the standard finger-print of injection to be measured, should be 0.80～1.00;
II. in the injection finger-print to be measured, non-total peak area must not surpass 10% of total peak area;
The odds ratio that each total peak area in the ratio of 40%～100% total peak area and the standard finger-print arranged in the injection finger-print III. to be measured, its difference must not be greater than ± 50%.
(ratio of total peak area is meant: as 1, calculate the ratio of each total fingerprint peaks area and object of reference peak area with the object of reference peak area)
Be preferably:
(1) preparation of need testing solution: precision takes by weighing lamp to be measured, and ejection preparation is not an amount of, adds methyl alcohol and makes the solution that every 1ml contains the 0.01-0.1mg Dipyridamole, filters with miillpore filter, gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is the 0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5) of pH value 3.5, gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 60 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 2～3;
(5) with the described method in (1)～(3) as in the injection to be measured based on the means of testing of the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with the contrast of injection finger-print to be measured and above-mentioned standard finger-print, in should meeting the following requirements partly or entirely:
I. calculate the similarity of the finger-print and the standard finger-print of injection to be measured, should be 0.90～1.00;
II. in the injection finger-print to be measured, non-total peak area must not surpass 5% of total peak area;
The odds ratio that each total peak area in the ratio of 60%～100% total peak area and the standard finger-print arranged in the injection finger-print III. to be measured, its difference must not be greater than ± 30%.
The method of quality control of described injection containing breviscapini and dipyridamole is characterized in that: the discrimination method of described injection is one or both a thin-layer chromatography discrimination method in scutellarin, the Dipyridamole:
Ejection preparation is not an amount of to get lamp to be measured, adds methyl alcohol or water or dissolve with ethanol and is diluted to suitable concn, as need testing solution; Other gets in scutellarin, the Dipyridamole reference substance one or both, adds methyl alcohol or ethanol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H thin layer plate or silica GF254 thin layer plate or polyamide film, with ethyl acetate or ethyl formate or methenyl choloride or methylene chloride-formic acid or glacial acetic acid-water 5～60: 0.5～5: 0.2～5 is developping agent, launch, take out, dry, spray is with aluminium choride-ethanol test solution, 60～150 ℃ are dried by the fire to spot colour developing clearly, put sight or uviol lamp 365nm or 254nm and detect down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, should show the fluorescence spot of same color.
Be preferably:
Ejection preparation is not an amount of to get lamp to be measured, adds dissolve with methanol and is diluted to suitable concn, as need testing solution; Other gets in scutellarin, the Dipyridamole reference substance one or both, adds methyl alcohol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-formic acid of 20: 2.5: 1.5-water is developping agent, launch, take out, dry, spray is with 3% aluminium choride-ethanol test solution, 105 ℃ were dried by the fire 5 minutes, put uviol lamp 365nm and detected down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
The method of quality control of described injection containing breviscapini and dipyridamole is characterized in that: the method for testing of described injection content is one or both a assay in scutellarin, the Dipyridamole:
Ejection preparation is not an amount of to get lamp to be measured, adds water or methyl alcohol or dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution; With scutellarin, one or both water or methyl alcohol or ethanolic solution are contrast in the Dipyridamole, adopt liquid phase chromatography, chromatographic column is a filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, acetonitrile or methyl alcohol-pH value is that 5.0～2.0 0.01mol/L～2mol/L biphosphate sodium water solution or pH value are that 5.0～2.0 0.01mol/L～2mol/L potassium dihydrogen phosphate aqueous solution or pH value are that 0.01mol/L～2mol/L sodium hydrogen phosphate aqueous solution of 5.0～2.0 or water or 0.1%～5% glacial acetic acid solution or 0.1%～5% formic acid solution or 0.02%～5% phosphoric acid solution are moving phase, the detection wavelength is one or several in 190～410nm scope, and column temperature is in 20～60 ℃ of scopes; Calculate with one point external standard method or calibration curve method, lamp to be measured not ejection preparation is unit quantity to be equivalent to every day with output, and content limit should be with the next item down or several:
(1) the per unit amount limit that contains scutellarin must not be less than 4.0mg;
(2) the per unit amount limit that contains Dipyridamole should be the 80-120% of preparation labelled amount.
Be preferably:
Ejection preparation is not an amount of to get lamp to be measured, adds dissolve with methanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution.With one or both methanol solutions in scutellarin, the Dipyridamole is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is the 0.02mol/L sodium dihydrogen phosphate of pH value 3.5, gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, lamp to be measured not ejection preparation is unit quantity to be equivalent to every day with output, and content limit should be with the next item down or several:
(1) the per unit amount limit that contains scutellarin must not be less than 8.0mg;
(2) the per unit amount limit that contains Dipyridamole should be the 90-110% of preparation labelled amount.
The method of quality control of described injection containing breviscapini and dipyridamole is characterized in that: all total contents that can survey composition of the Breviscapinun in the described injection, Dipyridamole and other account for more than 80% of total solid of deduction auxiliary material amount and amount of moisture in the preparation.
(total solid is meant: the weight of content deducts the weight behind auxiliary material amount and the amount of moisture)
Compared with prior art, the present invention's quality of the injection product made with Breviscapinun, Dipyridamole of perfect control more.The injection containing breviscapini and dipyridamole complicated component, if only with wherein one, two kind of composition illustrate its inherent quality, has certain one-sidedness, more can't judge the index components of its drug effect.Therefore the applicant has formulated the quality of comprehensively controlling injection based on the finger-print of Breviscapinun, Dipyridamole composition characteristics.But because the contained complex chemical composition of injection containing breviscapini and dipyridamole, the formulation of finger-print is caused interference, cause part finger-print feature instability, thus must control moving phase isochromatic spectrum condition, just can obtain good finger-print.That is to say that the lamp not finger-print of preparation is not that the finger-print simple superposition of Breviscapinun, Dipyridamole is just getable; Because two parts ingredient disturbing effect each other in the prescription, cause lamp not in the preparation finger-print characteristic peak of Breviscapinun, Dipyridamole change, adopt the conventional detection Breviscapinun or the liquid phase chromatogram condition of Dipyridamole, can't make the principal character composition in the preparation under a condition, go out the peak, and satisfy the Quality Control requirement of aspects such as degree of separation, has only the condition of the present invention of employing, select for use suitable parameter to carry out gradient elution, just can obtain exclusive, accurate, stable, workable desirable finger-print.
Under the liquid phase chromatogram condition acetonitrile or methyl alcohol-0.1% phosphate aqueous solution condition of the detection Breviscapinun of routine, Dipyridamole appearance time evening, and the peak type is poor; Under liquid phase chromatogram condition methyl alcohol-0.1% sodium hydrogen phosphate (phosphoric acid is regulated pH=4.6) of the detection Dipyridamole of routine, the scutellarin appearance time is too early, have overlapping with solvent peak, in order to increase the convenient and operability of fingerprint atlas detection method, we have investigated multiple condition, strive adopting a liquid-phase condition to detect characteristic components whole in the sample under the prerequisite that satisfies accuracy, specificity and stability, finally, obtained satisfied conclusion.Experiment showed, that assay of the present invention and fingerprint atlas detection method are more effective to the quality control of the injection product made with Breviscapinun, Dipyridamole, method precision, stability are all higher.
Experimental example 1 is based on the preparation of the finger-print of Breviscapinun, Dipyridamole composition characteristics
1, experimental apparatus, reagent and sample:
Reference substance: scutellarin: Nat'l Pharmaceutical ﹠ Biological Products Control Institute
2, chromatographic condition and system flexibility experiment:
2.1. the selection of chromatographic column:
In the research process, having selected conventional octadecylsilane chemically bonded silica for use is the liquid-phase chromatographic column of filling agent, tried out Zorbax, Inertsil ODS-3 respectively, Diamonsil ODS (is C18,4.6mm * 200mm, 5 μ m) chromatographic column of three kinds of trades mark, the result shows that the chromatographic column of three kinds of trades mark all can reach separating effect preferably, and wherein Diamonsil ODS chromatographic column separating effect is best, and post is imitated the highest.So finally selecting Diamonsil ODS chromatographic column (4.6mm * 200mm, 5 μ m) for use is the experimental study post.
2.2. the selection of moving phase:
Investigated (1) methyl alcohol-1% glacial acetic acid solution (30: 70) in the research process respectively, (2) acetonitrile-1% glacial acetic acid solution (17: 73), (3) acetonitrile-0.2% phosphoric acid solution (26: 74) (4) methyl alcohol-0.2% phosphoric acid solution (30: 70), (5) acetonitrile-0.02moL/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), solvent ratios is from 0 minute to 8 minutes, the ratio of acetonitrile rises to 30% by 18%, from 8 minutes to 15 minutes, the ratio of acetonitrile rises to 50% by 30%, from 15 minutes to 18 minutes, the ratio of acetonitrile reduces to 18% by 50%, and from 18 minutes to 25 minutes, the ratio of acetonitrile was 18% 5 kind of flow phase system.The result shows that peak shape is relatively poor under moving phase (1) condition, and it is less to go out the peak in 1 hour, goes out the peak after still having many components to be trapped in; Under moving phase (2) the moving phase condition, peak shape is relatively poor, separates bad; (3) under the condition, the peak hangover is serious, and it is incomplete to go out the peak; (4) under the condition, the peak hangover is serious, and it is incomplete to go out the peak; Under moving phase (5) condition, peak shape is better, goes out the peak fully and be evenly distributed, so finally selected.
2.3. detection wavelength determination:
In the research under acetonitrile-0.02moL/L sodium dihydrogen phosphate (25% phosphoric acid transfer pH=3.5) (gradient elution) moving phase condition, investigated the chromatographic peak situation under different-waveband typical wavelengths 203,230,250,283,335 respectively, the result shows, chromatographic peak is more under 283nm, peak shape is better, so finally select for use 283nm as detecting wavelength.
2.4. instrument, chromatographic column and integral parameter:
2.4.1 instrument parameter: selected liquid-phase chromatographic analysis field mainstream configuration and well behaved Agilent 1100 series of high efficiency liquid chromatographs for use, the Chemstation chromatographic work station.Chromatographic column is Diamonsil ODS (4.6mm * 200mm, 5 μ m); 30 ℃ of column temperatures, flow velocity 1.0ml/min.
2.4.2 integral parameter: Slope Sensitivity:1, peak width:0.05, smallest peaks area are 5% of object of reference (S) peak-to-peak area, minimum peak height be the S peak-to-peak high 5%.So setting can be avoided some very calculating of the chromatographic peak of small size (unimodal area accounts for total peak area less than 0.5%), guarantees the correlativity with object of reference simultaneously.
3, the preparation of need testing solution:
It is an amount of to get injection containing breviscapini and dipyridamole to be measured, adds methyl alcohol and makes the solution that every 1ml contains the 0.04mg Dipyridamole, filters with miillpore filter, gets subsequent filtrate as need testing solution.
4, the preparation of object of reference solution:
Scutellarin is one of Breviscapinun main active, its integral area in finger-print proportion more greatly and more stable, therefore selected scutellarin is as object of reference.
5, finger-print and technical parameter:
Formulate standard finger-print according to 10 batch samples, test sample finger-print and standard finger-print are compared, similarity is all between 0.90～1.00.
The thin-layer chromatography discrimination method of scutellarin, Dipyridamole in experimental example 2 injection containing breviscapini and dipyridamoles:
Feature for outstanding injection containing breviscapini and dipyridamole, selected scutellarin, Dipyridamole as its feature spot, but owing to have more and scutellarin, the Dipyridamole structure is close, polarity is similar composition in the injection, usual terms is difficult to reach requirements for quality control, so we have screened following thin layer plate and unfolding condition launches scutellarin, Dipyridamole:
| Condition | Problem |
| Normal hexane-benzene-ethyl formate-formic acid (5-5-10-4) silica gel H thin layer plate | Reference substance is expanded to the forward position |
| Ethyl acetate-benzene-acetate (5-4-3) silica gel H thin layer plate | Reference substance does not separate, and feminine gender has interference |
| Ethyl acetate-benzene-formic acid (5-4-3) polyamide membrane | Reference substance does not separate, and feminine gender has interference |
| Chloroform-ethyl acetate-methyl alcohol (7-2-4) silica GF254 thin layer plate | Reference substance does not separate, and feminine gender has interference |
| Toluene-ethyl acetate (8-1) silica gel H thin layer plate | Reference substance does not separate, and feminine gender has interference |
| Methyl alcohol-ethyl acetate-formic acid (10-5-0.5) silica gel g thin-layer plate | Reference substance is expanded to the forward position |
| Ethyl acetate-formic acid-water (20-2.5-1.5) silica gel g thin-layer plate | It is clear to separate, and the moderate feminine gender of Rf value is noiseless |
Through screening, determined with the silica gel g thin-layer plate to be stationary phase, ethyl acetate-formic acid-water (20-2.5-1.5) is developping agent, and with this understanding, the Rf value of scutellarin, Dipyridamole is moderate, and it is clear to separate with other spot, negative noiseless.
The content assaying method of scutellarin, Dipyridamole in experimental example 3 injection containing breviscapini and dipyridamoles
1 instrument and reagent
(1) key instrument:
High performance liquid chromatograph 1100series Agilent
High performance liquid chromatograph LC-2010AHT SHIMADZU
Electronic analytical balance BP211D SARTORIUS
Ultrasonic washing instrument KQ250DB Kunshan ultrasonic instrument company limited
(2) reagent:
Scutellarin Nat'l Pharmaceutical ﹠ Biological Products Control Institute
Dipyridamole Nat'l Pharmaceutical ﹠ Biological Products Control Institute
Methyl alcohol is analyzed pure Beijing logical wide fine chemistry industry company
Acetonitrile chromatographically pure CALEDON
The pure Beijing Chemical Plant of phosphoric acid top grade
Sodium dihydrogen phosphate is analyzed the pure Shanghai City glad chemical reagent work of shaking
2 chromatographic conditions
Chromatograph: SHIMADZU LC-2010ATH
Chromatographic column: Diamonsil (TM) C18,250 * 4.6mm, 5 μ m;
Moving phase: acetonitrile-0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred PH=3.5), gradient elution (seeing the following form)
| Time (min) | Acetonitrile (%) | Buffer salt solution |
| 0 8 15 18 25 | 18 30 50 18 18 | 82 70 50 82 82 |
Detect wavelength: 283nm; Column temperature: 30 ℃;
Flow velocity: 1.0ml/min; Sample size: 10 μ l.
The test of 3 system suitabilities
The preparation precision of reference substance solution takes by weighing scutellarin, the Dipyridamole reference substance is an amount of, adds methyl alcohol and makes the solution that every 1ml contains scutellarin 0.06mg, Dipyridamole 0.05mg, shakes up, promptly.
This product is got in the preparation of need testing solution, and mixing is therefrom got about 20mg, the accurate title, decide, and puts in the 10ml measuring bottle, adds an amount of sonicated of methyl alcohol (power 250W, frequency 33KHz) makes dissolving, take out, put, add methyl alcohol to scale to room temperature, shake up, filter with miillpore filter (0.45 μ m), get subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Obtain scutellarin, Dipyridamole, mixing reference substance, test sample chromatogram according to above-mentioned condition, its number of theoretical plate is pressed the scutellarin peak and is calculated greater than 3500.In the sample reference substance peak separate with close peak clear fully, degree of separation is all greater than 1.5.
The test of 4 negative ELIMINATION OF ITS INTERFERENCE, takes by weighing other auxiliary material in the prescription ratio and makes also mensuration of negative control product solution with need testing solution with method except that Breviscapinun and Dipyridamole for investigating the mensuration whether auxiliary material disturbs scutellarin and Dipyridamole.The result shows that negative sample is noiseless to the assay of scutellarin and Dipyridamole.
The investigation precision of 5 linear relationships is measured scutellarin, Dipyridamole mixed solution (every 1ml contains scutellarin 0.6288mg, Dipyridamole 0.5404mg respectively) 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, split in the 5ml measuring bottle, it is fixed to scale to add methyl alcohol, shake up, the therefrom accurate respectively 10 μ l that draw, inject liquid chromatograph, measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).Amount (μ g) with scutellarin, Dipyridamole is a horizontal ordinate respectively, and peak area is that ordinate is figure, the drawing standard curve.
The scutellarin linear relationship
| Numbering | Scutellarin amount (μ g) | Conversion back (μ g) | Peak area |
| 1 | 0.25152 | 0.242365 | 324484.5 |
| 2 | 0.50304 | 0.484729 | 644884.5 |
| 3 | 0.75456 | 0.727094 | 973386.5 |
| 4 | 1.00608 | 0.969459 | 1285592 |
| 5 | 1.25760 | 1.211823 | 1613625 |
Regression equation: y=1328159.51x+2697.69
Related coefficient: γ=0.9999
The result shows that scutellarin linear relationship within 0.242365 μ g～1.211823 μ g scopes is good.
The Dipyridamole linear relationship
| Numbering | Dipyridamole amount (μ g) | Peak area |
| 1 | 0.21616 | 438641.5 |
| 2 | 0.43232 | 880820 |
| 3 | 0.64848 | 1305267.5 |
| 4 | 0.86464 | 1747140 |
| 5 | 1.08080 | 2184614 |
Regression equation: y=2016621.78x+3817.10
Related coefficient: γ=0.9999
The result shows that Dipyridamole linear relationship within 0.21616 μ g～1.08080 μ g scopes is good.
Through calculating, scutellarin, Dipyridamole typical curve were the straight line of initial point, therefore selected one point external standard method to measure the content of scutellarin, Dipyridamole in the injection containing breviscapini and dipyridamole.
The accurate absorption of 6 reference substance solution precision and stability test scutellarin, the every 1ml of Dipyridamole reference substance mixed solution contain scutellarin 0.06288mg, Dipyridamole 0.05404mg respectively) 10 μ l, inject liquid chromatograph, the record peak area is measured at 0,2,6,10,24 hour sample introduction respectively.
Reference substance solution stability test result
| Time (h) | 0 | 2 | 6 | 10 | 24 | Mean value | RSD(％) |
| Scutellarin | 798057 | 790693 | 800924 | 793218 | 797310 | 796040 | 0.51 |
| Dipyridamole | 1098659 | 1100803 | 1092907 | 1100952 | 1094139 | 1097492 | 0.34 |
The result shows that reference substance solution precision is good, and is stable in 24 hours.
Accurate need testing solution (2.017mg/ml) the 10 μ l that draw of 7 need testing solution stability tests inject liquid chromatograph, and the record peak area is measured at 0,2,6,10,24 hour sample introduction respectively.
Need testing solution stability test result
| Time (h) | 0 | 2 | 6 | 10 | 24 | Mean value | RSD(％) |
| Scutellarin | 831295 | 835542 | 842908 | 846211 | 831659 | 837523 | 0.80 |
| Dipyridamole | 1078448 | 1083496 | 1106949 | 1107380 | 1084206 | 1092096 | 1.28 |
The result shows that need testing solution is good at 24 hours internal stabilities.
8 replica tests are got content under this product content uniformity item, and mixing is therefrom got about 20mg (totally 5 parts), and precision claims fixed, split in the 10ml measuring bottle, by operating under preparation of text need testing solution and the mensuration item.
Replica test
| Numbering | The scutellarin peak area | Scutellarin content (mg/ props up) | The Dipyridamole peak area | Dipyridamole content (mg/ props up) |
| 1 | 854668 | 4.909 | 1108045 | 4.143 |
| 2 | 812223 | 4.764 | 1048109 | 4.002 |
| 3 | 811394 | 4.869 | 1045012 | 4.083 |
| 4 | 816596 | 4.824 | 1052872 | 4.049 |
| 5 | 815282 | 4.833 | 1051625 | 4.058 |
Scutellarin average content=4.84mg/ props up; RSD=1.00%;
Dipyridamole average content=4.07mg/ props up; RSD=1.13%.
The result shows that repeatability is good.
Content under this product content uniformity item is got in the test of 9 average recoveries, and mixing is therefrom got about 10mg (totally 6 parts), splits in the 10ml measuring bottle; Precision takes by weighing scutellarin reference substance 15.72mg, and Dipyridamole reference substance 13.51mg puts in the 25ml measuring bottle altogether, add an amount of sonicated of methyl alcohol (power 250W, frequency 33KHz) and make dissolving, take out, put to room temperature, add methyl alcohol to scale, shake up, therefrom precision is measured 0.5ml (totally 6 parts), split in the above-mentioned 10ml measuring bottle, add an amount of sonicated of methyl alcohol (power 250W, frequency 33KHz) and make dissolving, take out, put to room temperature, add methyl alcohol to scale, shake up, filter with miillpore filter (0.45 μ m), the accurate subsequent filtrate 10 μ l that draw, inject liquid chromatograph, measure, promptly.
The content of scutellarin, Dipyridamole is respectively in the injection containing breviscapini and dipyridamole: 3.0215%, 2.5409%.The test of scutellarin average recovery
Scutellarin average recovery rate=96.54%, RSD=1.48%;
| Numbering | Test sample weighing (mg) | Pure product amount (mg) in the test sample | Scutellarin addition (mg) | Pure product addition (mg) | The amount of recording (mg) | The recovery (%) |
| 1 | 9.71 | 0.2934 | 0.3144 | 0.3030 | 0.5840 | 95.91 |
| 2 | 9.64 | 0.2913 | 0.3144 | 0.3030 | 0.5808 | 95.54 |
| 3 | 9.79 | 0.2958 | 0.3144 | 0.3030 | 0.5889 | 96.73 |
| 4 | 10.14 | 0.3064 | 0.3144 | 0.3030 | 0.5991 | 96.60 |
| 5 | 9.40 | 0.2840 | 0.3144 | 0.3030 | 0.5846 | 99.21 |
| 6 | 10.19 | 0.3079 | 0.3144 | 0.3030 | 0.5965 | 95.25 |
The test of Dipyridamole average recovery
| Numbering | Test sample weighing (mg) | Pure product amount (mg) in the test sample | Dipyridamole addition (mg) | The amount of recording (mg) | The recovery (%) |
| 1 | 9.71 | 0.2467 | 0.2702 | 0.5197 | 101.04 |
| 2 | 9.64 | 0.2449 | 0.2702 | 0.5206 | 102.04 |
| 3 | 9.79 | 0.2488 | 0.2702 | 0.5182 | 99.70 |
| 4 | 10.14 | 0.2576 | 0.2702 | 0.5230 | 98.22 |
| 5 | 9.40 | 0.2388 | 0.2702 | 0.5162 | 102.66 |
| 6 | 10.19 | 0.2589 | 0.2702 | 0.5271 | 99.26 |
Dipyridamole average recovery rate=100.49%, RSD=1.71%;
10 sample sizes are measured and are got the not middle test agent of freeze-dried powder (5mg Dipyridamole/bottle) of three batches of lamps, press the described method of text and handle, and measure.
The assay result
| Lot number | Scutellarin (mg/ props up) | Dipyridamole (mg/ props up) |
| A collection of | 4.813 | 4.039 |
| Two batches | 4.808 | 4.031 |
| Three batches | 4.800 | 4.014 |
According to The above results draft this lamp not in the freeze-dried powder content of scutellarin be not less than 4mg/ and prop up, the content of Dipyridamole is that 3.6-4.4mg/ props up.
Embodiment:
Embodiment 1: the number of patent application of submitting according to the applicant is " 200410022790X ", and name is called the preparation of the technology preparation of " a kind of Chinese medicine preparation for the treatment of cardiovascular and cerebrovascular disease and preparation method thereof "; Adopt the liquid phase chromatography test lamp not in the parenteral solution based on the finger-print of Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: it is an amount of that precision is measured test sample, adds methyl alcohol and make the solution that every 1ml contains the 0.04mg Dipyridamole, filters with miillpore filter, gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 60 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 2;
(5) with the described method in (1)～(3) as in the parenteral solution to be measured based on the means of testing of the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with parenteral solution finger-print to be measured and the contrast of above-mentioned standard finger-print, should meet the following requirements: calculate the similarity of the finger-print and the standard finger-print of freeze-dried powder to be measured, should be 0.90～1.00.
Embodiment 2: adopt the liquid phase chromatography test lamp not in the powder pin based on the finger-print of Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: it is an amount of that precision takes by weighing freeze-dried powder to be measured, adds ethanol and make the solution that every 1ml contains the 0.04mg Dipyridamole, filters with miillpore filter, gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02mol/L potassium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 60 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 2;
(5) with the described method in (1)～(3) as in the freeze-dried powder to be measured with means of testing based on the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with freeze-dried powder finger-print to be measured and the contrast of above-mentioned standard finger-print, should meet the following requirements:
Calculate the similarity of the finger-print and the standard finger-print of freeze-dried powder to be measured, should be 0.90～1.00.
Embodiment 3: adopt the liquid phase chromatography test lamp not in the parenteral solution based on the finger-print of Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: precision is measured and is got parenteral solution to be measured, adds water, shakes up, and makes the solution that every ml contains Dipyridamole 0.08mg, filters with miillpore filter, gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: it is an amount of that precision takes by weighing Dipyridamole, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is a methyl alcohol, Mobile phase B is 0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 35% by 22% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 55% by 35%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 22% by 55%, from 18 minutes to 60 minutes, the ratio of mobile phase A was 22%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 2;
(5) with the described method in (1)～(3) as in the parenteral solution to be measured with means of testing based on the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with the contrast of injection finger-print to be measured and above-mentioned standard finger-print, in should meeting the following requirements partly or entirely:
I. calculate the similarity of the finger-print and the standard finger-print of parenteral solution to be measured, should be 0.80～1.00;
II. in the parenteral solution finger-print to be measured, non-total peak area must not surpass 5% of total peak area;
III. in the parenteral solution finger-print to be measured in the ratio of Dipyridamole and scutellarin peak area and the standard finger-print odds ratio of each total peak area, its difference must not be greater than ± 30%.
Embodiment 4: the thin-layer chromatography discrimination method of scutellarin, Dipyridamole in the freeze-dried powder:
It is an amount of to get freeze-dried powder to be measured, adds dissolve with methanol and is diluted to suitable concn, as need testing solution; Other gets scutellarin, Dipyridamole reference substance, adds methyl alcohol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-formic acid-water (20: 2.5: 1.5) is developping agent, launch, take out, dry, spray is with 3% aluminium choride-ethanol test solution, 105 ℃ were dried by the fire 5 minutes, put uviol lamp 365nm and detected down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
Embodiment 5: the thin-layer chromatography discrimination method of scutellarin, Dipyridamole in the freeze-dried powder:
It is an amount of to get freeze-dried powder to be measured, adds dissolve with ethanol and is diluted to suitable concn, as need testing solution; Other gets scutellarin, Dipyridamole reference substance, adds ethanol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively on same silica gel H thin layer plate, with ethyl acetate-formic acid-water (16: 1: 1) is developping agent, launch, take out, dry, spray is with 3% aluminium choride-ethanol test solution, 105 ℃ were dried by the fire 5 minutes, put uviol lamp 365nm and detected down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
Embodiment 6: the thin-layer chromatography discrimination method of scutellarin, Dipyridamole in the parenteral solution:
Get parenteral solution 1ml, to evaporating dish, water bath method, residue methyl alcohol 1ml makes dissolving, as need testing solution; Other gets scutellarin, Dipyridamole reference substance, adds ethanol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively on same silica gel H thin layer plate, with ethyl formate-glacial acetic acid-water (20: 2: 1) is developping agent, launch, take out, dry, spray is with 1% aluminium choride-ethanol test solution, 105 ℃ were dried by the fire 5 minutes, put uviol lamp 365nm and detected down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
Embodiment 7: the assay of scutellarin, Dipyridamole in the freeze-dried powder (every bottle contains Dipyridamole 4mg):
It is an amount of to get freeze-dried powder to be measured, adds dissolve with methanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution.With scutellarin, Dipyridamole methanol solution is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, content limit should be following several:
(1) the per unit amount limit that contains scutellarin must not be less than 8.0mg
(2) to contain the limit of Dipyridamole be the 90-110% of preparation labelled amount to the per unit amount.
Embodiment 8: the assay of scutellarin, Dipyridamole in the freeze-dried powder:
It is an amount of to get freeze-dried powder to be measured, adds dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution.With scutellarin, Dipyridamole ethanolic solution is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02mol/L potassium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of stream merit phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, freeze-dried powder to be measured is unit quantity to be equivalent to every day with output, and content limit should be following several:
(1) the per unit amount limit that contains scutellarin must not be less than 4.0mg
(2) the per unit amount limit that contains Dipyridamole should be the 80-120% of preparation labelled amount.
Embodiment 9: the assay of scutellarin, Dipyridamole in the parenteral solution:
Get parenteral solution 1ml to be measured, put in the 10ml measuring bottle, add water and decide to shake up, filter, get subsequent filtrate as need testing solution to scale.With scutellarin, Dipyridamole methanol solution is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is a methyl alcohol, Mobile phase B is 0.02mol/L potassium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.0), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 35% by 22% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 55% by 35%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 22% by 55%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 22%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, parenteral solution to be measured is unit quantity to be equivalent to every day with output, and content limit should be following several:
(1) the per unit amount limit that contains scutellarin must not be less than 4.0mg;
(2) the per unit amount limit that contains Dipyridamole should be the 85-115% of preparation labelled amount.
Embodiment 10: lamp is the assay of the middle scutellarin of parenteral solution (every contains Dipyridamole 4mg), Dipyridamole not:
It is an amount of to get parenteral solution to be measured, adds dissolve with methanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution.With scutellarin, Dipyridamole methanol solution is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is 0.02mol/L sodium dihydrogen phosphate (25% phosphoric acid is transferred pH=3.5), gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, content limit should be following several:
(1) the per unit amount limit that contains scutellarin must not be less than 8.0mg;
(2) to contain the limit of Dipyridamole be the 90-110% of preparation labelled amount to the per unit amount.
Claims (8)
1. the method for quality control of an injection containing breviscapini and dipyridamole, it is characterized in that: this method comprises following all or part of content:
(1) finger-print of injection containing breviscapini and dipyridamole test comprises the finger-print based on Breviscapinun, Dipyridamole composition characteristics;
(2) the differential test method of all or part of composition in scutellarin, the Dipyridamole;
(3) content test method of all or part of composition in scutellarin, the Dipyridamole.
2. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 1, it is characterized in that this method comprises following finger-print:
Adopt the finger-print of liquid phase chromatography test based on Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: ejection preparation is not an amount of to get lamp to be measured, adds water or methyl alcohol or dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: get a kind of in an amount of scutellarin, the Dipyridamole, water or methyl alcohol or dissolve with ethanol are settled to suitable concn, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler: moving phase is that acetonitrile or methyl alcohol-pH value are that 5.0～2.0 0.01mol/L～2mol/L biphosphate sodium water solution or pH value are that 5.0～2.0 0.01mol/L～2mol/L potassium dihydrogen phosphate aqueous solution or pH value are 5.0～2.0 0.01mol/L～2mol/L sodium hydrogen phosphate aqueous solution or water or 0.1%～5% glacial acetic acid solution or 0.1%～5% formic acid solution or 0.02%～5% phosphoric acid solution, gradient elution, flow velocity is 0.5～2.0ml/min, the detection wavelength is one or several in 190～410nm scope, and column temperature is in 20～60 ℃ of scopes;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 1～5;
(5) with the described method in (1)～(3) as in the injection to be measured based on the means of testing of the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with the finger-print of injection to be measured and the contrast of above-mentioned standard finger-print, in should meeting the following requirements partly or entirely:
I. calculate the similarity of the finger-print and the standard finger-print of injection to be measured, should be 0.80～1.00;
II. in the injection finger-print to be measured, non-total peak area must not surpass 10% of total peak area;
The odds ratio that each total peak area in the ratio of 40%～100% total peak area and the standard finger-print arranged in the injection finger-print III. to be measured, its difference must not be greater than ± 50%.
3. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 2, it is characterized in that this method comprises following finger-print:
Adopt the finger-print of liquid phase chromatography test based on Breviscapinun, Dipyridamole composition characteristics:
(1) preparation of need testing solution: precision takes by weighing lamp to be measured, and ejection preparation is not an amount of, adds methyl alcohol and makes the solution that every 1ml contains the 0.01-0.1mg Dipyridamole, filters with miillpore filter, gets subsequent filtrate as need testing solution;
(2) preparation of object of reference solution: it is an amount of that precision takes by weighing scutellarin, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, as object of reference solution;
(3) chromatographic condition: the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is the 0.02mol/L sodium dihydrogen phosphate of pH value 3.5, gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 60 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃;
(4) formulation of standard finger-print: with said method as the means of testing of formulating based on the finger-print of Breviscapinun, Dipyridamole composition characteristics; According to 10 batches or 10 batches of collection of illustrative plates that above test sample is measured, formulate standard finger-print, be benchmark with the retention time of the object of reference chromatographic peak determined, calculate the relative retention time of other total chromatographic peak, in the described standard finger-print, total peak has 2～3;
(5) with the described method in (1)～(3) as in the injection to be measured based on the means of testing of the finger-print of Breviscapinun, Dipyridamole composition characteristics, the finger-print of preparation testing sample;
(6) with the contrast of injection finger-print to be measured and above-mentioned standard finger-print, in should meeting the following requirements partly or entirely:
I. calculate the similarity of the finger-print and the standard finger-print of injection to be measured, should be 0.90～1.00;
II. in the injection finger-print to be measured, non-total peak area must not surpass 5% of total peak area;
The odds ratio that each total peak area in the ratio of 60%～100% total peak area and the standard finger-print arranged in the injection finger-print III. to be measured, its difference must not be greater than ± 30%.
4. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 1, it is characterized in that:
The discrimination method of described injection is one or both a thin-layer chromatography discrimination method in scutellarin, the Dipyridamole:
Ejection preparation is not an amount of to get lamp to be measured, adds methyl alcohol or water or dissolve with ethanol and is diluted to suitable concn, as need testing solution; Other gets in scutellarin, the Dipyridamole reference substance one or both, adds methyl alcohol or ethanol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 1～30 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate or silica gel H thin layer plate or silica GF254 thin layer plate, with ethyl acetate or ethyl formate or methenyl choloride or methylene chloride-formic acid or glacial acetic acid-water 5～60: 0.5～5: 0.2～5 is developping agent, launch, take out, dry, spray is with aluminium choride-ethanol test solution, 60～150 ℃ are dried by the fire to spot colour developing clearly, put daylight or uviol lamp 365nm or 254nm and detect down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, should show the fluorescence spot of same color.
5. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 4, it is characterized in that:
The discrimination method of described injection is:
One or both thin-layer chromatography discrimination method in scutellarin, the Dipyridamole: ejection preparation is not an amount of to get lamp to be measured, adds dissolve with methanol and is diluted to suitable concn, as need testing solution; Other gets in scutellarin, the Dipyridamole reference substance one or both, adds methyl alcohol respectively and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Adopt the disclosed thin-layered chromatography test of Chinese Pharmacopoeia appendix, draw each 10 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-formic acid of 20: 2.5: 1.5-water is developping agent, launch, take out, dry, spray is with 3% aluminium choride-ethanol test solution, 105 ℃ were dried by the fire 5 minutes, put uviol lamp 365nm and detected down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
6. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 1, it is characterized in that:
The method of testing of described injection content is one or both a assay in scutellarin, the Dipyridamole:
Ejection preparation is not an amount of to get lamp to be measured, adds water or methyl alcohol or dissolve with ethanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution; With scutellarin, one or both water or methyl alcohol or ethanolic solution are contrast in the Dipyridamole, adopt liquid phase chromatography, chromatographic column is a filling agent with octadecylsilane chemically bonded silica or eight alkyl silane bonded silica gels or dialkyl silane bonded silica gel, acetonitrile or methyl alcohol-pH value is that 5.0～2.0 0.01mol/L～2mol/L biphosphate sodium water solution or pH value are that 5.0～2.0 0.01mol/L～2mol/L potassium dihydrogen phosphate aqueous solution or pH value are that 0.01mol/L～2mol/L sodium hydrogen phosphate aqueous solution of 5.0～2.0 or water or 0.1%～5% glacial acetic acid solution or 0.1%～5% formic acid solution or 0.02%～5% phosphoric acid solution are moving phase, the detection wavelength is one or several in 190～410nm scope, and column temperature is in 20～60 ℃ of scopes; Calculate with one point external standard method or calibration curve method, lamp to be measured not ejection preparation is unit quantity to be equivalent to every day with output, and content limit should be with the next item down or several:
(1) the per unit amount limit that contains scutellarin must not be less than 4.0mg;
(2) the per unit amount limit that contains Dipyridamole should be the 80-120% of preparation labelled amount.
7. according to the method for quality control of the described injection containing breviscapini and dipyridamole of claim 6, it is characterized in that:
The method of testing of described injection content is one or both a assay in scutellarin, the Dipyridamole:
Ejection preparation is not an amount of to get lamp to be measured, adds dissolve with methanol or dilution, shakes up, and filters, and gets subsequent filtrate as need testing solution; With one or both methanol solutions in scutellarin, the Dipyridamole is contrast, adopts liquid phase chromatography, and the chromatographic column octadecylsilane chemically bonded silica is a filling agent, and the chromatographic column adopting octadecylsilane chemically bonded silica is a filler; Mobile phase A is an acetonitrile, Mobile phase B is the 0.02mol/L sodium dihydrogen phosphate of pH value 3.5, gradient elution, solvent ratios are that the ratio of mobile phase A rose to 30% by 18% from 0 minute to 8 minutes, from 8 minutes to 15 minutes, the ratio of mobile phase A rises to 50% by 30%, and from 15 minutes to 18 minutes, the ratio of mobile phase A reduced to 18% by 50%, from 18 minutes to 25 minutes, the ratio of mobile phase A was 18%; Flow velocity is 1.0ml/min, and the detection wavelength is 283 ± 2nm, and column temperature is 30 ℃; Calculate with one point external standard method, lamp to be measured not ejection preparation is unit quantity to be equivalent to every day with output, and content limit should be with the next item down or several:
(1) the per unit amount limit that contains scutellarin must not be less than 8.0mg;
(2) the per unit amount limit that contains Dipyridamole should be the 90-110% of preparation labelled amount.
8. according to the method for quality control of claim 6 or 7 described injection containing breviscapini and dipyridamoles, it is characterized in that: all total contents that can survey composition of the scutellarin in the described injection, Dipyridamole and other account for more than 80% of total solid of deduction auxiliary material amount and amount of moisture in the preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006102000393A CN100480697C (en) | 2005-02-07 | 2006-01-17 | Quality control method for injection containing breviscapini and dipyridamole |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200510003014.X | 2005-02-07 | ||
| CN200510003014 | 2005-02-07 | ||
| CNB2006102000393A CN100480697C (en) | 2005-02-07 | 2006-01-17 | Quality control method for injection containing breviscapini and dipyridamole |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1808114A true CN1808114A (en) | 2006-07-26 |
| CN100480697C CN100480697C (en) | 2009-04-22 |
Family
ID=36840128
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006102000393A Expired - Fee Related CN100480697C (en) | 2005-02-07 | 2006-01-17 | Quality control method for injection containing breviscapini and dipyridamole |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100480697C (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102038728A (en) * | 2009-10-26 | 2011-05-04 | 贵阳医学院 | Quality control method for erigeron breviscapus (Vant.) hand-mazz. |
| CN104849362A (en) * | 2015-03-17 | 2015-08-19 | 中山市中智药业集团有限公司 | Erigeron breviscapus wall-breaking decoction pieces fingerprinting common model construction and quality detection method thereof |
| CN110988234A (en) * | 2019-10-24 | 2020-04-10 | 昆明龙津药业股份有限公司 | Quality control method of erigeron breviscapus medicinal material |
-
2006
- 2006-01-17 CN CNB2006102000393A patent/CN100480697C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102038728A (en) * | 2009-10-26 | 2011-05-04 | 贵阳医学院 | Quality control method for erigeron breviscapus (Vant.) hand-mazz. |
| CN104849362A (en) * | 2015-03-17 | 2015-08-19 | 中山市中智药业集团有限公司 | Erigeron breviscapus wall-breaking decoction pieces fingerprinting common model construction and quality detection method thereof |
| CN110988234A (en) * | 2019-10-24 | 2020-04-10 | 昆明龙津药业股份有限公司 | Quality control method of erigeron breviscapus medicinal material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100480697C (en) | 2009-04-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108918700B (en) | Method for simultaneously detecting statin side chain and enantiomer impurities thereof | |
| CN1712051A (en) | Quality control of Gongyanping preparation | |
| CN1920554A (en) | Method for measuring OMEGA3 unsaturated fatty acid ester in seal oil | |
| CN1621836A (en) | Quality controlling method for pulse restoring injection | |
| CN1808114A (en) | Quality control method for injection containing breviscapini and dipyridamole | |
| CN1916602A (en) | Capillary electrophoresis method for detecting medications and poisons in blood and urine at the same time | |
| CN1824238A (en) | Quality control method of medicinal preparation for treating lypemania | |
| CN1255678C (en) | Method for measuring content of active constitutent of 'Siji-Sanhuang' capsule | |
| CN1857551A (en) | Quality control method for Chinese medicine preparation | |
| CN1261762C (en) | Quality determination method for external cultivated bezoar | |
| CN1785295A (en) | Quality control method of cbinese medicinal preparation | |
| CN1973855A (en) | Motherwort injection | |
| CN1785267A (en) | Quality control method of compound gallblader freeflow solid preparation | |
| CN1758060A (en) | Determination method of effective component in aliphatic oil | |
| CN1857620A (en) | Quality control method for visual fatigue treating medicine preparation | |
| CN1785219A (en) | Quality control method of composite prepn. of dry mango tree leaves extract | |
| CN1840118A (en) | Compound preparation for treating bronchitis, its preparation method and quality control method | |
| CN1857406A (en) | Quality control method for Chinese medicine preparation | |
| CN1823946A (en) | Quality control method of child speen supporting oral liquid preparation | |
| CN101066437A (en) | Quality control method for compound cantharis oral liquid | |
| CN1857422A (en) | Quality control method for Chinese medicine preparation | |
| CN1745803A (en) | Longxuejie extract with alpha-glycosidase inhibiting activity and its quality control | |
| CN1827147A (en) | Dispersible tablet with gastrodia tuber for treating headache, its preparation and quality control method | |
| CN1939502A (en) | Ginseng freeze-drying powdery injection and its quality control | |
| CN1785232A (en) | Method for quality control of Qianbai biyan solid prepn. for treating rhinitis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: BEIJING LIUSHENGHE MEDICAL TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: YUNYANXICHUANG MEDICINAL SCIENCE AND TECHNOLOGY DEVELOPMENT CO., LTD., GUIYANG C Effective date: 20110413 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 550001 A-12/F, ZHICHENG BUILDING, NO. 3, YAN AN EAST ROAD, GUIYANG CITY, GUIZHOU PROVINCE TO: 100070 ROOM 317, LIFE SCIENCE INCUBATION CENTER, NO. 8, HANGFENG ROAD, FENGTAI SCIENCE CITY, FENGTAI DISTRICT, BEIJING |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20110413 Address after: 100070 room 317 of life science incubation center, 8 Feng Feng Road, Fengtai Science City, Beijing, Fengtai District Patentee after: Beijing Liushenghe Medical Technology Co.,Ltd. Address before: 550001, A-12 building, Zhicheng building, 3 East Yanan Road, Guiyang, Guizhou Patentee before: Yunyanxichuang Medicinal Science and Technology Development Co., Ltd., Guiyang C |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090422 Termination date: 20150117 |
|
| EXPY | Termination of patent right or utility model |