CN101377475A - Quality control method suitable for solancarpine, solamargine and preparation thereof - Google Patents
Quality control method suitable for solancarpine, solamargine and preparation thereof Download PDFInfo
- Publication number
- CN101377475A CN101377475A CNA2007101313499A CN200710131349A CN101377475A CN 101377475 A CN101377475 A CN 101377475A CN A2007101313499 A CNA2007101313499 A CN A2007101313499A CN 200710131349 A CN200710131349 A CN 200710131349A CN 101377475 A CN101377475 A CN 101377475A
- Authority
- CN
- China
- Prior art keywords
- solution
- solasonine
- solamargine
- reference substance
- preparation
- 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.)
- Pending
Links
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention relates to a quality control method which is suitable for solasonine, solamargine and preparations thereof. For directly determining the content of the solasonine and the solamargine in solanum nigrum, an inventor gets the solasonine and the solamargine separated from solanum nigrum medicinal materials to manufacture the reference substance of the solasonine and the reference substance of the solamargine which have high purity reaching above 98 percent. The invention gropes and selects the optimal technical parameters of chemical identification, thin layer identification and content determination, as well as can correctly and rapidly reflect the existence and the content of the solasonine or the solamargine in raw materials and the preparations, thus the quality of the raw materials and the preparations are efficiently controlled.
Description
Technical field
The present invention relates to the method for quality control of a kind of medicine effective ingredient or its preparation, particularly a kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof.
Background technology
Black nightshade is the dry aerial parts of plant of Solanaceae black nightshade Solanum nigrum L, and bitter, little sweet, cold in nature is returned lung, liver, stomach warp.Function is clearing heat and detoxicating, dispersing swelling and dissipating binds, anti-inflammatory diuresis.Cure mainly sore furuncle carbuncle and painful swelling, urinary tract infections, difficult urination, tumour etc.Alkaloid such as contained solasonine, solamargine is its main active in the black nightshade herb, has anticancer isoreactivity, this the two aglycon is a solasodine, alkaloids content generally realizes by measuring solasodine in the mensuration black nightshade at present, need be hydrolyzed, complicated operation, and can not directly react the solasonine in medicinal material, raw material and the preparation and the content of solamargine.
Below be the structural formula of solasonine (solasonine):
Below be the structural formula of solamargine (solamargine):
Summary of the invention
Technical matters to be solved by this invention is at the deficiencies in the prior art, and a kind of content that can more directly measure its effective ingredient, simple to operation, the method for quality control that is applicable to solasonine, solamargine and preparation thereof that can effectively control drug quality are provided.
" solasonine, solamargine and preparation thereof " of the present invention comprising: solasonine raw material, solamargine raw material, solasonine preparation, solamargine preparation and be pharmaceutical preparation of effective ingredient or the like with solasonine and solamargine.Described preparation comprises solid pharmaceutical preparation and liquid preparation, and wherein solid pharmaceutical preparation comprises capsule, tablet, dripping pill, soft capsule, granule and freeze drying powder injection etc., and liquid preparation comprises oral liquid, parenteral solution, injection emulsion etc.
Technical matters to be solved by this invention is to realize by following technical scheme.The present invention is a kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, is characterized in, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 201～210nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.01mg～20mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 0.02～10mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control, be characterized in, the organic solvent that is used for moving phase is acetonitrile or tetrahydrofuran or lower alcohol, the perhaps mixed solution of acetonitrile and lower alcohol arbitrary proportion, the perhaps mixed solution of acetonitrile and tetrahydrofuran arbitrary proportion, the perhaps mixed solution of lower alcohol and tetrahydrofuran arbitrary proportion; Described lower alcohol comprises methyl alcohol and isopropyl alcohol.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, the buffer salt solution that is used for moving phase comprises phosphate, citrate or acetate, and its pH scope is between 6.5～11, as disodium phosphate soln or the ammonium acetate solution of 0.0005～0.1mol/L.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, the volume ratio of organic solvent and buffer salt solution is 20～40:80～60 in the moving phase.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With acetonitrile: 0.002mol/L Na
2HPO
4Solution (35:65) is moving phase, and wavelength is 203nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through phosphorus pentoxide dry 48 hours solasonine or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.8mg among every 1ml, promptly;
The freeze-dried powder of solasonine or solamargine is got in the preparation of need testing solution, and accurate the title decides, and it is an amount of to add pure water, makes dissolving, makes the solution that every 1ml contains 2mg, promptly;
Accurate respectively reference substance solution, each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
The inventor adopt high-efficient liquid phase chromatogram technology measure solasonine in solasonine, solamargine raw material and the preparation and (or) content of solamargine, the standard of solasonine, solamargine raw material and the preparation that testing result and inventor are worked out contrasts, raw material up to specification and preparation just allow to enter next process procedure, with the quality that guarantees preparation and safe and effective.In the research that high-efficient liquid phase chromatogram technology of the present invention is measured, the inventor at first adopts uv absorption full wavelength scanner technology, detected the uv absorption situation of solasonine, solamargine, the uv absorption wavelength of having determined solasonine, solamargine is 201～210nm, and optimum uv absorption wavelength is 203nm.Then moving phase is screened, discovery is with acetonitrile or tetrahydrofuran or lower alcohol (as methyl alcohol, isopropyl alcohol), the perhaps mixed solution of acetonitrile and lower alcohol arbitrary proportion, the perhaps mixed solution of acetonitrile and tetrahydrofuran arbitrary proportion, the perhaps mixed solution of lower alcohol and tetrahydrofuran arbitrary proportion can make in the chromatographic peak solasonine or solamargine and other impurity reach baseline separation; But peak shape is relatively poor, and peak shape is asymmetric, hangover; In order to improve the chromatographic peak peak shape, make detection more accurate, the inventor has carried out further optimization to moving phase, adds a certain proportion of buffer salt solution, improves stability and system's separating effect of system.Introduce buffer salt solution by screening, make the high performance liquid chromatography peak separating effect of gained good, chromatographic peak peak shape symmetry, no conditions of streaking, thus guaranteed the testing result accuracy.The inventor has adopted the preferred technique condition of screening to test, and the result shows under these technical conditions that every high-efficient liquid phase chromatogram technology index is all up to specification, can be used in solasonine.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control, be characterized in, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated make dissolving, add methyl alcohol to scale, make the solution of 0.05mg/ml～10mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 0.05mg/ml～10mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 0.01mg～20mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 12～17cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, in the thin layer discrimination method, described developping agent is selected from: normal butyl alcohol: glacial acetic acid: water (4:1:5) upper strata; Chloroform: methyl alcohol: water (2:1:0.1); Ethyl acetate: ethanol: water (1:1:0.1); Perhaps chloroform: methyl alcohol: water (2.5:1:0.05).
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, in the thin layer discrimination method, described developer is selected from: the phosphoric acid solution of 0.5g/L paraformaldehyde, perhaps 10% ethanol solution of sulfuric acid, perhaps 5% vanillic aldehyde concentrated sulfuric acid solution, perhaps 5% bismuth potassium iodide solution.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control, be characterized in, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated make dissolving, add methyl alcohol to scale, make the solution of 0.5mg/ml～4mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to 9, with ethyl acetate extraction, solvent evaporated, residue is made the solution of 0.5mg/ml～4mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch 15cm, take out, dry, in the phosphoric acid solution (the 0.5g paraformaldehyde is dissolved in 1L phosphoric acid) of spray with paraformaldehyde, 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
In the thin layer Study on Identification of solasonine raw material and preparation, solamargine raw material and preparation, the researchist of the present patent application has made this thin layer by oneself and has differentiated necessary solasonine, solamargine reference substance, grope developping agent and developer that multiple thin layer is differentiated, selected optimum expansion and color condition.Test findings shows, with solasonine, the corresponding position of solamargine reference substance chromatogram on, solasonine, solamargine raw material and preparation have all detected the spot of same color, blank solvent is noiseless.
Technical matters to be solved by this invention can also further realize by following technical scheme.Above-described method of quality control is characterized in, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 0.01mg/ml～100mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 0.01mg/ml～100mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 1～1000 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 0.01mg/ml～100mg/ml in (1), perhaps get 1ml and be diluted to 1～1000 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 0.01mg/ml～100mg/ml, perhaps get 1ml and be diluted to 1～1000 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Solasonine and solamargine are alkaloid compound, by multiple alkaloid identification test, researchist of the present invention finds that chemistry differentiates down under (1)～(3), and solasonine and solamargine significant reaction can be used as the characteristic chemistry discrimination method of these two kinds of compounds.
It below is the quality testing control experiment of solamargine raw material that the inventor did and preparation.
(1). instrument and reagent
Instrument: Agilentl 100 high performance liquid chromatographs comprise quaternary pump, DAD detecting device, Agilentl 100 chem workstations; Mettler AE 240 electronic balances (100,000/).
Chromatographic column: Zobax Eclipse XDB C
18, 5 μ m, 4.6mm * 150mm.
Moving phase: acetonitrile: 0.01mol/L Na
2HPO
4Solution (40:60).
Flow velocity: 1ml/min.
Measure wavelength: 203nm.
Column temperature: 30 ℃.
Reagent: acetonitrile is a chromatographically pure, and water is double distilled water, and it is pure that other reagent is analysis.
(2). reference substance
Solasonine, solamargine are self-control.
(3). the preparation of need testing solution
Get this product content under the content uniformity item, mixing is got 20mg, and accurate the title decides, and it is an amount of to add pure water, makes dissolving, makes the solution that every 1ml contains 2.0mg, promptly.
(4). blank test
Former preparation prescription removes nightshade total alkali, is mixed with aqueous solution by concentration, gets blank sample, handles sample and stratographic analysis according to content determination, and the blank sample chromatogram does not have Interference Peaks on solasonine, the corresponding retention time of solamargine.
(5). linear relationship is investigated
Get solasonine 25.12mg, solamargine reference substance 25.03mg, add methyl alcohol and make the solution that every 1ml contains solasonine 1.00mg, solamargine 1.00mg respectively, promptly get standard reserving solution.Draw respectively in 0.5ml, 1ml, 2ml, 4ml, 6ml, 8ml standard reserving solution to the 6 10ml measuring bottle, add methyl alcohol respectively to scale.Accurate each the 10 μ l sample introduction of above-mentioned solution of drawing are measured, and (μ g) is horizontal ordinate with the reference substance sample size, and the peak area integrated value is an ordinate drawing standard curve, calculates regression equation.The result shows that solasonine is good linear relationship (seeing Table 1) in 0.5024 μ g～10.048 μ g scopes; Solamargine is good linear relationship (seeing Table 2) in 0.5006 μ g～10.012 μ g scopes.
Table 1 solasonine linear relationship is investigated
Table 2 solamargine linear relationship is investigated
(6). the precision experiment
Sample thief need testing solution (lot number 20050609) repeats sample introduction 5 times continuously, and each 2 μ l measure solasonine, solamargine peak area value, and the result shows, instrument precision good (seeing Table 3).
Table 3 Precision test result
(7). stability experiment
Sample thief need testing solution (lot number 20050609) respectively at 0,1,2,4,8,16,24 hour sample introduction 2 μ l, is measured solasonine, solamargine peak area value.The result shows that need testing solution was at 24 hours internal stabilities good (seeing Table 4).
Table 4 stability test result
(8). repeated experiment
Precision takes by weighing this product an amount of (lot number 20050609), according to 6 parts of need testing solutions of " injection black nightshade quality standard (draft) " preparation, measures its content (each sample size 2 μ l) respectively.The result shows, this law repeatability good (seeing Table 5).
Table 5 replica test result
(9). the average recovery test
Precision takes by weighing the injection black nightshade (lot number 20050609) of known content, and accurate a certain amount of solasonine, the solamargine reference substance solution of adding prepares sample as stated above, measure, and with following formula calculate recovery rate.The result shows that this law has the good recovery (seeing Table 6,7).
Table 6 sample recovery rate test findings (solasonine)
Annotate:
Table 7 sample recovery rate test findings (solamargine)
Annotate:
The present invention is for the content of solasonine, solamargine in the more direct mensuration black nightshade, the inventor separates from the black nightshade medicinal material and obtains solasonine, solamargine, make purity and reached highly purified solasonine reference substance and solamargine reference substance more than 98%, by a series of research, obtained can be used in the method for quality control of solasonine or solamargine raw material and preparation.
The present invention has prepared highly purified reference substance, grope and selected the technical parameter of best chemistry discriminating, thin layer discriminating and assay, can reflect the existence and the content of solasonine in raw material and the preparation or solamargine quickly and accurately, thereby effectively control the quality of raw material and preparation.
Embodiment
Further technical solution of the present invention is described below,, and does not constitute restriction right of the present invention so that those skilled in the art further understand the present invention.
Embodiment 1.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With acetonitrile: 0.002mol/L Na
2HPO
4Solution (35:65) is moving phase, and wavelength is 203nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through phosphorus pentoxide dry 48 hours solasonine or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.8mg among every 1ml, promptly;
The freeze-dried powder of solasonine or solamargine is got in the preparation of need testing solution, and accurate the title decides, and it is an amount of to add pure water, makes dissolving, makes the solution that every 1ml contains 2mg, promptly;
Accurate respectively reference substance solution, each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 2.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 201nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.01mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 0.02mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 3.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 210nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 20mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 10mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 4.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 206nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 100mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 1mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 5.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 205nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.1mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 0.1mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 6.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 207nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.5mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 0.5mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 7.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 208nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 1mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 1mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 8.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 202nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 5mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 5mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 9.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 204nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 15mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 8mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 10.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 206nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 10mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 3mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Embodiment 11.In embodiment 2 described method of quality control, the organic solvent that is used for moving phase is an acetonitrile.
Embodiment 12.In embodiment 3 described method of quality control, the organic solvent that is used for moving phase is a tetrahydrofuran.
Embodiment 13.In embodiment 4 described method of quality control, the organic solvent that is used for moving phase is a lower alcohol.
Embodiment 14.In embodiment 5 described method of quality control, the organic solvent that is used for moving phase is methyl alcohol or isopropyl alcohol.
Embodiment 15.In embodiment 6 described method of quality control, the organic solvent that is used for moving phase is the mixed solution of acetonitrile and lower alcohol arbitrary proportion.
Embodiment 16.In embodiment 7 or 8 described method of quality control, the organic solvent that is used for moving phase is the mixed solution of acetonitrile and tetrahydrofuran arbitrary proportion.
Embodiment 17.In embodiment 9 or 10 described method of quality control, the organic solvent that is used for moving phase is the mixed solution of lower alcohol and tetrahydrofuran arbitrary proportion.
Embodiment 18.In embodiment 13 or 15 or 17 described method of quality control, described lower alcohol is methyl alcohol or isopropyl alcohol.
Embodiment 19.In embodiment 2 or 3 described method of quality control, the buffer salt solution that is used for moving phase is a phosphate, and its pH is 6.5.
Embodiment 20.In embodiment 4 or 5 described method of quality control, the buffer salt solution that is used for moving phase is a citrate, and its pH is 11.
Embodiment 21.In embodiment 6 or 7 described method of quality control, the buffer salt solution that is used for moving phase is an acetate, and its pH is 9.
Embodiment 22.In embodiment 8 or 9 described method of quality control, the buffer salt solution that is used for moving phase is the disodium phosphate soln of 0.0005mol/L.
Embodiment 23.In embodiment 10 or 11 described method of quality control, the buffer salt solution that is used for moving phase is the ammonium acetate solution of 0.1mol/L.
Embodiment 24.In embodiment 12 or 13 described method of quality control, the buffer salt solution that is used for moving phase is the disodium phosphate soln of 0.001mol/L.
Embodiment 25.In embodiment 14 or 15 described method of quality control, the buffer salt solution that is used for moving phase is the ammonium acetate solution of 0.01mol/L.
Embodiment 26.In embodiment 16 or 17 described method of quality control, the buffer salt solution that is used for moving phase is the disodium phosphate soln of 0.05mol/L.
Embodiment 27.In embodiment 18 described method of quality control, the buffer salt solution that is used for moving phase is disodium phosphate soln or the ammonium acetate solution of 0.015mol/L.
Embodiment 28.In any one described method of quality control of embodiment 2-5, the volume ratio of organic solvent and buffer salt solution is 20:80 in the moving phase.
Embodiment 29.In any one described method of quality control of embodiment 6-10, the volume ratio of organic solvent and buffer salt solution is 20:60 in the moving phase.
Embodiment 30.In any one described method of quality control of embodiment 15-20, the volume ratio of organic solvent and buffer salt solution is 40:80 in the moving phase.
Embodiment 31.In any one described method of quality control of embodiment 21-24, the volume ratio of organic solvent and buffer salt solution is 40:60 in the moving phase.
Embodiment 32.In any one described method of quality control of embodiment 25-27, the volume ratio of organic solvent and buffer salt solution is 30:70 in the moving phase.
Embodiment 33.In any one described method of quality control of embodiment 1-10, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 0.05mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 0.05mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 0.01mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 12cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 34.In any one described method of quality control of embodiment 11-20, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 10mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 10mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 20mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 17cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 35.In any one described method of quality control of embodiment 21-28, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 1mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 1mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 1mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 14cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 36.In any one described method of quality control of embodiment 29-32, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 0.1mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 0.1mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 10mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 16cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 37.In any one described method of quality control of embodiment 1-10, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 0.5mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to 9, with ethyl acetate extraction, solvent evaporated, residue is made the solution of 0.5mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch 15cm, take out, dry, spray is with the phosphoric acid solution of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 38.In any one described method of quality control of embodiment 11-20, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 4mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to 9, with ethyl acetate extraction, solvent evaporated, residue is made the solution of 4mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch 15cm, take out, dry, spray is with the phosphoric acid solution of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 39.In any one described method of quality control of embodiment 21-32, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated, make dissolving, add methyl alcohol to scale, make the solution of 2mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to 9, with ethyl acetate extraction, solvent evaporated, residue is made the solution of 2mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch 15cm, take out, dry, spray is with the phosphoric acid solution of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 40.In embodiment 33 described method of quality control, described developping agent is a normal butyl alcohol: glacial acetic acid: water (4:1:5) upper strata.
Embodiment 41.In embodiment 34 described method of quality control, described developping agent is a chloroform: methyl alcohol: water (2:1:0.1).
Embodiment 42.In embodiment 35 described method of quality control, described developping agent is an ethyl acetate: ethanol: water (1:1:0.1).
Embodiment 43.In embodiment 36 described method of quality control, described developping agent is a chloroform: methyl alcohol: water (2.5:1:0.05).
Embodiment 44.In embodiment 33 or 34 described method of quality control, described developer is the phosphoric acid solution of 0.5g/L paraformaldehyde.
Embodiment 45.In embodiment 35 or 36 described method of quality control, described developer is 10% ethanol solution of sulfuric acid.
Embodiment 46.In embodiment 40 or 41 described method of quality control, described developer is 5% vanillic aldehyde concentrated sulfuric acid solution.
Embodiment 47.In embodiment 42 or 43 described method of quality control, described developer is 5% bismuth potassium iodide solution.
Embodiment 48.In any one described method of quality control of embodiment 1-10, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 0.01mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 0.01mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 1 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 0.01mg/ml in (1), perhaps get 1ml and be diluted to 1 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 0.01mg/ml, perhaps get 1ml and be diluted to 1 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Embodiment 49.In any one described method of quality control of embodiment 11-20, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 100mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 100mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 1000 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 100mg/ml in (1), perhaps get 1ml and be diluted to 1000 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 100mg/ml, perhaps get 1ml and be diluted to 1000 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Embodiment 50.In any one described method of quality control of embodiment 21-30, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 1mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 1mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 500 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 1mg/ml in (1), perhaps get 1ml and be diluted to 500 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 1mg/ml, perhaps get 1ml and be diluted to 500 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Embodiment 51.In any one described method of quality control of embodiment 31-40, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 0.5mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 0.5mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 200 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 0.5mg/ml in (1), perhaps get 1ml and be diluted to 200 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 0.5mg/ml, perhaps get 1ml and be diluted to 200 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Embodiment 52.In any one described method of quality control of embodiment 41-47, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 50mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 50mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 800 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 50mg/ml in (1), perhaps get 1ml and be diluted to 800 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 50mg/ml, perhaps get 1ml and be diluted to 800 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Embodiment 53.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With acetonitrile: 0.002mol/L Na
2HPO
4Solution (35:65) is moving phase, and wavelength is 203nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through phosphorus pentoxide dry 48 hours solasonine, solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.8mg among every 1ml, promptly;
The freeze-dried powder of solasonine or solamargine is got in the preparation of need testing solution, and accurate the title decides, and it is an amount of to add pure water, makes dissolving, makes the solution that every 1ml contains 2mg, promptly;
Accurate respectively reference substance solution, each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Its discrimination method is as follows: get the freeze-dried powder 10mg of solasonine or solamargine, put in the 2ml measuring bottle, add methyl alcohol 2ml, sonicated made dissolving in 20 minutes, added methyl alcohol to scale, as need testing solution.Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch about 15cm, take out, dry, spray is with the phosphoric acid solution (the 0.5g paraformaldehyde is dissolved in the 1L phosphoric acid) of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 54.A kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With acetonitrile: methyl alcohol: 0.002mol/L Na
2HPO
4Solution (30:5:65) is moving phase, and wavelength is 203nm.Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through phosphorus pentoxide dry 48 hours solasonine, solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.8mg among every 1ml, promptly;
It is an amount of that this product is got in the preparation of need testing solution, adds methyl alcohol and make the solution that every 1ml contains 2mg, promptly.
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Its thin layer discrimination method is as follows: get this product 10mg, put in the 2ml measuring bottle, add methyl alcohol 2ml, sonicated made dissolving in 20 minutes, added methyl alcohol to scale, as need testing solution; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch about 15cm, take out, dry, spray is with the phosphoric acid solution (the 0.5g paraformaldehyde is dissolved in the 1L phosphoric acid) of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Embodiment 55.Embodiment 53 or 54 described method of quality control, its chemical discrimination method is as follows:
(1) gets this product 20mg, put in the 10ml measuring bottle, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, get 2ml, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get solution 2ml in (1), splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get solution 2ml in (1), splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Claims (10)
1, a kind of method of quality control that is applicable to solasonine, solamargine and preparation thereof is characterized in that, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With the organic solvent that contains buffer salt solution is moving phase, and wavelength is 201～210nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through the solasonine of phosphorus pentoxide drying or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.01mg～20mg among every 1ml, promptly;
It is an amount of that solasonine or solamargine or its preparation are got in the preparation of need testing solution, and the arbitrary proportion mixed solution that adds methyl alcohol or water or methyl alcohol and water is made the solution that every 1ml contains 0.02～10mg, promptly;
Accurate respectively reference substance solution, each the 2 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
2, method of quality control according to claim 1, it is characterized in that, the organic solvent that is used for moving phase is acetonitrile or tetrahydrofuran or lower alcohol, the perhaps mixed solution of acetonitrile and lower alcohol arbitrary proportion, the perhaps mixed solution of acetonitrile and tetrahydrofuran arbitrary proportion, the perhaps mixed solution of lower alcohol and tetrahydrofuran arbitrary proportion; Described lower alcohol comprises methyl alcohol and isopropyl alcohol.
3, method of quality control according to claim 1, it is characterized in that, the buffer salt solution that is used for moving phase comprises phosphate, citrate or acetate, and its pH scope is between 6.5～11, as disodium phosphate soln or the ammonium acetate solution of 0.0005～0.1mol/L.
4, method of quality control according to claim 1 is characterized in that, the volume ratio of organic solvent and buffer salt solution is 20～40:80～60 in the moving phase.
5, method of quality control according to claim 1 is characterized in that, its content assaying method is as follows:
Chromatographic condition and system suitability test octadecylsilane chemically bonded silica are filling agent; With acetonitrile: 0.002mol/L Na
2HPO
4Solution (35:65) is moving phase, and wavelength is 203nm; Theoretical cam curve is calculated by the solasonine peak and is not less than 3000;
The preparation precision of reference substance solution takes by weighing through phosphorus pentoxide dry 48 hours solasonine or solamargine reference substance an amount of, adds methyl alcohol respectively and makes the solution that contains 0.8mg among every 1ml, promptly;
The freeze-dried powder of solasonine or solamargine is got in the preparation of need testing solution, and accurate the title decides, and it is an amount of to add pure water, makes dissolving, makes the solution that every 1ml contains 2mg, promptly;
Accurate respectively reference substance solution, each the 5 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
6, according to any one described method of quality control among the claim 1-5, it is characterized in that, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated make dissolving, add methyl alcohol to scale, make the solution of 0.05mg/ml～10mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to alkalescence, with normal butyl alcohol or ethyl acetate or chloroform or dichloromethane extraction, solvent evaporated, residue is made the solution of 0.05mg/ml～10mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 0.01mg～20mg product solution in contrast; Draw solasonine need testing solution and solasonine reference substance solution, perhaps draw solamargine need testing solution and solamargine reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, launch 12～17cm with developping agent, take out, dry, spray is with chromogenic reagent solution, and 105 ℃ to be heated to spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
7, method of quality control according to claim 6 is characterized in that, described developping agent is selected from: normal butyl alcohol: glacial acetic acid: water (4:1:5) upper strata; Chloroform: methyl alcohol: water (2:1:0.1); Ethyl acetate: ethanol: water (1:1:0.1); Perhaps chloroform: methyl alcohol: water (2.5:1:0.05).
8, method of quality control according to claim 6 is characterized in that, described developer is selected from: the phosphoric acid solution of 0.5g/L paraformaldehyde, perhaps 10% ethanol solution of sulfuric acid, perhaps 5% vanillic aldehyde concentrated sulfuric acid solution, perhaps 5% bismuth potassium iodide solution.
9, according to any one described method of quality control among the claim 1-5, it is characterized in that, its thin layer discrimination method is as follows: get solasonine or solamargine or its solid pharmaceutical preparation is put in the volumetric flask, add methyl alcohol, jolting or sonicated make dissolving, add methyl alcohol to scale, make the solution of 0.5mg/ml～4mg/ml, as need testing solution; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, transfer pH value to 9, with ethyl acetate extraction, solvent evaporated, residue is made the solution of 0.5mg/ml～4mg/ml, as need testing solution with dissolve with methanol; Other gets solasonine, solamargine reference substance, adds dissolve with methanol respectively and is diluted to solution that every 1ml contains 2mg product solution in contrast; Draw each 5 μ l of above-mentioned test sample and reference substance solution, put respectively in same be on the silica G plate of bonding agent with the sodium carboxymethyl cellulose, with chloroform: methyl alcohol: water (2.5:1:0.05) is developping agent, launch 15cm, take out, dry, spray is with the phosphoric acid solution of paraformaldehyde, and 105 ℃ to be heated to the spot colour developing clear; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
According to any one described method of quality control among the claim 1-5, it is characterized in that 10, its chemical discrimination method is as follows:
(1) gets solasonine or solamargine or its solid pharmaceutical preparation, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and make dissolving, and be diluted to scale, make the solution of 0.01mg/ml～100mg/ml; Perhaps get the liquid preparation of solasonine or the liquid preparation of solamargine, put in the volumetric flask, add pH and be 3 watery hydrochloric acid and be diluted to scale, make the solution of 0.01mg/ml～100mg/ml; Get above-mentioned solution 2ml, perhaps get 1ml and be diluted to 1～1000 times, splash into the potassium iodide iodine test solution, the rufous amorphous sediment occurs;
(2) get the solution 2ml of 0.01mg/ml～100mg/ml in (1), perhaps get 1ml and be diluted to 1～1000 times, splash into bismuth potassium iodide test solution, the Chinese red precipitation occurs;
(3) get the solution 2ml of (1) 0.01mg/ml～100mg/ml, perhaps get 1ml and be diluted to 1～1000 times, splash into 10% silico-tungstic acid test solution, the canescence amorphous sediment occurs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101313499A CN101377475A (en) | 2007-08-27 | 2007-08-27 | Quality control method suitable for solancarpine, solamargine and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101313499A CN101377475A (en) | 2007-08-27 | 2007-08-27 | Quality control method suitable for solancarpine, solamargine and preparation thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101377475A true CN101377475A (en) | 2009-03-04 |
Family
ID=40421120
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101313499A Pending CN101377475A (en) | 2007-08-27 | 2007-08-27 | Quality control method suitable for solancarpine, solamargine and preparation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101377475A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8614196B2 (en) | 2011-05-12 | 2013-12-24 | G & E Herbal Biotechnology Co., Ltd. | Treatment and/or prevention of inflammation and cutaneous photodamage and photoprotection of the skin with a water-soluble extract from plant of Solanum genus |
| CN108205033A (en) * | 2018-01-04 | 2018-06-26 | 吉林师范大学 | A kind of method of solanine and black nightshade polysaccharide in synchronous extracting and developing, measure black nightshade Chinese olive |
| CN110501200A (en) * | 2019-09-23 | 2019-11-26 | 吉林师范大学 | A kind of black nightshade effective ingredient that can be recycled is extracted and separation method |
-
2007
- 2007-08-27 CN CNA2007101313499A patent/CN101377475A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8614196B2 (en) | 2011-05-12 | 2013-12-24 | G & E Herbal Biotechnology Co., Ltd. | Treatment and/or prevention of inflammation and cutaneous photodamage and photoprotection of the skin with a water-soluble extract from plant of Solanum genus |
| CN108205033A (en) * | 2018-01-04 | 2018-06-26 | 吉林师范大学 | A kind of method of solanine and black nightshade polysaccharide in synchronous extracting and developing, measure black nightshade Chinese olive |
| CN110501200A (en) * | 2019-09-23 | 2019-11-26 | 吉林师范大学 | A kind of black nightshade effective ingredient that can be recycled is extracted and separation method |
| CN110501200B (en) * | 2019-09-23 | 2022-05-03 | 吉林师范大学 | Method for extracting and separating effective components of recyclable black nightshade |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106940355B (en) | A kind of brufen, the detection method of its sodium salt and its preparation in relation to substance | |
| CN111122724B (en) | Method for analyzing acarbose and related substances | |
| Mukker et al. | HPLC method with fluorescence detection for the quantitative determination of flaxseed lignans | |
| CN101377475A (en) | Quality control method suitable for solancarpine, solamargine and preparation thereof | |
| CN108037209B (en) | Liquid chromatography analysis method of ticagrelor chiral intermediate | |
| Basavaiah et al. | Simple high-performance liquid chromatographic method for the determination of acyclovir in pharmaceuticals | |
| Bharate et al. | Spectrophotometric and Chromatographic determination of acetylsalicylic acid and Caffeine in pure and in tablet dosage form | |
| Arayne et al. | A new RP-HPLC method for analysis of mebeverine hydrochloride in raw materials and Table (ts | |
| CN106706769B (en) | Separation and determination method of empagliflozin and optical isomer thereof | |
| Bhandage et al. | Extractive spectrophotometric determination of omeprazole in pharmaceutical preparations | |
| Gumustas et al. | Simple, sensitive and reliable LC-DAD method of gemifloxacin determination in pharmaceutical dosage forms | |
| WO2009155755A1 (en) | Method for determining the contents of oligosaccharides in morinda officinalis | |
| CN110057943A (en) | A kind of efficient liquid phase chromatographic analysis detection method of Amlodipine Besylate Tablet intermediate | |
| Braimah et al. | Impurity profiling of paracetamol dosage forms used in maiduguri metropolis | |
| Rivai et al. | Development and validation of bisacodyl analysis method in tablet with absorbance method and area under curves method in ultraviolet spectrophotometry | |
| CN104237399A (en) | Method for detecting ambrisenta crud materials and content of ambrisenta active ingredients in preparations | |
| CN109251161A (en) | The preparation method of 2- bisulfite tryptophan | |
| El-Rabbat et al. | A validated spectrofluorometric assay for the determination of certain macrolide antibiotics in pharmaceutical formulations and spiked biological fluids | |
| Montero et al. | Determination of chloramphenicol by coupling a continuous reduction system to an atomic-absorption spectrometer | |
| Tzouganaki et al. | Development and Validation of an HPLC Method for the Determination of the macrolide antibiotic Clarithromycin using Evaporative Light Scattering Detector in raw materials and Pharmaceutical Formulations | |
| CN112816570B (en) | Method for detecting azithromycin related substances | |
| Sevim et al. | Validation of high performance liquid chromatographic and spectrophotometric methods for the determination of the antiparkinson agent pramipexole dihydrochloride monohydrate in pharmaceutical products | |
| Breda et al. | Determination of 3'-deamino-3'-[2 (S)-methoxy-4-morpholinyl] doxorubicin, a new morpholinyl anthracycline, in plasma by high-performance liquid chromatography with fluorescence detection | |
| CN109251162A (en) | Tryptophan derivative and application thereof | |
| CN101865857A (en) | Method for detecting chlorpheniramine maleate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20090304 |