CN101264226A - Quality detecting method of diabetes treating medicine - Google Patents
Quality detecting method of diabetes treating medicine Download PDFInfo
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Abstract
The invention discloses a quality detecting method of medicine for curing diabetes, belonging to the field of Chinese herbal medicine quality detection, comprising steps such as the discriminating and checking of the medicine for curing diabetes and the detecting of extracts and content. The quality detecting method realizes the discrimination of astragaloside, puerarin, sarsasapogenin, American ginseng, ginsenoside Rb<SUB>1</SUB> and Rg<SUB>1</SUB> through thin layer chromatography, and at the same time adopts the dual-wavelength scanning method to accurately detect the content of ginsenoside Re; at least 0.10mg American ginseng should be contained in each grain calculated by ginsenoside Re (C<SUB>48</SUB>H<SUB>82</SUB>O<SUB>18</SUB>). The quality detecting method has the advantages of easy operation, accurate and advanced performance and good linear relationship, reproducibility, precision, stability and recovery rate, can effectively control the quality of the product and can ensure the therapeutic effects of the product.
Description
Technical field
The invention belongs to medicine quality detection technique field, relate to a kind of quality determining method of Chinese patent medicine, be specifically related to a kind of quality determining method for the treatment of the medicine of diabetes.
Background technology
In the prior art, application number is 200310115012.0, name is called the patent of " a kind of medicine for the treatment of diabetes and preparation method thereof ", a kind of medicine that is used for the treatment of diabetes and preparation method thereof is disclosed, pulverize, extract concentrated, precipitate with ethanol by processes such as Radix Panacis Quinquefolii, the Radix Astragali, Rhizoma Dioscoreae, Radix Rehmanniae, Fructus Corni, Fructus Lycii, Radix Ophiopogonis, the Rhizoma Anemarrhenae, Radix Trichosanthis, Fructus Schisandrae Chinensis, Galla Chinensis, Radix Puerariaes, the oral solid formulation that operations such as drying under reduced pressure, pulverizing mixing are made.This product has supplementing QI and nourishing YIN, the effect of strengthening spleen, tonifying kidney, characteristics such as evident in efficacy, as to have no side effect.Be the detection by quantitative index with ginsenoside Re in the Radix Panacis Quinquefolii in the standard, once carried out assay with high performance liquid chromatography, but owing to disturb excessive, can't operate, treat the content of ginsenoside Re in the medicine of diabetes so adopt the dual wavelength tlc determination, this method have sensitivity, accurately, characteristics such as favorable reproducibility, as the content assaying method of control said preparation quality.
Summary of the invention
The objective of the invention is for overcoming disadvantages of background technology, and provide a kind of easy and simple to handle, sensitive, accurately, favorable reproducibility, the quality determining method of the medicine of treatment diabetes that can more effective control product quality.
To achieve these goals, the technical solution used in the present invention is: a kind of quality determining method for the treatment of the medicine of diabetes is characterized in that this method may further comprise the steps:
(1) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Get the astragaloside reference substance, add methanol and make the solution that every 1mL contains the 1mg astragaloside, in contrast product solution; According to the thin layer chromatography test, draw reference substance solution 5 μ L, need testing solution 10 μ L, putting respectively on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-methanol-water, and the percent by volume of chloroform, first alcohol and water is 65: 35: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; Under ultra-violet lamp, inspect, show the fluorescence speckle of same color;
(2) get the medicine 4g that treats diabetes, add methanol 20～30mL, supersound process 10～15 minutes filters, and filtrate evaporate to dryness, residue add methanol 5mL dissolving, as need testing solution; Other gets the puerarin reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg puerarin, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-ethyl acetate-methanol-water is developing solvent, the percent by volume of chloroform, ethyl acetate, first alcohol and water is 15: 40: 22: 10, launch, and take out, dry the back and smoke half an hour, under ultra-violet lamp, inspect with ammonia; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(3) get the medicine 2g that treats diabetes, add water 20～30mL, the slight fever dissolving filters, filtrate adds hydrochloric acid 1mL, and reflux 50～70 minutes is put coldly, adds chloroform 20～30mL and extracts, divide and get chloroform liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, as need testing solution; Other gets the Sarsasapogenin reference substance, adds methanol and makes the solution that every 1mL contains the 1mg Sarsasapogenin, in contrast product solution; According to the thin layer chromatography test, draw each 20～30 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-acetone is developing solvent, benzene: the percent by volume of acetone is 9: 1, launches, and takes out, dry, spray is with the mixed liquor of concentration 8% vanillin ethanol solution and sulfuric acid solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; The percent by volume of described concentration 8% vanillin ethanol solution and sulfuric acid solution is 0.5: 5;
(4) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution.Get Radix Panacis Quinquefolii control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10～20mL again, supersound process 10～20 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution; Other gets ginsenoside Rb
1, Re and Rg
1, add methanol and make every 1mL and contain 1mg ginsenoside Rb
1, Re and Rg
1Solution, product solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution and reference substance solution each 5 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-ethyl acetate-methanol-water is developing solvent, chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color; With the corresponding position of reference substance chromatograph on, show the speckle of same color.
(5) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Get Radix Ginseng control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10～20mL again, supersound process 10～20 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution.Test according to thin layer chromatography, draw need testing solution 10 μ L, control medicinal material solution 5 μ L, putting respectively on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-ethyl acetate-methanol-water, and chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, it is clear to be heated to the speckle colour developing, puts respectively under daylight and the ultra-violet lamp and inspects; In the test sample chromatograph, must not show and the complete corresponding to speckle of control medicinal material;
(6) get the medicine 3g that treats diabetes, the accurate title, decide, and adds 65-75% alcoholic solution 80-120mL, and according to the hot dipping mensuration under the ethanol-soluble extractives algoscopy item, extractum must not be less than 34.0%;
(7) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Other gets ginsenoside Re's reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg ginsenoside Re, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ L, reference substance solution 4 μ L, 6 μ L, respectively the cross point is on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-methanol-water, and chloroform: methanol: the percent by volume of water is 65: 35: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, takes out, on lamellae, cover onesize glass plate, use immobilization with adhesive tape on every side, scan wavelength according to thin layer chromatography: λ
S=525nm, λ
R=700nm measures test sample trap integrated value and reference substance trap integrated value, calculates, promptly.
The concentration of ethanol solution of sulfuric acid is 10% described in above-mentioned steps (1), (4), (5) and (7).
Lower floor's solution described in above-mentioned steps (1), (2), (4), (5) and (7) is lower floor's solution of placing below 10 ℃ 12 hours.
Being heated to the speckle colour developing heating-up temperature in clear described in above-mentioned steps (1), (3), (4), (5) and (7) is 105 ℃.
The present invention compared with prior art has the following advantages: the present invention has realized astragaloside, puerarin, Sarsasapogenin, Radix Panacis Quinquefolii and ginsenoside Rb by thin layer chromatography
1, Re and Rg
1Differentiate that adopt the accurate mensuration of dual-wavelength scanning to ginsenoside Re's content simultaneously, method is easy and simple to handle, accurately advanced, linear relationship, repeatability, precision, stability, the response rate are all better, can effectively control the quality of product, guarantee the product curative effect.
Description of drawings
Fig. 1 is ginsenoside Re's standard curve sketch map among the present invention.
A-peak area integrated value among the figure, B-point sample amount (μ g).
The specific embodiment
The present invention will be further described below in conjunction with embodiment, but protection scope of the present invention is not limited only to following examples.
Embodiment
(1) production technology
Get application number and be 200310115012.0 recipe quantity medical material, Radix Panacis Quinquefolii is pulverized, and crosses 80 mesh sieves; All the other flavour of a drug add 15 times of water gagings and soak after 4 hours, decoct twice, 1.5 hours for the first time, 1 hour for the second time, merging filtrate, filtrate is concentrated into the thick paste that relative density is 1.15～1.20 (80 ℃), add ethanol and make that to contain alcohol amount be 60%, left standstill 24 hours, and filtered filtrate recycling ethanol, and be concentrated into the extractum that relative density is 1.15～1.20 (80 ℃), drying under reduced pressure is pulverized, and crosses 100 mesh sieves.With above-mentioned Radix Panacis Quinquefolii medicated powder mix homogeneously, encapsulated, make 1000, promptly.
(2) quality determining method
Differentiate: (1) gets the medicine 4g of treatment diabetes, and accurate the title decides, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 2 hours discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 20mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, its consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2 times, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Get the astragaloside reference substance, add methanol and make the solution that every 1mL contains the 1mg astragaloside, in contrast product solution; Test according to thin layer chromatography, draw reference substance solution 5 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-methanol-water is developing solvent, the percent by volume of chloroform, first alcohol and water is 65: 35: 10, launch, take out, dry, spray is 10% ethanol solution of sulfuric acid with concentration, 105 ℃ to be heated to speckle colour developing clear, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; Under ultra-violet lamp, inspect, show the fluorescence speckle of same color;
(2) get the medicine 4g that treats diabetes, add methanol 20mL, supersound process 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 5mL dissolving, as need testing solution; Other gets the puerarin reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg puerarin, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-ethyl acetate-methanol-water is developing solvent, the percent by volume of chloroform, ethyl acetate, first alcohol and water is 15: 40: 22: 10, launch, and take out, dry the back and smoke half an hour, under ultra-violet lamp, inspect with ammonia; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(3) get the medicine 2g of treatment diabetes, add water 20mL, slight fever dissolving filters, and filtrate adds hydrochloric acid 1mL, and reflux 60 minutes is put coldly, adds chloroform 20mL and extracts, and divides and gets chloroform liquid, and evaporate to dryness, residue add methanol 1mL makes dissolving, as need testing solution; Other gets the Sarsasapogenin reference substance, adds methanol and makes the solution that every 1mL contains the 1mg Sarsasapogenin, in contrast product solution; Test according to thin layer chromatography, draw each 20～30 μ L of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is developing solvent with benzene-acetone, and the percent by volume of benzene, acetone is 9: 1, launch, take out, dry, spray is 8% the vanillin ethanol solution and the mixed liquor of sulfuric acid solution with concentration, 105 ℃ to be heated to speckle colour developing clear, and the percent by volume of 8% vanillin ethanol solution and sulfuric acid solution is 0.5: 5; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(4) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 2 hours discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 20mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, its consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2 times, each consumption is 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution.Get Radix Panacis Quinquefolii control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10mL again, supersound process 10 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution; Other gets ginsenoside Rb
1, Re and Rg
1, add methanol and make every 1mL and contain 1mg ginsenoside Rb
1, Re and Rg
1Solution, product solution in contrast; Test according to thin layer chromatography, draw each 5 μ L of control medicinal material solution and reference substance solution, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-ethyl acetate-methanol-water is developing solvent, chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10, launch, take out, dry, spray is 10% ethanol solution of sulfuric acid with concentration, and 105 ℃ to be heated to the speckle colour developing clear, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color; With the corresponding position of reference substance chromatograph on, show the speckle of same color.
Check: (5) get the medicine 4g of treatment diabetes, and accurate the title decides, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 2 hours discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 20mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, its consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2 times, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution.Get Radix Ginseng control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10mL again, supersound process 10 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution.Test according to thin layer chromatography, draw need testing solution 10 μ L and above-mentioned control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-ethyl acetate-methanol-water is developing solvent, chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10 launch, take out, dry, spray is 10% ethanol solution of sulfuric acid with concentration, 105 ℃ to be heated to speckle colour developing clear, puts respectively under daylight and the ultra-violet lamp and inspect; In the test sample chromatograph, must not show and the complete corresponding to speckle of control medicinal material;
Extract content is measured: (6) get the medicine 3g of treatment diabetes, and accurate the title decides, and adds 70% alcoholic solution 100mL, measure according to the hot dipping under the ethanol-soluble extractives algoscopy item, and extract content must not be less than 34.0%;
Ginsenoside Re's assay: (7) get the medicine 4g of treatment diabetes, and accurate the title decides, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 2 hours discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 20mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, its consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2 times, each consumption is 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Other gets ginsenoside Re's reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg ginsenoside Re, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ L, reference substance solution 4 μ L, 6 μ L, the cross point is on same silica gel g thin-layer plate respectively, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-methanol-water is developing solvent, chloroform: methanol: the percent by volume of water is 65: 35: 10, launches, and takes out, dry, spray is 10% ethanol solution of sulfuric acid with concentration, and 105 ℃ to be heated to the speckle colour developing clear, takes out, on lamellae, cover onesize glass plate, use immobilization with adhesive tape on every side, scan wavelength according to thin layer chromatography: λ
S=525nm, λ
R=700nm measures test sample trap integrated value and reference substance trap integrated value, calculates, promptly.
(3) methodological study
1, negative control test: in application number is 200310115012.0 patent prescription ratio, and according to a kind of preparation technology who treats the medicine of diabetes, preparation does not contain the negative control sample of Radix Panacis Quinquefolii, makes negative control solution by the need testing solution compound method again.Other gets ginsenoside Re's reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg ginsenoside Re, in contrast product solution; Test according to thin layer chromatography, draw need testing solution 10 μ L, reference substance solution 4 μ L, 6 μ L, the cross point is on same silica gel g thin-layer plate respectively, and lower floor's solution of placing 12 hours below 10 ℃ with chloroform-methanol-water is developing solvent, chloroform: methanol: the percent by volume of water is 65: 35: 10, launch, take out, dry, the record chromatogram.The result proves: negative control solution is noiseless to the need testing solution chromatograph, and this chromatographic condition can be used as this product assay specificity chromatographic condition.
2, standard curve preparation: get ginsenoside Re's reference substance, add methanol and make the solution of 0.58mg/mL, draw 2,4,6,8,10 μ L points respectively on same silica gel g thin-layer plate, lower floor's solution of placing 12 hours below 10 ℃ with chloroform-methanol-water is developing solvent, chloroform: methanol: the percent by volume of water is 65: 35: 10, launch, take out, dry.Spray is 10% ethanol solution of sulfuric acid with concentration, puts 105 ℃ and is heated to clear spot, tests according to thin layer chromatography.Recording the peak area integrated value, is vertical coordinate with the peak area integrated value, and the point sample amount is an abscissa production standard curve, sees Table 1 and accompanying drawing 1.
The existing sexual relationship data of table 1 ginsenoside Re
Regression equation: y=324.7+6822.6X, r=0.9994.
The result shows: the ginsenoside Re shows good linear relationship between 1.04-5.2 μ g, but straight line is not by initial point, so adopt the external standard two-point method to measure content.
3, stability experiment: accurate ginsenoside Re's reference substance solution 3 μ L that draw, put on silica gel g thin-layer plate, launch, take out, dry colour developing.Every the 15min run-down, continuous sweep 5 times, the RSD that measures for 5 times is 0.94% as a result, shows: colour developing back this product measurement result in 1 hour is basicly stable.See table 2 for details.
Table 2 stability experiment data
4, precision experiment
(1) precision in the plate: accurate absorption reference substance solution 3 μ L (5 parts), put respectively on lamellae, launch, take out, to dry, spray is with the ethanol solution of sulfuric acid of concentration 10%, and colour developing scans, and the results are shown in Table 3.
Table 3 precision experimental data
The result shows that precision is higher.
(2) precision between plate: the accurate respectively reference substance solution 3 μ L that draw, put respectively on five blocks of different plates, launch the back sweep measuring, the results are shown in Table 4.
The different plate precision of table 4 experimental result
The result shows that precision is better.
5, repeatability experiment: get the same batch sample of this product (lot number: 950629), press 5 parts of text content assaying method samplings, measure content, the results are shown in Table 5.
Table 5 repeatability experimental result
6, response rate experiment:
Precision takes by weighing the about 2g of this product (lot number 950629, content 0.674mg/g), and other adds ginsenoside Re 1.0-1.4mg, and press the text content assaying method and measure, calculate recovery rate, average recovery rate is 97.81% as a result, RSD is 2.25%.See table 6 for details.
Table 6 response rate experimental result
Experiment shows that the response rate is higher.
7, ten batch sample assays: get ten batch samples, press the experiment of text content assaying method, the average content that records ten batch samples is the 0.18mg/ grain, presses floating of average content 20%, the regulation content limit is the 0.10mg/ grain, and ten batch sample extractum knot is measured fruit and seen Table 10.
Table 10 ten batch sample assay data
| Lot number | Content (mg/ grain) | Lot number | Content (mg/ grain) |
| 20000607 | 0.21 | 20010306 | 0.16 |
| 20000608 | 0.23 | 20010812 | 0.16 |
| 20000609 | 0.19 | 20010813 | 0.17 |
| 20010304 | 0.18 | 20010814 | 0.18 |
| 20010305 | 0.17 | 20020122 | 0.16 |
Claims (4)
1, a kind of quality determining method for the treatment of the medicine of diabetes is characterized in that this method may further comprise the steps:
(1) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Get the astragaloside reference substance, add methanol and make the solution that every 1mL contains the 1mg astragaloside, in contrast product solution; According to the thin layer chromatography test, draw reference substance solution 5 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-methanol-water is developing solvent, the percent by volume of chloroform, first alcohol and water is 65: 35: 10, launches, and takes out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; Under ultra-violet lamp, inspect, show the fluorescence speckle of same color;
(2) get the medicine 4g that treats diabetes, add methanol 20～30mL, supersound process 10～15 minutes filters, and filtrate evaporate to dryness, residue add methanol 5mL dissolving, as need testing solution; Other gets the puerarin reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg puerarin, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-ethyl acetate-methanol-water is developing solvent, the percent by volume of chloroform, ethyl acetate, first alcohol and water is 15: 40: 22: 10, launch, and take out, dry the back and smoke half an hour, under ultra-violet lamp, inspect with ammonia; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
(3) get the medicine 2g that treats diabetes, add water 20～30mL, the slight fever dissolving filters, filtrate adds hydrochloric acid 1mL, and reflux 50～70 minutes is put coldly, adds chloroform 20～30mL and extracts, divide and get chloroform liquid, evaporate to dryness, residue add methanol 1mL makes dissolving, as need testing solution; Other gets the Sarsasapogenin reference substance, adds methanol and makes the solution that every 1mL contains the 1mg Sarsasapogenin, in contrast product solution; According to the thin layer chromatography test, draw each 20～30 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with benzene-acetone is developing solvent, benzene: the percent by volume of acetone is 9: 1, launches, and takes out, dry, spray is with the mixed liquor of concentration 8% vanillin ethanol solution and sulfuric acid solution, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; The percent by volume of described concentration 8% vanillin ethanol solution and sulfuric acid solution is 0.5: 5;
(4) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution.Get Radix Panacis Quinquefolii control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10～20mL again, supersound process 10～20 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution; Other gets ginsenoside Rb
1, Re and Rg
1, add methanol and make every 1mL and contain 1mg ginsenoside Rb
1, Re and Rg
1Solution, product solution in contrast; Test according to thin layer chromatography, draw control medicinal material solution and reference substance solution each 5 μ L, need testing solution 10 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-ethyl acetate-methanol-water is developing solvent, chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color; With the corresponding position of reference substance chromatograph on, show the speckle of same color;
(5) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Get Radix Ginseng control medicinal material 1g, add water 1mL, soak into, add water saturated n-butyl alcohol 10～20mL again, supersound process 10～20 minutes filters, and reclaims n-butyl alcohol to doing, and residue adds methanol 1mL makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw need testing solution 10 μ L, control medicinal material solution 5 μ L, put respectively on same silica gel g thin-layer plate, lower floor's solution with chloroform-ethyl acetate-methanol-water is developing solvent, chloroform: ethyl acetate: methanol: the percent by volume of water is 15: 40: 22: 10 launch, and take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, puts respectively under daylight and the ultra-violet lamp and inspects; In the test sample chromatograph, must not show and the complete corresponding to speckle of control medicinal material;
(6) get the medicine 3g that treats diabetes, the accurate title, decide, and adds 65-75% alcoholic solution 80-120mL, and according to the hot dipping mensuration under the ethanol-soluble extractives algoscopy item, extractum must not be less than 34.0%;
(7) get the medicine 4g that treats diabetes, the accurate title, decide, and puts in the apparatus,Soxhlet's, it is an amount of to add diethyl ether, and reflux 1.5-2.5 hour, discards ether solution, medicinal residues are flung to ether, and it is an amount of to add methanol, and reflux, extract, is to colourless, reclaim methanol, residue adds water 15-25mL makes dissolving, is transferred in the separatory funnel, with water saturation n-butanol extraction five times, consumption is followed successively by 25mL, 20mL, 20mL, 20mL and 15mL, merge extractive liquid, is with ammonia solution washing 2-3 time, each consumption 15mL, discard water layer, reclaim n-butyl alcohol to doing, residue adds methanol makes dissolving, quantitatively be transferred in the 10mL measuring bottle, add methanol and be diluted to scale, shake up, as need testing solution; Other gets ginsenoside Re's reference substance, adds methanol and makes the solution that every 1mL contains the 0.5mg ginsenoside Re, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ L, reference substance solution 4 μ L, 6 μ L, respectively the cross point is on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-methanol-water, and chloroform: methanol: the percent by volume of water is 65: 35: 10, launch, take out, dry, spray is with ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing, takes out, on lamellae, cover onesize glass plate, use immobilization with adhesive tape on every side, scan wavelength according to thin layer chromatography: λ
S=525nm, λ
R=700nm measures test sample trap integrated value and reference substance trap integrated value, calculates, promptly.
2, a kind of quality determining method for the treatment of the medicine of diabetes according to claim 1 is characterized in that the concentration of ethanol solution of sulfuric acid described in step (1), (4), (5) and (7) is 10%.
3, a kind of quality determining method for the treatment of the medicine of diabetes according to claim 1 is characterized in that lower floor's solution described in step (1), (2), (4), (5) and (7) is lower floor's solution of placing below 10 ℃ 12 hours.
4, a kind of quality determining method for the treatment of the medicine of diabetes according to claim 1, it is characterized in that being heated to described in step (1), (3), (4), (5) and (7) heating-up temperature of speckle colour developing in clear is 105 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102614378A (en) * | 2012-04-26 | 2012-08-01 | 陕西方舟制药有限公司 | Yin nourishing and blood sugar lowering Chinese medicinal composition and preparation method as well as detection method thereof |
| CN104792652A (en) * | 2015-05-02 | 2015-07-22 | 浙江大学 | Multi-index rapid detection method for radix astragali |
| CN105277648A (en) * | 2015-09-29 | 2016-01-27 | 河北中医学院 | Multi-information gradient thin layer discriminating method of Radix Puerariae medicinal material and water extract product thereof |
| CN106353446A (en) * | 2016-08-12 | 2017-01-25 | 上海黄海制药有限责任公司 | Identification and content measuring method of climacteric-syndrome-soothing granules |
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| CN1299742C (en) * | 2003-11-19 | 2007-02-14 | 王锦刚 | Medicine for treating diabetes, and its prepn. method |
| CN100335897C (en) * | 2005-11-02 | 2007-09-05 | 中国药科大学 | Quality detection method for compound prepn of red sage and notoginseng |
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2008
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102614378A (en) * | 2012-04-26 | 2012-08-01 | 陕西方舟制药有限公司 | Yin nourishing and blood sugar lowering Chinese medicinal composition and preparation method as well as detection method thereof |
| CN104792652A (en) * | 2015-05-02 | 2015-07-22 | 浙江大学 | Multi-index rapid detection method for radix astragali |
| CN104792652B (en) * | 2015-05-02 | 2017-07-25 | 浙江大学 | A multi-indicator rapid detection method for Radix Astragali |
| CN105277648A (en) * | 2015-09-29 | 2016-01-27 | 河北中医学院 | Multi-information gradient thin layer discriminating method of Radix Puerariae medicinal material and water extract product thereof |
| CN106353446A (en) * | 2016-08-12 | 2017-01-25 | 上海黄海制药有限责任公司 | Identification and content measuring method of climacteric-syndrome-soothing granules |
| CN109541117A (en) * | 2018-12-21 | 2019-03-29 | 青海普兰特药业有限公司 | A kind of pharmaceutical composition object detecting method for moistening lung |
| CN110794080A (en) * | 2019-11-20 | 2020-02-14 | 刘圣梅 | Method for detecting quality of medicine for treating colitis |
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