CN102749299A - Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation - Google Patents
Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation Download PDFInfo
- Publication number
- CN102749299A CN102749299A CN2012102434608A CN201210243460A CN102749299A CN 102749299 A CN102749299 A CN 102749299A CN 2012102434608 A CN2012102434608 A CN 2012102434608A CN 201210243460 A CN201210243460 A CN 201210243460A CN 102749299 A CN102749299 A CN 102749299A
- Authority
- CN
- China
- Prior art keywords
- preparation
- solution
- adds
- accurate
- astragaloside
- 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
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a quality detection method of a traditional Chinese medicine composition medlar thirst quenching preparation. By adopting the method, the quality standards on the conventional traditional Chinese medicine composition medlar thirst quenching capsules are correspondingly improved. The method comprises the step of determining the content of plant polysaccharides in the preparation by adopting ultraviolet-visible spectrophotometry method and determining the content of astragaloside in astragalus by adopting a high-performance liquid chromatography method. The quality detection method has the beneficial effects that the methods for determining the contents of the plant polysaccharides and the astragaloside in the medlar thirst quenching preparation are added on the basis of the original quality standard qualitative identification, so that the quality detection of the medlar thirst quenching preparation is relatively accurate and reliable, and the safety, the effectiveness and the quality controllability of the preparation are ensured.
Description
Technical field
The present invention relates to a kind of quality determining method of Chinese medicine composition, particularly a kind of quality determining method of Chinese medicine composition matrimony vine preparation for diabetes belongs to medical technical field.
Background technology
The matrimony vine wasting-thirst capsule is that gold is scolded the exclusive formulation product of Tibetan medicine incorporated company, records in Chinese patent drug provincial standard rising national standard part standard numbering: WS-10701 (ZD-0701)-2002.The effect that have supplementing qi and nourishing yin, promotes the production of body fluid to quench thirst is mainly used in and quenches one's thirst due to the deficiency of both qi and yin, and type ii diabetes is seen above-mentioned disease person.Type ii diabetes there is obvious curative effects, effectively control of blood sugar, the also effectively generation of control of diabetes complication.
The matrimony vine wasting-thirst capsule is to be the capsule preparations that bulk drug makes with bright sea-buckthorn 110g, the sub-160g of fresh fructus lycii, root bark of Chinese wolf-berry 125g, hawthorn 50g, Chinese yam 135g, Fructus Hordei Germinatus 65g, Radix Astragali 115g; Its preparation method is: bright sea-buckthorn is after pressing juice, concentrating, and freeze drying becomes powder; Fresh fructus lycii is pulverized postlyophilization with colloid mill and is become powder; The five tastes such as all the other root barks of Chinese wolf-berry, boiling twice, 2 hours for the first time, 1.5 hours for the second time, collecting decoction; Filter, filtrating is concentrated into the clear cream that relative density is 1.05 ~ 1.10 (50 ℃), and spraying drying powder-forming adds sea-buckthorn, matrimony vine freeze-dried powder; Mixing incapsulates, and processes 1000, promptly gets.
At present; Matrimony vine wasting-thirst capsule proper mass standard has only the item of discriminating; The fruit of Chinese wolfberry, Chinese yam, Astragaloside IV have been carried out the thin-layer chromatography qualitative identification; Have no the assay index, can not effectively control the quality of said preparation, also do not find report in the prior art other quality determining method aspects of matrimony vine wasting-thirst capsule.Therefore, cause the quality testing of matrimony vine wasting-thirst capsule preparation not accurate enough reliable, quality standard is badly in need of improving.
Summary of the invention
The objective of the invention is provides a kind of quality determining method of Chinese medicine composition matrimony vine preparation for diabetes for overcoming the deficiency of above-mentioned prior art.
Summary of the invention
The present invention has carried out corresponding raising to existing Chinese medicine composition matrimony vine preparation for diabetes quality standard; On the basis of primary standard, increased the content assaying method of plant polyose in the preparation; The content assaying method of Astragaloside IV has further been guaranteed the safe and effective, quality controllable of said preparation.
The term explanation:
The matrimony vine wasting-thirst capsule is the nomenclature of drug of national standard for traditional Chinese medicines compilation (Chinese patent drug provincial standard rising national standard part) record.
The matrimony vine preparation for diabetes comprises the matrimony vine wasting-thirst capsule and with other preparations of matrimony vine wasting-thirst capsule bulk drug formulation.
For realizing above-mentioned purpose, the present invention adopts following technical proposals:
A kind of quality determining method of matrimony vine preparation for diabetes; The various preparations of this matrimony vine preparation for diabetes for processing by the pharmacy conventional method by the bright sea-buckthorn of bulk drug, fresh fructus lycii, the root bark of Chinese wolf-berry, hawthorn, Chinese yam, Fructus Hordei Germinatus, the Radix Astragali; It is characterized in that it comprises adopts UV-VIS spectrophotometry that plant polyose in the preparation is carried out the method for assay and to adopting high performance liquid chromatography Astragaloside IV in the Radix Astragali to be carried out the method for assay.
The assay of said plant polyose is with the DEXTROSE ANHYDROUS reference substance; The preparation standard curve; The phenolsulfuric acid method is handled sample, uses ultraviolet-visible spectrophotometer under the 490nm wavelength, to measure absorbance, reads the weight that contains glucose the sample solution from typical curve; Calculate, promptly get the content of plant polyose in the matrimony vine preparation for diabetes.
The assay of said plant polyose is specifically:
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got Chinese medicine composition matrimony vine preparation for diabetes content or the about 0.1 ~ 0.3g of its preparation meal, and accurate the title decides, and adds 80% ethanol 100ml; Reflux l hour; Filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1ml that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
The assay of said Astragaloside IV is to be contrast with the Astragaloside IV reference substance; With the octadecylsilane chemically bonded silica is filling agent; With volume parts than (30 ~ 40): the acetonitrile-water of (70 ~ 60) is a moving phase; According to high effective liquid chromatography for measuring, calculate with external standard two-point method logarithmic equation, promptly get the content of Astragaloside IV in the matrimony vine wasting-thirst capsule.
Serve as preferred than 35:65 wherein with the acetonitrile-water volume parts.
The assay of said Astragaloside IV is specifically:
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 30 ~ 40:70 ~ 60; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
It is an amount of that the Astragaloside IV reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.3 ~ 0.4mg, promptly gets;
Preparation of the test solution composition wolfberry diabetes medicine preparations or preparations small study contents, take 4 ~ 6g, accurately weighed, set stoppered conical flask, precision methanol 40 ~ 60ml, weighed and the weight heated to reflux for 0.5 to 1.5 hours, let cool, then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue micro heat to dissolve 20ml of water, water-saturated Oscillating vibration butanol extract five times, each 20ml, combined n-butanol solution, sufficiently washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, n-butanol solution was concentrated to dryness, the residue was dissolved in methanol and set the volume to 5ml volumetric flask, shake, filtration, the filtrate obtained, that is, too;
Accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing of determination method, need testing solution 10 ~ 20 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
Said preparation is that bulk drug consists of bright sea-buckthorn 110 weight portions, fresh fructus lycii 160 weight portions, the root bark of Chinese wolf-berry 125 weight portions, hawthorn 50 weight portions, Chinese yam 135 weight portions, Fructus Hordei Germinatus 65 weight portions, the Radix Astragali 115 parts by weight of Chinese traditional medicine compositions, and quality determining method comprises:
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got Chinese medicine composition matrimony vine preparation for diabetes content or the about 0.2g of its preparation meal, and accurate the title decides, and adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
Plant polyose content is with glucose (C in Chinese medicine composition matrimony vine wasting-thirst capsule content of the present invention or its preparation
6H
12O
6) must not count and be less than 76mg/g.
The assay of Astragaloside IV
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Chinese wolfberry Diabetes formulation compositions or formulations finely content, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux 1 hour, let cool, then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue in 20ml of water micro heat dissolve the water-saturated butanol vibration Oscillating extracted five times, each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml flask, shake, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
Astragaloside IV (C in Chinese medicine composition matrimony vine wasting-thirst capsule content of the present invention or its preparation
41H
68O
14) content must not be less than 0.5mg/g.
Said preparation is meant gets Chinese medicine composition matrimony vine wasting-thirst capsule bulk drug, presses common process, adds conventional auxiliary material and is prepared into clinical acceptable any formulation, comprises pill, micropill, dripping pill, tablet, capsule, particle, medicine materical crude slice or dispersing tablet.
The unit corresponding relation of weight portion described in this instructions and parts by volume is g/ml or kg/l.
Matrimony vine preparation for diabetes prescription is: bright sea-buckthorn, fresh fructus lycii, the root bark of Chinese wolf-berry, hawthorn, Chinese yam, Fructus Hordei Germinatus, the Radix Astragali.Main ingredient in the matrimony vine side of being wherein, in the matrimony vine LBP-X can raise immunity, reducing blood lipid, protection liver, anti-oxidant, anti-ageing, antitumor, be principal ingredient.Astragalus polyose has antiviral, antitumor, anti-ageing, radioresistance, anti-stress, effect such as anti-oxidant in the Radix Astragali.Simultaneously Fructus Hippophae polysaccharide, root bark of Chinese wolf-berry polyoses content are all quite abundant, and pharmacological action is clear and definite.So the content of total polysaccharides helps the control of whole medicine security, validity and quality homogeneity in the control said preparation.
Astragaloside IV has and can improve cardio-pulmonary function in the Radix Astragali, strengthens cardiac contractile force, and hemangiectasis brings high blood pressure down, and improves skin blood circulation and nutrition condition; And can protect liver, prevent that liver glycogen from reducing, and to CAH, has good action.The Radix Astragali also has inducement interferon and the effect of transferring body's immunity, can suppress virus breeding and tumor growth.Enhance immunity power increases energy, and antifatigue makes sudden change, protects the liver, and suppresses the effect of osteoclast.The content of Astragaloside IV helps the control of preparation security, validity and quality homogeneity in the control preparation.Contain a large amount of saponins materials during matrimony vine is quenched one's thirst, its similar performance be difficult for to be measured, and this law is selected effective constituent Astragaloside IV in the Radix Astragali, and preferred through condition set up method of quality control.
This law uses ultraviolet spectrophotometry that polysaccharide composition in the preparation is carried out integral body control; Use specificity, the strong HPLC method of repeatability that Astragaloside IV has been carried out quality control simultaneously; The quality of fraction preparation from different perspectives, the curative effect that matrimony vine is quenched one's thirst is guaranteed.
Chinese medicine composition matrimony vine wasting-thirst capsule proper mass standard has only the item of discriminating, has no the assay index, makes that the quality testing of matrimony vine wasting-thirst capsule preparation is not accurate enough reliable, and quality standard has much room for improvement.The present invention has carried out corresponding raising to existing Chinese medicine composition matrimony vine wasting-thirst capsule quality standard; Adopt the phenolsulfuric acid colourimetry that the plant polyose in the said preparation has been carried out assay, adopt high performance liquid chromatography that the Astragaloside IV in the said preparation has been carried out detection by quantitative.The present invention has increased the content assaying method of plant polyose, Astragaloside IV in the matrimony vine wasting-thirst capsule on the basis of primary standard, guaranteed the safe and effective, quality controllable of said preparation.Simultaneously this law also can be used for other preparations of matrimony vine wasting-thirst capsule, like matrimony vine particle, matrimony vine wasting-thirst capsule, the matrimony vine sheet etc. of quenching one's thirst of quenching one's thirst.
Description of drawings
Fig. 1 is the glucose linear relationship chart;
Fig. 2 is an Astragaloside IV reference substance solution high-efficient liquid phase chromatogram;
Fig. 3 is a Radix Astragali control medicinal material solution high-efficient liquid phase chromatogram;
Fig. 4 is a Chinese medicine composition bark of ash bone-knitting capsule Radix Astragali need testing solution high-efficient liquid phase chromatogram;
Fig. 5 is that Chinese medicine composition bark of ash bone-knitting capsule lacks Radix Astragali negative control solution high-efficient liquid phase chromatogram;
Fig. 6 is the Astragaloside IV linear relationship chart.
Wherein, the horizontal ordinate of Fig. 2, Fig. 3, Fig. 4, Fig. 5 is the time, unit: minute (Minutes); Ordinate is a voltage, unit: volt (Volts).
Embodiment
Below in conjunction with embodiment the present invention is done detailed elaboration, but be not limited to the embodiment of these concrete records.The Chinese medicine composition matrimony vine wasting-thirst capsule that is detected is scolded Tibetan medicine medicine company incorporated company product for the Qinghai gold and is sold.
Experimental example 1: the assay experiment of plant polyose
1. instrument, reagent and confession test agent
Instrument: Tianjin, island UV-2450 ultraviolet-visible spectrophotometer; Tianjin, island AuW220D electronic balance.
Reference substance: DEXTROSE ANHYDROUS reference substance (Nat'l Pharmaceutical & Biological Products Control Institute), lot number: 110833-200503.
Sample: matrimony vine wasting-thirst capsule (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company), lot number: 20110301,20110302,20110303.
2. the preparation of reference substance solution
Get DEXTROSE ANHYDROUS reference substance 25mg, the accurate title, decide, and puts in the 250ml measuring bottle, adds water and dissolve in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
3. detect the selection of wavelength
Take by weighing reference substance solution 1ml, after the colour developing of phenolsulfuric acid system, make full wavelength scanner at 400nm ~ 600nm respectively, confirm maximum absorption wavelength with ultraviolet-visible spectrophotometer.Can know that according to ultraviolet absorpting spectrum reference substance solution is after the colour developing of phenolsulfuric acid system, in 400nm ~ 600nm wavelength coverage interscan, absorption maximum is at 490nm, so selected 490nm is for detecting wavelength.
4. the investigation of the preparation of typical curve and linear relationship
Precision is measured reference substance solution (104.08 μ g/ml) 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml, each accurate phenol solution 1m1 that adds; Shake up, the accurate rapidly sulfuric acid 5ml that adds shakes up; Color development at room temperature 30 minutes is blank with the reagent corresponding, the photograph UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010); Wavelength at 490nm is measured absorbance, is ordinate with the absorbance, and concentration is horizontal ordinate; The drawing standard curve gets regression equation A=0.0205C+0.0093, coefficient R=0.9998.The result shows that glucose is in 2.60 μ g/ml ~ 13.01 μ g/ml scopes, and absorbance (A) is good with reference substance concentration (C) linear relationship.The result sees Fig. 1 and table 1.
Table 1 polysaccharide determination linear relationship is investigated the result
5. sample solution is measured
Get the about 0.2g of Chinese medicine composition matrimony vine wasting-thirst capsule content meal, the accurate title, decide, and adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
6. precision test
Precision is measured glucose reference substance solution 0.5ml6 part, add water respectively and mend to 2.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds ", measure absorbance in accordance with the law.Measure the result, RSD=0.96% (n=6).The result shows that instrument precision is good.See table 2.
Table 2 polysaccharide determination Precision test result
7. replica test
Get with a collection of matrimony vine wasting-thirst capsule (product batch number: 20110301) the about 0.2g of content meal, accurate claim surely, totally 6 parts, the method under measuring by sample solution prepares sample solution, measures absorbance, the content of plant polyose in the calculation sample in accordance with the law.Measure the result, the content mean value of plant polyose is 94.37mg/g in the sample, RSD=1.18% (n=6).The result shows that this analytical approach repeatability is good.The result sees table 3.
Table 3 polysaccharide determination replica test result
8. recovery test
Precision takes by weighing the matrimony vine wasting-thirst capsule sample (product batch number: 20110301) 6 parts with a collection of known plants polyoses content; Each accurate glucose reference substance that adds; The method of measuring under the item by sample solution prepares sample solution; Measure absorbance, the content of plant polyose in the calculation sample, calculate recovery rate in accordance with the law.Measure the result, the assay average recovery rate of plant polyose is 97.70% in the sample, RSD=1.43% (n=6).The result shows that it is accurate that this assay method is measured the result.The result sees table 4.
Table 4 polysaccharide determination recovery test result
9. sample determination
Get three batches of Chinese medicine composition matrimony vine wasting-thirst capsules, measure and calculate plant polyose content.The result sees table 5.
Table 5 polysaccharide determination result
Experimental example 2: the assay experiment of Astragaloside IV
1. instrument, reagent and confession test agent
Instrument: high performance liquid chromatograph: the L-2100 of Hitachi pump, the detecting device L-2400 of Hitachi detecting device; Alltech 2000ES EISD; Tianjin, island AuW220D electronic balance.
Reference substance: Astragaloside IV reference substance (Nat'l Pharmaceutical & Biological Products Control Institute), lot number: 11078-200613.
Sample: matrimony vine wasting-thirst capsule (the Qinghai gold is scolded Tibetan medicine medicine company incorporated company), lot number: 20110301,20110302,20110303.
2. moving phase is selected
Studied respectively under methanol-water system, the acetonitrile-water system moving phase condition; The separating effect of Astragaloside IV chromatographic peak; It is moving phase that the result finds with the acetonitrile-water system, and the peak shape of Astragaloside IV chromatographic peak is better, the acetonitrile-water volume parts than 30 ~ 40:70 ~ 60 scopes in; Astragaloside IV all can reach chromatographic resolution effect preferably, is excellent with the acetonitrile-water volume parts than 35:65 wherein.
3. reference substance solution preparation
It is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets.
4. need testing solution preparation
By method under the assay item need testing solution is detected.Press the method under the item for preparing of need testing solution, investigated under Different Extraction Method, different extraction solvent, the difference extraction time condition extraction effect of Astragaloside IV respectively.Content with Astragaloside IV in every gram medicine is that index is confirmed method for distilling, extracted solvent, extraction time.The result sees table 6, table 7 and table 8.
Table 6 method for distilling is investigated test findings
Table 7 extracts solvent and investigates test findings
Table 8 extraction time investigation test findings
The result shows: the content of the measured Astragaloside IV of refluxing extraction is the highest, so the selective extraction method is a refluxing extraction; The content of the measured Astragaloside IV of methanol extraction is the highest, so the selective extraction solvent is a methyl alcohol; Extracted 1 hour and recorded in 1.5 hours the content basically identical of Astragaloside IV, so the selective extraction time is 1 hour.Preferred concrete grammar is following:
Chinese medicine composition wolfberry Diabetes Capsule contents and research fine, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux for 1 hour, let cool, then weighed and the weight of methanol to complement the weight loss, shake, filtration, the filtrate 25ml, concentrated to dryness, the residue 20ml water micro heat dissolve the water-saturated n-butanol extract vibration Oscillating five times, each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times per 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml volumetric flask, shake, filtration, the filtrate obtained, that is, too.
5. system suitability test and the negative investigation of disturbing
Press method under the assay item, accurate respectively Astragaloside IV reference substance solution, Radix Astragali control medicinal material solution, need testing solution, each 10 μ l of negative control solution of drawing inject liquid chromatograph, the record chromatogram.The result shows that the degree of separation that Astragaloside IV is adjacent chromatographic peak in reference substance, the test sample chromatogram is all greater than 1.5, and it is noiseless that negative control solution goes out the peak position at Astragaloside IV.The result sees Fig. 2, Fig. 3, Fig. 4 and Fig. 5.
6. the investigation of the preparation of typical curve and linear relationship
Precision is measured Astragaloside IV reference substance stock solution solution (693.6 μ g/ml) 1ml, 3ml, 5ml, 8ml, 10ml, puts respectively in the 10ml volumetric flask, and methyl alcohol is diluted to scale; Shake up, each accurate sample introduction 10 μ l is an ordinate with the logarithm (Log A) of peak area; The logarithm of reference substance concentration (Log C) is a horizontal ordinate; Carry out linear regression, get the Astragaloside IV regression equation: Log A=1.0021Log C+3.0032, related coefficient: R=0.9998.The result shows that Astragaloside IV is in 69.36 μ g/ml ~ 693.6 μ g/ml scopes, and the logarithm of Astragaloside IV peak area (Log A) is good with logarithm (Log C) linear relationship of reference substance concentration.The result sees table 9 and Fig. 6.
Table 9 Astragaloside IV linear relationship is investigated the result
7. precision test
The accurate respectively Astragaloside IV reference substance solution 10 μ l that draw inject liquid chromatograph, each METHOD FOR CONTINUOUS DETERMINATION 6 times, and the record peak area also calculates relative standard deviation.The result shows that instrument precision is good.See table 10.
Table 10 Astragaloside IV Precision test result
8. stability test
After the need testing solution preparation was accomplished, the accurate 10 μ l that draw injected liquid chromatograph, and the record peak area was whenever measured once at a distance from 2 hours later on, investigated 8 hours, and the record peak area also calculates relative standard deviation.The result shows, in 8 hours, to measure the result stable for Astragaloside IV in the need testing solution.The result sees table 11.
Table 11 Astragaloside IV stability test result
9. replica test
Get with a collection of matrimony vine wasting-thirst capsule sample (product batch number: 20110301) 3g, accurate claim fixed, totally 6 parts; The method for preparing under the item by need testing solution prepares need testing solution; The accurate respectively 10 μ l that draw inject liquid chromatograph, the content of Astragaloside IV in the calculation sample.The result shows that this analytical approach repeatability is good.The result sees table 12.
Table 12 Astragaloside IV replica test result
10. recovery test
Precision takes by weighing matrimony vine wasting-thirst capsule sample with a collection of known Astragaloside content, and (product batch number: 20110301) 6 parts, each accurate Astragaloside IV reference substance that adds is measured its content, calculate recovery rate.The result shows that it is accurate that this assay method is measured the result.The result sees table 13.
Table 13 Astragaloside IV recovery test result
11. sample determination
Get three batches of Chinese medicine composition matrimony vine wasting-thirst capsules, measure and calculate Astragaloside content.The result sees table 14.
Table 14 Astragaloside content is measured the result
Following embodiment all can realize the described effect of above-mentioned experimental example.
Embodiment 1: the quality determining method of matrimony vine wasting-thirst capsule
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got the about 0.2g of Chinese medicine composition matrimony vine wasting-thirst capsule content meal, and accurate the title decides, and adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
Plant polyose content is with glucose (C in the Chinese medicine composition matrimony vine wasting-thirst capsule content of the present invention
6H
12O
6) must not count and be less than 76mg/g.
The assay of Astragaloside IV
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Chinese medicine composition wolfberry Diabetes Capsule contents and research fine, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux for 1 hour, let cool, then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue in 20ml of water micro heat dissolve the water-saturated n-butanol extract five Oscillating Vibration times, each 20ml, combined n-butanol solution, sufficiently washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, n-butanol solution was concentrated to dryness, the residue was dissolved in methanol 5ml volumetric flask and dilute to the , shake, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
Astragaloside IV (C in the Chinese medicine composition matrimony vine wasting-thirst capsule content of the present invention
41H
68O
14) content must not be less than 0.5mg/g.
Embodiment 2: the quench one's thirst quality determining method of particle of matrimony vine
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got the Chinese medicine composition matrimony vine about 0.2g of particle meal that quenches one's thirst, and accurately claims surely, adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
Chinese medicine composition matrimony vine of the present invention is quenched one's thirst in the particle plant polyose content with glucose (C
6H
12O
6) must not count and be less than 76mg/g.
The assay of Astragaloside IV
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Chinese medicine composition wolfberry diabetes research fine particles, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux for 1 hour, let cool then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue 20ml water micro heat dissolve the water-saturated n-butanol extract vibration Oscillating five times, Each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml volumetric flask, shake uniform, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
The Chinese medicine composition matrimony vine of the present invention Astragaloside IV (C in the particle that quenches one's thirst
41H
68O
14) content must not be less than 0.5mg/g.
The described Chinese medicine composition matrimony vine particle of quenching one's thirst is meant with matrimony vine wasting-thirst capsule bulk drug fill a prescription bright sea-buckthorn 110g, the sub-160g of fresh fructus lycii, root bark of Chinese wolf-berry 125g, hawthorn 50g, Chinese yam 135g, Fructus Hordei Germinatus 65g, Radix Astragali 115g; Add conventional auxiliary material, according to the granule of common process preparation.
Embodiment 3: the quality determining method of matrimony vine diabetes pill
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got the about 0.2g of Chinese medicine composition matrimony vine diabetes pill meal, and accurate the title decides, and adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
Plant polyose content is with glucose (C in the Chinese medicine composition matrimony vine diabetes pill of the present invention
6H
12O
6) must not count and be less than 76mg/g.
The assay of Astragaloside IV
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Chinese medicine composition wolfberry Xiao Ke Wan small study, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux for 1 hour, let cool then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue 20ml water micro heat dissolve the water-saturated n-butanol extract vibration Oscillating five times, Each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml volumetric flask, shake uniform, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
Astragaloside IV (C in the Chinese medicine composition matrimony vine diabetes pill of the present invention
41H
68O
14) content must not be less than 0.5mg/g.
Described Chinese medicine composition matrimony vine diabetes pill is meant with matrimony vine wasting-thirst capsule bulk drug fill a prescription bright sea-buckthorn 110g, the sub-160g of fresh fructus lycii, root bark of Chinese wolf-berry 125g, hawthorn 50g, Chinese yam 135g, Fructus Hordei Germinatus 65g, Radix Astragali 115g; Add conventional auxiliary material, according to the pill of common process preparation.
Embodiment 4: the quench one's thirst quality determining method of sheet of matrimony vine
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets (containing DEXTROSE ANHYDROUS 0.1mg among every 1ml).
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml; Each accurate phenol solution 1m1 that adds shakes up, the accurate rapidly sulfuric acid 5ml that adds; Shake up, color development at room temperature 30 minutes is blank with the reagent corresponding; According to UV-VIS spectrophotometry (" appendix VA of Chinese pharmacopoeia version in 2010), measure absorbance in the wavelength of 490nm, be ordinate with the absorbance; Concentration is horizontal ordinate, the drawing standard curve.
Determination method is got the Chinese medicine composition matrimony vine about 0.2g of sheet meal that quenches one's thirst, and accurately claims surely, adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid.Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight (mg) that contains glucose the need testing solution, calculate, promptly get from typical curve.
Chinese medicine composition matrimony vine of the present invention is quenched one's thirst in the sheet plant polyose content with glucose (C
6H
12O
6) must not count and be less than 76mg/g.
The assay of Astragaloside IV
Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2010 D);
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Diabetes wolfberry Chinese medicine composition finely slice, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux for 1 hour, let cool then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue 20ml water micro heat dissolve the water-saturated n-butanol extract vibration Oscillating five times, Each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml volumetric flask, shake uniform, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
The Chinese medicine composition matrimony vine of the present invention Astragaloside IV (C in the sheet that quenches one's thirst
41H
68O
14) content must not be less than 0.5mg/g.
The described Chinese medicine composition matrimony vine sheet of quenching one's thirst is meant with matrimony vine wasting-thirst capsule bulk drug fill a prescription bright sea-buckthorn 110g, the sub-160g of fresh fructus lycii, root bark of Chinese wolf-berry 125g, hawthorn 50g, Chinese yam 135g, Fructus Hordei Germinatus 65g, Radix Astragali 115g; Add conventional auxiliary material, according to the tablet of common process preparation.
Claims (7)
1. the quality determining method of a matrimony vine preparation for diabetes; The various preparations of this matrimony vine preparation for diabetes for processing by the pharmacy conventional method by the bright sea-buckthorn of bulk drug, fresh fructus lycii, the root bark of Chinese wolf-berry, hawthorn, Chinese yam, Fructus Hordei Germinatus, the Radix Astragali; It is characterized in that it comprises adopts UV-VIS spectrophotometry that plant polyose in the preparation is carried out the method for assay and adopts high performance liquid chromatography Astragaloside IV in the Radix Astragali to be carried out the method for assay.
2. quality determining method as claimed in claim 1 is characterized in that, the assay of said plant polyose is with the DEXTROSE ANHYDROUS reference substance; The preparation standard curve; The phenolsulfuric acid method is handled sample, uses ultraviolet-visible spectrophotometer under the 490nm wavelength, to measure absorbance, reads the weight that contains glucose the sample solution from typical curve; Calculate, promptly get the content of plant polyose in the matrimony vine preparation for diabetes.
3. quality determining method as claimed in claim 1 is characterized in that, the assay of said plant polyose specifically:
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets;
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml, each accurate phenol solution 1m1 that adds; Shake up, the accurate rapidly sulfuric acid 5ml that adds shakes up color development at room temperature 30 minutes; With the reagent corresponding is blank, according to UV-VIS spectrophotometry, measures absorbance in the wavelength of 490nm; With the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve;
Determination method is got Chinese medicine composition matrimony vine preparation for diabetes content or the about 0.1 ~ 0.3g of its preparation meal, and accurate the title decides, and adds 80% ethanol 100ml; Reflux 1 hour; Filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid; Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight that contains glucose the need testing solution, calculate, promptly get from typical curve.
4. quality determining method as claimed in claim 1; It is characterized in that the assay of said Astragaloside IV is to be contrast with the Astragaloside IV reference substance, is filling agent with the octadecylsilane chemically bonded silica; With volume parts than (30 ~ 40): the acetonitrile-water of (70 ~ 60) is a moving phase; According to high effective liquid chromatography for measuring, calculate with external standard two-point method logarithmic equation, promptly get the content of Astragaloside IV in the matrimony vine wasting-thirst capsule.
5. quality determining method as claimed in claim 1 is characterized in that, the assay of said Astragaloside IV specifically:
According to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 30 ~ 40:70 ~ 60; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
It is an amount of that the Astragaloside IV reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.3 ~ 0.4mg, promptly gets;
Preparation of the test solution composition wolfberry diabetes medicine preparations or preparations small study contents, take 4 ~ 6g, accurately weighed, set stoppered conical flask, precision methanol 40 ~ 60ml, weighed and the weight heated to reflux for 0.5 to 1.5 hours, let cool, then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue micro heat to dissolve 20ml of water, water-saturated Oscillating vibration butanol extract five times, each 20ml, combined n-butanol solution, sufficiently washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, n-butanol solution was concentrated to dryness, the residue was dissolved in methanol and set the volume to 5ml volumetric flask, shake, filtration, the filtrate obtained, that is, too;
Accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing of determination method, need testing solution 10 ~ 20 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
6. quality determining method as claimed in claim 1; It is characterized in that; Said preparation is that bulk drug consists of bright sea-buckthorn 110 weight portions, fresh fructus lycii 160 weight portions, the root bark of Chinese wolf-berry 125 weight portions, hawthorn 50 weight portions, Chinese yam 135 weight portions, Fructus Hordei Germinatus 65 weight portions, the Radix Astragali 115 parts by weight of Chinese traditional medicine compositions, and its quality determining method is:
The assay of plant polyose
DEXTROSE ANHYDROUS reference substance 25mg is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 250ml measuring bottle, adds water and dissolves in right amount, is diluted to scale, shakes up, and promptly gets;
The preparation precision of typical curve is measured reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, puts respectively in the tool plug test tube, adds water respectively and mends to 2.0ml, each accurate phenol solution 1m1 that adds; Shake up, the accurate rapidly sulfuric acid 5ml that adds shakes up color development at room temperature 30 minutes; With the reagent corresponding is blank, according to UV-VIS spectrophotometry, measures absorbance in the wavelength of 490nm; With the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve;
Determination method is got Chinese medicine composition matrimony vine preparation for diabetes content or the about 0.2g of its preparation meal, and accurate the title decides, and adds 80% ethanol 100ml, and reflux l hour, filter while hot, filter residue and filter wash with hot 80% ethanol 30ml gradation, discard 80% ethanol liquid; Filter residue is put in the flask together with filter paper, adds water 100ml, and reflux 2 hours filters, with small amount of thermal water washing filter while hot; Merging filtrate and washing lotion are put coldly, move in the 250ml measuring bottle, are diluted with water to scale, shake up; Precision is measured 1ml, puts in the tool plug test tube, adds water 1.0ml, the sighting target directrix curve prepare the method under the item, from " each accurate phenol solution 1m1 that adds "; Measure absorbance in accordance with the law, read the weight that contains glucose the need testing solution, calculate, promptly get from typical curve;
The assay of Astragaloside IV
According to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With the acetonitrile-water is moving phase; The mobile phase volume portion rate is 35:65; EISD detects; Number of theoretical plate calculates by the Astragaloside IV peak should be not less than 4000;
The preparation of reference substance solution: it is an amount of to get the Astragaloside IV reference substance, and accurate the title decides, and adds methyl alcohol and processes the solution that every 1ml contains 0.35mg, promptly gets;
Preparation of the test solution: Chinese wolfberry Diabetes formulation compositions or formulations finely content, take 5g, accurately weighed, set stoppered conical flask, precision methanol 50ml, weighed and the weight was heated to reflux 1 hour, let cool, then weighed and the weight of methanol to complement the weight loss, shaking, filtration, the filtrate 25ml, concentrated to dryness, the residue in 20ml of water micro heat dissolve the water-saturated butanol vibration Oscillating extracted five times, each 20ml, combined butanol solution, thoroughly washed with ammonia solution 2 times, each 40ml, discard the liquid ammonia, butanol solution was concentrated to dryness, the residue was dissolved in methanol and dilute to 5ml flask, shake, filtration, the filtrate obtained, that is, too;
Determination method: accurate respectively reference substance solution 10 μ l, the 20 μ l of drawing, need testing solution 10 μ l inject liquid chromatograph, measure, and calculate with external standard two-point method logarithmic equation, promptly get.
7. like each described quality determining method of claim 1-6; It is characterized in that; Said preparation is meant gets Chinese medicine composition matrimony vine wasting-thirst capsule bulk drug; Press common process, add conventional auxiliary material and be prepared into clinical acceptable any formulation, comprise pill, micropill, dripping pill, tablet, capsule, particle, medicine materical crude slice or dispersing tablet.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102434608A CN102749299A (en) | 2012-07-13 | 2012-07-13 | Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102434608A CN102749299A (en) | 2012-07-13 | 2012-07-13 | Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102749299A true CN102749299A (en) | 2012-10-24 |
Family
ID=47029663
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012102434608A Pending CN102749299A (en) | 2012-07-13 | 2012-07-13 | Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102749299A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439449A (en) * | 2013-07-25 | 2013-12-11 | 广东远大药业有限公司 | Detection method of medicine used for nourishing lung and activating blood |
| CN103520585A (en) * | 2013-10-22 | 2014-01-22 | 山东阿如拉药物研究开发有限公司 | Medlar thirst-quenching medicine composition and preparation method as well as application thereof |
| CN103550630A (en) * | 2013-11-08 | 2014-02-05 | 山东阿如拉药物研究开发有限公司 | Medlar thirst-eliminating chewable tablet as well as preparation method and application thereof |
| CN106769295A (en) * | 2016-12-08 | 2017-05-31 | 成都中医药大学 | The extracting method and detection method of lamiophlomis rotata polysaccharide |
| WO2020228708A1 (en) * | 2019-05-14 | 2020-11-19 | Hong Kong Baptist University | Chromophore-labeled oligosaccharide markers and methods of use thereof |
| CN112161849A (en) * | 2020-08-28 | 2021-01-01 | 回音必集团安徽制药有限公司 | High-efficiency smashing device for donkey-hide gelatin blood-nourishing particles and inspection process thereof |
| WO2021047394A1 (en) * | 2019-09-12 | 2021-03-18 | 北方民族大学 | Method for analyzing root-bark of chinese wolfberry compound structure by molecular networking |
| CN115792017A (en) * | 2022-12-06 | 2023-03-14 | 中国中医科学院眼科医院 | Quality detection method of traditional Chinese medicine composition for reducing recurrence of herpes simplex viral keratitis |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1899361A (en) * | 2006-07-14 | 2007-01-24 | 贵州信邦制药股份有限公司 | Zhenqi medicinal composition and its preparation |
| CN1931236A (en) * | 2005-09-14 | 2007-03-21 | 蔡军 | Medicine composition of red sage and rhodiola root |
| CN1931217A (en) * | 2005-09-14 | 2007-03-21 | 蔡军 | Medicine composition of gingko leaf and rhodiola root |
| CN1981857A (en) * | 2005-12-05 | 2007-06-20 | 黄振华 | Medicinal composition of milkvetch root, chinaroot greenbrier and Hong Jingtian and its making method |
| CN102266492A (en) * | 2011-07-22 | 2011-12-07 | 安徽康顺名贵中草药产业开发有限公司 | Method for processing dendrobium huoshanense plant dry product |
-
2012
- 2012-07-13 CN CN2012102434608A patent/CN102749299A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1931236A (en) * | 2005-09-14 | 2007-03-21 | 蔡军 | Medicine composition of red sage and rhodiola root |
| CN1931217A (en) * | 2005-09-14 | 2007-03-21 | 蔡军 | Medicine composition of gingko leaf and rhodiola root |
| CN1981857A (en) * | 2005-12-05 | 2007-06-20 | 黄振华 | Medicinal composition of milkvetch root, chinaroot greenbrier and Hong Jingtian and its making method |
| CN1899361A (en) * | 2006-07-14 | 2007-01-24 | 贵州信邦制药股份有限公司 | Zhenqi medicinal composition and its preparation |
| CN102266492A (en) * | 2011-07-22 | 2011-12-07 | 安徽康顺名贵中草药产业开发有限公司 | Method for processing dendrobium huoshanense plant dry product |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439449A (en) * | 2013-07-25 | 2013-12-11 | 广东远大药业有限公司 | Detection method of medicine used for nourishing lung and activating blood |
| CN103439449B (en) * | 2013-07-25 | 2015-07-29 | 广东远大药业有限公司 | A kind of detection method of medicine of invigorating blood circulation for tonifying lung |
| CN103520585A (en) * | 2013-10-22 | 2014-01-22 | 山东阿如拉药物研究开发有限公司 | Medlar thirst-quenching medicine composition and preparation method as well as application thereof |
| CN103520585B (en) * | 2013-10-22 | 2015-12-09 | 山东金诃药物研究开发有限公司 | A kind of Fructus Lycii is quenched one's thirst pharmaceutical composition and its preparation method and application |
| CN103550630A (en) * | 2013-11-08 | 2014-02-05 | 山东阿如拉药物研究开发有限公司 | Medlar thirst-eliminating chewable tablet as well as preparation method and application thereof |
| CN103550630B (en) * | 2013-11-08 | 2015-07-01 | 山东金诃药物研究开发有限公司 | Medlar thirst-eliminating chewable tablet as well as preparation method and application thereof |
| CN106769295A (en) * | 2016-12-08 | 2017-05-31 | 成都中医药大学 | The extracting method and detection method of lamiophlomis rotata polysaccharide |
| WO2020228708A1 (en) * | 2019-05-14 | 2020-11-19 | Hong Kong Baptist University | Chromophore-labeled oligosaccharide markers and methods of use thereof |
| WO2021047394A1 (en) * | 2019-09-12 | 2021-03-18 | 北方民族大学 | Method for analyzing root-bark of chinese wolfberry compound structure by molecular networking |
| CN112161849A (en) * | 2020-08-28 | 2021-01-01 | 回音必集团安徽制药有限公司 | High-efficiency smashing device for donkey-hide gelatin blood-nourishing particles and inspection process thereof |
| CN112161849B (en) * | 2020-08-28 | 2024-05-07 | 回音必集团安徽制药有限公司 | Donkey-hide gelatin blood-nourishing granule efficient smashing device and detection process thereof |
| CN115792017A (en) * | 2022-12-06 | 2023-03-14 | 中国中医科学院眼科医院 | Quality detection method of traditional Chinese medicine composition for reducing recurrence of herpes simplex viral keratitis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102749299A (en) | Quality detection method of traditional Chinese medicine composition medlar thirst quenching preparation | |
| CN100402053C (en) | Method for quality control of traditional Chinese medicine prepns. | |
| CN100418563C (en) | Quality control method of Chinese medicinal preparation | |
| CN103381217A (en) | Six-flavor hematinic capsule, its quality control method and application thereof | |
| CN102749401B (en) | Inspection method of traditional Chinese medicine composition twenty-five-ingredient lung disease preparation | |
| CN101574506B (en) | Method for preparing pingxiao preparation | |
| CN101966223A (en) | Fingerprint detection method for compound wintercreeper preparation | |
| CN101695520B (en) | Preparation method of medicament for treating diabetes | |
| CN101708208B (en) | Detection method of capsule preparation for treating painful swollen joint | |
| CN102590424B (en) | Quality detection method of compound houttuynia cordata mixture | |
| CN103285306A (en) | Preparation method and detection method of traditional Chinese medicine composition for benefiting Qi and tonifying kidney | |
| CN103239467B (en) | Agaricus blazei polysaccharide and preparation method thereof | |
| CN101843667B (en) | Shuanghuanglian medicinal composition and preparation method thereof | |
| CN103006878A (en) | Fresh rehmannia root total glycoside extract, preparation method and application thereof | |
| CN102539599B (en) | Method for detecting liver-enhancing medicine | |
| CN102068598B (en) | Quality control method of Yangrong Baicao Wan for treating irregular menses caused by hemophthisis | |
| CN101874852A (en) | Quality detecting method of restorex plus extract capsule | |
| CN108226325A (en) | Roripa montana gives birth to the method for building up of arteries and veins oral liquid composition finger-print | |
| CN102357131B (en) | Method for preparing astragalus polysaccharide glucose-lowering capsule | |
| CN101428130A (en) | Traditional Chinese medicine preparation for treating pancreatitis, preparation method and quality standard thereof | |
| CN102008541B (en) | Method for simultaneously detecting three main active ingredients in sugar-free type compound wintercreeper preparation | |
| CN107648424A (en) | A kind of preparation method of Lycium-rehmannia preparation | |
| CN109771463A (en) | Artemisia rupestris extract and its preparation method and application | |
| CN103575823A (en) | Detection method of 8 chemical components in Tangminling preparation | |
| CN101953978A (en) | Heart-soothing and lipid-lowering tablet medicine quality standard detecting method |
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: 20121024 |