CN101269097B - Alstonia-leaf medicinal materials and medicament testing method for its preparation - Google Patents
Alstonia-leaf medicinal materials and medicament testing method for its preparation Download PDFInfo
- Publication number
- CN101269097B CN101269097B CN2008100583404A CN200810058340A CN101269097B CN 101269097 B CN101269097 B CN 101269097B CN 2008100583404 A CN2008100583404 A CN 2008100583404A CN 200810058340 A CN200810058340 A CN 200810058340A CN 101269097 B CN101269097 B CN 101269097B
- Authority
- CN
- China
- Prior art keywords
- alstonia
- leaf
- preparation
- galactose
- medicinal material
- 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.)
- Active
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a medicinal material of alstonia-leaf and the drug detecting method of the pharmaceutical preparations of alstonia-leaf. The invention is characterized in that a thin-layer chromatography is adopted to detect the effective ingredients of kaempferol-3-O-Beta-D-galactose-(2-1)-O-Beta-D-xylopyranoside and quercetin-3-O-Beta-D-galactose-(2--1)-O-Beta-D- xylopyranoside of the alstonia-leaf and the pharmaceutical preparations of alstonia-leaf qualitatively. Spectrophotometry is adopted to realize a quality analysis and control of the alstonia-leaf and the pharmaceutical preparations of alstonia-leaf. The invention overcomes the deficiencies that the prior art can only detect the alkaloid components yet fail to detect a type of more important active ingredient flavonoid glycoside in the alstonia-leaf and the pharmaceutical preparations of alstonia-leaf . The method of the invention can precisely detect flavonoid glycoside active-ingredient compound 1, flavonoid glycoside active-ingredient compound 2 and the contents in the alstonia-leaf and the pharmaceutical preparations of alstonia-leaf. The method can be used for evaluating the quality of alstonia-leaf and detecting the quality analysis of the pharmaceutical preparations of various types adopting alstonia-leaf as the medicinal ingredient, such as granules, tablets, capsules, oral liquid and so on. The method is directly applicable to an actual production process and a quality analysis of the product.
Description
Technical field
The present invention relates to Pharmaceutical Analysis detection technique field, specifically the drug test method of alstonia-leaf medicinal material and preparation thereof.
Background technology
Cornus controversa Alstonia Scholaris (L.) R.Bro. is an apocynaceae plant, the south of Yunnan that mainly distributes, aboundresources.Alstonia-leaf is the natural drug of multi-national use, is used for the treatment of respiratory diseases such as cough with lung heat, phlegm are many, bronchitis, medicinal material for " Yunnan Province's drug standards ", " Chinese pharmacopoeia is recorded.Utilize alstonia-leaf to be raw material, developed formulation products such as alstonia-leaf granule, tablet, capsule, oral liquid.
Chemical research for alstonia-leaf, the bright PhD dissertation of Zhu Wei " Primary Study of five kinds of resources of medicinal plant chemistry ", Zhu Hua PhD dissertation " chemistry of five kinds of medicinal plants and active component research " has carried out systematic study and literature review to alkaloid component in (Kunming Inst. of Botany, Chinese Academy of Sciences, calendar year 2001,2006).Aspect the drug quality analysis, " Yunnan Province's drug standards " are only differentiated with simple alkaloid precipitation reaction the alstonia-leaf medicinal material, no assay project; Health Chinese medicine standard promulgated by the ministries or commissions of the Central Government alstonia-leaf particle does not have the physics and chemistry test item, and the lamp stand blade adopts acid base titration to survey alkaloid, and conversion is index components picrinine content, but the specificity of assay method is poor, can not adapt to the technical requirement of the modernization of Chinese medicine.
For improving alstonia-leaf medicinal material and quality of the pharmaceutical preparations analytical technique thereof, some research report or related application have been arranged.Zhu Guangrong application " application of picrinine in the control alstonia-leaf medicinal material and the quality of the pharmaceutical preparations thereof " patent (is authorized, CN100356172C); Kunming Zhenhua Pharmacy Co., Ltd. is studied the detection method of alstonia-leaf main alkaloid composition picrinine, patent (open, CN 1975412A) that application " is measured the efficient liquid-phase chromatography method of alkali picrinine content "; The Kunming plant research is applied for " medicine of treatment diseases concerned with respiratory and its production and application " (openly, CN101084951A), " medicine and the application thereof of treatment diseases concerned with respiratory " patent (open, CN 101084894A); Nat'l Pharmaceutical ﹠ Biological Products Control Institute's application " a kind of Chinese medicine composition for the treatment of cough, asthma " patent (open, CN101028323A).Research and patent that these are relevant all just relate to the alkaloids composition that contains in the alstonia-leaf, especially the analytical approach of principal ingredient alkali picrinine and application in drug quality control thereof.
Summary of the invention
The purpose of this invention is to provide a kind of analyzing and testing alstonia-leaf medicinal material and be effective constituent compound 1 and 2 method in the various preparations produced of raw material with the alstonia-leaf.This method is with compound 1,2 in thin-layer chromatography qualitative detection alstonia-leaf medicinal material and the preparation thereof, and realizes quality analysis and control to alstonia-leaf medicinal material and preparation thereof with its content of spectrophotometry.
The inventor further studies confirm that alstonia-leaf, alstonia-leaf also contains more flavonoid glycoside composition except that alkaloid component, and find there are two flavonoid glycoside compositions that content is higher first: Kaempferol-3-O-β-D-galactose-(2--1)-O-β-D-xyloside, Quercetin-3-O-β-D-galactose-(2--1)-O-β-D-xyloside (to call compound 1,2 in the following text, its structure is seen accompanying drawing 1).Through pharmacological testing proof compound 1 and 2 is alstonia-leaf and preparation anti-inflammatory thereof, cough-relieving, analgesic effective constituent.Existence and content to compound 1 and 2 are analyzed and are detected, and can be used as the important indicator of the control alstonia-leaf and the quality of the pharmaceutical preparations thereof.
(1) detect compound 1 and 2 in alstonia-leaf medicinal material and the preparation thereof with thin-layered chromatography: with the silica G is adsorbent, is developping agent with the mixed solution of ethyl acetate-butanone-formic acid-water; The volume ratio of the contained four kinds of solvents of developping agent is an ethyl acetate: butanone: formic acid: water=5: 3～5: 0.5～1: 0.5～1.Its best proportioning is an ethyl acetate: butanone: formic acid: water=5: 3: 1: 1.
Wherein, the ethyl acetate in the developping agent can be used C
1-C
4Fatty acid and C
1-C
4The ester that forms of fatty alcohol substitute, butanone can be used C
3-C
6Lower aliphatic ketone substitute, formic acid can be used C
2-C
6Lower fatty acid substitute.
(2) with the content of compound 1 in spectrophotometry alstonia-leaf medicinal material and the preparation thereof and 2: in the presence of sodium nitrite, be developer with the aluminium nitrate, absorption value is measured in 510nm in the colour developing back in diluted sodium hydroxide solution, and its content is relatively determined with the absorption value of 2 standard solution that mix under identical color condition with compound 1 with absorption value.
Above-described preparation is to be the various preparations that raw material is produced with the alstonia-leaf, comprises alstonia-leaf granule, tablet, capsule, oral liquid etc.
Compound 1 extracts from alstonia-leaf with 2 available phytochemistry method and separates, and measures structure with spectroscopic analysis methods, proves its pharmacological action with zoopery.Spectroscopic data such as table one, structural formula are seen accompanying drawing 1.
Table one compound 1 and 2 main spectroscopic data
Compound 1 and 2 pharmacologically active prove:
(1) influence of compound 1 and 2 pairs of ammoniacal liquor induced mice coughs
Choose 18～22g mouse, on glass plate, push 25% strong aqua 0.04mL with 300mL glass beaker left-hand thread simultaneously rapidly in vessel, observe the cough number of times in the mouse 2min, typical case's cough actor is the qualified animal of screening more than 3 times to occur in the 2min.Get 60 of the qualified mouse of screening, male and female half and half are divided into 6 groups at random by sex, body weight, 10 every group.Each treated animal is pressed table two dosage respectively and is irritated stomach 1 time, and pure water is held in blank group filling etc.30min puts into mouse one by one and uses 300mL glass beaker left-hand thread on the glass plate after irritating stomach, pushes 25% strong aqua 0.04mL simultaneously in vessel rapidly, the cough number of times of mouse in the record 2min, and result and statistical study see Table two.
The result shows that the high dose of codeine phosphate group (positive drug), compound 1,2 all can obviously reduce the cough number of times that strong aqua causes mouse; Compound 1,2 has antitussive action.
Table two compound 1 and the 2 pairs of strong aquas cause the influence of mouse cough
(2) influence of compound 1 and 2 P-xylene induced mice auricle edemas
Choose 60 of 18～22g mouse, male and female half and half are divided into 6 groups at random by sex and body weight, 10 every group.Each treated animal is pressed table three dosage respectively and is irritated stomach 1 time, and pure water is held in blank group filling etc., and 30min evenly smears the caused by dimethylbenzene xylene inflammation by 0.1mL/ amount only in the mouse right ear two sides after administration.Mouse is put to death in dislocation behind the 1h, cuts ears along the auricle baseline, and two ears are stacked alignment,, weighs in downcutting auricle with the position homalographic with the 9mm card punch, calculates swelling degree and inhibiting rate according to following formula, and result and statistical study see Table three.
Swelling degree=auris dextra sheet weight (causing scorching side)-left auricle weight (normal side)
Inhibiting rate=[(blank group auricle swelling degree-other respectively organize auricle swelling degree)/blank group auricle swelling degree] * 100%
The result shows, aspirin (positive drug), compound 1 high dose, compound are 2 low, high dose all can obviously suppress by the mice auricle swelling due to the dimethylbenzene; The strong antiinflammatory action of compound 1,2 tools.
Table three compound 1 and 2 P-xylene cause the influence of mice auricle swelling
(3) compound 1 and 2 pairs of influences that the phenol red secretion of mouse tracheae is discharged
Accurately take by weighing the phenol red solution that is formulated as variable concentrations, measure absorption value (OD) at the 546nm place with photometer, (μ g/mL) is horizontal ordinate with phenol red concentration, and the OD value is an ordinate drawing standard curve.
Select 60 of 18～22g mouse for use, male and female half and half are divided into 6 groups at random by sex and body weight, 10 every group.Each treated animal is pressed table four dosage gastric infusion respectively 1 time, pure water is held in blank group fillings etc., phenol red according to 0.1mL/10g body weight lumbar injection 0.5% respectively behind administration 30min, behind 30min, take off cervical vertebra again and put to death animal, peel off the tracheae surrounding tissue, cut one section tracheae down to the bronchus crotch from thyroid cartilage, put into the test tube that fills 2mL physiological saline, add 0.1mL NaOH (1mol/L) again, sonicated 5min, centrifuging is drawn supernatant and is measured the OD value at wavelength 546nm place.Calculate the phenol red concentration of discharge in the mouse tracheal secretion according to typical curve, result and statistical study see Table four.
The result shows, the phenol red excretion amount that ammonium chloride (positive drug), compound 1 high dose, compound are 2 low, high dose all can obviously increase the mouse tracheae; Compound 1,2 has phlegm-dispelling functions.
The influence of table Four Modernizations compound 1 and the phenol red discharge of 2 pairs of mouse tracheaes
(4) compound 1 and 2 Dichlorodiphenyl Acetates cause the influence of mouse writhing pain
Choose 60 of 18～22g mouse, male and female half and half are divided into 6 groups at random by sex and body weight, 10 every group.Each treated animal is pressed table five dosage respectively and is irritated stomach 1 time, and pure water is held in blank group filling etc.After irritating stomach 45min, inject with the amount mouse peritoneal of 0.1mL/10g with 0.6% glacial acetic acid normal saline solution respectively, observe and the writhing response number of times of record test mice in 10min, calculate inhibiting rate according to following formula, result and statistical study see Table five.
Inhibiting rate=[(blank group is turned round body number of times-test group and turned round the body number of times)/blank group is turned round the body number of times] * 100%
The result shows, aspirin (positive drug), compound are 1,2 low, high dose all can significantly suppress the acetic acid induced mice turns round body pain; Compound 1,2 has analgesic activity.
Table five compound 1 and 2 Dichlorodiphenyl Acetates cause the influence of mouse writhing pain
The present invention has overcome art methods only can detect alkaloids composition in alstonia-leaf and the preparation thereof, and do not detect the weak point of another kind of even more important active component flavonoid glycoside, a kind of qualitative identification and quantitative analysis method that can accurately detect in alstonia-leaf medicinal material and the preparation thereof two flavonoid glycoside effective constituent compounds 1 and 2 is provided, can be used for accurately estimating the alstonia-leaf quality of medicinal material, also can be used for detecting with the alstonia-leaf is various preparations (the alstonia-leaf granule that raw material is produced, tablet, capsule, oral liquid etc.) quality analysis can directly apply to the quality analysis of actual production process and product.
Description of drawings
Fig. 1 is the chemical structural formula of compound 1 and 2.
Fig. 2 is a different places of production alstonia-leaf medicinal material thin-layer chromatographic analysis collection of illustrative plates.
Fig. 3 is the typical curve of spectrophotometry content of the present invention.
Embodiment
Following specific embodiment can further specify the present invention and application thereof, but does not constitute the restriction of the scope that claim of the present invention is protected.Its principle of work and process are:
1. thin-layer chromatography qualitative detection: the compound of same structure, on same chromatographic sheet, launch with same mobile phase solvent, should show consistent chromatographic behavior; Directly on chromatographic sheet, launch the back contrast with reference substance and detected sample,, can determine whether there be the chemical constitution consistent in the sample with reference substance according to whether identical spot occurring in the position identical with reference substance.
The gordian technique main points of thin layer chromatography are at different products, different compositions, by systematic research and comparison, screen, determine to have the developping agent system of optimal separation effect.
2. spectrophotometric detection by quantitative: the flavone compound that contains in the alstonia-leaf, in the presence of sodium nitrite, produce complex reaction with aluminium nitrate reagent, there is absorption maximum the colour developing back at 510nm in diluted sodium hydroxide solution, its absorption value and content are linear, can relatively determine its content with the absorption value of standard solution under identical color condition according to its absorption value.
Embodiment 1: the analyzing and testing of alstonia-leaf medicinal material-1
1, compound 1 with 2 separate and structure is identified
(1) extracts separation
Cornus controversa medicinal material 50kg pulverizes the back and uses 95% alcohol reflux, and decompression and solvent recovery gets medicinal extract.Alcohol-extracted extract adds water and stirs and use ethyl acetate, extracting n-butyl alcohol successively after loosing, and reclaims normal butyl alcohol and partly obtains the flavonoid glycoside potpourri.The flavonoid glycoside potpourri is eluant, eluent enrichment flavonoid glycoside through the decolouring of MCI post with methyl alcohol.The flavonoid glycoside of enrichment separates repeatedly through the sephadex lh-20 chromatogram again, is eluant, eluent with methyl alcohol, in conjunction with conventional purification process such as recrystallizations, obtains compound 1 and 2.
(2) structure is identified
Compound 1: yellow powder, be soluble in methyl alcohol, water, be insoluble to chloroform, ethyl acetate etc., but dissolve in dilute alkaline soln.214～217 ℃ of fusing points.Determine that by physics and chemistry and spectrum test the structure of compound 1 is Kaempferol-3-O-β-D-galactose-(2--1)-O-β-D-xyloside.
Compound 2: yellow powder, be soluble in methyl alcohol, water, be insoluble to chloroform, ethyl acetate etc., but dissolve in dilute alkaline soln.215～219 ℃ of fusing points.Determine that by physics and chemistry and spectrum test the structure of compound 2 is Quercetin-3-O-β-D-galactose-(2--1)-O-β-D-xyloside.
2, compound 1 and 2 thin-layer chromatography qualitative detection
(1) preparation of reference substance and sample solution
Take by weighing respectively from alstonia-leaf that to extract the flavonoid glycoside compound 1 and 2 that separates an amount of, make the solution of about 1mg/mL, be reference substance solution with dissolve with ethanol.The alstonia-leaf that dries was pulverized 40 eye mesh screens, got powder 2g and added the 20mL alcohol solvent, soaked sonicated 30min extraction after 1 hour, filtered, and filtrate is reclaimed solvent and got ethanol extract.Ethanol extract filters with the weak aqua ammonia 20mL dissolving of pH=8.5; Filtrate again with 20mL extracting n-butyl alcohol flavonoid glycoside, is reclaimed butanol solution with the watery hydrochloric acid pH=4 that neutralizes, and residue 1mL dissolve with methanol is the sample solution of confession thin-layer chromatographic analysis.
(2) thin-layer chromatographic analysis of alstonia-leaf sample and judgement
With the silica G plate is adsorbent, presses 5: 3: 1 with ethyl acetate-butanone-formic acid-water: 1 volume ratio preparation developping agent; Analyze 17 parts of alstonia-leaf medicinal material samples that compare from 15 places of production, thin-layer chromatogram is seen accompanying drawing 2.Different place of production alstonia-leaf medicinal materials (sample 1-10,12-13,15-16) all can detect compound 1 and 2; 2 parts of alstonia-leaf medicinal materials that go mouldy fully (sample 11,14) fail to detect compound 1 and 2; 1 part of alstonia-leaf medicinal material (sample 17) that part is gone mouldy detects a spot of compound 1 and 2.
Result according to the thin-layer chromatography qualitative detection judges: alstonia-leaf medicinal material sample 1-10,12-13,15-16 are qualified medicinal materials, and sample 11,14,17 (go mouldy or partly go mouldy) is a medicinal material inferior.
2, compound 1 and 2 assay
(1) preparation of reference substance and sample solution
Accurately take by weighing compound 1 and 2 each 12.5mg, place the 50mL measuring bottle, add dissolve with methanol and be settled to scale, promptly get the reference substance solution that concentration is 0.5mg/mL, standby.
The alstonia-leaf pulverizing medicinal materials is crossed 60 eye mesh screens, and precision takes by weighing 1g, puts in the 100mL round-bottomed flask, accurately adds ethanol 50mL, claims decide weight, soaks after 1 hour, and refluxing extraction 1 hour is put coldly, supplies the weight that subtracts mistake with ethanol, filtration, and it is standby to get subsequent filtrate.The accurate extract 10mL decompression recycling ethanol of drawing, residue adds the weak aqua ammonia 20mL dissolving of pH=8.5, filters, and subsequent filtrate is standby.
(2) typical curve
Accurately draw 0.5mg/mL reference substance solution 0,1,2,3,4,5,6,7mL respectively, put in the 25mL measuring bottle, replenish an amount of purified water respectively to 10mL.The sodium nitrite solution 1mL that each adds 5% shakes up; Place after 6 minutes, add 10% aluminium nitrate 1mL, shake up; Place after 6 minutes, add 4% NaOH 10mL again, after shaking up, be settled to scale with purified water; Placing after 15 minutes, is reference with the blank solution, measures its absorbance A, data such as table six at the 510nm place.The data regression treatment gets regression equation: A=6.3036 * C+0.0182, and r=0.9999, typical curve see accompanying drawing 3.
The linear relationship of table six spectrophotometry content
| Sample volume mL | 0.5 | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| Solution concentration mg/mL | 0.01 | 0.02 | 0.04 | 0.06 | 0.08 | 0.10 | 0.12 | 0.14 |
| Absorbance A | 0.083 | 0.144 | 0.268 | 0.394 | 0.526 | 0.650 | 0.774 | 0.900 |
(3) sample determination
Accurately draw each sample solution 2mL of alstonia-leaf, put in the 25mL measuring bottle, add the 8mL purified water, add 5% sodium nitrite solution 1mL, shake up; Place after 6 minutes, add 10% aluminium nitrate 1mL, shake up; Place after 6 minutes, add 4% NaOH 10mL again, after shaking up, be settled to scale with purified water; Placing after 15 minutes, is reference with the blank solution, measures its absorbance A at the 510nm place.
The sample absorbance A of measuring, establishing criteria curve calculation flavonoid glycoside compound 1 and 2 total content.The determination data of the alstonia-leaf medicinal material in the different places of production sees Table seven.
The content of flavonoid glycoside compound in the alstonia-leaf medicinal material of the different place of production of table seven
| Sample number into spectrum | Content % | Sample number into spectrum | Content % | Sample number into | Content % | |
| 1 # | 3.458 | 7 # | 2.485 | 13 # | 4.616 | |
| 2 # | 2.750 | 8 # | 4.604 | 14 # | 0.440 |
| 3 # | 5.268 | 9 # | 2.666 | 15 # | 2.811 |
| 4 # | 2.656 | 10 # | 2.785 | 16 # | 5.247 |
| 5 # | 2.378 | 13 # | 0.436 | 17 # | 0.737 |
| 6 # | 5.118 | 12 # | 2.096 |
Testing result shows, the effective component content of different alstonia-leaf medicinal materials is differentiated, the flavonoid glycoside compound content of alstonia-leaf medicinal material sample 1-10,12-13,15-16 is between 2.096%～5.118%, and is best in quality with the sample 3,6,8,13,16 that content is higher.And go mouldy or the content of the sample 11,14,17 that goes mouldy of part only between 0.436%～0.7378%, belong to medicinal material inferior.The The qualitative analysis that the quantitative analysis results of assay and thin-layer chromatography detect has consistance.The content of flavonoid glycoside has been represented the quality of alstonia-leaf quality of medicinal material, can be directly used in objective evaluation alstonia-leaf quality of medicinal material.
Embodiment 2: the analyzing and testing of alstonia-leaf medicinal material-2
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: ethyl acetate-butanone-formic acid-water=5: 5: 1: 1.Testing result: judge consistent with embodiment 1.
Embodiment 3: the analyzing and testing of alstonia-leaf medicinal material-3
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: ethyl acetate-butanone-formic acid-water=5: 3: 0.5: 1.Testing result: judge consistent with embodiment 1.
Embodiment 4: the analyzing and testing of alstonia-leaf medicinal material-4
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: ethyl acetate-butanone-formic acid-water=5: 3: 1: 0.5.Testing result: judge consistent with embodiment 1.
Embodiment 5: the analyzing and testing of alstonia-leaf medicinal material-5
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: butyl formate-acetone-acetate-water=5: 3: 1: 1.Testing result: judge consistent with embodiment 1.
Embodiment 6: the analyzing and testing of alstonia-leaf medicinal material-6
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: methyl butyl-pentanone-formic acid-water=5: 4: 1: 1.Testing result: judge consistent with embodiment 1.
Embodiment 7: the analyzing and testing of alstonia-leaf medicinal material-7
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: methyl butyl-pentanone-butyric acid-water=5: 5: 1: 1.Testing result: judge consistent with embodiment 1.
Embodiment 8: the analyzing and testing of alstonia-leaf medicinal material-8
Method is with embodiment 1, but the expansion solvent of thin-layer chromatographic analysis is changed into: ethyl acetate-butanone-n-caproic acid-water=5: 4: 1: 1.Testing result: judge consistent with embodiment 1.
Embodiment 9: the detection of alstonia-leaf particle
Alstonia-leaf particle 2g adds 20mL hot water and dissolves, and neutralizes behind the pH=4 again with 20mL extracting n-butyl alcohol flavonoid glycoside with watery hydrochloric acid, reclaims butanol solution, and residue 1mL dissolve with methanol is sample solution.Reference substance solution preparation, point sample, expansion, judgement are equal to embodiment 1, and the expansion solvent is ethyl acetate-butanone-formic acid-water=5: 3: 1: 1.The result is: can detect flavonoid glycoside compound 1 and 2 in the alstonia-leaf particle.
Precision takes by weighing 1g alstonia-leaf particle, and the accurate weak aqua ammonia 20mL that adds pH=8.5 dissolves, and filters, and accurately draws subsequent filtrate 2mL, puts in the 25mL measuring bottle, and following moisturizing, colour developing, mensuration are equal to embodiment 1.The total amount that contains flavonoid glycoside compound 1 and 2 in the alstonia-leaf particle is 2.17%.
Embodiment 10: the detection of alstonia-leaf capsule
Method is got the alstonia-leaf capsule 's content and is detected with embodiment 9, and the result is: can detect flavonoid glycoside compound 1 and 2 in the alstonia-leaf capsule; The total amount that contains flavonoid glycoside compound 1 and 2 in the alstonia-leaf capsule is 1.75%.
Embodiment 11: the detection of alstonia-leaf oral liquid
Method is got the alstonia-leaf oral liquid and is detected with embodiment 9, and the result is: can detect flavonoid glycoside compound 1 and 2 in the alstonia-leaf oral liquid; The total amount that contains flavonoid glycoside compound 1 and 2 in the alstonia-leaf oral liquid is 1.58%.
Embodiment 12: the detection of lamp stand blade
Method is with embodiment 9, detects after getting lamp stand blade levigation, and the result is: can detect flavonoid glycoside compound 1 and 2 in the lamp stand blade; The total amount that contains flavonoid glycoside compound 1 and 2 in the lamp stand blade is 2.32%.
Claims (4)
1. the drug test method of alstonia-leaf medicinal material and preparation thereof, it is characterized in that:, realize quality analysis and control alstonia-leaf medicinal material and preparation thereof with its content of spectrophotometry with the effective constituent Kaempferol-3-O-β-D-galactose of thin-layer chromatography qualitative detection alstonia-leaf medicinal material and preparation thereof-(2--1)-O-β-D-xyloside, Quercetin-3-O-β-D-galactose-(2--1)-O-β-D-xyloside; Wherein:
1) being to be adsorbent with the silica G in the thin-layered chromatography detection method, is developping agent with the mixed solution of ethyl acetate-butanone-formic acid-water; The volume ratio of the contained four kinds of solvents of developping agent is an ethyl acetate: butanone: formic acid: water=5: 3～5: 0.5～1: 0.5～1; Reference substance in the thin-layered chromatography detection method with the preparation of sample solution is: take by weighing respectively that the flavonoid glycoside compound Kaempferol that extraction separates from alstonia-leaf-3-O-β-D-galactose-(2--1)-O-β-D-xyloside and Quercetin-3-O-β-D-galactose-(2--1)-O-β-D-xyloside is an amount of, make the solution of about 1mg/mL with dissolve with ethanol, be reference substance solution; The alstonia-leaf that dries was pulverized 40 eye mesh screens, got powder 2g and added the 20mL alcohol solvent, soaked sonicated 30min extraction after 1 hour, filtered, and filtrate is reclaimed solvent and got ethanol extract; Ethanol extract filters with the weak aqua ammonia 20mL dissolving of pH=8.5; Filtrate again with 20mL extracting n-butyl alcohol flavonoid glycoside, is reclaimed butanol solution with the watery hydrochloric acid pH=4 that neutralizes, and residue 1mL dissolve with methanol is the sample solution of confession thin-layer chromatographic analysis;
2) spectrophotometric method of described assay is: be developer with the aluminium nitrate in the presence of sodium nitrite, absorption value is measured in 510nm in colour developing back in diluted sodium hydroxide solution, and its content with the absorption value of standard solution under identical color condition that the absorption value of sample solution and Kaempferol-3-O-β-D-galactose-(2--1)-O-β-D-xyloside, Quercetin-3-O-β-D-galactose-(2--1)-O-β-D-xyloside mixes relatively and determine; The used reference substance and the preparation of sample solution are: accurately take by weighing compound Kaempferol-3-O-β-D-galactose-(2--1)-O-β-D-xyloside and Quercetin-3-O-β-D-galactose-(2--1)-O-β-each 12.5mg of D-xyloside, place the 50mL measuring bottle, add dissolve with methanol and be settled to scale, promptly get the reference substance solution that concentration is 0.5mg/mL; The alstonia-leaf pulverizing medicinal materials is crossed 60 eye mesh screens, and precision takes by weighing 1g, puts in the 100mL round-bottomed flask, accurately adds ethanol 50mL, claims decide weight, soaks after 1 hour, and refluxing extraction 1 hour is put coldly, supplies the weight that subtracts mistake with ethanol, filtration, and getting subsequent filtrate is sample.
2. according to the drug test method of described alstonia-leaf medicinal material of claim 1 and preparation thereof, it is characterized in that: the volume ratio of the contained four kinds of solvents of developping agent is an ethyl acetate: butanone: formic acid: water=5: 3: 1: 1.
3. according to the drug test method of described alstonia-leaf medicinal material of claim 2 and preparation thereof, it is characterized in that: the ethyl acetate C in the developping agent
1-C
4Fatty acid and C
1-C
4The ester that forms of fatty alcohol substitute butanone C
3-C
6Lower aliphatic ketone substitute formic acid C
2-C
6Lower fatty acid substitute.
4. according to the detection method of described alstonia-leaf medicinal material of claim 2 and preparation thereof, it is characterized in that: described preparation is to be the various preparations that raw material is produced with the alstonia-leaf, comprises alstonia-leaf granule, tablet, capsule, oral liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100583404A CN101269097B (en) | 2008-04-30 | 2008-04-30 | Alstonia-leaf medicinal materials and medicament testing method for its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100583404A CN101269097B (en) | 2008-04-30 | 2008-04-30 | Alstonia-leaf medicinal materials and medicament testing method for its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101269097A CN101269097A (en) | 2008-09-24 |
| CN101269097B true CN101269097B (en) | 2010-12-29 |
Family
ID=40003488
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100583404A Active CN101269097B (en) | 2008-04-30 | 2008-04-30 | Alstonia-leaf medicinal materials and medicament testing method for its preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101269097B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101543505A (en) * | 2009-04-29 | 2009-09-30 | 香港科技大学 | Medicine for preventing and treating Alzheimer's disease and preparing method thereof |
| CN103120361B (en) * | 2012-10-16 | 2016-04-13 | 红塔烟草(集团)有限责任公司 | A kind of cutting tobacco leaves method based on Distribution Law of Chemical Composition |
| CN105218617A (en) * | 2015-09-30 | 2016-01-06 | 宋晓梅 | A kind of new triterpenoid and preparation method thereof and medicinal use |
| RU2641093C1 (en) * | 2016-12-01 | 2018-01-15 | федеральное государственное автономное образовательное учреждение высшего образования "Российский университет дружбы народов" (РУДН) | Method for wild camomile flowers identification |
| CN110361469A (en) * | 2019-07-17 | 2019-10-22 | 长春中医药大学 | A kind of Chinese medicine mildew detection method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100356172C (en) * | 2006-03-16 | 2007-12-19 | 朱光荣 | Use of picrinine in quality control of Dengtaiye medicine and its preparation |
-
2008
- 2008-04-30 CN CN2008100583404A patent/CN101269097B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100356172C (en) * | 2006-03-16 | 2007-12-19 | 朱光荣 | Use of picrinine in quality control of Dengtaiye medicine and its preparation |
Non-Patent Citations (1)
| Title |
|---|
| 杜国顺等.灯台叶中的非碱性成分.中国天然药物.2007,5(4), * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101269097A (en) | 2008-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101167788B (en) | Quality control method of traditional Chinese medicine 'zhenqi fuzheng' containing glossy privet fruit and radix astragali for strengthening the body resistance for aeipathia deficiency damage and qi | |
| CN100536870C (en) | Qulity control method for new compound isatis leaf preparation | |
| CN102579612B (en) | Method for extracting total alkaloid of aconitum soongaricum | |
| CN101269097B (en) | Alstonia-leaf medicinal materials and medicament testing method for its preparation | |
| CN102269751B (en) | Detection method of Liuweinengxiao preparation | |
| CN102879495B (en) | Antidotal capsule of Tibetan medicine compound and quality detection method of preparation of antidotal capsule | |
| CN101884746B (en) | Method for detecting capsules for treating cough with asthma | |
| CN101327247A (en) | Senna formulation and quality control method | |
| CN100369614C (en) | Quality control method of compound gallblader freeflow solid preparation | |
| CN101703610A (en) | Quality detection method of Qingnao antihypertensive tablet | |
| CN102749407B (en) | The content assaying method of rhizoma anemarrhenae saponin BII in a kind of Chinese medicine composition | |
| CN1939472B (en) | Detection method of compound preparation for treating diabetes | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN101396492A (en) | Quality control method for toxin ingredient, yunaconitine in Yuannan Hongyao capsule | |
| CN102058828A (en) | Medicinal composition and detection method for preparation thereof | |
| CN102062761A (en) | Quality control method of compound dextromethorphan hydrobromide preparation | |
| CN102058829B (en) | Medicine composition and detection method of preparations of medicine composition | |
| CN101716260A (en) | Quality control method of capsule preparation for treating dermatosis | |
| CN101632804B (en) | Quality control method for wind-dispelling heat-dissipating capsules | |
| CN102735787B (en) | Novel digestion promoting tablet and its relative preparation quality detection method | |
| CN101176755B (en) | Application of meletin-7-0-glycoside in mass control of cudrania tricuspidata or preparations thereof | |
| CN105168390A (en) | Anti-inflammation and detoxification preparation as well as preparation method and quality control method | |
| CN102188468B (en) | Preparation method of Xinnaoshutong tablet | |
| CN1846730A (en) | Chaihuang soft capsule preparing process and its quality control technology | |
| CN103169782A (en) | Stomach-regulating qi-coordinating decoction formula granule, and preparation method and detection method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20171019 Address after: High tech Zone Jinpu Ma Cheng Road 650503 Yunnan city of Kunming province No. 2899 Patentee after: Yunnan Phytopharmaceutical Co.,Ltd. Address before: 650034 Wang Jia dam 22, Yunnan, Kunming Patentee before: Kunming Zhenhua Pharmaceutical Factory Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Alstonia-leaf medicinal materials and medicament testing method for its preparation Effective date of registration: 20200423 Granted publication date: 20101229 Pledgee: Bank of Communications Ltd. Yunnan branch Pledgor: YUNNAN PHYTOPHARMACEUTICAL Co.,Ltd. Registration number: Y2020530000010 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20220718 Granted publication date: 20101229 Pledgee: Bank of Communications Ltd. Yunnan branch Pledgor: Yunnan Phytopharmaceutical Co.,Ltd. Registration number: Y2020530000010 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |