CN105175426A - Method of extracting and purifying bergenin from cissus pteroclada hayata - Google Patents
Method of extracting and purifying bergenin from cissus pteroclada hayata Download PDFInfo
- Publication number
- CN105175426A CN105175426A CN201510657480.3A CN201510657480A CN105175426A CN 105175426 A CN105175426 A CN 105175426A CN 201510657480 A CN201510657480 A CN 201510657480A CN 105175426 A CN105175426 A CN 105175426A
- Authority
- CN
- China
- Prior art keywords
- bergeninum
- concentrated
- ethanol
- macroporous resin
- bergenin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/13—Crystalline forms, e.g. polymorphs
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention discloses a method of extracting and purifying bergenin from cissus pteroclada hayata. The method comprises the steps that the cissus pteroclada hayata is smashed and screened, ethanol solution is added according to the mass-to-volume ratio of 1 to 8-20 for reflux extraction, solid-liquid separation is conducted to obtain filtrate, the filtrate is concentrated, macroporous resin is added into the concentrated solution for static adsorption, the macroporous resin subjected to adsorption is placed on a pillar, impurities are removed and elution is conducted with distilled water and ethanol solution in sequence, the eluant is collected and concentrated to obtain crude bergenin, and the crude bergenin is recrystallized with ethanol to obtain pure bergenin. According to the method, sources of raw materials are wide, price is low, production cost is low, production conditions are moderate, and pollution to the environment is light.
Description
Technical field
The present invention relates to a kind of method of extraction, purifying Bergeninum from Treebine Stem, belong to natural drug evaluation and exploration technology field.
Background technology
Treebine Stem (
cissusPterocladaHayata) belong to Vitaceae (
vitaceae) Cissus (
cissusLinn.) plant, because its stem, branch, petiole are sharp square and gain the name, the side of having another name called rattan, Herba Lycopodii, leech rattan etc.Treebine Stem is mainly distributed in the southern area such as Guangxi, Guangdong, Yunnan of China, aboundresources, its property is sour, puckery, sweet, flat, there is the effects such as promoting blood circulation to remove blood stasis, wind-damp dispelling, relaxing muscles and tendons network, anti-inflammatory, be widely used in rheumatic arthralgia, lumbar muscle strain, wound, the ailing treatment such as hemorrhage China is among the people, there is good efficacy.Such as Bose Region peasant directly mashes external application hemostasis with the fresh leaf of Treebine Stem; Treebine Stem is made pulvis and injection by Baise hospital, is used for the treatment of interior traumatic hemorrhage, duodenal hemorrhage, the diseases such as gastritis.Treebine Stem records in the ancient books and records such as " Guangxi herbal medicine ", " Medicinal Plants of Guangxi register ", " Chinese Pharmacopoeia " (version one in 1977), " Chinese medicine voluminous dictionary ", because of its abundance, cheap, curative effect is extensively remarkable, is a kind of national medicinal plant of characteristic having value of exploiting and utilizing.Although Treebine Stem has long medicinal history and experience Chinese Minority Nationalities is among the people, but due to the shortage of Treebine Stem chemical composition and pharmacological research, in the exploitation of Treebine Stem effective constituent, not yet there is the appearance of depths products, it is commercially still sold with the form of crude drug, cheap.The structure of Bergeninum:
Bergeninum has another name called brave youngster grass element, vakerin, and molecular formula is C
14h
16o
9, be a kind of natural compounds with Isocoumarin structure.Bergeninum and derivative thereof have anti-inflammatory, cough-relieving, analgesia, the multiple pharmacologically active such as antibacterial, anti-oxidant.Bergeninum records in each version of the Pharmacopoeia of the People's Republic of China as antisussive and expectorant agent, clinically for the treatment of the diseases such as bronchitis, pulmonary emphysema, gastritis, having good efficacy, is the natural drug that a kind of pharmacologically active is extensive, evident in efficacy, toxic side effect is low.
Due to the difficulty of chemosynthesis, Bergeninum is mainly with extraction from natural medicinal plant, method of purification preparation.At present, the report of Bergeninum extracting method is more, and less for the report of its purification process.The extracting method of Bergeninum mainly contains soxhlet extraction method, ultrasonic extraction, super critical extraction, organic solvent reflux extraction, alkali extraction method etc.But the method at present for purifying Bergeninum extract is difficult to mass-producing preparation.If silica gel column chromatography is the separation means that laboratory is commonly used, but silica gel can not reclaim, and eluent is the toxic organic solvents such as sherwood oil, ethyl acetate, chloroform, and cost is high, and operation easier is large, can not be used for industrialization preparation; Recrystallization method must be that the content of Bergeninum in solution to be crystallized could use up to time to a certain degree, common Bergeninum extract, not before other method enrichment, compares and is difficult to crystallization and cannot directly purifies with recrystallization method.Given this, current organic solvent extraction still rests on and obtains containing stage of Bergeninum crude extract, and this extraction method will be utilized to prepare Bergeninum sterling in enormous quantities, must develop can industrialization amplification, purification process that cost is low.
Summary of the invention
The object of the present invention is to provide and a kind of be applicable to industrialization and amplify, the method for raw materials medicine Bergeninum from Treebine Stem that cost is low.
The present invention is achieved through the following technical solutions.
From Treebine Stem, a method for extraction purification Bergeninum, comprises the steps:
(1) pulverized by Treebine Stem, sieving, is the ethanolic soln refluxing extraction that 1:8 ~ 20 add 40 ~ 60% according to mass volume ratio;
(2) solid-liquid separation obtains filtrate, filtrate is concentrated into 1/2 ~ 1/3 of its original volume, simultaneously recycling design;
(3) in concentrated solution, add macroporous resin Static Adsorption 6 ~ 8 hours, macroporous resin is selected from AB-8, D3520, D101;
(4) by absorption after macroporous resin upper prop, successively first with distilled water, 10% ethanolic soln removing impurity;
(5) with 20% ethanolic soln wash-out, collect 20% ethanol eluate;
(6) 20% ethanol eluate is concentrated, obtain Bergeninum crude product;
(7) Bergeninum crude product ethyl alcohol recrystallization is obtained Bergeninum sterling.
In above-mentioned steps (2), the method for solid-liquid separation is for filtering or centrifugation, and filtrate is concentrated into Bergeninum concentration 2mg/mL.
In above-mentioned steps (4), with the distilled water of 2 times of column volumes and 10% ethanol removing impurity of 2 times of column volumes.
In above-mentioned steps (5), with 20% ethanol elution of 7 times of column volumes.
In above-mentioned steps (6), described concentration method is vacuum concentration or membrane concentration.
In above-mentioned steps (7), the ethanol with 95% carries out recrystallization.
The invention has the advantages that: adopt the alcoholic extraction of 40 ~ 60%, have green non-poisonous, extraction yield is high, cost is low, the advantage such as easy to be recycled; Adopt macroporous resin purification and enrichment, have adsorptive capacity large, can reuse, be easy to industrialization amplification, low cost and other advantages, after macroporous resin purification, the crude product of Rhizome or herb of Purple Bergenia purity more than 80% can be obtained; To be Bergeninum crude product after macroporous resin purification, just can obtain the Bergeninum sterling meeting and use as bulk drug through ethanolic soln recrystallization; Raw material sources of the present invention are extensive, low price, and production cost is low, and working condition is gentle, and environmental pollution is little.
Accompanying drawing explanation
Fig. 1 is extraction and purification process schema of the present invention;
The static saturated adsorption rate of the different macroporous resin of Fig. 2;
The static desorption efficiency of the different macroporous resin of Fig. 3.
Embodiment
The invention will be further described by the following examples.
Embodiment 1
By the Treebine Stem powder of 500g, add 50% ethanolic soln, make the quality of Treebine Stem powder and the volume ratio of ethanol be 1:20, refluxing extraction 1 time, each 2h, centrifugal, filter, discard waste residue and collect and obtain filtrate, filtrate is concentrated, simultaneously recycling design.Be the concentrated solution of 2.0mg/mL by concentration, be by volume the ratio of 1:1 with macroporous resin quality, add in AB-8 macroporous resin and carry out Static Adsorption, adsorption time is 8h, then upper prop.First with the distilled water of 2 times of column volumes and 10% ethanol removing impurity of 2 times of column volumes, and then carry out wash-out with 7 times of column volume 20% ethanolic solns, collect 20% ethanol elution part.Elutriant is concentrated, obtains Bergeninum crude product 10.3g, after 95% ethyl alcohol recrystallization, obtain Bergeninum sterling 8.9g.
Embodiment 2
By the Treebine Stem powder of 1Kg, add 50% ethanolic soln, make the mass volume ratio of Treebine Stem powder and ethanol be 1:15, refluxing extraction 2 times, each 2h, centrifugal, filter, discard waste residue collect obtain filtrate, filtrate is concentrated, reclaims organic solvent simultaneously.Be the concentrated solution of 2.0mg/mL by concentration, be by volume the ratio of 1:1 with macroporous resin quality, add in D3520 macroporous resin and carry out Static Adsorption, adsorption time is 8h, then upper prop.First with the distilled water of 2 times of column volumes and 10% ethanol removing impurity of 2 times of column volumes.And then carry out wash-out with 7 times of column volume 20% ethanolic solns, collect 20% ethanol elution part.Elutriant is concentrated, obtains Bergeninum crude product 20.5g, after 95% ethyl alcohol recrystallization, obtain Bergeninum sterling 17.6g.
Embodiment 3
By the Treebine Stem powder of 10Kg, add 60% ethanolic soln, make the mass volume ratio of Treebine Stem powder and ethanol be 1:10, refluxing extraction 3 times, each 2h, centrifugal, filter, discard waste residue collect obtain filtrate, filtrate is concentrated, reclaims organic solvent simultaneously.Be the concentrated solution of 2.0mg/mL by concentration, be by volume the ratio of 1:1 with macroporous resin quality, add in D101 macroporous resin and carry out Static Adsorption, adsorption time is 8h, then upper prop.First with the distilled water of 2 times of column volumes and 10% ethanol removing impurity of 2 times of column volumes.And then carry out wash-out with 20% ethanolic soln, collect 20% ethanol elution part.Elutriant is concentrated, obtains Bergeninum crude product 200.6g, after 95% ethyl alcohol recrystallization, obtain Bergeninum sterling 169.7g.
The yield of Bergeninum crude product is about 2%, purity is detected all more than 80% through HPLC, Bergeninum sterling can be obtained after the ethyl alcohol recrystallization of 95%, yield is about 1.7%, because the content difference of Bergeninum in the Treebine Stem that Different sources, season gather is comparatively large, yield is higher, on the low side all belongs to normal phenomenon.The method that this law adopts the macroporous resin that can reuse to be combined with recrystallization prepares Bergeninum, has the advantage that operation is succinct, cost is low, be easy to industrialization amplification.
Embodiment 4 macroporous resin Study on selection
Macroporous resin has polarity, low-pole, the broad variety such as nonpolar, in the present invention, contriver have selected LSA-700(polarity), LSA-700B(polarity), D941(polarity), AB-8(low-pole), D3520(is nonpolar), D101(is nonpolar) these 7 kinds of macroporous resins as investigation object, determine by static and dynamic Status Adsorption and desorption the macroporous resin being suitable for Bergeninum in purifying Treebine Stem.
Staticadsorption experiment results as shown in Figure 2, visible LSA-700, LSA-700B, AB-8, D101, D941, D3520 six kinds of macroporous resins are after Static Adsorption 24h, all close to saturated adsorption, its Static Adsorption rate is all more than 90%, and wherein the static saturated adsorption rate of AB-8, D101, D3520 tri-kinds of resins is higher than other 3 kinds.Require in production that resin absorption amount is large and desorption rate is high, to ensure that effective constituent reclaims to greatest extent.Therefore contriver utilizes the desorption experiment of macroporous resin to carry out preferred resin further, its experimental result as shown in Figure 3.
As can be seen from Figure 3, above-mentioned six kinds of macroporous resins show obvious otherness when static desorb: wherein the desorption ability of D101, AB-8, D3520 resin is comparatively strong, all more than 80%, is suitable for the separation and purification of Bergeninum in Treebine Stem; And the desorption ability of LSA-700, LSA-700B, D941 resin is more weak, only have an appointment 30%, in purge process, unfavorable raw materials recovery, causes wastage of material, therefore does not select.
In addition, show that these three kinds of resins of D101, AB-8, D3520 are all suitable to the dynamic saturated extent of adsorption of Bergeninum, dynamic desorption rate through dynamic experiment, there is no notable difference, therefore contriver selects these 3 kinds of macroporous resins of D101, AB-8, D3520 for the separation and purification of Bergeninum in Treebine Stem.
The selection optimization of embodiment 5 alcohol concn and consumption
Adopt thin-layer chromatography (TLC) to detect composition in different concentration ethanol effluent liquid, the condition of developping agent is chloroform: ethyl acetate: methyl alcohol=5:4:2.5, its result is as shown in table 1.As shown in Table 1, distilled water and 10% ethanol elution macroporous resin time, there is no Bergeninum and washed out, be now mainly water-soluble impurity in elutriant; When elutriant is the ethanol of 20%, follows the tracks of according to TLC, find that there is Bergeninum by wash-out.Along with the increase of alcohol concn, the while that Rhizome or herb of Purple Bergenia being eluted, other oil-soluble impurities also flows out thereupon, be unfavorable for that the purifying of Bergeninum also causes waste of solvent, cost to increase, therefore elution profile when investigation alcohol concn is 20%, 40%, 60% further in subsequent experimental, its result is as shown in table 1,2,3.
As can be seen here, first use the water of 2 times of column volumes (BV), the ethanolic soln wash-out of 2 times of column volumes 10% can remove impurity, use the aqueous ethanolic solution wash-out of 7 times of column volumes 20% again, the Bergeninum crude product purity obtained is high, for AB-8, D101, D3520 tri-kinds of macroporous resins, be respectively 84.98%, 88.13%, 82.18%.
The optimization of embodiment 6 sample concentration
When extraction is complete, after carrying out solid-liquid separation, in extracting solution, the amount of Bergeninum is certain, but the degree direct influence that concentrates of extracting solution is to the concentration of Bergeninum in extracting solution, the concentration of Bergeninum is influential to macroporous resin adsorption, desorb, therefore contriver investigates the impact of sample concentration.Table 4,5,6 is experimental result when Determination of Bergenin is 39.94mg in extracting solution.
From above-mentioned experimental result, when sample concentration is 1.997mg/ml ≈ 2mg/ml, the adsorptive capacity of macroporous resin and desorption efficiency the best, therefore carry out loading (detecting by liquid phase chromatography) when the concentration being concentrated into Bergeninum is about 2mg/mL.
Macroporous resin can become large to the adsorption rate of Bergeninum in extracting solution along with the growth of adsorption time, contriver finds as more than adsorption time 8h in Staticadsorption experiment, adsorption rate rangeability is little, and considers the production cycle, therefore adsorption time is decided to be 8h.
Claims (6)
1. the method for extraction purification Bergeninum from Treebine Stem, is characterized in that comprising the steps:
(1) pulverized by Treebine Stem, sieving, is the ethanolic soln refluxing extraction that 1:8 ~ 20 add 40 ~ 60% according to mass volume ratio;
(2) solid-liquid separation obtains filtrate, filtrate is concentrated into 1/2 ~ 1/3 of its original volume, simultaneously recycling design;
(3) in concentrated solution, add macroporous resin Static Adsorption 6 ~ 8 hours, macroporous resin is selected from AB-8, D3520, D101;
(4) by absorption after macroporous resin upper prop, successively first with distilled water, 10% ethanolic soln removing impurity;
(5) with 20% ethanolic soln wash-out, collect 20% ethanol eluate;
(6) 20% ethanol eluate is concentrated, obtain Bergeninum crude product;
(7) Bergeninum crude product ethyl alcohol recrystallization is obtained Bergeninum sterling.
2. method according to claim 1, is characterized in that: in step (2), and the method for solid-liquid separation is for filtering or centrifugation, and filtrate is concentrated into Bergeninum concentration 2mg/mL.
3. method according to claim 1, is characterized in that: in step (4), with the distilled water of 2 times of column volumes and 10% ethanol removing impurity of 2 times of column volumes.
4. method according to claim 1, is characterized in that: in step (5), with 20% ethanol elution of 7 times of column volumes.
5. method according to claim 1, it is characterized in that: in step (6), described concentration method is vacuum concentration or membrane concentration.
6. method according to claim 1, is characterized in that: in step (7), the ethanol with 95% carries out recrystallization.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510657480.3A CN105175426B (en) | 2015-10-14 | 2015-10-14 | A kind of method of the extraction purification Bergenin from treebine stem |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510657480.3A CN105175426B (en) | 2015-10-14 | 2015-10-14 | A kind of method of the extraction purification Bergenin from treebine stem |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105175426A true CN105175426A (en) | 2015-12-23 |
| CN105175426B CN105175426B (en) | 2018-05-18 |
Family
ID=54897937
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510657480.3A Active CN105175426B (en) | 2015-10-14 | 2015-10-14 | A kind of method of the extraction purification Bergenin from treebine stem |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105175426B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106632373A (en) * | 2016-09-30 | 2017-05-10 | 陕西科技大学 | A D crystal form solid substance of bergenin, a preparing method thereof and uses of the substance |
| CN106749298A (en) * | 2016-11-18 | 2017-05-31 | 文县嘉诚农副产品有限公司 | A kind of Bergenin is extracted and process for purification |
| CN110272433A (en) * | 2018-08-12 | 2019-09-24 | 长沙爱扬医药科技有限公司 | A method of extracting quercitrin and Bergenin |
-
2015
- 2015-10-14 CN CN201510657480.3A patent/CN105175426B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| 李雯霏等: "四方藤中总香豆素的提取工艺研究", 《中药材》 * |
| 池翠云等: "瑶药四方藤化学成分研究", 《中药材》 * |
| 陈如意等: "HPLC测定四方藤中3种岩白菜素类化合物的含量", 《广西师范大学学报：自然科学版》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106632373A (en) * | 2016-09-30 | 2017-05-10 | 陕西科技大学 | A D crystal form solid substance of bergenin, a preparing method thereof and uses of the substance |
| CN106749298A (en) * | 2016-11-18 | 2017-05-31 | 文县嘉诚农副产品有限公司 | A kind of Bergenin is extracted and process for purification |
| CN106749298B (en) * | 2016-11-18 | 2019-03-29 | 文县嘉诚农副产品有限公司 | A kind of Bergenin extracts and refining methd |
| CN110272433A (en) * | 2018-08-12 | 2019-09-24 | 长沙爱扬医药科技有限公司 | A method of extracting quercitrin and Bergenin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105175426B (en) | 2018-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2020119001A1 (en) | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification | |
| CN102846784B (en) | Paederia scandens water extract, and preparation method and application thereof | |
| CN102976909B (en) | Method for extracting and purifying 6-gingerol from ginger | |
| CN101260131A (en) | Method for extracting iridoid active site and monomer from eucommia bark | |
| CN101037467B (en) | Method for separating purifying ursolic acid from loquat leaf | |
| CN105294628A (en) | Method for preparing flavonoid component by separating wild chrysanthemum flower | |
| CN101348474A (en) | Method for preparing salvianolic acid B and tanshinol from Salvia miltiorrhiza stem | |
| CN105175426A (en) | Method of extracting and purifying bergenin from cissus pteroclada hayata | |
| CN101747195B (en) | Separation and purifying method for DCQA (dicaffeoylquinic acid) component in jerusalem artichoke | |
| CN101525328B (en) | Method for extracting alpha-mangostin from mangosteen fruit peel | |
| CN107098942A (en) | A kind of method of kaempferia galamga glycosides in Subcritical Water Extraction radish leaves | |
| CN106496292A (en) | A kind of method for preparing 6 iridoid glycoside constituents from Fructus Gardeniae simultaneously | |
| CN104163754A (en) | Method for extraction and separating of high purity resveratrol from peanut root | |
| CN104844547B (en) | A kind of high efficiency extraction of barbaloin and grading purification method | |
| CN104402949B (en) | A kind of method that separation simultaneously prepares Syringin and oleuropein from lilac | |
| CN103467262A (en) | Method for preparing 9-oxonerolidol from camphor tree plants | |
| CN101412724B (en) | Method for extracting bilobalide compound from ginkgo leaf | |
| CN102250174A (en) | Method for extracting jasminin from winter jasmine leaves | |
| CN102145040A (en) | Technique for adsorbing and extracting effective part of folium orthosiphoni by employing macroporous resin | |
| CN106336440B (en) | The method of extraction separation oleanolic acid from olive growing leaves | |
| CN114634537A (en) | Preparation method and application of diterpene in gold leaves | |
| CN107722080A (en) | A kind of method that ursin is extracted in the leaf from purple bergenia herb | |
| CN106986904A (en) | A kind of Gastrodin extracted in the anesthesia stem from day and preparation method thereof | |
| CN102389456A (en) | Method for extracting isodon japonica var.galaucocalyx total diterpenoids or Glaucocalyxin A | |
| CN1730027A (en) | Method for preparing anti-hepatitis active part from swertia main pharmaceutical plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |